HPLC POST-COLUMN DERIVATIZATION ENVIRONMENTAL FOOD AND FEED ANALYSIS PRODUCT TESTING SOLUTIONS PRODUCT CATALOG 15 /16

Transcription

1 PRODUCT CATALOG 15 /16 HPLC POST-COLUMN DERIVATIZATION ENVIRONMENTAL FOOD AND FEED ANALYSIS PRODUCT TESTING SOLUTIONS INSTRUMENTS / COLUMNS / METHODS / REAGENTS

2 TABLE OF CONTENTS INTRODUCTION 3 Customer Service and Technical Support 3 How to Place an Order 4 Factory Repairs and Limited Warranty 5 Instrument Service Plans INSTRUMENTS & HARDWARE, 6 Pinnacle PCX Derivatization Instrument 9 Vector PCX Derivatization Instrument 11 UVE Photochemical Reactor CHROMATOGRAMS, COLUMNS & GUARDS 13 Columns & Guards Catalog Information 14 Cation-exchange Columns Catalog Information 14 Reversed-Phase Columns Catalog Information 15 Cation-exchange GARD Column Protection System 15 Lithium Amino Acids 21 Sodium Amino Acids 25 Carbamate Pesticides 29 Glyphosate Herbicide Analysis Column 30 Alkion Column 34 Mycotox Mycotoxins 35 Polyether Antibiotics REAGENTS, DILUENTS & SAMPLE PREPARATION SOLUTIONS 36 TRIONE Ninhydrin Reagent 37 o-phthaladehyde 37 Naphthalene-2,3-Dicarboxaldehyde 37 Vanillin 38 p-dimethylaminobenzaldehyde (DMAB) 38 Thiofluor 38 o-phthalaldehyde Diluents 39 Hydrolysis Reagent for Carbamate Pesticide Analysis CB Hydrolysis Reagent for Carbamate Pesticide Analysis CB Hypochlorite Diluent for Glyphosate Herbicide Analysis GA Samples and Standards Diluents (Li220 & Na220) 40 Solutions for Sample Preparation for Amino Acid Analysis (SERAPREP and URIPREP) 41 ChlorAC Buffer 41 Restore 41 Extraction Solution, Aminoglycoside ELUANTS 42 Sodium-based Solutions for Amino Acid Analysis 42 Lithium-based Solutions for Amino Acid Analysis 42 Glyphosate Herbicide Analysis 42 Potassium-based Solutions for ALKION Column for Aminoglycoside Antibiotics & Streptomycin 42 Biogenic Amines/Polyamines Analysis 42 Muti-Residue Mycotoxin Analysis 42 Pure Water TEST MIXTURES & STANDARDS 43 Calibration Standards and Test Mixtures PRODUCT TESTING SOLUTIONS 45 Artificial Perspiration 47 Artificial Saliva 47 Artificial Urine APPLICATION KITS min High-Efficiency Collagen Hydrolysate min High-Efficiency Protein Hydrolysate min High-Efficiency Oxidized Feed Hydrolysate 48 Standard 60-min Protein Hydrolysate 48 High-Efficiency 55-min Protein Hydrolysate 48 High-Efficiency 55-min Collagen Hydrolysate min Oxidized Feed Hydrolysate min High-Efficiency Physiologic Fluid/Native Sample 49 Standard 185-min Physiologic Fluid/Native Sample 49 High-Efficiency 120-min Physiologic Fluid/Native Sample 49 Temperature Gradient, Physiologic Fluid/ Native Sample Kit 50 Carbamate Pesticides, 23+ Compounds 50 Carbamate Pesticides, EPA Method Carbamate Pesticides, AOAC Method Carbamate Pesticides, AOAC Method Glyphosate Herbicide, EPA Method 547, AOAC Method Biogenic Amines/Polyamines 50 Aminoglycoside Antibiotics 50 Multi-Residue Mycotoxins 50 Aflatoxins 50 Polyether Antibiotics REPLACEMENT COMPONENTS, FITTINGS & TUBING 51 Pinnacle PCX Flow Diagram Two-Pump System 52 Pinnacle PCX Flow Diagram One-Pump System 53 Vector PCX Flow Diagram 54 Pinnacle PCX Reagent Pump and Valve Components 54 Vector PCX Reagent Pump Components 54 Pinnacle PCX Replacement Reactor Cartridge 54 Vector PCX Substitute Reactor Volumes or Replacement Reators 54 Reagent Reservoirs & Cap Assemblies 54 Pressure Regulators, Valves & Guages 55 Flow Restrictors 55 Filters & Frits 55 Tees & Unions 55 Tubings, Plastic 56 Fittings, Plastic 56 Fittings, Type 316 Stainless Steel 56 Miscellaneous Accessories CLEAN-UP COLUMNS 57 Immunoaffinity Sample Clean-up Columns 59 DONeX 60 Glyphosate and AMPA Sample Clean-up Cartridges METHOD ABSTRACTS AMINO ACIDS minute Amino Acid Analysis of Hydrolyzed Samples (380.1) Min Amino Acid Analysis of Physiological Samples (382) 63 Amino Acids Analysis of Cell Culture Media (371) 65 Amino Acids Analysis of Monoclonal Antibodies (373) 67 Amino Acids Analysis of According to European Pharmacopoeia 8.0. (391) 71 Amino Acids Analysis Sports Drinks and Supplements (393) MYCOTOXINS 75 Clean-Up and Determination of Aflatoxins in Peanuts and Peanut Butter (215) 77 Clean-up and Analysis of Aflatoxins and Ochratoxin A in Herbs and Spices (218) 79 Ochratoxin A in Red and White Wine with OtaCLEAN and AcceCLEAN (220) 81 Multi-Residue Mycotoxin Analysis (203.1) 83 Multi-Residue Mycotoxin Analysis of Dry Distillers Grains (208) CONTAMINATES AND RESIDUE 85 Bromate Analysis (120) 87 Nitrite and Nitrate Analysis (121) 89 Simultaneous Determination of Nitrite & Nitrate in Processed Foods (123) 91 Sensitive and Selective Analysis of Nitrite and Nitrate in Drinking Water by Ion Chromatography (IC) with Post-Column Derivatization and UV/VIS Detection (129)

3 INTRODUCTION 93 Highly Sensitive Analysis of Chromium (VI) in Drinking Water by Ion Chromatography (IC) with Post-Column Derivatization and UV/VIS Detection (388) 95 Expanded HPLC Method for N-Methyl Carbamates (112) 97 Analysis of N-Methyl Carbamate pesticides in food (472) 99 Glyphosate and AMPA Analysis in Crops (206) 101 Glyphosate Analysis in Soy Beans, Corn and Sunflower Seeds (207) 103 Analysis of Glufosinate and Glyphosate in Water by HPLC with Post-Column Derivatization (296) 105 Analysis of Paralytic Shellfish Toxins in Bivalve Mollusks using HPLC Method with Post-column Derivatization and Fluorescence Detection (105.3) 107 Analysis of Biogenic Amines (102) 109 Polyether Antibiotics (104) 110 Analysis of Polyether Antibiotics in Animal Feeds by HPLC with Post-column Derivatization (264) 112 Aminoglycoside Antibiotics in Feed (205) 114 Analysis of Formaldehyde (325) FOOD / FEED / SUPPLEMENTS 115 Analysis of L-Theanine in Tea by HPLC With Post-column Derivatization (378) 117 Analysis of Antioxidants in Foods and Dietary Supplements Using HPLC With Post-column Derivatization (231) 119 Analysis Of Vitamin B1 In Foods And Dietary Supplements By Hplc With Post- Column Derivatization And Fluorescence Detection (233) 121 Analysis of Bioavailable Niacin (Vitamin B3) by HPLC with Postcolumn Photochemical Derivatization in Foods and Supplements (237) 123 Simultaneous Analysis of Vitamines B1, B2, B3 and B6 in Protein Powders and Supplements (239) 125 Sulfonamide Artificial Sweetners in Food Products (124) 126 Analysis of Sugars in Feeds (223) 128 Analysis of Voglibose in Pharmaceutical Formulations (327) 130 Analysis of Alendronic Acid in Pharmaceutical Formulations (328) 131 Analysis of Amino Acids Supplements and Sports Drink (393) 135 GENERAL TERMS AND CONDITIONS 136 DISTRIBUTORS CUSTOMER SERVICE & TECHNICAL SUPPORT Pickering Laboratories has earned the reputation for strong Technical Support in HPLC post-column technology: chemistry, columns, reagents, and derivatization instruments. We start with assisting our customers in identifying the best HPLC configuration required for a successful integration of our HPLC post-column system. Then we continue our support through and beyond the warranty period to ensure our customers chromatograms reflect current standards of quality and reproducibility. HOW TO CONTACT US For Customer Service or Technical Support contact: sales@pickeringlabs.com support@pickeringlabs.com / Fax: HOW TO PLACE AN ORDER Please contact us directly to place an order: orders@pickeringlabs.com / Pickering Laboratories, Inc Space Park Way Mountain View, CA When ordering, please provide the following: Complete shipping and billing addresses Catalog number and quantity of each item Brief product description, as found in Ordering Information Purchase order number (and contract number, if applicable) Name and telephone number of person calling Confirmation required Special instructions, e.g., package markings, mode of transit if other than UPS If ordering a complete post-column system, please specify the brand of HPLC with which it will be operating. We will advise you if any modifications to the HPLC are necessary. From European countries, please contact our European distributor: LCTech GmbH Postfach/P.O. Box 1360 D Dorfen, Germany Telefon: +49(0) Telefax: +49(0) info@lctech.de Internet: From non-european countries, please contact Pickering Laboratories directly. We will provide the names of companies which distribute our products. TERMS A minimum order is not required Net 30 days for established accounts, or prepayment in U.S. dollars, or Letter-of-Credit We accept MasterCard & VISA, Discover Card or American Express Prices are Ex-Works Seller s Factory, Mountain View, California U.S. Destinations & Canada: Freight charges are prepaid and added to the invoice. Shipments are via UPS- Ground unless otherwise specified. Destinations Outside of the U.S.: New Accounts: Before a credit-line has been established, new account orders must be pre-paid. Freight charges are collect. Established Accounts in Good Standing: Freight charges are collect. Unless otherwise specified, shipments overseas will be via consolidated air-freight to minimize costs. 3 PRODUCT CATALOG PICKERING LABORATORIES 15 /16

4 INTRODUCTION Prices and specifications are subject to change without prior notice. installation & training We encourage you to take advantage of Pickering Laboratories individualized two or three-day installation and training courses. For an all-inclusive fee we can provide post-column system installation and training at your site by an experienced chemist from Pickering Laboratories or it s representatives. The training includes performing analyses, routine maintenance and troubleshooting, and chromatogram interpretation. Some of our authorized distributors offer their own training and installation services. Whenever you buy installation and training by Pickering Laboratories, please call Technical Support early to schedule our visit to your site. This is especially important if you buy installation through a distributor or HPLC Company because we need to hear all the details directly from you. Individually tailored courses are also given at our facility in Mountain View, California. Please telephone or with your requirements and we will advise cost and schedule. PRODUCT RETURNS Products can be returned if unopened. A restocking fee of 15 % of list price will be applied. Before returning a product for any reason, please: Phone or requesting a Returned Goods Authorization Number. Provide a Purchase Order Number for product returns intended for service and repair. Package the product so that it will arrive undamaged. Use the original packing if possible. If in doubt, please call. Pickering Laboratories cannot assume responsibility for goods which arrive damaged in shipment. Return the completed Health & Safety Declaration with the items intended for service or repair (See page 5 for details.) FACTORY REPAIRS Pickering offers factory repair for it s instruments (Pinnacle PCX and Vector PCX only). We will re-build or replace components as necessary in our judgment, then test the instrument to make certain that the original problem has been solved. For repairs, we will charge for shipping, parts and labor (current shop rate). What is Required: Obtain a Return Goods Authorization Number (RGA) and a Health and Safety Declaration form. When the instrument is returned it must have the RGA noted on the outside of the box and a completed Health and Safety Declaration, with the instrument. To expedite your repair we request the instrument is returned with a purchase order for the cost to evaluate the system (current shop rate). What is Done: Upon arrival we will evaluate your system to generate an estimated cost for repairs. Once we have your approval we will complete the repair. We will replace damaged parts, pressure test the instrument, and then test the instrument to confirm the system meets the products original manufacturer s specification. What you get: The minimum parts replaced are: Restrictor(s), Seal(s) and Filters. If we test the instrument to determine that it meets the original manufacturer s specification, we will warranty the instrument for 1 year on all the parts that were replaced. If you request that we do not complete this test we will not warranty our work. FIELD SERVICE Because of our limited personnel, we prefer to resolve problems without site visits, but when necessary we will schedule a site visit with you. Please call for a quotation. On-site service entails an initial evaluation on the first day, with repairs potentially extending to the second day to allow for shipment of parts. The shipment of parts will be included in the invoice. On-site repairs costs will include travel time, air fare, hotel, car rentals, expenses, plus parts and labor. LIMITED WARRANTY Instruments Pickering Laboratories, Inc., (Pickering) Instruments are warranted to be free of defects in material and workmanship under normal installation, use, and maintenance, for a period of one year from the date of delivery to the customer. Pickering will replace or repair, without cost, any defective items. Expendable items such as check valves, pistons, piston seals, and filters are excluded from this warranty. In addition, physical damage, poor-quality reagent- and sample-induced damage, and instrument damage due to customer s misuse are not covered by this warranty. Analytical Columns Pickering s Analytical Columns are warranted for 90 days to be free of defects in materials and workmanship under normal installation, use, and maintenance, for the warranted time beginning from the date of delivery to the customer. Pickering will replace the Analytical Column under warranty if found defective in material or workmanship. However, the warranty is void if the Analytical Column was damaged due to customer s misuse.

5 INTRODUCTION How to Obtain Warranty Service If there is a problem with your Instrument or Analytical Column within the warranty period, immediately notify Pickering or your local authorized representative. If the Instrument or Analytical Column was not purchased directly from Pickering, please contact the vendor where it was purchased. Any Instrument, part of the Instrument, or Analytical Column returned to Pickering for examination or repair shall have Pickering s prior approval (call for a Returned Goods Authorization Number) and be shipped prepaid by the Customer. Return transportation will be at Pickering s expense if the Instrument, part of the Instrument, or Analytical Column is found to be defective and under warranty. HEALTH & SAFETY DECLARATION Instruments and columns must be decontaminated and cleaned prior to returning to Pickering. To protect and ensure the safety of Pickering personnel, customer must properly flush instrument of any and all hazards (e.g. reactive chemicals, blood borne pathogens). Please contact Pickering to obtain a Health and Safety Declaration Form (B-0107) describing return instructions and customer responsibilities. Failure to do so will result in delay of service and/or additional charges. INSTRUMENT & SERVICE PLANS We encourage you to consider including a Service Plan to match the quality and reliability you have come to expect from Pickering Laboratories. These offerings provide factory trained engineers experienced in HPLC and post-column technologies to service your Pickering post-column derivatization instruments. Each plan has the features and benefits most requested by customers to offer the best value and effectiveness for today s busy laboratories. When you sign-up for the Pickering Laboratories Service Plans you are assuring the continued quality and reliability of your post-column instrument as well as your analysis. BENEFITS OF SERVICE AGREEMENTS Free Software Upgrades Installation of all required performance upgrades PM/Service performed by Pickering Laboratory trained service specialists Fixed Annual Cost No Surprises Unlimited Technical/Analytical Support Multi-Unit Service Discounts Available 5 COMPARISON OF SERVICE PLANS FEATURES FACTORY WARRANTY WARRANTY UPGRADE** DEPOT SERVICE ELITE SERVICE* Telephone support to isolate and resolve technical problems Monday - Friday 8AM - 5PM Excluding Holidays Monday - Friday 8AM - 5PM Excluding Holidays Monday - Friday 8AM - 5PM Excluding Holidays Monday - Friday 8AM - 5PM Excluding Holidays Recertification Not included Included Included Included Site visits At prevailing rates Two (including PM) At prevailing rates Two (including PM) Preventative Maintenance At prevailing rates One One One Freight Charges Not Included Included Included Included Parts and Labor Included Included Included Included Service Plans offered for the Pickering Laboratories Pinnacle PCX and Vector PCX post-column derivatization instruments. Pre-service inspection is required for instruments that have an expired manufacturers warranty or service contracts. *The Elite Service plan provides a total of two service calls that will cover parts, labor and travel for the period of one year. Included is unlimited telephone support from Pickering Technical Service Engineers and Applications Specialists. **Applies to new systems only. For more information and quotations: Please contact Pickering Laboratories headquarters Technical Support at support@pickeringlabs.com or PRODUCT CATALOG PICKERING LABORATORIES 15 /16

6 INSTRUMENTS & HARDWARE PINNACLE PCX DERIVATIZATION INSTRUMENT Part of the complete integrated system of instruments, chemicals, columns, methods and support from Pickering Laboratories. The Pinnacle PCX Delta Series is an optimized HPLC post-column derivatization system for analysis of Amino Acids, Carbamates, Mycotoxins, Antibiotics and many other applications. This instrument is the culmination of Pickering Laboratories 30 years of experience in post-column analysis and instrument manufacturing. Each component is specifically designed to optimize the sensitivity and selectivity of post-column analysis. Only Pickering Laboratories offers the complete package of chemicals, columns, methods and post-column systems. Because each part of the method is designed to work together, Pickering Laboratories can offer the extraordinary promise that the analysis is guaranteed to work for the intended application. The Pinnacle PCX reflects the ease of use, reliability and ruggedness you have come to expect from Pickering Laboratories. SYSTEM DESIGN ADVANCEMENTS RESULT IN OPTIMIZED ANALYSIS: The electronic syringe pump provides true pulse-free flow for superior sensitivity and consistency. The pump cylinder and head is made from a single piece of inert ceramic for durability and non-reactivity. Electronic valves eliminate troublesome check valves and allow automated pump flushing. The quick-change reactor cartridge makes application switching easy and replacements quick and inexpensive. The column oven utilizes circulating air for consistency of heating and quick cooling. Column oven temperature gradient programming improves separation and run times. Pinnacle PCX is the only post-column system with this feature. Inert flow path extends system life and reduces maintenance. The PCX control software allows for precise control of the reagent delivery and conservation. Works with any HPLC system. RoHS compliant

7 INSTRUMENTS & HARDWARE FEATURE HIGHLIGHT The Pinnacle PCX has a column oven capable of programmable temperature gradients. This feature is useful in Amino Acid Analysis by enabling additional flexibility when optimizing methods. Using the temperature gradient program allows for improved separation, shortened analysis time and fine-tuned methods for specific compound detection. The Chromatogram in Figure 1 demonstrates some of the clear advantages of the temperature gradient program. Compared to the standard method without the temperature gradient the total run time is 42 % shorter. Additionally, separation is improved. With the temperature gradient program, you are able to focus on different areas of the chromatogram for your specific compound of interest o C 21 o 65 C FIGURE 1 High-efficiency Lithium-based Analysis for Physiological samples using temperature gradient 34 oc : min 1 Phosphoserine 2 Taurine 3 Phosphoethanolamine 4 Urea 5 Aspartic acid 6 Hydroxyproline 7 Threonine 8 Serine 9 Asparagine 10 Glutamic acid 11 Glutamine 12 Sarcosine 13 a-aminoadipic acid 14 Proline 15 Glycine 16 Alanine 17 Citrulline 18 a-amino-n-butyric acid 19 Valine 20 Cystine 21 Methionine 22 Allo-Isoleucine 23 Cystathionine 24 Isoleucine 25 Leucine 26 Tyrosine 27 Phenylalanine 28 b-alanine 29 b-amino-i-butyric acid 30 Homocystine 31 g-aminobutyric acid 32 Tryptophan 33 Ethanolamine 34 Ammonia 35 Hydroxylysines 36 Ornitine 37 Lysine 38 1-Methylhistidine 39 Histidine 40 3-Methylhistidine 41 Anserine 42 Carnosine 43 Arginine 44 Glucosaminic Acid* 45 2-Aminoethyl-cysteine (AEC)* *Internal Standard PINNACLE PCX SPECIFICATIONS Dimensions: (h x w x d): 54 x 26.7 x 46.4 cm (21.25 x 10.5 x inches) Weight: 54.4 lbs (24.7 Kg) Duplex Electrical: V, 50/60 Hz, 1.7 A, 200 W or V, 50/60 Hz, 0.8 A, 200 W Mains voltage ± 10 % of nominal Installation (over voltage) category II, pollution degree 2 Indoor use only Environmental: Altitude up to 6500 ft Ambient Temperature C Relative humidity up to 80 % at 31 C REAGENT PUMPS True pulse-free syringe pump Single piece ceramic barrel Completely inert flow path Maximum operating pressure 500 psi Programmable flow rate Flow range: 50 µl/minute to 1500 µl/minute Flow Accuracy 3 % Flow Precision 0.5 % RSD Refill cycle of 60 seconds Automatic piston wash Automatic reagent flush cycle No check valves PRODUCT CATALOG PICKERING LABORATORIES 15 /16

8 INSTRUMENTS & HARDWARE CONTROL Computer controlled Windows PC with XP, Vista or Windows 7 USB connection or network REACTOR Heated reactor for temperature from 5 C above ambient to 130 C Easy replacement reactor coil cartridges Range of reactor dwell volumes: 0.1 ml to 3.5 ml Reaction coil withstands up to 42 bar (600 psi) inlet pressure at 130 C Thermal safety switch limits temperature to 150 C to prevent damage Fast response SAFEGUARDS In-line check valve: Prevents reagent back flow into the column when HPLC pressure drops Post-column system over pressure: A pre-calibrated relief valve opens at 35 bar (500 psi) to prevent rupture of the post-column reactor tubing in the event of down-stream blockage Back-pressure regulator: Applies 7 bar (100 psi) to the detector flow cell outlet (waste) to prevent detector noise and precipitation due to out-gassing or boiling COLUMN HEATER AND REACTOR CONTROLLER Heater accepts up to 6 or 8 mm OD (0.25 or 0.31 inch) x mm in length (column and guard) Programmable temperature gradient Power cooling capability Temperature holds within ± 0.4 C from the set point. Can be set with 1 C resolution from 5 C above ambient to 75 C Easy column access INSTRUMENT PACKAGE AND FLOW PATH Advanced fluidics valve management system Easy access to internal components Standard fittings Post-column pressure relief valve Side panels easily removed for service access Integrated reagent reservoir tray Corrosion proof pan and panels DISPLAY Backlit LCD Real time temperature and pressure display System status icons Simple system control interface GAS PRESSURE MANIFOLD AND REGULATOR Panel mount manifold Regulator maintains 0.3 bar (3-5 psi) on reagent reservoirs with 3-5 bar (45-75 psi) source pressure Safety pressure relief valve opens at 1 bar (14 psi) Manifold has two 1/4-28 tubing connections Gas lines with anti-syphon valves PRESSURIZED REAGENT RESERVOIR One liter capacity (2 L reservoirs available) Maintained under inert gas pressure to inhibit oxidation of oxygen-sensitive reagents Valve built into reservoir cap permits sparging during reagent preparation Reagent reservoirs fitted with 3.1 mm (1/8 ) OD, oxygen-impermeable Air Barrier tubing for oxygen-sensitive reagents PINNACLE PCX CATALOG INFORMATION CATALOG NO. APPLICATION REAGENT PUMP REACTOR VOLUME VOLTAGE Amino Acids (OPA detection), Aminoglycoside Antibiotics, Biogenic Amines, Polyamines Amino Acids (TRIONE Ninhydrin detection), Ochratoxin A, Formaldehyde, Bromate Analysis Single 0.15 ml 120 V 240 V Single 0.5 ml 120 V 240 V Mycotoxins including Aflatoxins & Fumonisins Analysis Single Knitted 1.4 ml 120 V 240 V Carbamate Pesticides, Glyphosate Herbicide, Streptomycin, (OPA detection) Dual 0.5 ml 120 V 240 V Paralytic Shellfish Toxins Dual Knitted 1.0 ml Trichothecene Mycotoxins Dual Knitted 1.2 & 1.6 ml Polyether Antibiotics Dual Knitted 1.4 ml Voglibose Analysis Single Knitted 3.5 ml Chromium V Analysis Single Knitted 1.0 ml Alprostadil Analysis Single Knitted 2.0 ml 120 V 240 V 120 V 240 V 120 V 240 V 120 V 240 V 120 V 240 V 120 V 240 V NOTE: Please contact Pickering for assistance in configuring a custom Pinnacle PCX that will meet your specific analysis needs. All configurations include gas manifold, accessories, spares, reservoir(s), power cord and operation manual.

