Lipid Analysis by Evaporative Light Scattering Detection

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1 Grace Davison Discovery Sciences Lipid Analysis by Evaporative Light Scattering Detection Brochure #A

2 High Performance Analysis of Polar and Non-Polar Lipid Introduction The composition of the lipid fraction in fats and oils influences the nutritional value of the food and is important for health related issues such as digestion, heart disease, and more. Triglycerides are a major component of the non-polar lipid fraction. This is usually very complex due to various chain lengths and different degrees of saturation. Phospholipids and sphingolipids have a positive influence on cholesterol levels and are also used for their emulsifying qualities. Combine the benefits of Evaporative Light Scattering Detection with Alltech s HPLC columns for sensitive, efficient, gradient-compatible separations of both polar and non-polar lipid fractions. References Analysis of Phospho- and Sphingolipids in Dairy Products by a New HPLC Method, R. Rombaut, J. V. Camp and K. Dewettinck - Ghent University, Belgium, published in J. Dairy Sci. :-. Triglyceride Analysis of Edible Oils by HPLC-ELSD, R. Rombaut, I. Foubert, V. Van Hoed, K. Degrave, R. Verhé and K. Dewettinck - Ghent University, Belgium, to be published. Benefits of Evaporative Light Scattering Detection (ELSD) for Lipid Analysis More sensitive than RI and low-wavelength UV Gradient compatible Eliminates need for derivatization Solvent choice not limited by UV-cutoff The Evaporative Light Scattering Detector (ELSD) is a universal HPLC detector that detects any compound less volatile than the mobile phase to low nanogram levels. It has many advantages over RI and low-wavelength UV detection for lipid analyses. Since most lipid compounds are not good UV absorbers, they must be monitored at low wavelengths. This can be hindered by the mobile phase solvent s UV cutoff and poor baseline stability. With ELSD, the mobile phase evaporates before detection, so it is blind to the mobile phase s spectroscopic properties and compositional changes. Although refractive index (RI) detection is universal in nature, its lack of sensitivity and gradient incompatibility make it less than ideal for lipid analysis. Evaporation of the mobile phase before detection makes the ELSD gradient compatible for improved resolution and faster separations. In addition, ELSDs can be as much as 0 to 000 times more sensitive than RI detectors. How does Evaporative Light Scattering Detection work? ELSDs detect any compound less volatile than the mobile phase using a simple three-step process:. Nebulization Column effluent passes through a needle and mixes with nitrogen gas to form a dispersion of droplets.. Evaporation Droplets pass through a heated drift tube where the mobile phase evaporates, leaving a fine mist of dried sample particles in solvent vapor.. Detection The sample particles pass through a cell and scatter light from a laser beam. The scattered light is detected, generating a signal.

3 HPLC Columns for Triglycerides and Polar Lipids Alltima HP C Hi-Load Columns for Triglycerides Excellent peak symmetry Superior retention and loading capacity Stable from ph -0 Extremely low bleed Alltima HP C Hi-Load columns use high purity, low metal, ph stable silica for high resolution with no tailing and little to no bleed. The high carbon load, combined with strong organic mobile phases, enhance triglyceride solubility and performance with ELSD or MS detection. Alltima HP C Hi- Load columns are ideal for analysis of complex hydrophobic mixtures. Prevail Silica for Polar Lipids High capacity and resolution of polar lipids Long column life Stable from ph - Prevail silica s high surface area and high purity increase capacity and resolution of polar lipids while maintaining excellent peak shape. Choose µm particles for high efficiencies, and a variety of column formats for speed and ELSD and MS compatibility. Prevail Silica is ideal for analysis of polar lipids in dairy products. Olive Oil Lecithin, 0.mg/mL. LLO. OOLn. PLnO. LOO. PoOO. PLO. PoOP. OOO. POO+SOL 0. POP. SOO. POS+SLS. PPS. C 0. PA. PE. PI. LPE. PC. LPC 0 0 Column: Alltima HP C Hi-Load, µm, 0 x.0mm (Part No. 0) Gradient: Time: 0 %B: Flow Rate: 0.mL/min Column Temp: 0 C Column: Prevail Silica, µm, 0 x.0mm (Part No. ) Mobile Phase: A: Chloroform B: Methanol C: Triethylamine buffer ph Gradient: Time: 0 %B: 0 %C: Flow Rate: 0.mL/min Column Temp: 0 C

4 Triglycerides Butter Coconut Oil 0 0. CCLa. LaLaO. C. LaMM. CLaLa 0. LaMO. CCM. LaMP/MMM. LaLaLa. MMP/LaPP/MPO. CLaM. C. LaLaM 0 Column: Alltima HP C Hi-Load, µm, 0 x.0mm (Part No. 0) Gradient: Time: 0 %B: Flow Rate: 0.mL/min Column Temp: 0 C Column: Alltima HP C Hi-Load, µm, 0 x.0mm (Part No. 0) Gradient: Time: 0 %B: Flow Rate: 0.mL/min Column Temp: 0 C Cocoa Butter Corn Oil 0 0. C. PLO. PLP. POO. PLS. POP. SOO/PPP. SLS. POS 0. PPS. SOS. PSS. SOA. C. C. LLLn. LLL. LLO. PLL. LOO. PLO. PLP. OOO 0. SLO. POO. POP. SOO/PPP C 0 0 Column: Alltima HP C Hi-Load, µm, 0 x.0mm (Part No. 0) Gradient: Time: 0 %B: Flow Rate: 0.mL/min Column Temp: 0 C Column: Alltima HP C Hi-Load, µm, 0 x.0mm (Part No. 0) Gradient: Time: 0 %B: Flow Rate: 0.mL/min Column Temp: 0 C

