Jack C. Cappozzo Separation Science Webinar March 12, 2013
Research/Analytical Purpose Adulteration (economical/food safety) Authenticity (economical/quality) Quality Sensory Analytical Instrumentation
Adulteration Nineteen Arrested Over Alleged Olive Oil Fraud (Spain, Olive Oil Times) Spanish Police Say Palm, Avocado, Sunflower Was Passed Off as Olive Oil (Spain, Olive Oil Times) Two Plead Guilty in Olive Oil Fraud Scheme, Sentenced to Two Years in Jail (Spain, 2011, Olive Oil Times) Toxic Oil Syndrome (TOS), Spain 1981
Adulteration Quality Health Nutritional Loss Illness Economics
Economics At 10% adulteration (soybean oil), the producer saves about $0.20/Kg commodity pricing Metric Ton of Olive Oil.@$2900 Metric Ton.$200 3 M Metric Tons in Spain 10% = 300,000 Metric Tons @ $60 million (in Savings) Retail 10% oil (100 Kg) savings $5000-10,000
Analytical Tests Needed for EVOO/OO 23 tests (reduce that to about 20 tests) $$$$$$$$
Olive Oil (Extra Virgin) (Standards of Identity) Sterol composition and content Fatty acid composition Saturated fatty acid content in position 2 of the triglyceride Unsaponifiable material Wax content Erythrodiol and uvaol ECN 42 content UV Light Absorbency Moisture and Volatiles Metal Traces Hydrocarbons: stigmastadiene, stigmasta-3, 5 diene, and campesta-3, 5 diene Trans fatty acid isomer Total aliphatic alcohols DNA characterization Color Aspect Free Acidity Peroxide Value Insoluble Impurities Flash Point
Olive Oil (Standards of Identity) Sensory Halogenated Solvents Additives Heavy Metals & Pesticide Residues
Olive Oil (IFSH Focus) Technology for Olive Oil Authenticity & Adulteration Rapid Technology for Olive Oil Quality (Sensory) Collaborators Agilent Alpha Mos AOCS LC-MS/MS and QTOF GC-MS ICP-MS Electronic Nose (FID) Statistical Analysis Software Goal Rapid Techniques to relate to sensory quality (Rapid) Technology to confirm Authenticity and Adulteration
Technology LC-Q-TOF Electronic Nose/GC-MS PCA Statistical Mapping
Mass Spectroscopy LC-MS-TOF (Q- TOF) LC-MS/MS
LC-QTOF Sample Prep and Chromatographic Parameters Method: 5 ml oil diluted with 5 ml hexane extracted twice with 10 ml methanol, evaporated and reconstituted in 50% methanol in water. (Polar Fraction) Chromatographic separation achieved on a Poroshell 120 EC-C18 2.7 µm column kept at 45 o C Gas temp was 325 o C with a flow of 10 L/min Nebulizer was 45 psig Analysis was conducted in positive ionization mode Injection volume 5 µl The mobile phase was a gradient of 0.1% acetic acid in water and acetonitrile starting at 5% ACN and increasing to 100% ACN over 25 min with a post time of 3 min.
Italian Olive Oil/Soybean Oil
Overlay
Oil Analysis Using QTOF Technology Chromatograms Produced Using MFE (Molecular Feature Extraction) and Contain Several Thousands of Compounds Olive Oil from Palestine Soybean Oil
Mass Profiler Professional MPP software allows multivariance statistical analysis of MS data Data can be filtered based on set parameters and organized into entity lists.
Mass, Retention Time
PCA Italian Olive Oil
Olive Oil Varieties
PCA Analysis of Olive Oils and Olive Oils Combined with Vegetable Oils
Venn Diagram of Olive Oils and Olive Oils Overlap with Soybean Specific entity lists can be displayed in many different ways. An example is A Venn diagram representing similarities and differences among various groups of samples.
ID Browser for Identification of Compounds included in Entity Lists An entity list can be also identified based on available databases and libraries using the ID browser