9 INSTRUMENTS & HARDWARE VECTOR PCX DERIVATIZATION INSTRUMENT Vector PCX serves as another post-column choice ideal for application-specific methods. Vector PCX provides the selectivity and sensitivity required for most standard post-column applications while being reliable and easy to use. Since Vector PCX does not have a column oven it is important to use the HPLC column oven to ensure stable column temperature and prevent retention time drifts and separation problems. VECTOR PCX FEATURES Inert pumps PEEK liquid manifolds Integrated gas manifold Easy access and monitoring of fluidics Simple control interface (no computer required) Reduced bench space profile Matched with Pickering s Chromatographic Grade reagents, eluants and columns, the Vector PCX instrument is a solid choice for laboratories standard post-column needs. VECTOR PCX SPECIFICATIONS Dimensions: (h x w x d): 43 x 21.6 x 41.2 cm (17 x 8.75 x 16 inches) Weight: 27.6 lbs (12.5 kg) Duplex Electrical: V, 50/60 Hz 1.7 A, 200 W or V, 50/60 Hz, 0.8 A, 200 W Mains voltage ± 10 % of nominal Installation (over voltage) category II, pollution degree 2 Indoor use only Environmental: Altitude up to 6,500 ft Ambient temperature ºC Relative humidity up to 80 % at 31 ºC REAGENT PUMPS Independently adjustable, low-pulsation Adjustable from 0.05 to 2.00 ml/minute against back-pressures of up to 2000 psi Flow Accuracy 3 % Flow Precision 0.5 % RSD Sapphire pistons Liquid ends, including check valve housing, PEEK PEEK bypass/purge valves for each pump located on front of instrument panel Automatic piston wash FLOW PATH Independent pressure transducer for each pump 210 bar ( psi) Diamond-packed restrictors, matched to flow rate and viscosity of reagents PEEK bypass/purge valves Replaceable reagent filter PEEK mixing manifold REACTOR Heater reactor controls at ± 0.4 C for temperatures from 5 C above ambient to 130 C Range of reactor dwell volumes, 0.1 ml to 3.5 ml Reaction coil withstands up to 42 bar (600 psi) inlet pressure at 130 C LCD display of actual temperature or set point Thermal safety switch limits temperature to 150 C to prevent damage GAS PRESSURE MANIFOLD AND REGULATOR Panel mounted manifold Regulator maintains 0.3 bar (3-5 psi) on reagent reservoirs with 3-5 bar (45-75 psi) source pressure Safety pressure relief valve opens at 1 bar (14 psi) Manifold has two 1/4-28 tubing connections Gas line with anti-siphon valve PRESSURIZED REAGENT RESERVOIR One liter capacity (2 L reservoirs available) Maintained under inert gas pressure to inhibit oxidation of oxygensensitive reagents Valve built into reservoir cap permits sparging during reagent preparation Reagent reservoirs fitted with 3.1 mm (1/8 ) OD, oxygen-impermeable Air Barrier tubing for oxygen-sensitive reagents and/or with 3.1 mm (1/8 ) OD FEP tubing. 9 PRODUCT CATALOG PICKERING LABORATORIES 15 /16

10 INSTRUMENTS & HARDWARE SAFEGUARDS A pressure switch installed between LC (eluant) pump and sample injector turns off power to reagent pumps and reactor when the eluant pump pressure drops to 30 bar (425 psi), ensuring that reagent will not flow upstream and damage the analytical column. Low eluant pressure can result from power failure, eluant pump malfunciton, automatic or intentional shut-down, or an empty reservoir. The Vector PCX will not restart automatically. Post-column system over pressure: A pre-calibrated relief valve opens at 35 bar (500 psi) to prevent rupture of the post-column reactor tubing in the event of down-stream blockage Back-pressure regulator: Applies 7 bar (100 psi) to the detector flow cell outlet (waste) to prevent detector noise and precipitation due to out-gassing or boiling VECTOR PCX CATALOG INFORMATION CATALOG NO. APPLICATION REAGENT PUMP REACTOR VOLUME VOLTAGE Amino Acids (OPA detection), Aminoglycoside Antibiotics, Biogenic Amines, Polyamines Amino Acids (TRIONE Ninhydrin detection), Ochratoxin A, Formaldehyde, Bromate Single 0.15 ml 120 V 240 V Single 0.5 ml 120 V 240 V Aflatoxins Analysis Single Knitted 1.4 ml 120 V 240 V Carbamate Pesticides, Glyphosate Herbicide, Streptomycin, (OPA detection) Dual 0.5 ml 120 V 240 V Paralytic Shellfish Toxins Dual Knitted 1.0 ml Trichothecene Mycotoxins Dual Knitted 1.2 ml and 1.6 ml Polyether Antibiotics Inert PEEK Dual Knitted 1.4 ml Voglibose Analysis Single Knitted 3.5 ml 120 V 240 V 120 V 240 V 120 V 240 V 120 V 240 V Streptomycin Dual 0.5 ml 120 V 240 V NOTE: Please contact Pickering for assistance in configuring a custom Vector PCX that will meet your specific analysis needs. All configurations include gas manifold, accessories, spares, reservoir(s), power cord and operation manual.

11 INSTRUMENTS & HARDWARE UVE PHOTOCHEMICAL REACTOR Detection Enhancement for Aflatoxins, Phenylurea Pesticides, Barbiturates and Other Compounds Photochemical derivatization is a simple, inexpensive and flexible technique that improves sensitivity and selectivity of detection for a broad range of analytes. Among the applications for the photochemical reactor are analysis of Aflatoxins in foods, Phenylurea Pesticides in water and Barbiturates in biological samples. Photochemical derivatization also allows identification of closely related compounds such as polyphenols. Pickering Laboratories Multi-residue Mycotoxins method for DON, Aflatoxins, Fumonisins, Ochratoxin A and Zearalenone employs photochemical derivatization for Aflatoxins allowing detection at sub-ppb levels. The photochemical reactor has a 254 nm lamp and a knitted reactor coil. 11 FEATURE HIGHLIGHTS 254 nm UV low pressure lamp with cooled reflector tube Long term stability of lamp and coil High light transmission Robust steel housing to meet laboratory requirements Special designed fluorocarbon coil Photochemical post-column derivatization of Aflatoxins in a special reactor loop with UV light Result: clear peaks Comparable to electrochemical derivatization with Cobra Cell (DG Joint Research Center of the European Commission in the Institute for Health and Consumer Protection) AOAC accepted methodology Standard reactor volume is 1.0 ml PRODUCT CATALOG PICKERING LABORATORIES 15 /16

12 INSTRUMENTS & HARDWARE UV DERIVATIZATION RESULTS IN CLEAR PEAKS FOR AFLATOXINS Note the short run time: B1 elutes at 9.5 Min 10,000 UVE HPLC 9000 FLD 8000 B G2 Easy Handling: Simply place the UVE between your HPLC device and detector, switch it on - ready to use! Intensity G1 B1 What actually happens? Aflatoxins B1 and G1 are transformed to stable fluorescent derivatives resulting in clear peaks Without UVE : Low response for G1 and B1 Min 10, B G UVE SPECIFICATIONS Intensity G2 B2 CE CERTIFIED UV LAMP REACTOR COIL DIMENSIONS POWER INPUT Yes 254 nm Special 14.5 x 8.5 x 27 cm 50 W 3000 WEIGHT 3 kg Min CATALOG NO UVE CATALOG INFORMATION DESCRIPTION UVE - Photochemical Reactor, 1mL,120 V UVE - Photochemical Reactor, 1mL, 240 V UVE - Reactor coil, 1mL knitted Aflatoxin Analysis. Other coil sizes available on request UV replacement lamp With UVE : High response and no band spreading

13 CHROMATOGRAMS, COLUMNS & GUARDS Pickering Laboratories columns and guards are intended for specific applications that require post-column derivatization. This technology guarantees detection of certain classes of compounds at very low concentrations amino acids, carbamate pesticides and polyamines, for example. Each column is packed and tested to separate the target compounds according to Pickering s chromatographic quality control standards and published analytical method. The following acceptance criteria apply to all of Pickering s columns: With guard column installed, produce a specified chromatogram of the compounds in a standard test mixture. Separate the compounds in the established order, with specified resolution of critical pairs. Operate within the specified range of back pressure. Be free of contaminating material which can cause baseline artifacts. Only after all criteria have been met can the column s serial number and label be applied. Quite simply, the column is guaranteed to produce a chromatogram for its intended application if it is operated according to the conditions and methods prescribed by Pickering Laboratories. About Guard Columns While it is true that any of our columns may be run without a guard, the practical consequence is a shorter column lifetime. Pickering sells a variety of appropriate guard columns to protect our analytical columns. Additionally, GARD provides protection against contamination for cation-exchange applications without affecting column efficiency and it is far less expensive to replace than an analytical column. See page 14 for more information about our GARD column protection system. 13 PRODUCT CATALOG PICKERING LABORATORIES 15 /16

14 CHROMATOGRAMS, COLUMNS & GUARDS CATION-EXCHANGE COLUMNS & GUARDS CATALOG INFORMATION CATALOG NO. DESCRIPTION LITHIUM AMINO ACID ANALYSIS COLUMNS T T High-efficiency Lithium Cation-exchange Column 4.0 x 100 mm & amino acid test mixture 70 min High-efficiency Lithium Ion-exchange Column, 4.6 x 75 mm & amino acid test mixture Standard Lithium Cation-exchange Column 3.0 x 250 mm & amino acid test mixture T T SODIUM AMINO ACID ANALYSIS COLUMNS High-efficiency Sodium Cation-exchange for protein and collagen hydrolysates Column mm & amino acid test mixture High-efficiency Sodium Cation-exchange for protein, collagen and oxidized feed hydrolysates Column mm & amino acid test mixture 30-minute High-efficiency Sodium Cation-exchange Column mm & amino acid test mixture Standard Sodium Cation-exchange Column mm & amino acid test mixture GLYPHOSATE COLUMNS Cation-exchange Column for Glyphosate analysis, mm & Glyphosate test mixture GARD COLUMN PROTECTION SYSTEM (FOR USE WITH ANY CATION-EXCHANGE COLUMN) Cation-exchange GARD Assembly: Includes holder and 2 replaceable GARDs Replacement Cation-exchange GARDs (2/PK) GARD Holder ALKION COLUMN ALKION Cation-exchange column, K + form, mm ALKION Guard column, K + form, mm REVERSED-PHASE COLUMNS & GUARDS CATALOG INFORMATION CATALOG NO. DESCRIPTION CARBAMATE PESTICIDE COLUMNS Carbamate Column C 8, expanded resolution, mm & Carbamate Test Mixture Carbamate Column C 8, high resolution/capacity, mm & Carbamate Test Mixture Carbamate Column C 18, rapid analysis, mm & Carbamate Test Mixture POLYETHER ANTIBIOTICS Polyether reversed-phase column, mm MYCOTOX COLUMN MYCOTOX reversed-phase column, mm GUARDS FOR STANDARD REVERSED-PHASE COLUMNS 18ECG001 18ECG002 Guard Cartridge Holder with 3 cartridges Guard Cartridges, 2/pk

15 CHROMATOGRAMS, COLUMNS & GUARDS CATION-EXCHANGE GARD COLUMN PROTECTION SYSTEM Cation-exchange GARD uses a proprietary material to prevent matrix compounds from passing through (and thereby protecting the analytical column), but allows the analytes of interest to pass unimpeded through the GARD and onto the analytical column. The GARD significantly prolongs column life without band spreading or added pressure. We demonstrated, by means of a performance comparison for Amino Acid Analysis, that the use of a GARD will effectively protect the analytical column, will be more cost-effective for the laboratory, is easy to change, is universal to cation-exchange applications, and most importantly has zero band spreading. Wth GARD Column protection system from Pickering Laboratories the same cation-exchange GARD can be used for nearly all cation-exchange applications. GARD COLUMN PROTECTION SYSTEM (FOR USE WITH ANY CATION-EXCHANGE COLUMN) CATALOG NO. DESCRIPTION Cation-exchange GARD Assembly: Includes holder and 2 replaceable GARDs Replacement Cation-exchange GARDs (2/PK) GARD Holder 15 LITHIUM AMINO ACID ANALYSIS COLUMNS Pickering Laboratories specialize in manufacturing of cation-exchange columns for amino acid analysis. No other techniques, including reversed-phase chromatography, have been shown to match post-column ion-exchange methods in quantitation and reproducibility. Advantages of this method, such as absence of matrix interferences, are especially important in the analysis of native samples. Lithium columns and buffers systems have high selectivity and are perfect for physiological fluids and food analysis. Post-column Conditions For Amino Acids Analysis: Reagent: Trione Reactor: 130 C, 0.5 ml Reagent Flow Rate: 0.3 ml/min Detection: UV-Vis Detector: λ=570 nm for primary amino acids λ=440 nm for secondary amino acids or Reagent: 300 mg of OPA, 2 g Thiofluor, 3 ml of 30 % Brij 35 solution in 950 ml of OD104 Reactor: 45 C, 0.15 ml Reagent Flow Rate: 0.3 ml/min Detection: Fluorometer: λ ex 330 nm, λ em 465 nm The recommended gradient conditions are subject to change without notice. This may happen because of lotspecific changes in the columns, or improvements in the overall method. The recommended gradient for the column will always be included in the column package, and it supersedes the information in this catalog. Use the program recommended on the column data sheet for the initial testing. The column oven temperature programming gives additional flexibility when optimizing methods. Using a temperature gradient allows the user to improve separation, shorten analysis time and fine-tune the method for detecting compounds of interest. PRODUCT CATALOG PICKERING LABORATORIES 15 /16

16 CHROMATOGRAMS, COLUMNS & GUARDS 70-MIN HIGH-EFFICIENCY LITHIUM CATION-EXCHANGE COLUMN (4.6 X 75 mm) CATALOG NUMBER T USE WITH CATION-EXCHANGE GARD COLUMN PROTECTION SYSTEM USE FOR PHYSIOLOGICAL SAMPLES TEMPERATURE GRADIENT HPLC PROGRAM TIME % Li365 % Li375 % RG003 % HPLC FLOW RATE: 0.55 ml/min, INITIAL TEMP.: 34 ºC INJECTION VOLUME: 10 µl of 0.25 µmole/ml Std. 1 COLUMN OVEN PROGRAM TIME TEMP ºC For more information see Method Abstract MA 382 (page 77) and Pickering Amino Acids Brochure : min 1 Phosphoserine 2 Taurine 3 Phosphoethanolamine 4 Urea 5 Aspartic acid 6 Hydroxyproline 7 Threonine 8 Serine 9 Asparagine 10 Glutamic acid 11 Glutamine 12 Sarcosine 13 a-aminoadipic acid 14 Proline 15 Glycine 16 Alanine 17 Citrulline 18 a-amino-n-butyric acid 19 Valine 20 Cystine 21 Methionine 22 Allo-Isoleucine 23 Cystathionine 24 Isoleucine 25 Leucine 26 Tyrosine 27 Phenylalanine 28 b-alanine 29 b-amino-i-butyric acid 30 Homocystine 31 g-aminobutyric acid 32 Tryptophan 33 Ethanolamine 34 Ammonia 35 Hydroxylysines 36 Ornitine 37 Lysine 38 1-Methylhistidine 39 Histidine 40 3-Methylhistidine 41 Anserine 42 Carnosine 43 Arginine 44 Glucosaminic Acid* 45 2-Aminoethyl-cysteine (AEC)* *Internal Standard NOTE: This method utilizes column temperature gradient. Use Pinnacle PCX column oven or HPLC column oven with temperature gradient capabilities.

17 CHROMATOGRAMS, COLUMNS & GUARDS HIGH-EFFICIENCY LITHIUM CATION-EXCHANGE COLUMN (4.0 X 100 mm) CATALOG NUMBER T USE WITH CATION-EXCHANGE GARD COLUMN PROTECTION SYSTEM USE FOR PHYSIOLOGICAL SAMPLES AND COMPLEX MATRICES TEMPERATURE ISOCRATIC Phosphoserine 2 Taurine 3 Phosphoethanolamine 4 Urea 5 Aspartic acid 6 Hydroxyproline 7 Threonine 8 Serine 9 Asparagine 10 Glutamic acid 11 Glutamine 12 Sarcosine 13 α Aminoadipic acid 14 Proline 15 Glycine 16 Alanine 17 Citrulline 18 α Amino-n-butyric acid 19 Valine 20 Cystine 21 Methionine 22 Cystathionine 23 Isoleucine 24 Leucine 25 Norleucine 26 Tyrosine 27 Phenylalanine 28 β Alanine 29 β Amino-i-butyric acid 30 Homocystine 31 γ Aminobutyric acid 32 Tryptophan 33 Ethanolamine 34 Hydroxylysines 35 Ammonia 36 Creatinine 37 Ornithine 38 Lysine 39 Histidine 40 3-Methyl-histidine 41 1-Methyl-histidine 42 Carnosine 43 Anserine 44 α Amino β guanidinopropionic acid 45 Arginine 17 CONDITIONS STEP TIME (Min) INTERVAL Li275 % Li750 % RG003 % COMMENT Inject Isocratic Linear Gradient Linear Gradient Isocratic Linear Gradient Isocratic Step Change Re-equilibration FLOW RATE: 0.35 ml/min, COLUMN TEMP: 37 ºC, INJECTION VOLUME: 10 µl of 0.25 µmole/ml Std. For more information see Pickering Amino Acids Brochure. PRODUCT CATALOG PICKERING LABORATORIES 15 /16

18 CHROMATOGRAMS, COLUMNS & GUARDS HIGH-EFFICIENCY LITHIUM CATION-EXCHANGE COLUMN (4.0 X 100 mm) CATALOG NUMBER T USE WITH CATION-EXCHANGE GARD COLUMN PROTECTION SYSTEM USE FOR PHYSIOLOGICAL FLUIDS TEMPERATURE GRADIENT CONDITIONS STEP TIME (Min) % Li365 % Li375 % RG003 % COMMENT Inject Isocratic Linear Gradient Linear Gradient Isocratic Step Gradient Isocratic Step Gradient Isocratic Step Gradient Equilibration HPLC FLOW RATE: 0.4 ml/min, INITIAL TEMP.: 34 ºC INJECTION VOLUME: 10 µl of 0.25 µmole/ml Std. COLUMN OVEN PROGRAM TIME (Min) TEMPERATURE ºC min 1 Phosphoserine 2 Taurine 3 Phosphoethanolamine 4 Urea 5 Aspartic acid 6 Hydroxyproline 7 Threonine 8 Serine 9 Asparagine 10 Glutamic acid 11 Glutamine 12 Sarcosine 13 a-aminoadipic acid 14 Proline 15 Glycine 16 Alanine 17 Citrulline 18 a-amino-n-butyric acid 19 Valine 20 Cystine 21 Methionine 22 Cystathionine 23 Isoleucine 24 Leucine 25 Tyrosine 26 Phenylalanine 27 b-alanine 28 b-amino-i-butyric acid 29 Homocystine 30 g-aminobutyric acid 31 Tryptophan 32 Ethanolamine 33 Ammonia 34 Hydroxylysines 35 Ornitine 36 Lysine 37 1-Methylhistidine 38 Histidine 39 3-Methylhistidine 40 Anserine 41 Carnosine 42 Arginine 43 Glucosaminic Acid* *Internal Standard NOTE: This method utilizes column temperature gradient. Use Pinnacle PCX column oven or HPLC column oven with temperature gradient capabilities. For more information see Pickering Amino Acids Brochure.

19 CHROMATOGRAMS, COLUMNS & GUARDS STANDARD LITHIUM CATION-EXCHANGE COLUMN (3.0 X 250 mm) CATALOG NUMBER USE WITH CATION-EXCHANGE GARD COLUMN PROTECTION SYSTEM USE FOR PHYSIOLOGICAL FLUIDS, COMPLEX MATRICES TEMPERATURE ISOCRATIC min 1 Phosphoserine 2 Taurine 3 Phosphoethanolamine 4 Urea 5 Aspartic acid 6 Hydroxyproline 7 Threonine 8 Serine 9 Asparagine 10 Glutamic acid 11 Glutamine 12 Sarcosine 13 α-aminoadipic acid 14 Proline 15 Glycine 16 Alanine 17 Citrulline 18 α-amino-n-butyric acid 19 Valine 20 Cystine 21 Methionine 22 Cystathionine 23 Isoleucine 24 Leucine 25 Norleucine 26 Tyrosine 27 Phenylalanine 28 β-alanine 29 β-amino-i-butyric acid 30 Homocystine 31 γ-aminobutyric acid 32 Tryptophan 33 Ethanolamine 34 Hydroxylysines 35 Ammonia 36 Creatinine 37 Ornithine 38 Lysine 39 Histidine 40 3-Methyl-histidine 41 1-Methyl-histidine 42 Carnosine 43 Anserine 44 α-amino-β- guanidino propionic acid 45 Arginine CONDITIONS TIME (Min) Li275 % Li750 % RG003 % FLOW RATE: 0.3 ml/min, COLUMN TEMP.: 40 ºC, INJECTION VOLUME: 10 µl of 0.25 µmole/ml Std. PRODUCT CATALOG PICKERING LABORATORIES 15 /16

20 Valine Norleucine CHROMATOGRAMS, COLUMNS & GUARDS HIGH-EFFICIENCY LITHIUM CATION-EXCHANGE COLUMN (4.0 X 100 mm) CATALOG NUMBER T USE WITH CATION-EXCHANGE GARD COLUMN PROTECTION SYSTEM USE FOR PKU/MSUD SCREENING OF PHYSIOLOGICAL FLUIDS TEMPERATURE ISOCRATIC a-amino-n-butyric Acid Cystathionine Methionine Isoleucine Leucine Tyrosine Phenylalanine min CONDITIONS STEP TIME (Min) INTERVAL Li275 % Li750 % RG003 % COMMENT Inject Linear Gradient Step Gradient Isocratic Step Gradient Re-equilibration HPLC FLOW RATE: 0.35 ml/min, COLUMN TEMP.: 38 ºC INJECTION VOLUME: 10 µl of 0.25 µmole/ml Std. LITHIUM COLUMNS CATALOG INFORMATION CATALOG NO. DESCRIPTION T 70-min High-efficiency Lithium Ion-exchange Column, 4.6 x 75 mm T High-efficiency Lithium Cation-exchange Column 4.0 x 100 mm & amino acid test mixture Standard Lithium Cation-exchange Column 3.0 x 250 mm & amino acid test mixture GARD COLUMN PROTECTION SYSTEM (FOR USE WITH ANY CATION-EXCHANGE COLUMN) CATALOG NO. DESCRIPTION Cation-exchange GARD Assembly: Includes holder and 2 replaceable GARDs Replacement Cation-exchange GARDs (2/PK) GARD Holder

21 CHROMATOGRAMS, COLUMNS & GUARDS SODIUM AMINO ACID ANALYSIS COLUMNS Ion-exchange chromatography followed by post-column derivatization has been a method of choice for amino acid analysis for many years. Pickering ion-exchange columns allow you to obtain consistent results with sensitivity, stability, selectivity and speed. Sodium columns and buffers systems are designed for amino acids analysis of hydrolyzed samples. Post-column Conditions For Amino Acids Analysis: Reagent: Trione Reactor: 130 C, 0.5 ml Reagent Flow Rate: 0.3 ml/min Detection: UV-Vis Detector: λ=570 nm for primary amino acids λ=440 nm for secondary amino acids or Reagent: 300 mg of OPA, 2 g Thiofluor, 3 ml of 30 % Brij 35 solution in 950 ml of OD104 Reactor: 45 C, 0.15 ml Reagent Flow Rate: 0.3 ml/min Detection: Fluorometer: λ ex 330 nm, λ em 465 nm The recommended gradient conditions are subject to change without notice. This may happen because of lotspecific changes in the columns, or improvements in the overall method. The recommended gradient for the column will always be included in the column package, and it supersedes the information in this catalog. Use the program recommended on the column data sheet for the initial testing. The column oven temperature programming gives additional flexibility when optimizing methods. Using a temperature gradient allows the user to improve separation, shorten analysis time and fine-tune the method for detecting compounds of interest. 21 PRODUCT CATALOG PICKERING LABORATORIES 15 /16

22 CHROMATOGRAMS, COLUMNS & GUARDS 30-MIN HIGH-EFFICIENCY SODIUM CATION-EXCHANGE COLUMN (4.6 X 110 mm) CATALOG NUMBER T USE WITH CATION-EXCHANGE GARD COLUMN PROTECTION SYSTEM USE FOR PROTEIN AND OXIDIZED FEEDS HYDROLYSATE TEMPERATURE GRADIENT Fig 1. Chromatogram of protein hydrolysate standard 1 Aspartic Acid 2 Threonine 3 Serine 4 Glutamic Acid 5 Proline 6 Glycine 7 Alanine 8 Cystine Valine 10 Methionine 11 Isoleucine 12 Leucine 13 Norleucine 14 Tyrosine 15 Phenylalanine 16 Histidine Lysine 18 Tryptophan 19 Ammonia 20 Arginine 21 Cysteic Acid 22 Methionine Sulfone METHOD FOR PROTEIN HYDROLYSATE SAMPLES TIME Na315 % Na425 % Na640 % RG011 % HPLC FLOW RATE: 0.6 ml/min INITIAL TEMP.: 46 ºC INJECTION VOLUME: 10 ml OF 0.25 mmole/ml STD. For more information see Method Abstract (page 76) and Pickering Amino Acids Brochure. COLUMN OVEN PROGRAM TIME TEMP ºC min Fig 2. Chromatogram of oxidized feeds hydrolysate standard METHOD FOR OXIDIZED FEEDS HYDROLYSATE SAMPLES TIME Na270 % Na425 % Na640 % RG011 % HPLC FLOW RATE: 0.6 ml/min INITIAL TEMP.: 55 ºC INJECTION VOLUME: 10 ml OF 0.25 mmole/ml STD. COLUMN OVEN PROGRAM TIME TEMP ºC Aspartic Acid 2 Threonine 3 Serine 4 Glutamic Acid 5 Proline 6 Glycine 7 Alanine 8 Cystine 9 Valine 10 Methionine 11 Isoleucine 12 Leucine 13 Norleucine 14 Tyrosine 15 Phenylalanine 16 Histidine 17 Lysine 18 Tryptophan 19 Ammonia 20 Arginine 21 Cysteic Acid 22 Methionine Sulfone For more information see Method Abstract (page 76) and Pickering Amino Acids Brochure.

23 CHROMATOGRAMS, COLUMNS & GUARDS HIGH-EFFICIENCY CATION-EXCHANGE COLUMN (4.0 X 150 mm) CATALOG NUMBER T USE WITH GARD COLUMN PROTECTION SYSTEM USE FOR OXIDIZED HYDROLYSATE, PROTEIN HYDROLYSATE, COLLAGEN HYDROLYSATE TEMPERATURE ISOCRATIC Oxidized Hydrolysate 22 min CONDITIONS FOR OXIDIZED SAMPLES T TIME (Min) Na270 % Na740 % RG011 % FLOW RATE: 0.4 ml/min, COLUMN TEMP.: 50 ºC, INJECTION VOLUME: 10 ml OF 0.25 mmole/ml STD. 3 Aspartic Acid 4 Threonine 5 Serine 6 Glutamic Acid 7 Proline 8 Glycine 9 Alanine 11 Valine 13 Isoleucine 14 Leucine 16 Phenylalanine 18 Lysine 19 Ammonia 20 Histidine 22 Arginine 23 Cysteic Acid 24 Methionine Sulfone Protein Hydrolysate Collagen Hydrolysate min min 1 Methionine-D,L,-Sulfoxide 2 trans-4-hydroxy-l-proline 3 Aspartic Acid 4 Threonine 5 Serine 6 Glutamic Acid 7 Proline 8 Glycine 9 Alanine 10 Cystine 11 Valine 12 Methionine 13 Isoleucine 14 Leucine 15 Tyrosine 16 Phenylalanine 17 D,L & allo-hydroxylysine 18 Lysine 19 Ammonia 20 Histidine 21 Tryptophan 22 Arginine CONDITIONS FOR PROTEIN & COLLAGEN HYDROLYSATE SAMPLES TIME (Min) Na315 % Na740 % RG011 % FLOW RATE: 0.4 ml/min, COLUMN TEMP.: 48 ºC, INJECTION VOLUME: 10 ml OF 0.25 mmole/ml STD. PRODUCT CATALOG PICKERING LABORATORIES 15 /16

24 CHROMATOGRAMS, COLUMNS & GUARDS STANDARD SODIUM CATION-EXCHANGE COLUMN (3.0 X 250 mm) CATALOG NUMBER USE WITH CATION-EXCHANGE GARD COLUMN PROTECTION SYSTEM USE FOR PROTEIN HYDROLYSATE TEMPERATURE ISOCRATIC Protein Hydrolysate min 3 Aspartic acid 4 Threonine 5 Serine 6 Glutamic acid 7 Proline 8 Glycine 9 Alanine 10 Cystine 11 Valine 12 Methionine 13 Isoleucine 14 Leucine 15 Tyrosine 16 Phenylalanine 17 Hydroxylysines 18 Lysine 19 Ammonia 20 Histidine 21 Tryptophan 22 Arginine SODIUM COLUMNS CATALOG INFORMATION CATALOG NO. DESCRIPTION T 30-minute High-efficiency Sodium Cation-exchange Column mm & amino acid test mixture T High-efficiency Sodium Cation-exchange for protein, collagen and oxidized feed hydrolysates Column mm & amino acid test mixture Standard Sodium Cation-exchange Column mm & amino acid test mixture CONDITIONS FOR PROTEIN HYDROLYSATE SAMPLES TIME (Min) Na328 % Na740 % RG011 % FLOW RATE: 0.3 ml/min, COLUMN TEMP.: 48 ºC, INJECTION VOLUME: 10 ml OF 0.25 mmole/ml STD. GARD COLUMN PROTECTION SYSTEM (FOR USE WITH ANY CATION-EXCHANGE COLUMN) CATALOG NO. DESCRIPTION Cation-exchange GARD Assembly: Includes holder and 2 replaceable GARDs Replacement Cation-exchange GARDs (2/PK) GARD Holder

25 CHROMATOGRAMS, COLUMNS & GUARDS CARBAMATE PESTICIDE ANALYSIS COLUMNS Pickering Laboratories carbamate columns are guaranteed to produce the separation of carbamate residues, specified by EPA 531.1, EPA and AOAC methods. Post-column conditions for pesticide analysis: Reagent 1: Hydrolysis reagent CB130 or CB130.2 Reagent 2: 100 mg of OPA, 2 g Thiofluor in 950 ml of CB910 Reactor 1: 100 C, 0.5 ml Reactor 2: ambient, 0.1 ml Reagents flow rate: 0.3 ml/min Detection: Fluorometer: λ ex 330 nm, λ em 465 nm The recommended gradient conditions are subject to change without notice. This may happen because of lotspecific changes in the columns, or improvements in the overall method. The recommended gradient for the column will always be included in the column package, and it supersedes the information in this catalog. HIGH RESOLUTION / CAPACITY CARBAMATE COLUMN (4.6 X 250 mm), C 8, 5 µm 4 Response [mv] ng of Carbamates in 10 µl Time [min] 1 Aldicarb sulfoxide 2 Aldicarb sulfone 3 Oxamyl 4 Methomyl 5 3-Hydroxycarbofuran 6 Aldicarb 7 Propoxur 8 Carbofuran 9 Carbaryl 10 1-Naphthol 11 Methiocarb 12 BDMC CONDITIONS FOR AQUEOUS SAMPLES TIME (Min) WATER % MeOH % FLOW RATE: 1 ml/min, COLUMN TEMP.: 42 ºC, INJECTION VOLUME: Up to 400 ml CONDITIONS FOR METHANOLIC SAMPLES TIME (Min) WATER % MeOH % FLOW RATE: 1 ml/min, COLUMN TEMP.: 42 ºC, INJECTION VOLUME: 10 ml PRODUCT CATALOG PICKERING LABORATORIES 15 /16

26 CHROMATOGRAMS, COLUMNS & GUARDS RAPID ANALYSIS CARBAMATE COLUMN (4.6 X 150 mm), 5 µm CONDITIONS FOR AQUEOUS SAMPLES TIME (Min) WATER % MeOH % FLOW RATE: 1 ml/min, COLUMN TEMP.: 42 ºC, INJECTION VOLUME: Up to 250 ml ng of Carbamates in 150 µl min CONDITIONS FOR METHANOLIC SAMPLES TIME (Min) WATER % MeOH % FLOW RATE: 1 ml/min, COLUMN TEMP.: 42 ºC, INJECTION VOLUME: 10 ml min 1 Aldicarb sulfoxide 2 Aldicarb sulfone 3 Oxamyl 4 Methomyl 5 3-Hydroxycarbofuran 6 Aldicarb 25 ng of Carbamates in 10 µl 7 Propoxur 8 Carbofuran 9 Carbaryl 10 1-Naphthol 11 Methiocarb 12 BDMC (internal standard)

27 CHROMATOGRAMS, COLUMNS & GUARDS EXPANDED RESOLUTION CARBAMATE COLUMN (4.0 X 250 mm), 5 µm 4 25 ng of Carbamates in 10 µl CONDITIONS FOR WATER/METHANOL GRADIENT TIME (Min) WATER % MeOH % FLOW RATE: 0.8 ml/min, COLUMN TEMP.: 50 ºC, INJECTION VOLUME: 10 ml for methanolic and up to 400 ml for aqueous samples min CONDITIONS FOR WATER/ACETONITRILE GRADIENT TIME (Min) WATER % ACN % FLOW RATE: 0.8 ml/min, COLUMN TEMP.: 50 ºC, INJECTION VOLUME: 10 ml for methanolic and up to 400 ml for aqueous samples 0 40 ng of Carbamates in 400 µl min 27 1 Aldicarb sulfoxide 2 Aldicarb sulfone 3 Oxamyl 4 Methomyl 5 3-Hydroxycarbofuran 6 Aldicarb 7 Propoxur 8 Carbofuran 9 Carbaryl 10 1-Naphthol 11 Methiocarb 12 BDMC (internal standard) PRODUCT CATALOG PICKERING LABORATORIES 15 /16

28 CHROMATOGRAMS, COLUMNS & GUARDS SEPARATION OF 23 CARBAMATES USING COLUMN (4.0 X 250 mm) CONDITIONS FOR WATER/METHANOL GRADIENT TIME (Min) WATER % MeOH % FLOW RATE: 0.8 ml/min, COLUMN TEMP.: 50 ºC, INJECTION VOLUME: 10 ml for methanolic and up to 400 ml for aqueous samples 1 Aldicarb sulfoxide 2 Aldicarb sulfone 3 Oxamyl 4 Methomyl 5 3-Hydroxy 6 Aldicarb 7 Propoxur 8 Carbofuran 9 Carbaryl 10 Methiocarb 11 BDMC internal standard 12 Butocarboxim sulfoxide 13 Butocarboxim sulfone 14 Ethiofencarb sulfoxide 15 Ethiofencarb sulfone 16 Butocarboxim 17 Ethiofencarb 18 Thiofanox sulfoxide 19 Thiofanox sulfone 20 Methiocarb sulfoxide 21 Methiocarb sulfone 22 3-Ketocarbofuran 23 Thiofanox N 1-Naphthol N min CATALOG NO. CARBAMATE COLUMNS CATALOG INFORMATION DESCRIPTION Carbamate Column, expanded resolution, C 8, mm & Carbamate Test Mixture, 2.5 µg/ml, 1.5 ml Carbamate Column, high resolution/capacity, C 8, mm & Carbamate Test Mixture, 2.5 µg/ml, 1.5 ml Carbamate Column, rapid analysis, C 18, mm & Carbamate Test Mixture, 2.5 µg/ml, 1.5 ml 18ECG001 18ECG002 Guard Cartridge Holder with 3 guard cartridges Guard Cartridges, 2/pk.

29 CHROMATOGRAMS, COLUMNS & GUARDS GLYPHOSATE HERBICIDE ANALYSIS COLUMN Pickering Laboratories cation-exchange Glyphosate column is designed and tested for analysis of Glyphosate and its primary metabolite Aminomethylphosphonic acid (AMPA). This short, isocratic method provides separation of the peaks of interest. Reproducible performance is guaranteed run-to-run and column-to-column. Post-column Conditions For Glyphosate Analysis: Reagent 1: Oxidizing reagent 100 µl of 5 % Sodium Hypochlorite in 950 ml of GA116 Reagent 2: 100 mg of OPA, 2 g Thiofluor in 950 ml of GA104 Reactor 1: 36 C, 0.5 ml Reactor 2: ambient, 0.1 ml Reagents Flow Rate: 0.3 ml/min Detection: Fluorometer: λ ex 330 nm, λ em 465 nm GLYPHOSATE COLUMN (4.0 X 150 mm) K + CATION-EXCHANGE. TO BE USED WITH CATION-EXCHANGE GARD COLUMN PROTECTION SYSTEM Glyphosate AMPA min Glyphosate Test Mixture, 2.5 µg/ml For more information see Method Abstract MA 206 and MA 207 (pages 93 and 95). CONDITIONS TIME (Min) K200 % RG019 % FLOW RATE: 0.4 ml/min, COLUMN TEMP.: 55 ºC, INJECTION VOLUME: 10 ml GLYPHOSATE COLUMNS CATALOG INFORMATION CATALOG NO. DESCRIPTION Cation-exchange Column for Glyphosate analysis, mm & Glyphosate test mixture, 2.5 µg/ml, 1.5 ml CATALOG NO GARD COLUMN PROTECTION SYSTEM (FOR USE WITH ANY CATION-EXCHANGE COLUMN) DESCRIPTION Cation-exchange GARD Assembly: Includes holder and 2 replaceable GARDs Replacement Cation-exchange GARDs (2/PK) GARD Holder PRODUCT CATALOG PICKERING LABORATORIES 15 /16

30 CHROMATOGRAMS, COLUMNS & GUARDS ALKION COLUMN The stationary phase in the ALKION column is a rigid, non-porous polymeric phase that is surface sulfonated. The two modes of separation are ion-exchange and moderate partitioning. The low capacity of the ion-exchange resin makes it an ideal phase for the separation of strongly basic and positively charged compounds. The reversed-phase character allows for discrimination between closely related species. The unique properties of this column allow for its use in a wide range of applications. ALKION COLUMN ANALYSIS OF BIOGENIC AMINES Histamine Putrescine Cadaverine Spermidine Chromatogram of fish sauce sample min Post-column Conditions: Reagent: 300 mg of OPA, 2 g Thiofluor, 3 ml of 30 % Brij 35 in 950 ml of OD104 Reactor: 45 C, 0.15 ml Reagent Flow Rate: 0.3 ml/min Detection: Fluorometer: λ ex 330 nm, λ em 465 nm CONDITIONS TIME (Min) K600 % K563 % K130 % FLOW RATE: 0.8 ml/min, COLUMN TEMP.: 45 ºC, INJECTION VOLUME: 10 ml Putrescine Cadaverine Spermidine Spermine Tyramine Histamine min Chromatogram of calibration standard, 100 um Histamine Putrescine Cadaverine Spermidine Chromatogram of fish sample min

31 CHROMATOGRAMS, COLUMNS & GUARDS ALKION COLUMN ANALYSIS OF AMINOGLYCOSIDE ANTIBIOTICS IN FEEDS Post-column Conditions: Reagent: 300 mg of OPA, 2g Thiofluor, 3 ml of 30 % Brij 35 in 950 ml of OD104 Reactor: 45 C, 0.15 ml Reagent Flow Rate: 0.3 ml/min Detection: Fluorometer: λ ex 330 nm, λ em 465 nm Apramycin CONDITIONS FOR APRAMYCIN ANALYSIS TIME (Min) % % % FLOW RATE: 0.8 ml/min, COLUMN TEMP.: 40 ºC, INJECTION VOLUME: 10 ml min 31 Gentamicin Complex CONDITIONS FOR GENTAMICIN ANALYSIS TIME (Min) % % % FLOW RATE: 0.8 ml/min, COLUMN TEMP.: 40 ºC, INJECTION VOLUME: 10 ml min Neomycin CONDITIONS FOR NEOMYCIN ANALYSIS TIME (Min) % % % FLOW RATE: 0.8 ml/min, COLUMN TEMP.: 40 ºC, INJECTION VOLUME: 10 ml min PRODUCT CATALOG PICKERING LABORATORIES 15 /16

32 CHROMATOGRAMS, COLUMNS & GUARDS CONDITIONS FOR SEPARATION OF APRAMYCIN GENTAMICIN AND NEOMYCIN TIME (Min) % % % FLOW RATE: 0.8 ml/min, COLUMN TEMP.: 40 ºC, INJECTION VOLUME: 10 ml Neomycin Apramycin Gentamicin Complex min

33 CHROMATOGRAMS, COLUMNS & GUARDS ALKION COLUMN ANALYSIS OF STREPTOMYCIN Post-column Conditions: Reagent 1: 0.75 N Sodium Hydroxide Reactor 1: ambient, 0.1 ml Reagent 2: 0.6 % Ninhydrin Reactor 2: 60 C, 0.5 ml Reagents Flow Rate: 0.3 ml/min Detection: Fluorometer: λ ex 395 nm, λ em 500 nm or Reagent 1: Oxidizing Reagent: 100 µl of 5 % Sodium Hypochlorite in GA116 Reactor 1: 50 C, 0.5 ml Reagent 2: 300 mg of OPA, 2 g Thiofluor, 3 ml of 30 % Brij 35 solution in 950 ml of OD104 Reactor 2: ambient, 0.1 ml Detection: Fluorometer: λ ex 330 nm, λ em 465 nm CONDITIONS TIME (Min) % % % FLOW RATE: 0.8 ml/min, COLUMN TEMP.: 40 ºC, INJECTION VOLUME: up to several ml 33 Streptomycin min ALKION COLUMN CATALOG INFORMATION CATALOG NO. DESCRIPTION ALKION Cation-exchange column, K + form, mm ALKION Guard column, K + form, mm PRODUCT CATALOG PICKERING LABORATORIES 15 /16

34 CHROMATOGRAMS, COLUMNS & GUARDS MYCOTOX MYCOTOXINS ANALYSIS COLUMN AFLATOXINS ANALYSIS Aflatoxins are naturally occurring toxins belonging to the class Mycotoxins. They are produced by fungi and occur in peanuts, peanut meal, cotton-seed meal, wheat, milk and many other foods and feeds. The most important feature of the post-column method described below is that all six Aflatoxins are detectable at the same fluorescence emission wavelength in a single isocratic HPLC analysis. LC Conditions: Flow Rate: 1.0 ml/min, column temperature 42 C, injection volume 10 µl Mobile Phase: Methanol/acetonitrile/water: (22:22:56), isocratic Injection: 10 µl in Methanol 5 ng B 1 & G 1.5 ng B 2 & G ng M 1 & M 2 B1 Post-column Conditions: Reagent: I mg/l in water Reactor: 90 C, 1.4 ml Reagent Flow Rate: 0.4 ml/min Detection: Fluorometer: λ ex 365 nm, λ em 430 nm For more information see Method Abstract MA 215, MA 208, MA and MA 218 (pages 64, 72, 70 and 66). G1 B2 G2 M2 M1 With derivitization Without derivitization min MYCOTOX COLUMN CATALOG INFORMATION CATALOG NO. DESCRIPTION MYCOTOX Reversed-phase column, C 18, mm 18ECG001 Guard Cartridge Holder with 3 cartridges 18ECG002 Guard Cartridges, 2/pk.