Table of representative 38 compounds identified in EVOO. Over 200 compounds identified. Name Formula Mass RT 12-amino-octadecanoic acid C18 H37 N O2 299.282 21.911 2-Oxo-4E-hexenoic acid C6 H8 O3 128.0478 1.657 Vanillin C8H8O3 152.0473 1.298 3,4-bis(methylene)-Hexanedioic acid C8 H10 O4 170.0577 1.657 2,3-Dihydroxy-3-methylvaleric acid C6 H12 O4 148.0733 4.995 Oleamide C18 H35 N O 281.2714 21.911 2,2-dimethyl-4-pentenoic acid C7 H12 O2 128.0831 8.676 (R)-3-methyl-2-oxo-Pentanoic acid C6 H10 O3 130.0628 3.276 2,2-dimethyl-3-butenoic acid C6 H10 O2 114.0691 3.919 2-Hydroxycinnamic acid C9 H8 O3 164.0465 10.404 3-oxo-4-pentenoic acid C5 H6 O3 114.031 3.61 (5R)-5-Hydroxyhexanoic acid C6 H12 O3 132.079 16.458 (R)-3-methyl-2-oxo-Pentanoic acid C6 H10 O3 130.0627 5.296 2-Octenedioic acid C8 H12 O4 172.0729 2.709 (5R)-5-Hydroxyhexanoic acid C6 H12 O3 132.079 15.79 2-keto-n-caprylic acid C8 H14 O3 158.0938 1.716 2,2-dimethyl-4-pentenoic acid C7 H12 O2 128.0825 10.621 Isovalerylglucuronide C11 H18 O8 278.1018 4.495 2E,4E,6E,8E-Decatetraenedioic acid C10 H10 O4 194.0573 14.721 11-amino-undecanoic acid C11 H23 N O2 201.1724 1.248 2-ethyl-3E-hexenoic acid C8 H14 O2 142.0984 1.949 12-amino-dodecanoic acid C12 H25 N O2 215.1878 2.166 6,8-dihydroxy-octanoic acid C8 H16 O4 176.1056 9.803 2,2-dimethyl-4-pentenoic acid C7 H12 O2 128.083 10.32 13-methyl-pentadecanoic acid C16 H32 O2 256.2393 0.931 10,12,14-octadecatrienoic acid C18 H30 O2 278.2254 20.984 3,4,5-Trimethoxycinnamic acid C12 H14 O5 238.0826 15.314 10-oxo-5,8-decadienoic acid C10 H14 O3 182.093 10.629 3,6-dioxo-decanoic acid C10 H16 O4 200.1035 10.629 2S-amino-pentanoic acid C5 H11 N O2 117.0795 6.372 5-oxo-7-octenoic acid C8 H12 O3 156.0798 11.306 (+)-catechin C15 H14 O6 290.0809 8.108 Tyrosol C8H10O2 138.0681 11.306 (+)-pinoresinol C20H22O6 358.1416 12.684 Syringic acid C9H10O5 198.0536 10.195 Caffeic acid C9H8O4 180.0423 9.752 Elenolic acid C11 H14 O6 242.0788 4.578 p-coumaric acid C9H8O3 164.0465 10.404
Name Formula Mass RT 12-amino-octadecanoic acid C18 H37 N O2 299.282 21.911 2-Oxo-4E-hexenoic acid C6 H8 O3 128.0478 1.657 Vanillin C8H8O3 152.0473 1.298 3,4-bis(methylene)-Hexanedioic acid C8 H10 O4 170.0577 1.657 2,3-Dihydroxy-3-methylvaleric acid C6 H12 O4 148.0733 4.995 Oleamide C18 H35 N O 281.2714 21.911 2,2-dimethyl-4-pentenoic acid C7 H12 O2 128.0831 8.676 (R)-3-methyl-2-oxo-Pentanoic acid C6 H10 O3 130.0628 3.276 2,2-dimethyl-3-butenoic acid C6 H10 O2 114.0691 3.919 2-Hydroxycinnamic acid C9 H8 O3 164.0465 10.404 3-oxo-4-pentenoic acid C5 H6 O3 114.031 3.61 (5R)-5-Hydroxyhexanoic acid C6 H12 O3 132.079 16.458 (R)-3-methyl-2-oxo-Pentanoic acid C6 H10 O3 130.0627 5.296 2-Octenedioic acid C8 H12 O4 172.0729 2.709 (5R)-5-Hydroxyhexanoic acid C6 H12 O3 132.079 15.79 2-keto-n-caprylic acid C8 H14 O3 158.0938 1.716 2,2-dimethyl-4-pentenoic acid C7 H12 O2 128.0825 10.621 Isovalerylglucuronide C11 H18 O8 278.1018 4.495 2E,4E,6E,8E-Decatetraenedioic acid C10 H10 O4 194.0573 14.721 11-amino-undecanoic acid C11 H23 N O2 201.1724 1.248 2-ethyl-3E-hexenoic acid C8 H14 O2 142.0984 1.949 12-amino-dodecanoic acid C12 H25 N O2 215.1878 2.166 6,8-dihydroxy-octanoic acid C8 H16 O4 176.1056 9.803 2,2-dimethyl-4-pentenoic acid C7 H12 O2 128.083 10.32 13-methyl-pentadecanoic acid C16 H32 O2 256.2393 0.931 10,12,14-octadecatrienoic acid C18 H30 O2 278.2254 20.984 3,4,5-Trimethoxycinnamic acid C12 H14 O5 238.0826 15.314 10-oxo-5,8-decadienoic acid C10 H14 O3 182.093 10.629 3,6-dioxo-decanoic acid C10 H16 O4 200.1035 10.629 2S-amino-pentanoic acid C5 H11 N O2 117.0795 6.372 5-oxo-7-octenoic acid C8 H12 O3 156.0798 11.306 (+)-catechin C15 H14 O6 290.0809 8.108 Tyrosol C8H10O2 138.0681 11.306 (+)-pinoresinol C20H22O6 358.1416 12.684 Syringic acid C9H10O5 198.0536 10.195 Caffeic acid C9H8O4 180.0423 9.752 Elenolic acid C11 H14 O6 242.0788 4.578 p-coumaric acid C9H8O3 164.0465 10.404 Blue=phenolic compounds Green=fatty acids
PCA
3-D PCA
PCA Analysis of Italian (P) Greek (G) Olive Oils and Soybean Oil(B). Cluster analysis based on five sensors with the highest discrimination power.
Palestinian Olive Oil Adulterated with 2%, 5% and 10% Soybean Oil
IFSH Direction Oxidation correlated to Analytical Methods PV, FFA, Sensory correlated to Analytical Methods Continue with Adulteration and Authenticity ICP-MS HS-GC-MS Statistical Analysis PCA Mass Profiler Professional
IFSH Institute for Food Safety & Health (IFSH) Bedford Park, IL National Center for Food Safety and Technology (NCFST) At the Illinois Institute of Technology (IIT) Chicago 50:50 IIT:FDA Collaborative Model of Industry:Academia:FDA Food Safety Chemistry, Process Engineering, Microbiology (molecular biology) Nutrition (biological/clinical) In existence for 23 years
Thank you Questions?