35 CHROMATOGRAMS, COLUMNS & GUARDS POLYETHER ANTIBIOTICS ANALYSIS COLUMN Polyether antibiotics, such as Monensin, Narasin and Salinomycin, found in raw material, premix, liquid supplements and final feeds, are best quantified using HPLC with post-column derivatization. Because of selectivity of the post-column reaction, almost no sample clean-up is needed. LC Conditions: Flow Rate: 0.7 ml/min, column temperature 40 C, injection volume 10 µl Mobile Phase: 90:10 of Methanol / 5 % Acetic Acid in water, isocratic Feed Extract Post-column Conditions (non-metallic post-column derivatization system is required): Reagent 1: Concentrated Sulfuric acid / Methanol (4:96 v/v) Reactor 1: Ambient, 0.1 ml Reagent 2: 60 g of Vanillin (or p-dimethylaminobenzaldehyde) in 950 ml of Methanol Reactor 2: 90 C, 1.4 ml Reagents Flow Rate: 0.3 ml/min NOTE: Using 2-reagent system for this application greatly extends life of reagents. One-reagent method and post-column derivatization system are also available. Detection: UV-Vis detector: Vanillin λ=520 nm, DMAB λ=450 nm 35 3 Standard min 1 Monensin B 2 Monensin A 3 Narasin A POLYETHER ANTIBIOTICS CATALOG INFORMATION CATALOG NO. DESCRIPTION Polyether reversed-phase column, C 18, mm 18ECG001 Guard Cartridge Holder with 3 cartridges 18ECG002 Guard Cartridges, 2/pk. PRODUCT CATALOG PICKERING LABORATORIES 15 /16

36 REAGENTS, DILUENTS & SAMPLE PREPARATION SOLUTIONS TRIONE NINHYDRIN REAGENT CHROMATOGRAPHIC GRADE A PREPARED REAGENT FOR AUTOMATED POST-COLUMN DERIVATIZATION OF PRIMARY AND SECONDARY AMINES TRIONE Ninhydrin reagent is specially formulated for amino acid analysis. It contains Ninhydrin, Hydrindantin (reduced Ninhydrin), a Lithium Acetate buffer, and Sulfolane, a water-miscible organic solvent. The solvent is necessary to maintain the solubility of both the Hydrindantin and the primary amine product, Ruhemann s Purple. The buffer is required because the reaction is ph dependent. The active ingredients Ninhydrin and Hydrindantin are required for proper development of secondary and primary amines, respectively. TRIONE is so stable that is does not require refrigeration, either in shipment, storage, or in the reservoir. Quantitation is consistent from the first to the last ml, so there is no waste. The high signal-to-noise ratio of TRIONE, when compared to DMSO-containing reagents, permits detection sensitivity to be increased with minimum increase in background noise a feature particularly appreciated at sample concentrations of <50 picomoles. Two Preparations are available to suit your usage and storage requirements: T100 AND T100C Pour into your reservoir and use; the ultimate in convenience with a minimum of handling 4-month* shelf life T200 Combine two solutions, swirl, and use 12-month* shelf life before mixing preparation; one month in the reservoir TRIONE NINHYDRIN REAGENT CATALOG INFORMATION CATALOG NO. DESCRIPTION QUANTITY T100 TRIONE Ninhydrin Reagent (4-month* shelf life) each ( 950 ml/bottle) T100C TRIONE Ninhydrin Reagent (4-month* shelf life) case of 4 (950 ml/bottle) T200 TRIONE Two-part Ninhydrin Reagent (12-month* shelf life before mixing) to prepare case of 4 (900 ml/bottle) * From date of manufacture

37 REAGENTS, DILUENTS & SAMPLE PREPARATION SOLUTIONS o-phthalaldehyde (OPA) CHROMATOGRAPHIC GRADE FOR AUTOMATED POST-COLUMN DERIVATIZATION OF PRIMARY AMINES Primary amines form highly fluorescent adducts when reacted with o-phthalaldehyde (OPA) and a mercaptan under basic conditions. The products of this reaction, 1-alkyl-2-thioalkyl-substituted isoindoles, exhibit optimal excitation at 330 nm and maximal emission at 465 nm. Pickering s OPA is specially prepared to meet the demanding requirements of high-sensitivity pre- and post-column HPLC derivatization methods. Our repurification process actually begins with commercially available 99 % material. The entire process is controlled to eliminate trace interfering contamination. Lot quality is verified by post-column HPLC using a high-sensitivity fluorescence detector. o-phthalaldehyde (OPA) CATALOG INFORMATION CATALOG NO. DESCRIPTION QUANTITY 0120 o-phthalaldehyde (OPA) each (5 g/bottle) NAPHTHALENE-2,3-DICARBOXALDEHYDE CHROMATOGRAPHIC GRADE FOR DERIVATIZATION OF PRIMARY AMINO GROUPS Naphthalene-2,3-dicarboxaldehyde reacts with primary amines in presence of a nucleophile (e.g., mercaptan) and forms highly fluorescent 1-Alkylbenz-2-thioalkyl[f]isoindole derivatives. This reagent is especially effective for post-column or precolumn derivatization of proteins, peptides and other large amines since the derivative is stable and doesn t internally quench. Naphthalene-2,3-dicarboxaldehyde can also be used as a reagent for chemiluminescence analysis. NAPHTHALENE-2,3-DICARBOXALDEHYDE CATALOG INFORMATION CATALOG NO. DESCRIPTION QUANTITY Naphthalene-2,3-dicarboxaldehyde each (500 mg/bottle) 37 VANILLIN 4-HYDROXY-3-METHOXYBENZALDEHYDE CHROMATOGRAPHIC GRADE FOR POST- COLUMN ANALYSIS OF POLYETHER ANTIBIOTICS Reagent formulation with Vanillin, Methanol and Sulfuric Acid allows specific post-column analysis of polyether antibiotics, such as Monensin, Narasin and Salinomycin. This reagent can also be used for the post-column derivatization of Sulfa Drugs. Polyether antibiotics are monitored at 520 nm and Sulfa Drugs at 400 nm. Chromatographic Grade Vanillin meets the exacting purity requirements for a post-column chemical, ensuring that detection at 520 nm will be free of reagent artifacts. Because it is sensitive to oxygen, Pickering Laboratories Vanillin is sealed under CO 2. VANILLIN CATALOG INFORMATION CATALOG NO. DESCRIPTION QUANTITY Vanillin each (30 g/bottle) Vanillin each (300 g/bottle) PRODUCT CATALOG PICKERING LABORATORIES 15 /16

38 REAGENTS, DILUENTS & SAMPLE PREPARATION SOLUTIONS p-dimethylaminobenzaldehyde (DMAB) CHROMATOGRAPHIC GRADE FOR POST-COLUMN ANALYSIS OF SULFA DRUGS IN FEED AND ANIMAL TISSUES p-dimethylaminobenzaldehyde (DMAB; Ehrlich s Reagent) reacts rapidly with many primary amines to form a complex with maximum absorbance at 450 nm. Reversed-phase LC followed by post-column derivatization is used to determine the levels of Sulfamethazine, Sulfathiazole and related compounds in feeds, premixes and animal tissues. Although these compounds have a UV chromophore, so do most of the other constituents in the complex sample matrix. Forming a colored derivative postcolumn allows shifting of the detection wavelength into the visible range thus greatly enhancing selectivity. p-dimethylaminobenzaldehyde (DMAB) CATALOG INFORMATION CATALOG NO. DESCRIPTION QUANTITY p-dimethylaminobenzaldehyde (DMAB) each (30 g/bottle) p-dimethylaminobenzaldehyde (DMAB) each (300 g/bottle) THIOFLUOR N,N-DIMETHYL-2-MERCAPTOETHYLAMINE HYDROCHLORIDE CHROMATOGRAPHIC GRADE FOR AUTOMATED POST-COLUMN DERIVATIZATION OF PRIMARY AMINES Primary amines form highly fluorescent adducts when reacted with o-phthalaldehyde (OPA) and a mercaptan under basic conditions. The products of this reaction, 1-alkyl-2-thioalkyl-subsitituted isoindoles, exhibit optimal excitation at 330 nm and maximal emission at 465 nm. Pickering s Thiofluor, a solid, nearly odorless nucleophile, is a superior substitute for 2-Mercaptoethanol in the preparation of OPA reagents. It forms a more stable reagent and a longer-lasting fluorophore with OPA than does 2-Mercaptoethanol, yet it has the same fluorescence properties. Unlike the volatile 2-Mercaptoethanol, Thiofluor will not migrate through the gas manifold and regulator of the OPA reagent pressurization system. Two grams of Thiofluor is equivalent to 1 ml of 2-Mercaptoethanol. THIOFLUOR CATALOG INFORMATION CATALOG NO. DESCRIPTION QUANTITY Thiofluor each (10 g/bottle) o-phthalaldehyde (OPA) DILUENT CHROMATOGRAPHIC GRADE FOR PREPARATION OF o-phthalaldehyde REAGENTS The derivatization of primary amines with o-phthalaldehyde and a mercaptan requires basic conditions. To ensure the optimum ph of the reaction it is important to use the correct diluent. Three application specific diluents are available : CB910 and GA104 for Carbamate and Glyphosate analyses respectively, and OD104 for analysis of amino acids and other amines. These borate buffers are produced from starting materials which are free of heavy metals and amines. As with most other products of Pickering Laboratories, the quality of the OPA Diluents is verified by actual post-column HPLC analysis. o-phthalaldehyde (OPA) DILUENT CATALOG INFORMATION CATALOG NO. DESCRIPTION QUANTITY CB910 o-phthalaldehyde (OPA) Diluent for Carbamate analysis case of 4 (950 ml/bottle) GA104 o-phthalaldehyde (OPA) Diluent for Glyphosate analysis case of 4 (950 ml/bottle) OD104 o-phthalaldehyde (OPA) Diluent for amino acids and amines analysis case of 4 (950 ml/bottle)

39 REAGENTS, DILUENTS & SAMPLE PREPARATION SOLUTIONS HYDROLYSIS REAGENT CB130 CHROMATOGRAPHIC GRADE This 0.05 M NaOH/C47 reagent is applied in the first stage of post-column Carbamate derivatization. At 100 ºC the separated carbamates are converted from urethanes to methylamines. The methylamines can then react with OPA and Thiofluor to form the characteristically fluorescent isoindole. Hydrolysis Reagent contains C47 which prevents the precipitation of insoluble minerals (e.g. Ca 2+, Mg 2+ ) from samples of hard drinking water and ground water. The C47 additive complexes these minerals and keeps them in solution. HYDROLYSIS REAGENT CB130.2 CHROMATOGRAPHIC GRADE The new Hydrolysis Reagent CB130.2 is formulated according to USEPA Method and contains M Sodium Hydroxide. This reagent is applied during the first stage of post-column derivatization for the analysis of Carbamate pesticides. Hydrolysis Reagent CB130.2 contains C47, which prevents the precipitation of insoluble minerals from samples of hard drinking and ground water, and protects your instruments from damage. HYDROLYSIS REAGENT CATALOG INFORMATION CATALOG NO. DESCRIPTION QUANTITY CB130 Hydrolysis Reagent for Carbamate analysis 0.05 M case of 4 (950 ml/bottle) CB130.2 Hydrolysis Reagent for Carbamate analysis M case of 4 (950 ml/bottle) 39 HYPOCHLORITE DILUENT GA 116 CHROMATOGRAPHIC GRADE FOR THE PREPARATION OF THE OXIDIZING REAGENT FOR POST-COLUMN GLYPHOSATE HERBICIDE ANALYSIS This diluent is used to prepare the oxidizing reagent required to convert Glyphosate to a primary amine prior to reacting it with OPA. The reagent is prepared by adding 100 µl of 5 % solution of Sodium Hyphochlorite to one 950 ml bottle of Diluent. The ph 11.6 diluent is formulated to ensure ph stability of the mixed stream of column effluent and oxidizing reagent. This diluent could also be used in fluorescence detection of other secondary amines. HYPOCHLORITE DILUENT CATALOG INFORMATION CATALOG NO. DESCRIPTION QUANTITY GA116 Hypochlorite Diluent for Glyphosate analysis case of 4 (950 ml/bottle) PRODUCT CATALOG PICKERING LABORATORIES 15 /16

40 REAGENTS, DILUENTS & SAMPLE PREPARATION SOLUTIONS Li220 & Na220 CHROMATOGRAPHIC GRADE SAMPLES AND STANDARDS DILUENTS FOR AMINO ACID ANALYSIS Use of these diluents is essential to ensuring reproducibility from injection to injection. They establish a uniform ph and ion concentration at the outset, regardless of the source and pre-treatment of the sample. The sample is maintained buffered and at optimum ph for sample storage and analysis. Use Li220 for prepared physiological samples and to dilute amino acid calibration standards for use with Lithium columns and buffers. Use Na220 for hydrolysate samples and to dilute amino acid calibration standards for use with Sodium columns and buffers. Li220 & Na220 CATALOG INFORMATION CATALOG NO. DESCRIPTION QUANTITY Li220 Lithium Diluent, ph 2.36 case of 4 (250 ml/bottle) Na220 Sodium Diluent, ph 2.20 case of 4 (250 ml/bottle) SERAPREP & URIPREP CHROMATOGRAPHIC GRADE REAGENTS FOR THE PREPARATION OF NATIVE SAMPLES FOR AMINO ACID ANALYSIS Preparation of samples such as serum, urine, other physiological fluids, plant extracts, foods and beverages requires control of ph and normality, and removal of soluble protein. The samples must be held to a narrow ph range between 2.1 and 2.5, and at the proper Lithium ion concentration to ensure reproducibility in the early part of the chromatogram. SERAPREP and URIPREP replace commonly used protein precipitation reagents such as Acetonitrile, Perchloric acid, and Picric acid, and eliminate the need for dialysis, ultrafiltration, and the repeated centrifugation steps, followed by ph adjustment. SERAPREP is used for preparing serum and other samples with high buffering capacity, e.g. sardine oil. URIPREP is used for urine and other samples with low buffering capacity, such as fruit juices, beer and wines. The efficiency of protein precipitation and need for post-centrifugation ph adjustment of the sample determine which reagent is best for your particular sample. SERAPREP & URIPREP CATALOG INFORMATION CATALOG NO. DESCRIPTION QUANTITY SP100 SERAPREP each (250 ml/bottle) UP100 URIPREP each (250 ml/bottle)

41 REAGENTS, DILUENTS & SAMPLE PREPARATION SOLUTIONS CHLORAC BUFFER CHROMATOGRAPHIC GRADE FOR PRESERVATION OF AQUEOUS CARBAMATE SAMPLES Trace analysis of Carbamate insecticide residues in drinking water is a mandated requirement in the U.S. Since several of the common Carbamates Carbaryl, Oxamyl, Hydroxycarbofuran are labile in water due to oxidation or hydrolysis, the samples and standards must be preserved in order to obtain valid results. ChlorAC Buffer from Pickering Laboratories is a highly purified Chromatographic Grade preservative. It is prepared from Monochloracetic Acid and Potassium Acetate to EPA specifications. ChlorAC is guaranteed to be free of co-eluting interferences for the analytes in EPA CHLORAC BUFFER CATALOG INFORMATION CATALOG NO. DESCRIPTION QUANTITY ChlorAC Buffer each (250 ml/bottle) RESTORE CHROMATOGRAPHIC GRADE FOR REMOVAL OF METAL ION CONTAMINATION AND RESTORATION OF PROPER ION BALANCE IN GLYPHOSATE ANALYTICAL COLUMNS AND GUARDS Glyphosate herbicide analysis by post-column HPLC according to US EPA Method 547 employs a cation-exchange column. Many polyvalent metal ions which may be present in the sample, especially Iron, will accumulate in the guard or analytical column. As little as 100 nmole of Ferric Iron, for example, will cause serious degradation of column performance; larger amounts can actually cause the Glyphosate peak to vanish completely. Glyphosate RESTORE rapidly removes Iron and all other polyvalent metals from the column and guard. In addition, it preserves the balance of K + /H + in the resin, thus avoiding a lengthy re-equilibration. 41 RESTORE CATALOG INFORMATION CATALOG NO. DESCRIPTION QUANTITY RESTORE, removes metal ions from Glyphosate column and guard each (250 ml/bottle) EXTRACTION SOLUTION AMINOGLYCOSIDE EXTRACTION SOLUTION CATALOG INFORMATION CATALOG NO. DESCRIPTION QUANTITY Aminoglycoside Extraction Solution 4 x 950 ml/bottle PRODUCT CATALOG PICKERING LABORATORIES 15 /16

42 ELUANTS CHROMATOGRAPHIC GRADE Pickering Laboratories eluants are manufactured under strictly controlled conditions to guarantee purity, stability and consistency for a reproducible high quality chromatogram. This quality standard guarantees the resulting chromatogram will be free of any noise and interference. The Lithium and Sodium eluants are not sensitive to oxidation and do not need refrigeration, either in storage or use. Degassing is not required. However, they should be protected from air to prevent contamination. Ambient air actually contains amines and amino acids that will dissolve in the low-ph eluants and will appear in the chromatograms. All buffers are packaged in cases of four 950 ml polyethylene bottles. The regenerants are packaged in single 950 ml bottles because of the small volumes used during each analysis. Sample diluents are packaged in cases of four 250 ml borosilicate glass bottles. CUSTOM ELUANTS We develop and manufacture custom lots of eluants for customers who require our Guaranteed Chemistry commitment of purity and certified use. Let us make some for you! Call for details. ELUANTS CATALOG INFORMATION CATALOG NO. DESCRIPTION QUANTITY Amino Acid Analysis, Sodium-Based Solutions: Na270 Sodium Eluant, ph 2.80 case of 4 (950 ml/bottle) Na315 Sodium Eluant, ph 3.15 case of 4 (950 ml/bottle) Na328 Sodium Eluant, ph 3.28 case of 4 (950 ml/bottle) Na425 Sodium Eluant, ph 4.25 case of 4 (950 ml/bottle) Na640 Sodium Eluant, ph 6.40 case of 4 (950 ml/bottle) Na740 Sodium Eluant, ph 7.40 case of 4 (950 ml/bottle) * Sodium Eluant, ph 3.15, case of 4 (950 ml/bottle) with 5 % Sulfolane, * Sodium Eluant, ph 3.33, case of 4 (950 ml/bottle) with 2.5 % Sulfolane, RG011 Sodium Column Regenerant each (950 ml/bottle) Na220 Sodium Diluent, ph 2.20, for calibration standards/hydrolyzed samples case of 4 (950 ml/bottle) Amino Acid Analysis, Lithium-Based Solutions: Li275 Lithium Eluant, ph 2.75 case of 4 (950 ml/bottle) Li280 Lithium Eluant, ph 2.75 case of 4 (950 ml/bottle) Li292 Lithium Eluant, ph 2.92 case of 4 (950 ml/bottle) Lithium Eluant, ph 2.80 case of 4 (950 ml/bottle) Li365 Lithium Eluant, ph 3.65 case of 4 (950 ml/bottle) Li375 Lithium Eluant, ph 3.75 case of 4 (950 ml/bottle) Li750 Lithium Eluant, ph 7.50 case of 4 (950 ml/bottle) RG003 Lithium Column Regenerant each (950 ml/bottle) Li220 Lithium Diluent, ph 2.36, for calibration standards/prepared samples case of 4 (250 ml/bottle) Glyphosate Herbicide Analysis: K200 Glyphosate Potassium case of 4 (950 ml/bottle) Phosphate Eluant RG019 Glyphosate Column Regenerant each (950 ml/bottle) Potassium-Based Solutions for ALKION Column for Aminoglycoside Antibiotics & Streptomycin: Potassium Phosphate Eluant, KO1 case of 4 (950 ml/bottle) Potassium Hydroxide Eluant, KO2 case of 4 (950 ml/bottle) Potassium Chloride Eluant, KO3 case of 4 (950 ml/bottle) Biogenic Amines/Polyamines Analysis: K563 Potassium Phosphate Eluant, ph 5.63 case of 4 (950 ml/bottle) K600 Potassium Phosphate Eluant, ph 6.00 case of 4 (950 ml/bottle) K130 Potassium Column Regenerant each (950 ml/bottle) Muti-Residue Mycotoxin Analysis: Sodium Phosphate Eluant, ph 3.3 case of 4 (950 ml/bottle) Pure Water: Ultra High Pure Water 4 x 950 ml/bottle * For use with column with serial numbers after 1314

43 TEST MIXTURES & STANDARDS CALIBRATION STANDARDS & TEST MIXTURES Quantitative and guaranteed Each lot tested chromatographically Chromatographically-pure starting components Pickering s Amino Acid Standards have a reputation worldwide for quality and reliability in all post-column systems and methods. Six Amino Acid Standard mixtures are available for a variety of applications. The standards are in 5 ml vials in an appropriate citrate buffer. Although they are stored frozen at the factory, Pickering s calibration standards remain stable when shipped at ambient temperatures. Upon receipt, however, it is important to place them into a freezer immediately, and store until ready for use. Test Mixtures are qualitative standards and intended to be used to establish an elution profile and for troubleshooting. TEST MIXTURES CATALOG INFORMATION CATALOG NO. DESCRIPTION CONSTITUENTS Amino Acid Test Mixture Concentration 0.25 µmole/ml in 0.01 NHCl, 5.0 ml L-Cysteic Acid L-Threonine L-Serine Carbamate Test Mixture Concentration 2.5 µg/ml in Methanal, 5.0 ml Aldicarb Aldicarb Sulfone Aldicarb Sulfoxide DMC (4-Bromo-3,5-dimethylphenyl-N-methylcarbamate) Carbaryl Carbofuran 3-Hydroxycarbofuran Methiocarb Methomyl 1-Naphthol Oxamyl Propoxur Glyphosate Test Mixture Concentration 2.5 µg/ml in water, 5.0 ml Glyphosate AMPA (Aminomethyl-phosphonic acid) 43 CALIBRATION STANDARDS & CATALOG INFORMATION CATALOG NO. DESCRIPTION P Native Sample Standard with Norleucine, in 0.2 N Lithium citrate buffer ph 2.36, 5 ml P Native Sample Standard without Norleucine, in 0.2 N Lithium citrate buffer ph 2.36, 5 ml Native Sample Standard for rapid-screen for PKU and MSUD, in 0.2 N Lithium citrate buffer ph 2.36, 5 ml C Collagen Hydrolysate Standard, in 0.2 N Sodium citrate buffer ph 2.20, 5 ml H Protein Hydrolysate Standard, in 0.2 N Sodium citrate buffer ph 2.20, 5 ml Oxidized Feed Hydrolysate Standard, in 0.2 N Sodium citrate buffer ph 2.20, 5 ml Native Sample Standard without Norleucine & Alpha-Amino-Beta-guanidinopropionic acid, in 0.2 N Lithium citrate buffer ph 2.36, 5 ml Native Sample Standard, basics, in 0.1 N HCl, 5 ml Native Sample Standard, acidics and neutrals, in 0.1 N HCl, 5 ml NOTE: See next page for constituents PRODUCT CATALOG PICKERING LABORATORIES 15 /16

44 TEST MIXTURES & STANDARDS CALIBRATION STANDARDS FOR AMINO ACID ANALYSIS CONSTITUENTS * * P P C H Beta-Alanine Alanine D,L-a-Amino-adipic acid Gamma-Amino butyric acid Alpha-Amino-n-butyric acid D,L,b-Amino-i-butyric acid Alpha-Amino-Beta- guanidinopropionic acid Ammonia Anserine Arginine Asparagine Aspartic acid Carnosine Citrulline Creatinine Cystathionine Cystine (1.25) Cysteic acid Ethanolamine Glutamic acid Glycine Histidine D,L-Homocystine L,L & allo-hydroxylysine 4-trans-L-Hydroxyproline (1.25) Isoleucine Leucine Lysine Methionine Methionine-D,L-sulfoxide Methionine-D,L-sulfone 1-Methyl-histidine 3-Methyl-histidine Norleucine Ornithine Phenylalanine Phosphoethanolamine Phosphoserine Proline (1.25) Sarcosine Serine Taurine Threonine Tryptophan Tyrosine Urea Valine NOTE: Concentration of all the constituents in the Amino Acid standards is 0.25 µmole/ml unless otherwise specified. *Concentration of all the constituents is 2.5 µmole/ml unless otherwise specified

45 PRODUCT TESTING SOLUTIONS ARTIFICIAL ECCRINE PERSPIRATION The Artificial Eccrine Perspiration we offer is a ready to use solution and the closest mimic to true human eccrine sweat. It consists of 19 amino acids, the seven most abundant minerals, and the four most abundant metabolites at a ph of 4.5. All concentrations closely match experimentally determined values for adult human eccrine sweat. Custom formulations are available on request. ARTIFICIAL ECCRINE PERSPIRATION LIST OF INGREDIENTS Uric Acid Urea METABOLITES Lactic Acid Ammonia MINERALS AMINO ACIDS Sodium Iron Phosphate Calcium Copper Sulfate Magnesium Potassium Zinc Chloride Glycine L-Histidine L-Serine (Largest amount) L-Alanine L-Isoleucine L-Threonine L-Arginine L-Leucine L-Tyrosine L-Asparagine L-Lysine L-Valine L-Aspartic acid L-Methionine Taurine L-Citrulline L-Glutamic acid L-Ornithine L-Phenylalanine Pickering Laboratories is now pleased to offer our Artificial Eccrine Perspiration in a larger format in addition to the 200 ml and 5 ml quantities, we now offer our stabilized formulation in a 950 ml volume, packaged in a 1 L plastic eluent bottle. Storage and Handling The stabilized solution is preserved with a fungicide and bactericide for two years of shelf life. The non-stabilized product is at a ph of 4.5 and kept frozen. ARTIFICIAL ECCRINE CATALOG INFORMATION CATALOG NO. DESCRIPTION QTY Artificial Eccrine Perspiration 200 ml Artificial Eccrine Perspiration - stabilized 200 ml Artificial Eccrine Perspiration - stabilized 5 ml Artificial Eccrine Perspiration - stabilized 950 ml Artificial Eccrine Perspiration, custom ph 200 ml Artificial Eccrine Perspiration, custom ph - stabilized 200 ml Pickering Laboratories created our Artificial Eccrine Perspiration to universally standardize across all industries; it is the only formula that can satisfy all test challenges. Although it is the most complete formulation available, we also offer the following industry-specific artificial perspiration formulations. ISO 3160 ARTIFICIAL PERSPIRATION CORROSION RESISTANCE FOR ALLOYS This is a ready to use solution that is used to determine corrosion resistance. The non-stabilized formulation is at ph 4.7 and is kept frozen. (Custom ph and stabilized versions available) ISO 3160 ARTIFICIAL PERSPIRATION CATALOG INFORMATION CATALOG NO. DESCRIPTION QTY ISO 3160 Artificial Perspiration 200 ml ISO 3160 Artificial Perspiration - stabilized 200 ml ISO 3160 Artificial Perspiration, custom ph 200 ml ISO 3160 Artificial Perspiration, custom ph - stabilized 200 ml ISO 105-B07 AND ISO 105-E04 ARTIFICIAL PERSPIRATION COLORFASTNESS TO LIGHT FOR FABRIC These are ready to use solutions that are used to test the colorfastness to light of a fabric saturated with either the acidic or alkaline solution. The ph of the acidic solution is 5.5 and that of the alkaline solution is 8.0. The non-stabilized product is kept frozen. (Custom ph and stabilized versions available) ISO 105-B07 AND ISO 105-E04 ARTIFICIAL PERSPIRATION CATALOG INFORMATION CATALOG NO. DESCRIPTION QTY ISO 105-B07 / ISO 105-E04 Artificial Perspiration, 200 ml ph ISO 105-B07 / ISO 105-E04 Artificial Perspiration, 200 ml ph ISO 105-B07 / ISO 105-E04 Artificial Perspiration, 200 ml ph stabilized ISO 105-B07 / ISO 105-E04 Artificial Perspiration, 200 ml ph stabilized ISO 105-B07 / ISO 105-E04 Artificial Perspiration, 200 ml acidic custom ph ISO 105-B07 / ISO 105-E04 Artificial Perspiration, 200 ml alkaline custom ph ISO 105-B07 / ISO 105-E04 Artificial Perspiration, 200 ml acidic custom ph - stabilized ISO 105-B07 / ISO 105-E04 Artificial Perspiration, alkaline custom ph - stabilized 200 ml 45 PRODUCT CATALOG PICKERING LABORATORIES 15 /16

46 PRODUCT TESTING SOLUTIONS AATCC TEST METHOD 15 ARTIFICIAL PERSPIRATION COLORFASTNESS TO PERSPIRATION FOR FABRIC This ready to use solution is used to determine the fastness of colored textiles to the effects of acid perspiration. The non-stabilized formulation at ph 4.3 and is kept frozen. (Custom ph and stabilized versions available) AATCC TEST METHOD 15 ARTIFICIAL PERSPIRATION CATALOG INFORMATION CATALOG NO. DESCRIPTION QTY AATCC Test Method 15 Artificial Perspiration 200 ml AATCC Test Method 15 Artificial Perspiration - stabilized 200 ml AATCC Test Method 15 Artificial Perspiration, custom ph 200 ml AATCC Test Method 15 Artificial Perspiration, custom ph - stabilized 200 ml ISO 1164 ARTIFICIAL PERSPIRATION COLORFASTNESS TO PERSPIRATION FOR LEATHER This ready to use solution is used to determine the fastness of leather to perspiration. The non-stabilized formulation at ph 8.0 and is kept frozen. (Custom ph and stabilized versions available) ISO 1164 ARTIFICIAL PERSPIRATION CATALOG INFORMATION CATALOG NO. DESCRIPTION QTY ISO 1164 Artificial Perspiration 200 ml ISO 1164 Artificial Perspiration- stabilized 200 ml ISO 1164 Artificial Perspiration, custom ph 200 ml ISO 1164 Artificial Perspiration, custom ph - stabilized 200 ml ISO ARTIFICIAL PERSPIRATION OPHTHALMIC OPTICS This ready to use solution is used to determine the effect of sweat on ophthalmic optics and spectacle frames. The nonstabilized formulation is kept frozen. (Custom ph and stabilized versions available) ISO ARTIFICIAL PERSPIRATION CATALOG INFORMATION CATALOG NO. DESCRIPTION QTY ISO Artificial Perspiration 200 ml ISO Artificial Perspiration- stabilized 200 ml ISO Artificial Perspiration, custom ph 200 ml ISO Artificial Perspiration, custom ph - stabilized 200 ml BS EN 1811:2011 ARTIFICIAL PERSPIRATION FOR RELEASE OF NICKEL This ready to use solution is used in the test method for release of Nickel from all post assembles which are inserted into pierced parts of the human body and articles intended to come into direct and prolonged contact with skin. This non-stabilized version is kept frozen. (Custom ph and stabilized versions available) BS EN 1811:2011 ARTIFICIAL PERSPIRATION CATALOG INFORMATION CATALOG NO. DESCRIPTION QTY BS EN 1811:2011 Artificial Perspiration 200 ml BS EN 1811:2011 Artificial Perspiration - stabilized 200 ml BS EN 1811:2011 Artificial Perspiration, custom ph 200 ml BS EN 1811:2011 Artificial Perspiration, custom ph - stabilized 200 ml DIN : ARTIFICIAL PERSPIRATION COLORFASTNESS OF ARTICLES FOR COMMON USE This ready to use solution is used in the test method to determine colorfastness of articles for common use to perspiration. This non-stabilized version is kept frozen. (Custom ph version available) DIN : ARTIFICIAL PERSPIRATION CATALOG INFORMATION CATALOG NO. DESCRIPTION QTY DIN : Artificial Perspiration 200 ml DIN : Artificial Perspiration - stabilized 200 ml DIN : Artificial Perspiration, custom ph 200 ml DIN : Artificial Perspiration, custom ph - stabilized 200 ml ANSI-BHMA A ARTIFICIAL PERSPIRATION FOR TESTING BUILDERS HARDWARE AND FINISHES This ready to use solution is used in the test method specified by the builder s hardware association. (Custom ph version available) ISO 1164 ARTIFICIAL PERSPIRATION CATALOG INFORMATION CATALOG NO. DESCRIPTION QTY ANSI-BHMA A Artificial Perspiration 200 ml ANSI-BHMA A Artificial Perspiration, custom ph 200 ml

47 PRODUCT TESTING SOLUTIONS FUSAYAMA/MAYER ARTIFICIAL SALIVA FOR TESTING OF PRODUCTS FOR CORROSION, COLORFASTNESS AND DISCOLORATION This ready-to-use solution closely resembles the mineral composition of natural saliva and is the most common media used for testing dental metal alloys. This formulation is at ph 4.9+/- 0.1 and should be stored refrigerated. (Custom formulations at different ph are available) AFNOR NF S ARTIFICIAL SALIVA FOR TESTING BIODEGRADABILITY OF DENTAL METAL ALLOYS Artificial Saliva is prepared according to AFNOR NF S standard procedure and is intended for testing biodegradability of dental metal alloys. The formulation consists of two parts that are mixed right before use. This minimizes changes in the solution during storage and allows for a longer shelf life. The ph of the solution after mixing is 7.8 +/ The Artificial Saliva should be stored refrigerated both before and after mixing. Premixed, ready-to-use solution is available upon request. DIN : ARTIFICIAL SALIVA TO DETERMINE COLORFASTNESS OF PRODUCTS INTENDED TO BE TAKEN INTO THE MOUTH Artificial Saliva is prepared according to DIN : standard procedure. DIN specifies the method to determine colorfastness of products intended to be taken into the mouth. The formulation consists of two parts that are mixed right before use. This minimizes changes in the solution during storage and allows for a longer shelf life. The solution should be stored refrigerated both before and after mixing. The ph of the solution after mixing is 6.8 +/ ARTIFICIAL SALIVA FOR PHARMACEUTICAL RESEARCH Artificial Saliva is formulated according to literature for pharmaceutical research such as studies of drug dissolution and drug delivery through oral mucosa. This is a ready to use formulation that should be stored refrigerated. The ph of the solution is 6.8 +/ ARTIFICIAL SALIVA FOR MEDICAL AND DENTAL RESEARCH This Artificial Saliva is formulated according to literature references for medical and dental research. This formulation has similar composition to commercially available products used to treat dry mouth and other conditions. This ready to use solution contains Sodium Carboxymethyl Cellulose to increase viscosity of the solution and make it behave similar to natural human saliva. This formulation can be stored at room temperature and has ph of / But this solution is only intended for product testing and research, and not for medical use. ARTIFICIAL SALIVA CATALOG INFORMATION CATALOG NO. DESCRIPTION QTY Fusayama / Meyer Artificial Saliva 200 ml AFNOR NF S Artificial Saliva 200 ml DIN : Artificial Saliva 200 ml Artificial Saliva for pharmaceutical research 200 ml Artificial Saliva for medical and dental research 200 ml DIN EN 1616:1999 ARTIFICIAL URINE FOR TESTING STERILE URETHRAL CATHETERS Artificial Urine is prepared according to DIN EN 1616:1999 standard procedure. DIN EN 1616 specifies the method to test sterile urethral catheters. This ready-to-use solution should be stored frozen to avoid bacteria growth. The ph of the solution is 6.6+/ ARTIFICIAL URINE FOR CORROSION TESTING OF UROLOGICAL IMPLANTS, STABILIZED This artificial urine is designed for testing metallic biomaterials used to produce urological implants and catheters. This convenient product is a ready to use solution. The formulation contains non-toxic preservative to avoid bacteria growth and can be stored at room temperature. The ph of the final solution is 6.0+/ (Custom formulations are also available) ARTIFICIAL URINE MEDIUM FOR GROWING UROLOGICAL PATHOGENS This ready-to-use solution closely resembles composition of human urine and can be used for clinical studies as well as for product testing. This formulation supports growth of wide range of urinary pathogens and it is also capable of forming crystals similar to these found in natural urinary tract infections. It can be used as negative controls in laboratory testing. ph of the Artificial Urine Medium is 6.5+/ This product is stored frozen. ARTIFICIAL URINE CATALOG INFORMATION CATALOG NO. DESCRIPTION QUANTITY DIN EN 1616:1999 Artificial Urine 200 ml Artificial Urine for Corrosion Testing of 200 ml Urological Implants, Stabilized Artificial Urine Medium for Growing Urological Pathogens 200 ml 47 PRODUCT CATALOG PICKERING LABORATORIES 15 /16

48 APPLICATION KITS AMINO ACID ANALYSIS KITS AMINO ACID ANALYSIS KITS CATALOG NO. DESCRIPTION MIN HIGH-EFFICIENCY COLLAGEN HYDROLYSATE KIT: T 30-Minute Sodium Cation-exchange Column 4.6 x 110 mm & amino acid test mixture Cation-exchange GARD assembly: Holder w/ 2 replaceable GARDs Na220 Sodium Diluent, ph 2.20, 4 x 250 ml Na315 Sodium Eluant, ph 3.15, case of 4 ( 950 ml/bottle) Na425 Sodium Eluant, ph 4.25, 4 x 950 ml Na640 Sodium Eluant, ph 6.40, 4 x 950 ml RG011 Sodium Column Regenerant, 950 ml T100C TRIONE Ninhydrin Reagent, 4 x 950 ml (4-month shelf life) C Sodium Calibration Standard, for collagen hydrolysate, 0.25 µmole/ml, 5 ml KIT IDENTICAL TO WITH T200 REPLACING T100C: T200 TRIONE Two-part Ninhydrin Reagent, prepares 4 x 900 ml (12-month shelf life) KIT IDENTICAL TO WITH ITEMS BELOW REPLACING T100C: OD104 OPA Diluent, 4 x 950 ml O120 o-phthalaldehyde, 5 g Thiofluor, 10 g (2 each per kit) MIN HIGH-EFFICIENCY PROTEIN HYDROLYSATE KIT: T 30-Minute Sodium Cation-exchange Column 4.6 x 110 mm & amino acid test mixture Cation-exchange GARD assembly: Holder w/ 2 replaceable GARDs Na220 Sodium Diluent, ph 2.20, 4 x 250 ml Na315 Sodium Eluant, ph 3.15, case of 4 ( 950 ml/bottle) Na425 Sodium Eluant, ph 4.25, 4 x 950 ml Na640 Sodium Eluant, ph 6.40, 4 x 950 ml RG011 Sodium Column Regenerant, 950 ml T100C TRIONE Ninhydrin Reagent, 4 x 950 ml (4-month shelf life) H Sodium Calibration Standard, for protein hydrolysate, 0.25 µmole/ml, 5 ml KIT IDENTICAL TO WITH T200 REPLACING T100C: T200 TRIONE Two-part Ninhydrin Reagent, prepares 4 x 900 ml (12-month shelf life) KIT IDENTICAL TO WITH ITEMS BELOW REPLACING T100C: OD104 OPA Diluent, 4 x950 ml O120 o-phthalaldehyde, 5 g Thiofluor, 10 g (2 each per kit) MIN HIGH EFFICIENCY OXIDIZED FEED HYDROLYSATE KIT: T 30-Minute Sodium Cation-exchange Column 4.6 x 110 mm & amino acid test mixture Cation-exchange GARD assembly: Holder w/ 2 replaceable GARDs Na220 Sodium Diluent, ph 2.20, 4 x 250 ml Na270 Sodium Eluant, ph 2.80, 4 x 950 ml Na425 Sodium Eluant, ph 4.25, 4 x 950 ml Na640 Sodium Eluant, ph 6.40, 4 x 950 ml RG011 Sodium Column Regenerant, 950 ml T100C TRIONE Ninhydrin Reagent, 4 x 950 ml (4-month shelf life) Sodium Calibration Standard, for oxidized feed hydrolysate, 0.25 µmole/ml, 5 ml KIT IDENTICAL TO WITH T200 REPLACING T100C: T200 TRIONE Two-part Ninhydrin Reagent, prepares 4 x 900 ml (12-month shelf life) KIT IDENTICAL TO WITH ITEMS BELOW REPLACING T100C: OD104 OPA Diluent, 4 x 950 ml O120 o-phthalaldehyde, 5 g Thiofluor, 10 g (2 each per kit) AT30SH STANDARD 60-MIN PROTEIN HYDROLYSATE KIT: Sodium Ion-exchange Column, 3.0 x 250 mm (with test mixture ) Cation-exchange GARD assembly: Holder w/ 2 replaceable GARDs Na328 Na740 T100C RG011 Na H Sodium Eluant, ph 3.28, 4 x 950 ml Sodium Eluant, ph 7.40, 4 x 950 ml TRIONE Ninhydrin Reagent, 4 x 950 ml (4-month shelf life) Sodium Column Regenerant, 950 ml Sodium Diluent, ph 2.20, 4 x 250 ml Sodium Calibration Standard, for protein hydrolysate, 0.25 µmole/ml, 5 ml KIT IDENTICAL TO AT30SH WITH T200 REPLACING T100C: T200 TRIONE Two-part Ninhydrin Reagent, prepares 4 x 900 ml (12-month shelf life) AO30SH KIT IDENTICAL TO AT30SH WITH ITEMS BELOW REPLACING T100C: OD104 OPA Diluent, 4 x950 ml O120 o-phthalaldehyde, 5 g Thiofluor, 10 g (2 each per kit) AT31FH HIGH-EFFICIENCY 55-MIN PROTEIN HYDROYSIS KIT: T Sodium Ion-exchange Column, 4.0 x 150 mm (with test mixture ) Cation-exchange GARD assembly: Holder w/ 2 replaceable GARDs Na315 Sodium Eluant, ph 3.15, 4 x 950 ml Na740 Sodium Eluant, ph 7.40, 4 x 950 ml T100C TRIONE Ninhydrin Reagent, 4 x 950 ml (4-month shelf life) RG011 Sodium Column Regenerant, 950 ml Na220 Sodium Diluent, ph 2.20, 4 x 250 ml H Sodium Calibration Standard, for protein hydrolysate, 0.25 µmole/ml, 5 ml KIT IDENTICAL TO AT31FH WITH T200 REPLACING T100C: T200 TRIONE Two-part Ninhydrin Reagent, prepares 4 x 900 ml (12-month shelf life) AO31FH KIT IDENTICAL TO AT31FH WITH ITEMS BELOW REPLACING A T100C: OD104 OPA Diluent, 4 x 950 ml O120 o-phthalaldehyde, 5 g Thiofluor, 10 g (2 each per kit) AT32FC HIGH-EFFICIENCY 55-MIN COLLAGEN HYDROLYSATE KIT: T Sodium Ion-exchange Column, 4.0 x 150 mm (with test mixture ) Cation-exchange GARD assembly: Holder w/ 2 replaceable GARDs Na315 Sodium Eluant, ph 3.15, 4 x 950 ml Na740 Sodium Eluant, ph 7.40, 4 x 950 ml T100C TRIONE Ninhydrin Reagent, 4 x 950 ml (4-month shelf life) RG011 Sodium Column Regenerant, 950 ml Na220 Sodium Diluent, ph 2.20, 4 x 250 ml C Sodium Calibration Standard, for collagen hydrolysate 0.25 µmole/ ml, Proline and Hydroxyproline, 1.25 µmole/ml, 5 ml KIT IDENTICAL TO AT32FC WITH T200 REPLACING T100C: T200 TRIONE Two-part Ninhydrin Reagent, prepares 4 x 900 ml (12-month shelf life) AO32FC KIT IDENTICAL TO AT32FC WITH ITEMS BELOW REPLACING T100C: OD104 OPA Diluent, 4 x 950 ml O120 o-phthalaldehyde, 5 g Thiofluor, 10 g (2 each per kit)

49 APPLICATION KITS MIN OXIDIZED FEED HYDROLYSATE KIT: T Sodium Ion-exchange Column, 4.0 x 150 mm (with test mixture ) Cation-exchange GARD assembly: Holder w/ 2 replaceable GARDs Na270 Na740 T100C RG011 Na220 Sodium Eluant, ph 2.80, 4 x 950 ml Sodium Eluant, ph 7.40, 4 x 950 ml TRIONE Ninhydrin Reagent, 4 x 950 ml (4-month shelf life) Sodium Column Regenerant, 950 ml Sodium Diluent, ph 2.20, 4 x 250 ml Sodium Calibration Standard, for oxidized feed hydrolysate, 0.25 µmole/ml, 5 ml KIT IDENTICAL TO WITH T200 REPLACING T100C: T200 TRIONE Two-part Ninhydrin Reagent, prepares 4 x 900 ml (12-month shelf life) KIT IDENTICAL TO W/ ITEMS BELOW REPLACING T100C OD104 OPA Diluent, 4 x 950 ml O120 o-phthalaldehyde, 5 g Thiofluor, 10 g, (2 each per kit) MIN HIGH-EFFICIENCY PHYSIOLOGIC FLUID/NATIVE SAMPLE KIT: T Lithium Ion-exchange Column, 4.6 x 75 mm (with test mixture ) Cation-exchange GARD assembly: Holder w/ 2 replaceable GARDs Lithium Eluant, ph 2.80, 4 x 950 ml Li220 Lithium Diluent, ph 2.20, 4 x 250 ml Li365 Lithium Diluent, ph 3.65, 4 x 950 ml Li375 Lithium Diluent, ph 3.75, 4 x 950 ml RG003 Lithium Column Regenerant, 950 ml Native Sample Standard without Norleucine & Alpha-Amino-Betaguanidinopropionic acid, in 0.2 N Lithium citrate buffer ph 2.36, 5 ml SP100 SERAPREP, 250 ml UP100 URIPREP, 250 ml T100C TRIONE Ninhydrin Reagent, 4 x 950 ml (4-month shelf life) KIT IDENTICAL TO WITH T200 REPLACING T100C: T200 TRIONE Two-part Ninhydrin Reagent, prepares 4 x 900 ml (12-month shelf life) KIT IDENTICAL TO WITH ITEMS BELOW REPLACING T100C: OD104 OPA Diluent, 4 x 950 ml O120 o-phthalaldehyde, 5 g Thiofluor, 10 g (2 each per kit) AT33SP STANDARD 185-MIN PHYSIOLOGIC FLUID/NATIVE SAMPLE KIT: Lithium Ion-exchange Column, 3.0 x 250 mm (with test mixture ) Cation-exchange GARD assembly: Holder w/ 2 replaceable GARDs Li275 Lithium Eluant, ph 2.75, 4 x 950 ml Li750 Lithium Eluant, ph 7.50, 4 x 950 ml T100C TRIONE Ninhydrin Reagent, 4 x 950 ml (4-month shelf life) RG003 Lithium Column Regenerant, 950 ml Li220 Lithium Diluent, ph 2.36, 4 x 250 ml P Lithium Calibration Standard, with norleucine, 0.25 µmole/ml, 5 ml SP100 SERAPREP, 250 ml UP100 URIPREP, 250 ml KIT IDENTICAL TO AT33SP WITH T200 REPLACING T100C: T200 TRIONE Two-part Ninhydrin Reagent, prepares 4 x 900 ml (12-month shelf life) AO33SP KIT IDENTICAL TO AT33SP WITH ITEMS BELOW REPLACING T100C: OD104 OPA Diluent, 4 x 950 ml O120 o-phthalaldehyde, 5 g Thiofluor, 10 g (2 each per kit) MIN HIGH-EFFICIENCY PHYSIOLOGIC FLUID/NATIVE SAMPLE KIT: T Lithium Ion-exchange Column, 4.0 x 100 mm (with test mixture ) Cation-exchange GARD assembly: Holder w/ 2 replaceable GARDs Li275 Lithium Eluant, ph 2.75, 4 x 950 ml Li750 Lithium Eluant, ph 7.50, 4 x 950 ml T100C TRIONE Ninhydrin Reagent, 4 x 950 ml (4-month shelf life) RG003 Lithium Column Regenerant, 950 ml Li220 Lithium Diluent, ph 2.36, 4 x 250 ml P Lithium Calibration Standard, with norleucine, 0.25 µmole/ml, 5 ml SP100 SERAPREP, 250 ml UP100 URIPREP, 250 ml KIT IDENTICAL TO WITH T200 REPLACING T100C: T200 TRIONE Two-part Ninhydrin Reagent, prepares 4 x 900 ml (12-month shelf life) KIT IDENTICAL TO WITH ITEMS BELOW REPLACING T100C: OD104 OPA Diluent, 4 x 950 ml O120 o-phthalaldehyde, 5 g Thiofluor, 10 g ( 2 each per kit) TEMPERATURE GRADIENT 90 MIN PHYSIOLOGIC FLUID/NATIVE SAMPLE KIT: T Lithium Ion-exchange Column, 4.0 x 100 mm (with test mixture ) Cation-exchange GARD assembly: Holder w/ 2 replaceable GARDs Lithium Eluant, ph 2.80, 4 x 950 ml Li220 Lithium Diluent, ph 2.20, 4 x 250 ml Li365 Lithium Diluent, ph 3.65, 4 x 950 ml Li375 Lithium Diluent, ph 3.75, 4 x 950 ml RG003 Lithium Column Regenerant, 950 ml Native Sample Standard without Norleucine & Alpha-Amino-Betaguanidinopropionic acid, in 0.2 N Lithium citrate buffer ph 2.36, 5 ml SP100 SERAPREP, 250 ml UP100 URIPREP, 250 ml T100C TRIONE Ninhydrin Reagent, 4 x 950 ml (4-month shelf life) KIT IDENTICAL TO WITH T200 REPLACING T100C: T200 TRIONE Two-part Ninhydrin Reagent, prepares 4 x 900 ml (12-month shelf life) KIT IDENTICAL TO WITH ITEMS BELOW REPLACING T100C: OD104 OPA Diluent, 4 x 950 ml O120 o-phthalaldehyde, 5 g Thiofluor, 10 g (2 each per kit) 49 PRODUCT CATALOG PICKERING LABORATORIES 15 /16

50 APPLICATION KITS PESTICIDE & HERBICIDE ANALYSIS KITS CATALOG NO. PESTICIDE & HERBICIDE ANALYSIS KITS DESCRIPTION CARBAMATE PESTICIDES ANALYSIS KIT, 23+ COMPOUNDS: ECG001 CB130 CB910 O120 Analytical Column, expanded resolution, C 8, mm (with Carbamate Test Mixture ) Guard Cartridge Holder with 3 guard cartridges Carbamate Hydrolysis Reagent, ml Carbamate OPA Diluent, ml o-phthalaldehyde, 5 g Thiofluor, 10 g (2 each per kit) ChlorAC Buffer, 250 ml CARBAMATE PESTICIDES ANALYSIS KIT, FOR EPA METHOD 531.1: ECG001 CB130 CB910 O120 Analytical Column, high resolution/capacity, C 8, mm (with Carbamate Test Mixture ) Guard Cartridge Holder with 3 guard cartridges Carbamate Hydrolysis Reagent, ml Carbamate OPA Diluent, ml o-phthalaldehyde, 5 g Thiofluor, 10 g (2 each per kit) ChlorAC Buffer, 250 ml CARBAMATE PESTICIDES ANALYSIS KIT, FOR EPA METHOD 531.2: ECG001 CB130.2 CB910 O120 Analytical Column, high resolution/capacity, C 8, mm (with Carbamate Test Mixture ) Guard Cartridge Holder with 3 guard cartridges Carbamate Hydrolysis Reagent, ml Carbamate OPA Diluent, ml o-phthalaldehyde, 5 g Thiofluor, 10 g (2 each per kit) ChlorAC Buffer, 250 ml CARBAMATE PESTICIDES ANALYSIS KIT, FOR AOAC METHOD : Analytical Column, rapid analysis/qc, C 18, mm (with Carbamate Test Mixture ) 18ECG001 Guard Cartridge Holder with 3 guard cartridges CB130 Carbamate Hydrolysis Reagent, ml CB910 Carbamate OPA Diluent, ml O120 o-phthalaldehyde, 5 g Thiofluor, 10 g (2 each per kit) ChlorAC Buffer, 250 ml GLYPHOSATE HERBICIDE ANALYSIS KIT, FOR EPA METHOD 547, AOAC METHOD : Cation-exchange Column, form, mm (with Glyphosate Test Mixture ) Cation-exchange GARD assembly: Holder w/ 2 replaceable GARDs K200 Glyphosate Potassium Phosphate Eluant, ml RG019 Glyphosate Column Regenerant, 950 ml GA104 Glyphosate OPA Diluent, ml GA116 Glyphosate Hypochlorite Diluent, ml O120 o-phthalaldehyde, 5 g Thiofluor, 10 g (2 each per kit) RESTORE, 250 ml OTHER POST-COLUMN ANALYSIS KITS OTHER POST-COLUMN ANALYSIS KITS CATALOG NO. DESCRIPTION BIOGENIC AMINES/POLYAMINES ANALYSIS KIT: ALKION Cation-exchange Column, K + form, mm ALKION Guard Column, K + form, mm Flow Restrictor, 300 psi, 10 cm K563 Potassium Phosphate Eluant, ph 5.63, ml K600 Potassium Phosphate Eluant, ph 6.00, ml K130 Potassium Column Regenerant, 950 ml OD104 OPA Diluent, ml O120 o-phthalaldehyde, 5 g Thiofluor, 10 g (2 each per kit) AMINOGLYCOSIDE ANTIBIOTICS ANALYSIS KIT: ALKION Cation-exchange Column, K + form, mm ALKION Guard Column, K + form, mm Flow Restrictor, 300 psi, 10 cm Potassium Phosphate Eluant, KO1, ml Potassium Hydroxide Eluant, KO2, ml Potassium Chloride Eluant, KO3, ml OD104 OPA Diluent, ml O120 o-phthalaldehyde, 5 g Thiofluor, 10 g (2 each per kit) Aminoglyside extraction solution 4 x 950 ml MULTI-RESIDUE MYCOTOXIN KIT: MYCOTOX Reversed-phase Column, C 18, mm 18ECG001 Guard Cartridge Holder with 3 guard cartridges OD104 OPA Diluent, ml O120 o-phthalaldehyde, 5 g Sodium Phosphate Eluant, ph Thiofluor, 10 g (2 each per kit) AFLATOXINS ANALYSIS KIT: MYCOTOX Reversed-phase Column, C 18, mm 18ECG001 Guard Cartridge Holder with 3 guard cartridges POLYETHER ANTIBIOTICS KIT: Polyether Column, C 18, mm 18ECG001 Guard Cartridge Holder with 3 cartridges Vanillin, 30 g (2 each per kit)

51 REPLACEMENT COMPONENTS, FITTINGS & TUBING PINNACLE PCX FLOW DIAGRAM TWO-PUMP SYSTEM Flow Diagram - Duplex Nut: Ferrule: Nut: Ferrule: Nut: Ferrule: COLUMN PUMP 2 FILTER TRANSDUCER VALVE R2 FLUSH R1 Nut: Ferrule: Nut: Ferrule: HEATED REACTOR AMBIENT REACTOR Nut: Ferrule: TRANSDUCER Nut: Ferrule: Coupler: FILTER Nut: Ferrule: Nut: Ferrule: Nut: Ferrule: Check Valve: Nut: Ferrule: FROM HPLC WASTE PUMP VALVE TO DETECTOR Nut: Ferrule: Nut: Ferrule: Union: Back Pressure Regulator: Nut: Ferrule: bold = kit PRODUCT CATALOG PICKERING LABORATORIES 15 /16

52 REPLACEMENT COMPONENTS, FITTINGS & TUBING PINNACLE PCX FLOW DIAGRAM ONE-PUMP SYSTEM Flow Diagram - Simplex Nut: Ferrule: R1 Nut: Ferrule: FLUSH Nut: Ferrule: COLUMN FILTER HEATED REACTOR Coupler: Nut: Ferrule: TRANSDUCER Nut: Ferrule: PUMP VALVE Nut: Ferrule: Nut: Ferrule: Nut: Ferrule: Nut: Ferrule: Check Valve: Nut: Ferrule: FROM HPLC WASTE TO DETECTOR Nut: Ferrule: Union: Back Pressure Regulator: bold = kit

53 REPLACEMENT COMPONENTS, FITTINGS & TUBING VECTOR PCX FLOW DIAGRAM (PEEK Tubing) (Liquid Manifold) (Filter Frit) from HPLC Column 500 psi Over Pressure Relief Valve A E HEATED REACTOR A E (Filter Frit) (Filter Manifold) RESTRICTOR B RESTRICTOR AMBIENT REACTOR µm filter E A E E 0.5 µm filter to Detector A (Pulse Dampener Rebuild Kit) PULSE DAMPENER C PULSE DAMPENER (Pulse Dampener Rebuild Kit) E E (Pressure Switch) to Injector from Pump E 425 psi E D B F PUMP 1 A (Vector Pump Components) D PUMP 2 D F Reagent Reservoir & Cap Assemblies BOTTLE BOTTLE A B Nut: Ferrule: Nut: Ferrule: C D Nut: Ferrule: Nut: Ferrule: E F (PEEK Tubing, 0.02 ID) PRODUCT CATALOG PICKERING LABORATORIES 15 /16

54 REPLACEMENT COMPONENTS, FITTINGS & TUBING PINNACLE PCX REAGENT PUMP AND VALVE COMPONENTS CATALOG NO Pinnacle PCX Pump Seal Kit DESCRIPTION Pinnacle PCX Pump Cylinder, Ceramic 70 ml Pinnacle PCX Pump Assembly, 70 ml with motor Pinnacle PCX Valve Face Assembly, liquid end Pinnacle PCX Valve Assembly, Valve Face and motor Pinnacle PCX Pressure Tranducer Assembly Pinnacle PCX Valve Maintenance Kit (Seals and Tools) Pinnacle PCX Valve Seal Tool Pinnacle PCX Valve Seals Kit Pinnacle Dead-head Kit for Transducer VECTOR PCX REAGENT PUMP COMPONENTS CATALOG NO. DESCRIPTION Pump Check Valve Kit PEEK (Inlet & Outlet) Seal Kit (Piston seal, Back-up O-ring, Diaphragm, Tool) Pulse Damper Rebuild Kit (Diaphragm, O-rings, Diaphragm seal, Seal tool, 4 Hex wrenches) Prime/Purge Valve PEEK Head Kit, Vector Pump, (head, check valves, piston and seal) Piston, Vector Pump Prime Purge Seal Kit PEEK Piston retainer bushing Ferrule, Peek 1/16 x S/T, 5 each Nut, Peek Long 1/16 x S/T, 5 each Check Valve Cartridge PEEK (2/pk) Vector PINNACLE PCX REPLACEMENT REACTOR CARTRIDGE CATALOG NO. DESCRIPTION Reactor Cartridge Coil Assembly ml 130 C Reactor Cartridge Coil Assembly ml 130 C Reactor Cartridge Coil Assembly ml 130 C Reactor Cartridge Knitted Assembly ml 130 C Reactor Cartridge Knitted Assembly ml 130 C Reactor Cartridge Knitted Assembly ml 80 C Reactor Cartridge Knitted Assembly ml 80 C Reactor Cartridge Knitted Assembly ml 80 C Reactor Cartridge Knitted Assembly ml & 1.6 ml 130 C Reactor Cooling Coil, Voglibose VECTOR PCX SUBSTITUTE REACTOR VOLUMES OR REPLACEMENT REACTORS CATALOG NO. DESCRIPTION Reactor Heater & Coil Assembly, 0.5 ml 130 C, 120 V Reactor Heater & Coil Assembly, 0.5 ml 130 C, 240 V Reactor Heater & Knitted Assembly, 1.0 ml, 130 C, 120 V Reactor Heater & Knitted Assembly, 1.0 ml, 130 C, 240 V Reactor Heater & Knitted Assembly, 1.4 ml, 130 C, 120 V Reactor Heater & Knitted Assembly, 1.4 ml, 130 C, 240 V Reactor Heater & Knitted Assembly, 1.2 & 1.6 ml, 130 C, 120 V Reactor Heater & Knitted Assembly, 1.2 & 1.6 ml, 130 C, 240 V Reactor Heater & Knitted Assembly, 2.0 ml, 130 C, 120 V Reactor Heater & Knitted Assembly, 2.0 ml, 130 C, 240 V Reactor Heater & Knitted Assembly, 2.8 ml, 130 C, 120 V Reactor Heater & Knitted Assembly, 2.8 ml, 130 C, 240 V Reactor Heater & Knitted Assembly, 3.0 ml, 130 C, 120 V Reactor Heater & Knitted Assembly, 3.0 ml, 130 C, 240 V Reactor Heater & Knitted Assembly, 3.5 ml, 130 C, 120 V Reactor Heater & Knitted Assembly, 3.5 ml, 130 C, 240 V OPA Ambient Reactor, in ID TFE Tubing REAGENT RESERVOIRS & CAP ASSEMBLIES CATALOG NO. DESCRIPTION Reservoir Bottle, safety-coated, 1 L Reservoir Bottle, safety-coated, 2 L Reservoir Assembly, includes 1 L bottle, and cap with integrated valve Cap Assembly for 1 L bottle, includes cap with integrated valve Cap Assembly for 2 L bottle, includes cap with integrated valve Reservoir Cap Cap for Flush Bottle GL-38, Black, Pinnacle PCX Cap for Wash Bottle GL-38, Black, Vector PCX & Pinnacle PCX Bottle, Piston Wash with cap, Pinnacle PCX & Vector PCX Bottle, System Flush with Cap, Pinnacle PCX NOTE: Tubing, nuts and ferrules sold separately PRESSURE REGULATORS, VALVES & GAUGES CATALOG NO. DESCRIPTION Pressure Regulator, 100 psi, 1/4-28 connection, for detector waste, Vector and Pinnacle PCX Gas Manifold Assembly, Vector PCX and Pinnacle PCX Inline Check Valve, Pinnacle PCX Check Valve, Reagent Cap

55 REPLACEMENT COMPONENTS, FITTINGS & TUBING FILTERS & FRITS CATALOG NO. DESCRIPTION Replacement Reagent 10 µm Frit Filter Frit, 2 µm Reagent Filter Filter Frit, 0.5 µm PEEK Vector 0.5 µm Filter Upgrade Kit TEES & UNIONS FLOW RESTRICTORS Pickering restrictors are tubes packed with sized diamond particles designed to provide a desired back-pressure, relative to a given flow rate and viscosity. The restrictor tube and fittings are type 316 stainless steel or PEEK. The diamond packing is chemically and chromatographically inert to all mobile phases. FEATURES Pressure-calibrated to flow rate and viscosity Diamond-packed tubes with nuts and ferrules Improve pump efficiency Essential pulse-dampening component Troubleshooting aid for HPLC systems CATALOG NO. DESCRIPTION Flow Restrictor, for OPA, Hypochlorite & Hydrolysis reagent, ml/min, 10 cm Flow Restrictor, TRIONE Ninhydrin reagent, ml/min, 6 cm Flow Restrictor, ml/min, 10 cm Flow Restrictor, ml/min, 6 cm Flow Restrictor, ml/min, 17 cm Flow Restrictor, ml/min, 20 cm Flow Restrictor, PEEK Tubing for Polyether antibiotics systems NOTE: Listed pressures are for a flow rate of 0.3 ml/min at ambient temperature with 100 % water and pulse-dampened flow. CATALOG NO. DESCRIPTION Union, holes, 1/16" Liquid Manifold Assembly with Relief Valve, Vector PCX & Pinnacle PCX Liquid T Manifold Assembly, Vector PCX & Pinnacle PCX TUBING, PLASTIC CATALOG NO. DESCRIPTION TFE Tubing, 1/16" OD 0.010" ID, 90 cm TFE Tubing, 1/16" OD x 0.016" ID, 90 cm TFE Tubing, 1/16" OD x 0.020" ID, 90 cm TFE Tubing, 1/16" OD 0.025" ID, 90 cm Inert PEEK Tubing, 1/16" OD 0.010" ID, 90 cm Inert PEEK Tubing, 1/16" OD 0.020" ID, 90 cm FEP (clear) Tubing, 1/8" OD 0.063" ID, 90 cm Air Barrier Tubing, 1/8" OD 0.063" ID, 90 cm C-flex Tubing, 0.25" OD 0.125" ID, 90 cm 55 PRODUCT CATALOG PICKERING LABORATORIES 15 /16

56 REPLACEMENT COMPONENTS, FITTINGS & TUBING FITTINGS, PLASTIC CATALOG NO. DESCRIPTION Nut, 1/4-28, 1/8" (Polypropylene) - Pack of Ferrule, 1/4-28, 1/8" (Tefzel) - Pack of Plug (Male), 1/4-28 (Delrin) Plug (Male), 10-32, 1/16" (Delrin) Nut, Fingertight, 1/16" (Kel-F) Nut, 1/16 (PEEK), Lite Touch - Pack of Ferrule, 1/16 (PEEK), Lite Touch - Pack of Removal Tool, for Lite-Touch Ferrule Nut, Male 1/4-28 1/16 tubing SST Ferrule, 1/4" SST Nut, Female PEEK 1/ Ferrule, 1/16 x 1/4-28 (PEEK), Pinnacle PCX - Pack of Nut, Short, 1/8 x 1/4-28 (PEEK), Pinnacle PCX - Pack of Nut, Short, 1/16 x 1/4-28 (PEEK), Pinnacle PCX - Pack of Ferrule, 1/8 x 1/4-28 (PEEK), Pinnacle PCX - Pack of Nut, Long, 1/8 x 1/4-28 (PEEK), Pinnacle PCX - Pack of Nut, Long, 1/16 x 1/4-28 (PEEK), Pinnacle PCX - Pack of 5 FITTINGS, TYPE 316 STAINLESS STEEL CATALOG NO. DESCRIPTION Nut (Male), Parker type, 10-32, 1/16" - Pack of Ferrule, Parker type, 1/16" - Pack of 5 MISCELLANEOUS ACCESSORIES CATALOG NO. DESCRIPTION Fuse, 3.15A Time Delay Fuse, 5A Time Lag Highbreak, ceramic, Pinnacle PCX Tubing Cutter, for 1/8 1/16 in. tubing Column Fittings Kit, 1/16" SST Heat-Exchanger for Reactor Coupler for Guard and Column Pinnacle PCX, Tubing Assembly, Valve-Pump (2 each) Pinnacle PCX, Tubing Assembly, Pump-Transducer (2 each) Pinnacle PCX, Tubing Assembly, Column out Manifold, Dual pump system Pinnacle PCX, Tubing Assembly, Column out Manifold, Single pump system Pinnacle PCX & Vector PCX, Tubing Assembly, Gas Manifold - Bottle Pinnacle PCX, Tubing Assembly, Valve-Manifold, Single pump system Pinnacle PCX, Tubing Assembly, Transducer Valve (2 each) Pinnacle PCX, Tubing Assembly, Valve Manifold (2 each) Dual pump system Pinnacle PCX, Tubing Assembly, Transducer Valve, Single pump system Manual, Pinnacle PCX operations Manual, Vector PCX operations Cable, RS232, Relay Cable Assembly Enable Switch, Vector USB Cable, USB ' Kit, PM Service Pinnacle, Single pump system Kit, PM Service Pinnacle, Dual pump system Kit, PM Service Vector Tubing kit Pinnacle, Dual pump system Tubing kit Pinnacle, Single pump system Over-pressure Relief Valve Cartridge The following trademarks are the property of the companies listed: ALKION Brij-35 ChlorAC Chromatographic Grade Hydrolysis Reagent C47 RESTORE Seraprep Tefzel Thiofluor TRIONE Uriprep Vespel Windows Pickering Laboratories, Inc. ICI Americas, Inc. Pickering Laboratories, Inc. Pickering Laboratories, Inc. Pickering Laboratories, Inc. Pickering Laboratories, Inc. Pickering Laboratories, Inc. E.I. DuPont Corporation Pickering Laboratories, Inc. Pickering Laboratories, Inc. Pickering Laboratories, Inc. E.I. DuPont Corporation Microsoft Corporation

57 CLEAN-UP COLUMNS AFLACLEAN, OTACLEAN, AFLA-OTACLEAN & DONEX CLEAN-UP COLUMNS IMMUNOAFFINITY & SPE SAMPLE CLEAN-UP COLUMNS Aflatoxins and Ochratoxin A are produced by fungi, e.g. Aspergillus and Penicillium species. Therefore both toxin types are found together in many foods and animal feeds, e.g. cereals. Deoxynivalenol, also known as Vomitoxin, is a metabolite of various molds of the genus Fusarium (F. colmorum, F. graminearum) and can often be found together with other mycotoxins. Of significant assistance is the clean-up of extracts by a combined immunoaffinity column (the Afla-OtaCLEAN column) for both Aflatoxins and Ochratoxin A in one step. The subsequent analysis may then be performed by HPLC with postcolumn derivatization or other techniques. 57 SIMULTANEOUS CLEAN-UP OF AFLATOXINS AND OCHRATOXIN A The combination sample clean-up column Afla-OtaCLEAN from LCTech is very tolerant towards many matrices and allows for a comprehensive clean-up of the aflatoxins B1, B2, G1 and G2 as well as Ochratoxin A. The high maximum capacity of 150 ng for Aflatoxin B1 and 200 ng for Ochratoxin A provides a wide measurement range. With a shelf-life of 1 year (from date of manufacture) at room temperature, the storage and use of the columns is very convenient. EXAMPLE RECOVERIES OF AFLATOXIN AND OCHRATOXINS MATRIX B1 B2 G1 G2 OTA Maize (Afla 10 ppb, OTA 14.3 ppb) 107 % 91 % 103 % 75 % 97 % Rice (Afla 10 ppb, OTA 14.3 ppb) 107 % 93 % 98 % 85 % 101 % Malt (Afla 10 ppb, OTA 14.3 ppb) 98 % 99 % 97 % 70 % 96 % Raisins (Afla 10 ppb, OTA 14.3 ppb) 99 % 106 % 101 % 69 % 97 % AFLATOXINS & OCHRATOXIN A SAMPLE CLEAN-UP COLUMN CATALOG INFORMATION CATALOG NO. DESCRIPTION QTY 1102 Alfa-OtaCLEAN sample clean-up column, 3 ml 25/pkg NOTE: Larger quantities available upon request SAMPLE CLEAN-UP COLUMN FOR AFLATOXIN ANALYSIS AflaCLEAN from LCTech was developed for the sample clean-up of foods, grains, feeds, etc. for aflatoxin analysis using HPLC post-column derivatization or other techniques. The AflaCLEAN column is selective for B1, B2, G1 and G2. The maximum loading capacity is 150 ng of Aflatoxin. The shelf-life is 2 years (from date of manufacture) at room temperature. Our cost-effective AflaCLEAN Select columns have a shelf-life of 9 months at 4 ºC. EXAMPLE RECOVERIES OF AFLATOXIN MATRIX B1 B2 G1 G2 Peanut Butter 95 % 98 % 93 % 84 % Peanuts 104 % 94 % 96 % 85 % Dried Distillers Grain 109 % 97 % 90 % 77 % Maize 101 % 98 % 103 % 80 % AFLATOXIN SAMPLE CLEAN-UP COLUMN CATALOG INFORMATION CATALOG NO. DESCRIPTION QTY AflaCLEAN sample clean-up column, 3 ml 25/pkg AflaCLEAN Select sample clean-up column, 3 ml 25/pkg AflaCLEAN Select sample clean-up column, 1 ml 25/pkg NOTE: Larger quantities available upon request PRODUCT CATALOG PICKERING LABORATORIES 15 /16

58 CLEAN-UP COLUMNS AFLACLEAN SELECT COLUMNS CATALOG NO. DESCRIPTION QTY AflaCLEAN Select sample clean-up column, 3 ml 25/pkg AflaCLEAN Select sample clean-up column, 3 ml 500/pkg AflaCLEAN Select sample clean-up column, 1 ml 25/pkg AflaCLEAN Select sample clean-up column, 1 ml 500/pkg Maximum loading capacity of 300 ng of Aflatoxin. Shelf life is 9 months at 4 ºC. AFLACLEAN SMART CARTRIDGES CATALOG NO. DESCRIPTION QTY AflaCLEAN Smart sample clean-up cartridges, 1 ml 100/pkg AflaCLEAN Smart sample clean-up cartridges, 1 ml 500/pkg This innovative design utilizes a syringe for processing and is optimized for faster elution using less solvent. Aflatoxin sample-cleanup for Hazelnut, Peanut, Pistachio, and Corn. Maximum loading of 100 ng of Aflatoxin. Shelf life is 6 months at 4 ºC. 160 mv Emission: 460 nm B1 400 mv 200 mv Ota A Emission: 440 nm G1 75 B G min Distillers grain spiked with 5 ppb total Aflatoxins min Peanut Butter spiked with 5 ppb total Aflatoxins min Distillers grain spiked with 5 ppb Ochratoxin A SAMPLE CLEAN-UP COLUMN FOR OCHRATOXIN ANALYSIS OtaCLEAN was developed for sample clean-up of foods, grains, feeds, etc. for Ochratoxin A analysis using HPLC or other techniques. The antibody employed possesses a very high Ochratoxin A specificity. This leads to exceptional chromatographic results without any interfering secondary signals and very high recovery rates. Independent of the complexity of the matrix, excellent results can be achieved. The maximum loading capacity is 200 ng Ochratoxin A. The shelf-life is 2 years (from date of manufacture) at room temperature mv Ota A EXAMPLE RECOVERIES OF OCHRATOXIN A MATRIX RECOVERY RATE Dried Distillers Grain 102 % Duran Wheat 92 % Coffee 100 % Red Wine 108 % Rye Bran 91 % Beer 96 % Grapes 91 % Horse Feed 94 % Wort 107 % OTACLEAN COLUMNS CATALOG NO. DESCRIPTION QTY OtaCLEAN sample clean-up column, 3 ml 25/pkg Afla-OtaCLEAN sample clean-up column, 3 ml 25/pkg OtaCLEAN sample clean-up column, 3 ml 500/pkg OtaCLEAN sample clean-up column, 1 ml 25/pkg OtaCLEAN sample clean-up column, 1 ml 500/pkg Maximum loading capacity of 200 ng of Ochratoxin A. Shelf life is 2 years at room temperature min Roasted Coffee spiked with 5 ppb Ochratoxin A OTACLEAN SMART CARTRIDGES CATALOG NO. DESCRIPTION QTY OtaCLEAN Smart cartridges 100/pkg OtaCLEAN Smart cartridges 500/pkg This innovative design utilizes a syringe for processing and is optimized for faster elution using less solvent. Shelf life is 6 months at 4 ºC.

59 CLEAN-UP COLUMNS DONEX Deoxynivalenol, also known as Vomitoxin, is a metabolite of various molds of the genus Fusarium (F. culmorum, F. graminearum) and can often be found on contaminated cereals (wheat, barley, oat). Generally the toxin is analyzed with HPLC/UV detection or alternatively with HPLC / post column derivatization / fluorescence detection or LC / MS. Whichever method is chosen, a good sample preparation extends life time of the analyzer and of the HPLC column and reduces interferences by matrix components. Moreover the running times of the HPLC system can be reduced for easy matrices from about 25 to 10 minutes by pre-cleaning. If HPLC/UV detection is used, sensitivity can be dramatically enhanced with larger sample volumes. The DONeX column has a matrix load of up to 4 grams. It is ideally suited to many different matrices, including corn, barley, wheat, animal feeds, muesli, and bread. DONeX COLUMNS CATALOG NO. DESCRIPTION QTY DONeX sample clean-up column 25/pkg DONeX sample clean-up column, 3 ml 500/pkg Maximum load capacity of 4 g of DON. No expiration date. Chromatogram of a cereal-based poultry feed sample, analyzed with HPLC/post column derivatization/fld, sample was spiked with 1 ppm DON, extracted and purified, evaporated sample (1 g) was resolved in 2 ml HPLC solvent, 40 μl were injected (0.02 gram matrix equivalents represent 20 ng DON injected) 59 DONeX COLUMN RECOVERIES MATRIX RECOVERIES [%] DON Bread 108 Corn 90 Chicken Feed 101 Distillers Grain 91 Oat Flakes 100 Pasta (Dried) 105 Poultry Feed 100 Rice 104 Rye 91 Wheat 100 Wheat Bran 99 * All matrices were spiked prior to the extraction. Spiking solution was incubated for at least 1 h with matrix before extraction was started. Calculation of recovery is based on subtraction of values obtained without spiking. Recoveries for Nivalenol are similar to those observed for Deoxynivalenol. PRODUCT CATALOG PICKERING LABORATORIES 15 /16

60 CLEAN-UP COLUMNS GLYPHOSATE AND AMPA SAMPLE CLEAN-UP CARTRIDGES Pickering SPE sample clean-up cartridges are used to treat samples prior to injection. The strong cation-exchange resin is composed of sulfonic acid functional groups attached to a styrene divinylbenzene copolymer lattice. The high selectivity and ruggedness provides a simple and universal method for removing matrix interference from the sample. The SPE sample clean-up cartridges are ideal for the cleanup of vegetables, fruits and crop samples in the analysis of Glyphosate and AMPA. Clean-up Methylene Chloride Partition: To 20 ml of aqueous extract add 15 ml methylene chloride. Shake for 2-3 min and centrifuge. To 4.5 ml of aqueous layer add 0.5 ml acidic modifier solution. Shake and centrifuge. SPE Clean-up: Prepare SPE cartridge (refer to product abstract PA210) Transfer 1 ml portions of SPE mobile phase (refer to method abstract MA206 for formulation) and discard the effluent. Elute analytes with 12 ml SPE mobile phase Concentration: Evaporate to dryness using rotary evaporator or a vacuum vortex-type evaporator or lyophilize Re-dissolve in 2 ml of the SPE mobile phase CATALOG NO. DESCRIPTION QTY SPE sample clean-up cartridges 50/pkg RECOVERIES FOR GLYPHOSATE AND AMPA MATRIX GLYPHOSATE AMPA Alfalfa 129 % 116 % Strawberry 84 % 82 % Broccoli 97 % 95 % Sample Preparation Extraction: Take 25 g of homogenous sample and add enough water (after estimation of moisture content) to make the total volume of water 125 ml. Blend and Centrifuge. Matrix Specific Modification: High water content reduce sample amount to 12.5 g High protein content add 100 µl HCl to 20 ml of extract, shake and centrifuge. High fat content perform the methylene chloride partition twice.

61 METHOD ABSTRACTS - AMINO ACIDS / 30-MINUTE AMINO ACID ANALYSIS OF HYDROLYZED SAMPLES Pickering Laboratories specializes in the manufacturing of cation-exchange columns and eluants for Amino Acid Analysis. No other technique has been shown to match the reproducibility and selectivity of ion-exchange analysis with post-column Ninhydrin detection. Nor is there a chromatography technique that provides as much information; the 570/440 nm signal ratio for each amino acid is a constant and so offers information about peak purity. Pinnacle PCX post-column derivatization system provides a unique opportunity to combine eluant gradient capabilities of modern HPLC instruments with column temperature gradients. As might be expected, this capability also reduces the analysis time. We introduce an accelerated Amino Acid Analysis method for hydrolyzed samples that utilizes temperature and eluant gradients. METHOD Analytical Conditions Column: High-efficiency Sodium cation-exchange column, 4.6 x 110 mm, Catalog number T Flow Rate: 0.6 ml/min Mobile Phase: See method Post-Column Conditions Post-Column System: Pinnacle PCX Reactor Volume: 0.5 ml Temperature: 130 C METHOD FOR OXIDIZED FEEDS HYDROLYSATE SAMPLES TIME Na270 % Na425 % Na640 % RG011 % COLUMN OVEN PROGRAM TIME TEMP ºC Reagent: Trione Ninhydrin reagent 16 Flow Rate: 0.3 ml/min Detection: UV/VIS 570 nm for primary amino acids 440 nm for secondary amino acids Injection Volume: 10 µl of 0.25 µ mole/ml STD min Fig 1. Chromatogram of oxidized feeds hydrolysate standard 1 Aspartic Acid 2 Threonine 3 Serine 4 Glutamic Acid 5 Proline 6 Glycine 7 Alanine 8 Cystine 9 Valine 10 Methionine 11 Isoleucine 12 Leucine 13 Norleucine 14 Tyrosine 15 Phenylalanine 16 Histidine 17 Lysine 18 Tryptophan 19 Ammonia 20 Arginine 21 Cysteic Acid 22 Methionine Sulfone PRODUCT CATALOG PICKERING LABORATORIES 15 /16

62 METHOD ABSTRACTS - AMINO ACIDS 382 / 70-MIN AMINO ACID ANALYSIS OF PHYSIOLOGICAL SAMPLES Pickering Laboratories specializes in the manufacturing of cation-exchange columns and eluants for Amino Acid Analysis. Post-column derivatization with Ninhydrin offers unmatched selectivity and reproducibility of the analysis for most challenging matrixes. Pinnacle PCX post-column derivatization system allows analysts to combine eluant gradient capabilities of modern HPLC instruments with column temperature gradients. We introduce new accelerated Amino Acids Analysis method for physiological samples that utilizes temperature and eluant gradients. COLUMN OVEN PROGRAM TIME TEMP ºC HPLC PROGRAM TIME % Li365 % Li375 % RG003 % METHOD Analytical Conditions Column: High-efficiency Lithium cation-exchange column, 4.6 x 75 mm, 5 µm Catalog number T Flow Rate: 0.55 ml/min Mobile Phase: , Li365, Li375, RG003 Post-column Conditions Post-column System: Pinnacle PCX Reactor Volume: 0.5 ml Temperature: 130 C Reagent: Trione Flow Rate: 0.5 ml/min Detection: UV/VIS 570 nm for primary amino acids 440 nm for secondary amino acids Injection Volume: 10 µl of 0.25 µ mole/ml STD : min 1 Phosphoserine 2 Taurine 3 Phosphoethanolamine 4 Urea 5 Aspartic acid 6 Hydroxyproline 7 Threonine 8 Serine 9 Asparagine 10 Glutamic acid 11 Glutamine 12 Sarcosine 13 a-aminoadipic acid 14 Proline 15 Glycine 16 Alanine 17 Citrulline 18 a-amino-n-butyric acid 19 Valine 20 Cystine 21 Methionine 22 Cystathionine 23 Isoleucine 24 Leucine 25 Tyrosine 26 Phenylalanine 27 b-alanine 28 b-amino-i-butyric acid 29 Homocystine 30 g-aminobutyric acid 31 Tryptophan 32 Ethanolamine 33 Ammonia 34 Hydroxylysines 35 Ornitine 36 Lysine 37 1-Methylhistidine 38 Histidine 39 3-Methylhistidine 40 Anserine 41 Carnosine 42 Arginine 43 Glucosaminic Acid* *Internal Standard

63 METHOD ABSTRACTS - AMINO ACIDS 371 / AMINO ACIDS ANALYSIS OF CELL CULTURE MEDIA Cell cultures are widely used to produce biopharmaceuticals and other biologically active compounds. The composition of the cell culture media affects the yield and structure of the desired products and must be carefully optimized. Cell culture media is typically composed of mixtures of amino acids, vitamins, carbohydrates, inorganic salts as well as different peptides, proteins and other compounds. As the cells grow, they consume nutrients and release target biopharmaceuticals as well as waste products. Amino Acids serve as the building blocks of proteins, as well as intermediates in many metabolic pathways. Amino Acids are typically added to cell culture media to provide nutritional requirements for the cells. Monitoring and adjusting Amino Acid composition is an essential part of optimizing the manufacturing process to ensure high quality and optimum yield of the final product. Amino Acids Analysis using cation-exchange chromatography with post-column Ninhydrin derivatization allows for easy determination of Amino Acid concentrations in many complex matrices, including cell culture media. The post-column method is very sensitive, reproducible and rugged. It has been and continues to be a method of choice for laboratories running biological samples, protein, peptides and foods analysis. Most chemical compounds present in the media do not interfere with analysis, so the majority of samples only need diluting with citric buffer and filtering before analysis. If serum is added to the media, then the proteins need to be precipitated using either Seraprep solution or ultrafiltration. Pickering Laboratories, Inc. offers the complete solution for Amino Acids Analysis, including post-column derivatization instruments, columns, eluants, reagents and standards. The Pinnacle PCX derivatization system has a programmable column oven to allow for shorter run times and easy method optimization. METHOD Analytical conditions Column: High-efficiency Lithium cation-exchange column, 4.6 x 75 mm, Catalog Number T Flow Rate: 0.55 ml/min Mobile Phase: See method in Table 1 Post-Column Conditions Post-column System: Pinnacle PCX Reactor Volume: 0.5 ml Reactor Temperature: 130 C Flow Rate: 0.3 ml/min Detection: UV/VIS 570 nm for primary amino acids, 440 nm for secondary amino acids Injection Volume: ul TABLE 1. HPLC PROGRAM TIME , % LI365, % LI375, % RG003, % TABLE 2. COLUMN OVEN PROGRAM TIME Temp, ºC Continues on next page PRODUCT CATALOG PICKERING LABORATORIES 15 /16

64 METHOD ABSTRACTS - AMINO ACIDS 371 continued from previous page Urea Phosphoethanolamine Glucosaminoc Acid Taurine Phosphoserine Aspartic Acid Hydroxyproline Threonine Serine Asparagine Glutamic Acid Glutamine Sarcosine Proline a-aminoadipic Acid Glycine Alanine Citrulline Valine a-amino-n-butyric Acid Cystine Methionine Allo-Isoleucine Isoleucine Cystathionine Leucine Tyrosine Phenylalanine b-alanine b-amino-i-butyric Acid Homocystine gamma-aminobutyric Acid Tryptophan Ethanolamine Ammonia Hydroxylysines Ornithine AEC Lysine 1-Methylhistidine Histidine 3-Methylhistidine Anserine Carnosine Arginine Fig. 1. Chromatogram of Amino Acids standard Aspartic Acid Threonine Serine Glutamic Acid Asparagine Proline Glycine Alanine Cystine Methionine Tyrosine Valine Isoleucine Leucine Phenylalanine Tryptophan Ammonia Lysine Histidine Arginine Fig. 2. Chromatogram of cell culture media sample

65 METHOD ABSTRACTS - AMINO ACIDS 373 / AMINO ACID ANALYSIS OF MONOCLONAL ANTIBODIES The peptide and protein based pharmaceuticals are a rapidly expanding class of therapeutical agents that are used to treat a wide variety of health conditions, including cancer, metabolic and auto-immune diseases, HIV and more. Biologic drugs, such as monoclonal antibodies, are derived from living organisms and are usually very expensive. As many biologics are coming off of patents, the market is ready for cost-saving biogenerics. But all proteins, including monoclonal antibodies, have complex structures that determine their function. Differences in structure would alter biological activity leading to changes in safety and efficacy of the drug. ICH Q6B is a guidance document that provides a set of internationally accepted specifications for biotechnological and biological products to support new marketing applications. It establishes the set of criteria to which a drug substance, drug product or material should conform to be considered acceptable for intended use. Determining Amino Acid composition following hydrolysis is listed in ICH Q6B as a way to characterize the protein and to confirm its identity by comparing with Amino Acid composition deduced from the gene sequence of the desired product. Amino Acids Analysis data is also used to accurately determine the protein content. The Amino Acids Analysis with post-column derivatization is a very sensitive, reproducible and rugged method and it has been a preferred approach for laboratories running biological samples, protein, peptides and foods analysis. Pickering Laboratories Inc. offers many Amino Acids Analysis products including postcolumn derivatization instruments, columns, eluants, reagents and standards. All products are designed to work together to deliver optimum results for any chosen sample. METHOD Analytical conditions Column: High-efficiency Sodium cation-exchange column, 4.6 x 110 mm, Catalog Number T Flow Rate: 0.6 ml/min Mobile Phase: See method in Table 1 Post-Column Conditions Post-column System: Pinnacle PCX Reactor Volume: 0.5 ml Reactor Temperature: 130 C Flow Rate: 0.3 ml/min Detection: UV/VIS 570 nm for primary amino acids, 440 nm for secondary amino acids Injection Volume: ul TABLE 1. HPLC PROGRAM TIME NA315, % NA425, % NA640, % RG011, % TABLE 2. COLUMN OVEN PROGRAM TIME Temp, ºC Continues on next page PRODUCT CATALOG PICKERING LABORATORIES 15 /16

66 METHOD ABSTRACTS - AMINO ACIDS 373 continued from previous page min Fig. 1. Chromatogram of Amino Acids standard min Fig. 2. Chromatogram of hydrolyzed sample of IL-17F monoclonal antibody 1 Aspartic Acid 2 Threonine 3 Serine 4 Glutamic Acid 5 Proline 6 Glycine 7 Alanine 8 Cystine 9 Valine 10 Methionine 11 Isoleucine Alanine 12 Leucine 13 Norleucine 14 Tyrosine 15 Phenylalanine 16 Histidine 17 Lysine 18 Tryptophan 19 Ammonia 20 Arginine 21 Cysteic Acid 22 Taurine ACKNOWLEDGEMENTS We would like to thank Frank N. Konstantinides and AminoAcids.com for their contribution to this project.

67 METHOD ABSTRACTS - AMINO ACIDS 391 / AMINO ACID ANALYSIS ACCORDING TO EUROPEAN PHARMACOPOEIA 8.0. The European Pharmacopoeia (Ph. Eur.) defines requirements for the qualitative and quantitative composition of medicines, as well as the tests to be carried out on medicines and on substances and materials used in their production. It covers active substances, excipients and preparations of chemical, animal, human or herbal origin, homoeopathic preparations and homoeopathic stocks, antibiotics, as well as dosage forms and containers. It also includes tests on biologicals, blood and plasma derivatives, vaccines and radiopharmaceutical preparations. The European Pharmacopoeia and its requirements are legally binding in the member states of the European Pharmacopoeia Convention and the European Union. All manufacturers of medicines or substances for pharmaceutical use therefore must apply the Ph. Eur. quality standards in order to be able to market and use these products in Europe. Amino Acids Analysis can be used for: - Identification tests on biopharmaceutical active ingredients (e.g. peptides, proteins) by means of amino acids composition analysis; - Impurities and related substances determination on APIs (Active Pharmaceutical Ingredients, e.g. free amino acids) and intermediates; - Single or total amino acids quantification in drug products, including markers determination in complex matrixes (e. g. phytopharmaceuticals). The following Ph. Eur. monographs have already officially introduced the Amino Acid Analysis method with post-column Ninhydrin derivatization as the analytical procedure required for the determination of the Ninhydrin-positive substances, and additional papers are expected to be published in upcoming months: - Cysteine HCl Monohydrate 01/2014: Isoleucine 07/2013: Leucine 07/2013: Lysine HCl 07/2013: Serine 01/2014: Proline 01/2014: Threonine 01/2014: Valine 01/2014: Arginine 07/2014:0806 Pickering Laboratories, Inc. offers a complete solution for Amino Acids Analysis according to European Pharmacopoeia 8.0. This includes the Pinnacle PCX post-column derivatization instrument, analytical columns and GARDs, buffers and Trione Ninhydrin reagent. The Pinnacle PCX is capable of performing column temperature gradients that allow easily modified conditions and improved run times and amino acids separations. The methods presented in this application note were optimized to comply with system suitability requirements of Pharmacopoeia 8.0 methods. Each Pharmacopoeia monograph describes the preparation of the test and reference solutions specific for each amino acid. The solutions are used for calculations of percentage contents, impurity levels as well as parameters of system suitability. Resolution of 1.5 is required between Leucine and Isoleucine peaks. Table 1 summarizes the solutions used in each monograph. 67 Table 1. Reference guide for Pharmacopoeia 8.0 methods REFERENCE SOLUTIONS, UG/ML Amino acids Test solutions ug/ml Cys CSSC Lys Ser Thr Val Ala Arg Leu Ile Ile and Leu Pro NH4 Valine each Proline each Leucine 600 and each Threonine each Serine each Lysine each Isoleucine each Arginine each Cysteine each Cys = Cysteine, CSSC = Cystine, Lys = Lysine, Ser = Serine, Thr = Threonine, Val = Valine, Ala = Alanine, Arg = Arginine, Ile = Isoleucine, Leu = Leucine, Pro = Proline, NH4 = ammonia Continues on next page PRODUCT CATALOG PICKERING LABORATORIES 15 /16

68 METHOD ABSTRACTS - AMINO ACIDS 391 continued from previous page For all amino acids, except Cysteine, Sodium-based and Lithiumbased methods are available. For Cysteine analysis, only Lithiumbased methods are suitable. Sodium-based methods have shorter run times and are preferable for all amino acids except Cysteine. Methods using High-efficiency Sodium column for analysis of following Amino Acids: Valine, Proline, Leucine, Isoleucine, Serine, Threonine, Lysine, Arginine Analytical Conditions Column: High-efficiency Sodium cation-exchange column, 4.6 x 110 mm, Catalog Number T Flow Rate: 0.6 ml/min Mobile Phase: See method in Table 2 Injection Volume: 50 ul Post-column Conditions Post-Column System: Pinnacle PCX Reactor Volume: 0.5 ml Reagent: Trione Reagent Temperature: 130 ºC Column Temperature: See method in Table 3 Flow Rate: 0.3 ml/min Detection: UV/VIS 570 nm for primary amino acids, 440 nm for secondary amino acids Table 2. HPLC program for column T TIME NA315 NA425 NA640 RG Table 3. Column oven program Threonine Serine Proline Alanine Cystine Valine Isoleucine Leucine Fig. 1: Sodium chromatogram of amino acids analyzed using Pharmacopeia 8.0 methods (3 ug/ml each, 50 ul injection). Fig. 2: Sodium chromatogram of alternative amino acids analyzed using Pharmacopeia 8.0 methods (3 ug/ml each, 50 ul injection). Fig. 3: Sodium chromatogram of amino acids used for calculations and system suitability check in Pharmacopeia 8.0 methods (refer to Table 1). Proline 1.2 ug/ml; Isoleucine 3 ug/ ml; Leucine 3 ug/ml; Ammonia 0.12 ug/ml. Injection volume 50 ul. Lysine Ammonia Arginine min Threonine Proline Valine Isoleucine Leucine Lysine Ammonia min Proline Methionine Isoleucine Leucine Phenylalanine Ammonia min Isoleucine TIME TEMP, C min Fig. 4: Sodium chromatogram of low level Isoleucine reference solution 0.12 ug/ml. Injection volume 50 ul

69 METHOD ABSTRACTS - AMINO ACIDS Threonine Alanine Valine Ammonia Isoleucine Leucine min Fig. 5: Sodium chromatogram of Isoleucine test solution 600 ug/ml (zoomed). Injection volume 50 ul Fig. 9: Sodium chromatogram of Lysine test solution 600 ug/ml (zoomed). Injection volume 50 ul Proline Cystine min Fig. 6: Sodium chromatogram of Proline test solution 600 ug/ml (zoomed). Injection volume 50 ul Threonine Valine Isoleucine Leucine Ammonia min Fig. 7: Sodium chromatogram of Leucine test solution 600 ug/ml (zoomed). Injection volume 50 ul Cystine Alanine Valine Lysine Ammonia min Isoleucine Leucine Ammonia Lysine Due to the poor separation of Cysteine and Proline peaks with the Sodium-based method, the Lithium High-efficiency column needs to be used for Cysteine analysis according to Pharmacopeia 8.0. The same Lithium method can be used for all other amino acids, though analysis time is longer than with the Sodium method. Methods using High-efficiency Lithium column for analysis of following amino acids: Cysteine, Valine, Proline, Leucine, Isoleucine, Serine, Threonine, Lysine, Arginine Analytical Conditions Column: High-efficiency Lithium cation-exchange column, 4.6 x 75 mm, Catalog Number T Flow Rate: 0.55 ml/min Mobile Phase: See method in Table 4 Injection Volume: 50 ul Post-column Conditions Post-Column System: Pinnacle PCX Reactor Volume: 0.5 ml Reagent: Trione Reagent Temperature: 130 ºC Column Temperature: See method in Table 5 Flow Rate: 0.3 ml/min Detection: UV/VIS 570 nm for primary amino acids, 440 nm for secondary amino acids min Fig. 8: Sodium chromatogram of Valine test solution 600 ug/ml (zoomed). Injection volume 50 ul Continues on next page PRODUCT CATALOG PICKERING LABORATORIES 15 /16

70 METHOD ABSTRACTS - AMINO ACIDS 391 continued from previous page Table 4. HPLC Program for column T TIME LI365 LI375 RG Table 5. Column oven program TIME TEMP, C Proline Cysteine Cystine Isoleucine Leucine Fig. 10: Lithium chromatogram of amino acids used as reference solutions for Cysteine analysis (3 ug/ml each, 50 ul injection). Ammonia min Threonine Proline Valine Methionine Isoleucine Leucine Phenylalanine Ammonia Lysine To make it easier to start using Pickering Laboratories methods, we offer chemistry kits that include: analytical column, GARD, buffers and reagents for Amino Acids Analysis. All parts of the kit could be ordered individually if needed. Please contact Pickering Laboratories if you have any questions regarding this application. PART NUMBER DESCRIPTION Minute High Efficiency Protein Hydrolysate Kit (for Sodium methods): T 30-Minute Sodium Cation-exchange Column 4.6 x 110 mm & amino acid test mixture Cation-exchange GARD assembly: Holder w/ 2 replaceable GARD Na220 Na315 Sodium Diluent, ph 2.20, 4 x 250 ml Sodium Eluant, ph 3.15, 4 x 950 ml Na425 Sodium Eluant, ph 4.25 Na640 Sodium Eluant, ph 6.40 RG011 T100C H Sodium Column Regenerant, 950 ml TRIONE Ninhydrin Reagent, 4 x 950 ml (4-month shelf life) Sodium Calibration Standard, for protein hydrolysate, 0.25 µmole/ml, 5 ml Kit Identical to with T200 replacing T100C: T200 TRIONE Two-part Ninhydrin Reagent, prepares 4 x 900 ml (12-month shelf life) minute Physiologic Fluid/Native Sample Kit (for Lithium methods): T Lithium Ion-exchange Column, 4.6 x 75 mm (with test mixture ) Cation-exchange GARD assembly: Holder w/ 2 replaceable GARD Lithium Eluant, ph 2.80, 4 x 950 ml Li220 Li365 Li375 RG SP100 UP100 T100C Lithium Diluent, ph 2.20, 4 x 250 ml Lithium Diluent, ph 3.65, 4 x 950 ml Lithium Diluent, ph 3.75, 4 x 950 ml Lithium Column Regenerant, 950 ml Lithium Calibration Standard, without Norleucine and AGPA, 0.25 µmole/ml, 5 ml SERAPREP, 250 ml URIPREP, 250 ml TRIONE Ninhydrin Reagent, 4 x 950 ml (3-month shelf life) Kit Identical to with T200 replacing T100C: T200 TRIONE Two-part Ninhydrin Reagent, prepares 4 x 900 ml (12-month shelf life) Pickering Laboratories will keep updating its Pharmacopeia 8.0 methods as new monographs are released. Please contact support@pickeringlabs.com for the latest methods and chromatograms min Fig. 11: Lithium chromatogram of amino acids analyzed using Pharmacopeia 8.0 methods (3 ug/ml each, 50 ul injection).

71 METHOD ABSTRACTS - AMINO ACIDS 393 / ANALYSIS OF AMINO ACIDS SUPPLEMENTS Amino acids are building blocks of proteins and essential amino acids need to be obtain from diet as they cannot be synthesized by our bodies. Amino acids supplements are popular among athletes and bodybuilders and are sold in the form of pills, powders and drinks. They usually also contain vitamins, electrolytes, herbs, flavorings and other additives. Analysis of these supplements is important to confirm the label claims and to maintain industry standards. Amino Acids Analysis with post-column derivatization is a very sensitive, reproducible and rugged method and it has been a preferred approach for laboratories running biological samples, protein, peptides and foods analysis. Pickering Laboratories Inc. products for Amino Acid Analysis include post-column derivatization instruments, columns, eluants, reagents and standards. All products are designed to work together to deliver optimum results for any chosen sample. The amino acids present in the product determine if Sodium or Lithium columns and buffers need to be used. Lithium columns have longer run time but have higher selectivity and are able to separate more amino acids than Sodium columns. Pickering Laboratories will help you to identify the best method for your Amino Acid Analysis. Sample preparation Grind 10 pills to fine powder before taking a sample for analysis. Mix all the powdered supplements thoroughly. Sample size depends on concentrations of amino acids present. To finely ground sample (0.05 g 0.2 g) add 100 ml of Na220. Mix well and sonicate for 10 min. Filter through 0.45 um filter and inject. METHOD FOR SODIUM COLUMN Analytical Conditions Column System: High-efficiency Sodium cation-exchange column, 4.6 x 110 mm, Catalog Number T Flow rate: 0.6 ml/min Mobile Phase: See method in Table 1 Post-column Conditions Post-Column System: Pinnacle PCX Reactor Volume: 0.5 ml Reactor Temperature: 130 ºC Column Temperature: See method in Table 2 Flow Rate: 0.3 ml/min Injection Volume: ul Table 1. HPLC Program for column T TIME NA315, % NA425, % NA640, % RG011, % Table 2. Column Oven Program TIME TEMP, C Continues on next page PRODUCT CATALOG PICKERING LABORATORIES 15 /16

72 METHOD ABSTRACTS - AMINO ACIDS 393 continued from previous page Aspartic Acid Threonine Serine Proline Glutamic Acid Valine Glycine Alanine Cystine Methionine Isoleucine Leucine Tyrosine Phenylalanine Histidine Lysine Tryptophan Ammonia Arginine min Fig. 1: Chromatogram of amino acids standard using sodium column T Threonine Valine Methionine Isoleucine Phenylalanine Histidine Lysine Arginine Leucine min Fig. 2: Chromatogram of a sample of commercial training supplement using sodium column T Citrulline Glutamine Valine Isoleucine Leucine min Fig. 3: Chromatogram of the a sample of sport recovery drink using sodium column T

73 METHOD ABSTRACTS - AMINO ACIDS Taurine Valine Methionine Isoleucine Leucine Alanine min Fig. 4: Chromatogram of a commercial sport performance drink using sodium column T METHOD FOR LITHIUM COLUMN Analytical Conditions Column: High-efficiency Lithium cation-exchange column, 4.6 x 75 mm, Catalog Number T Flow rate:.55 ml/min Mobile Phase: See method in Table 3 Injection Volume: 50 ul Post-column Conditions Post-Column System: Pinnacle PCX Reactor Volume: 0.5 ml Reagent: Trione Reactor Temperature: 130 ºC Column Temperature: See method in Table 4 Flow Rate: 0.3 ml/min Detection: UV/VIS 570 nm for primary amino acids, 440 nm for secondary amino acids Table 3. HPLC Program for column T TIME , % NA425, % NA640, % RG011, % Table 4. Column Oven Program TIME TEMP, C Continues on next page PRODUCT CATALOG PICKERING LABORATORIES 15 /16

74 METHOD ABSTRACTS - AMINO ACIDS 393 continued from previous page Threonine Glutamine Alanine Methionine Citrulline Valine Isoleucine Leucine Tyrosine Phenylalanine b-alanine Lysine Histidine Arginine Taurine Urea Phosphoethanolamine Glucosaminoc Acid Taurine Phosphoserine Hydroxyproline Sarcosine Aspartic Acid Threonine Serine Asparagine Glutamic Acid a-aminoadipic Acid Glutamine Proline Glycine Alanine Citrulline Valine a-amino-n-butyric Acid Cystine Methionine Allo-Isoleucine Cystathionine Isoleucine Leucine Tyrosine Phenylalanine b-alanine b-amino-i-butyric Acid Homocystine gamma-aminobutyric Acid Tryptophan Ethanolamine Ammonia Hydroxylysines Ornithine AEC Lysine 1-Methylhistidine Histidine 3-Methylhistidine Anserine Carnosine Arginine min Fig. 5: Chromatogram of amino acids standard using lithium column T min Fig. 6: Chromatogram of a pre-workout amino acid supplement drink using lithium column T.

75 METHOD ABSTRACTS - MYCOTOXINS 215 / CLEAN-UP AND DETERMINATION OF AFLATOXINS IN PEANUTS AND PEANUT BUTTER Aflatoxins occur naturally in peanuts, cottonseed, corn, and dried chili pepper as well as many mixed or processed foods and feeds. Of significant assistance is the cleanup of extracts by an Immunoaffinity column containing antibodies specific to the Mycotoxin of interest. We used a simple, sensitive and robust HPLC method with post-column photochemical derivatization and fluorescence detection to analyze Aflatoxins B1, B2, G1, G2 in peanut butter and ground peanuts. The UVE (LCTech, Germany) photochemical reactor requires no additional reagents and is easy to install between the HPLC column and FLD detector. This method and instrumentation allows for quick and interference-free detection of Aflatoxins at the low ppb level. PROJECT OVERVIEW As participants in an NIST study, we analyzed samples of peanuts and peanut butter (table 1, 2). Four other laboratories that use other HPLC methods for analysis of Aflatoxins participated in this study. Community results for peanuts are presented in table 2. The extracts were cleaned up using the AflaCLEAN (LCTech, Germany) Immunoaffinity columns for Aflatoxin B1, B2, G1, G2. For the handling of the columns, we used the AcceCLEAN automated system that processes three samples simultaneously. The prepared samples were analyzed by HPLC with post-column photochemical derivatization using the UVE photochemical reactor. METHOD Isolation of Aflatoxins B1, B2, G1, G2 Blend 20 g of sample at high speed with extraction solution (100 ml of Methanol/water 80/20, 50 ml of Hexane, 2 g NaCl) and filter through fluted paper. Dilute 14 ml of aqueous layer with 86 ml of PBS buffer (ph 7.2), filter and apply 11 ml of solution on AflaCLEAN Immunoafinity column. The toxins are eluted with 2 ml of Methanol and analyzed as described. Analytical Conditions Analytical Column: Mycotox (Pickering Laboratories, Inc), C 18, 4.6 x 250 mm HPLC Eluent: Sodium Phosphate buffer (Cat # )/Methanol/ Acetonitrile (57/28/15) Flow Rate: 1 ml/min Injection Volume: 30 µl FLD: Excitation 365 nm, Emission 430 nm Placement of UVE (Note: HPLC Model shown is to demonstrate placement of the UVE only. No endorsement expressed or implied. ) 75 Continues on next page PRODUCT CATALOG PICKERING LABORATORIES 15 /16

76 METHOD ABSTRACTS - MYCOTOXINS 215 continued from previous page RESULTS & DISCUSSION The 6-point calibration curves were built in a range of ppb for B1, ppb for B2 and G2, ppb for G1 with R 2 exceeding There were no matrix interferences present after the sample clean-up using the Immunoaffinity columns. The results for all Aflatoxins are in good agreement with certified NIST values and with the results obtained by other methods. a) B1 Table 1. Peanut butter (NIST SRM2387) control sample AFLATOXIN B1 AFLATOXIN B2 TOTAL AFLATOXINS Target value, ng/g 4.2 ± ± ± 0.5 Packet A, ng/g Packet B, ng/g Packet C, ng/g G2 G1 B2 Table 2. Ground peanut sample AFLATOXIN B1 AFLATOXIN B2 AFLATOXIN G1 AFLATOXIN G2 TOTAL AFLATOXINS Packet A, ng/g Packet B, ng/g Packet C, ng/g Mean, ng/g RSDr % Community results* NIST assessed value**, ng/g 7.47 ± ± ± ± ± 5.9 * Results from 5 participating laboratories are presented as (minimum reported value maximum reported value), ng/g ** ± 95 % confidence interval about the NIST assessed value b) B1 B2 min min Fig. 1 Chromatograms of a) ground peanuts; b) NIST SRM2387 peanut butter sample. All samples are part of NIST Exercise E (April 2010).

77 METHOD ABSTRACTS - MYCOTOXINS 218 / CLEAN-UP AND ANALYSIS OF AFLATOXINS AND OCHRATOXIN A IN HERBS AND SPICES The production of herbal supplements and spices is a fast growing industry. Unfortunately, the raw materials are often imported from countries that lack adequate quality control and whose weather conditions during the growing season, along with improper harvesting and storage practices, can cause toxic mold contamination. There are numerous reports on the presence of Mycotoxins in commercially available herbs and spices such as chamomile, black and white tea leaves, ginkgo leaves, paprika and cumin. We developed a simple, sensitive and robust HPLC method for analyzing Aflatoxins B1, B2, G1, G2 and Ochratoxin A in herbs and spices. AflaOTAClean Immunoaffinity columns contain antibodies specific for both classes of Mycotoxins and allow for fast and efficient sample clean-up. We used the AcceClean automated workstation, which processes three samples simultaneously. Post-column photochemical derivatization was used to increase the sensitivity of detection of Aflatoxins B1 and G1. The UVE (LCTech, Germany) photochemical reactor requires no additional reagents and is easy to install between the HPLC column and fluorescence detector. Ochratoxin A is a naturally fluorescent compound that does not require derivatization and can be analyzed together with all four Aflatoxins. METHOD Isolation of Aflatoxins B1, B2, G1, G2 and Ochratoxin A Mix 5 g of finely ground sample with extraction solution (25 ml of Methanol:water 80:20, 12.5 ml of Hexane, 0.5 g of NaCl) and shake on a mechanical shaker for 1 hour. Filter the extract through fluted paper. Dilute 14 ml of the aqueous layer with 86 ml of PBS buffer (ph 7.2), filter and apply 11 ml of solution to AflaOTAClean Immunoaffinity column at a flow rate of 2 ml/min. Wash the column with 10 ml of water at a flow rate of 2 ml/min. Elute the toxins with two 1 ml portions of Methanol at a flow rate of 0.3 ml/min. Allow 5 min before applying the second portion of the Methanol to ensure complete breaking of the antibody-toxin bond. 77 Analytical Conditions Analytical Column: Mycotox (Pickering Laboratories, Inc), C 18, 4.6x250 mm HPLC Eluent: Sodium Phosphate buffer (Cat # ), Methanol, Acetonitrile Flow Rate: 1 ml/min Injection Volume: 30 ul FLD: Excitation 365 nm, Emission 430 nm for Aflatoxins Excitation 335 nm, Emission 455 nm for Ochratoxin A HPLC GRADIENT TIME (Min) ,% METHANOL, % ACETONITRILE, % EQUILIBRATION TIME: 12 min Continues on next page PRODUCT CATALOG PICKERING LABORATORIES 15 /16

78 METHOD ABSTRACTS - MYCOTOXINS 218 continued from previous page RESULTS AND DISCUSSION The 6-point calibration curves were built in the range of ppb for B1, ppb for B2 and G2, ppb for G1, ppb for Ochratoxin A with R 2 exceeding The samples of echinacea, ginger and ginseng were purchased from a local herbal store. These samples were not naturally contaminated with Mycotoxins. There were no matrix interferences present after Immunoaffinity column clean-up. The samples were spiked with five Mycotoxins at two levels and processed, along with sample blanks, as described above. The recovery data for Aflatoxins B1, B2, G1, G2 and Ochratoxin A are presented in Tables 1-3. Table 1. Recovery Results for Echinacea Sample Mycotoxins Aflatoxin B1 Aflatoxin B2 Aflatoxin G1 Aflatoxin G2 Ochratoxin A Spike level, ng/g Recoveries, % Spike level, ng/g Recoveries, % G1 G2 B1 B2 B2 B1 Ochratoxin A Pic 2. Ginseng sample spiked with Mycotoxins. Aflatoxin B ng/g, Aflatoxin B ng/g, Aflatoxin G ng/g, Aflatoxin G ng/g, Ochratoxin A 5.05 ng/g. Mycotoxins Table 2. Recovery Results for Ginger Sample Aflatoxin B1 Aflatoxin B2 Aflatoxin G1 Aflatoxin G2 Ochratoxin A Spike level, ng/g Recoveries, % Spike level, ng/g Recoveries, % G2 G1 Ochratoxin A Table 3. Recovery Results for Ginseng Sample Mycotoxins Aflatoxin B1 Aflatoxin B2 Aflatoxin G1 Aflatoxin G2 Ochratoxin A Spike level, ng/g Recoveries, % Spike level, ng/g Recoveries, % Pic 3. Ginger sample spiked with Mycotoxins. Aflatoxin B ng/g, Aflatoxin B ng/g, Aflatoxin G ng/g, Aflatoxin G ng/g, Ochratoxin A 10.1 ng/g. G1 G2 B2 B1 Ochratoxin A min Pic 1. Ginseng sample blank Pic 4. Echinacea sample spiked with Mycotoxins. Aflatoxin B ng/g, Aflatoxin B ng/g, Aflatoxin G ng/g, Aflatoxin G ng/g, Ochratoxin A 5.05 ng/g. NOTE: The shift on the baseline at 17 min is due to changes in gradient conditions and detector settings.

79 METHOD ABSTRACTS - MYCOTOXINS 220 / OCHRATOXIN A IN RED AND WHITE WINE WITH OTACLEAN AND ACCECLEAN INTRODUCTION The Mycotoxin Ochratoxin A is found in many matrices, such as grapes. As red and white wines are produced from grapes, the mycotoxin can be found in the final product. Therefore, a maximum limit for Ochratoxin A in red, white and rosé wine, other wine and/or grape-must-based beverages was set at 2.0 μg/kg by Commission Regulation (EC) No. 123/2005. Thus, these beverages must be analyzed for their Mycotoxin content before they are sold on the market. The standard procedure for analysis of Ochratoxin A in matrices is to use immunoaffinity column cleanup. The cleanup may either be performed manually or much more convenient by an automated system. The following application note shows how wine samples can be processed with the AflaCLEAN (LCTech, Germany) immunoaffinity columns in combination with the fully automated sample preparation system AcceCLEAN producing high recovery rates and repeatability. CHEMICALS AND MATERIALS For the analysis, standard laboratory equipment such as beakers, funnels, filters, and stirrers are required. Furthermore, a centrifuge or a syringe with PVDF filters may be needed. The HPLC system should at least have an injection system (manually or automated), an isocratic pump, a column oven with guard and HPLC column, and a sensitive fluorescence detector. Using a post-column derivatization system further increases the sensitivity by a factor of 6 to 8, and the increases selectivity as interfering peaks are removed significantly. Extraction solution, 5 % NaHCO 3 with 1 % PEG 8000 (e. g. Sigma 4423) PBS buffer ph 7.2 Water (HPLC grade) Methanol (HPLC grade) Acetonitrile (HPLC grade) Glacial acetic acid p. a. Sodium hydroxide 1.0 N OTACLEAN COLUMNS 79 ACCECLEAN SYSTEM Continues on next page PRODUCT CATALOG PICKERING LABORATORIES 15 /16