5 Triglycerides Sunflower Oil 0 Sesame Seed Oil 0. LLL. LLO. PLL. LOO. LLS. PLO. OOO. SLO. POO LLL. LLO. LLP. OOL. POL. PPL. OOO. OOP. PPO 0. OOS Min. Column: Alltima HP C Hi-Load, µm, 0 x.0mm (Part No. 0) Gradient: Time: 0 %B: Flow Rate: 0.mL/min Column Temp: 0 C Column: Alltima HP C Hi-Load, µm, 0 x.mm (Part No. ) Gradient: Time: 0 %B: Flow Rate:.0mL/min Column Temp: 0 C Refined Sunflower Oil. C. LLL. LLO. PLL. LOO. LLS. PLO. OOO. SLO 0. POO. C 0 00 Soybean Oil 0. C. LLnLn. LLLn. LLL. OLnL. PLLn. LLO. OOLn. PLL 0. LOO. LLS. PLO. PLP. OOO. SLO. POO. POP. SOO/PPP. C 0 Column: Alltima HP C Hi-Load, µm, 0 x.0mm (Part No. 0) Gradient: Time: 0 %B: Flow Rate: 0.mL/min Column Temp: 0 C Column: Alltima HP C Hi-Load, µm, 0 x.0mm (Part No. 0) Gradient: Time: 0 %B: Flow Rate: 0.mL/min Column Temp: 0 C

6 Triglycerides. C. OLL. PLL. LOO. PLO. PLP. MOP. OOO. POO 0. PLS. POP. SOO/PPP. POS. PPS. SOS. C Palm Oil 0 Palm Kernel Stearin Fraction 0 0. CCLa. LaMM. C. LaMO. CLaLa 0. LaMP/MMM. CMM. MMO/LaPO. LaLaLa. MMP/LaPP/MPO. LaLaM. C. LaLaO 0 Column: Alltima HP C Hi-Load, µm, 0 x.0mm (Part No. 0) Gradient: Time: 0 %B: Flow Rate: 0.mL/min Column Temp: 0 C Column: Alltima HP C Hi-Load, µm, 0 x.0mm (Part No. 0) Gradient: Time: 0 %B: Flow Rate: 0.mL/min Column Temp: 0 C Palm Kernel Olein Fraction. CCLa. C. CLaLa. CCM. LaLaLa. CLaM. LaLaM. LaLaO. LaMM LaMO. LaMP/MMM. MMO/LaPO. MMP/LaPP/MPO. PLP. POO. POP. C Column: Alltima HP C Hi-Load, µm, 0 x.0mm (Part No. 0) Gradient: Time: 0 %B: Flow Rate: 0.mL/min Column Temp: 0 C Compound Translation Key A C La L Ln M O P S Triglycerides Arachidic Acid Caproic Acid Lauric Acid Linoleic Acid Linolenic Acid Myristic Acid Oleic Acid Palmitic Acid Stearic Acid Polar Lipids PS Phosphatidylserine PE Phosphatidylethanolamine PI Phosphatidylinositol PC Phosphatidylcholine PA Phosphatidic Acid LPE lyso-phosphatidylethanolamine LPC lyso-phosphatidylcholine GLUCCER Glucosylceramide LACCER Lactosylceramide SM Sphingomyelin SM Sphingomyelin U Unknown

7 Polar Lipids Polar Lipid Standards. GLUCCER. LACCER. PE. PI. PS. PC. SM. SM Acid Whey 0 0. LACCER. PA. PE. PI. PS. PC. SM. SM Column: Prevail Silica, µm, 0 x.0mm (Part No. ) Mobile Phase: A: Chloroform; B: Methanol; C: Triethylamine buffer ph Gradient: Time: 0 %B: 0 %C: Flow Rate: 0.mL/min Column Temp: 0 C Column: Prevail Silica, µm, 0 x.0mm (Part No. ) Mobile Phase: A: Chloroform; B: Methanol; C: Triethylamine buffer ph Gradient: Time: 0 %B: 0 %C: Flow Rate: 0.mL/min Column Temp: 0 C Gouda Cheese Butter. LACCER. U. PA. PE. PI. PS. PC. SM. SM 0. LPC 0 0. PE. PI. PS. PC. SM. SM 0 Column: Prevail Silica, µm, 0 x.0mm (Part No. ) Mobile Phase: A: Chloroform; B: Methanol; C: Triethylamine buffer ph Gradient: Time: 0 %B: 0 %C: Flow Rate: 0.mL/min Column Temp: 0 C Column: Prevail Silica, µm, 0 x.0mm (Part No. ) Mobile Phase: A: Chloroform; B: Methanol; C: Triethylamine buffer ph Gradient: Time: 0 %B: 0 %C: Flow Rate: 0.mL/min Column Temp: 0 C

8 Ordering Information Triglycerides Alltima HP Columns Alltima HP HPLC Columns Phase Format i.d. x Length Part No. C Hi-Load, Expedite MS. x 0mm µm Expedite MS. x 0mm LC/MS. x 0mm 0 LC/MS. x 00mm LC/MS. x 0mm Solvent-Reducer.0 x 0mm 0 Expedite MS. x 0mm Expedite MS. x 0mm Analytical. x 0mm Analytical. x 00mm Analytical. x 0mm Rocket.0 x mm Rocket.0 x mm 0 C Hi-Load, LC/MS. x 0mm µm LC/MS. x 0mm 00 Solvent-Reducer.0 x 0mm Solvent-Reducer.0 x 0mm Analytical. x 0mm Analytical. x 0mm Alltima HP All-Guard Cartridges* Phase i.d. x Length Qty Part No. C Hi-Load. x.mm 0.0 x.mm. x.mm 0 All-Guard Cartridge Holder (Includes Direct-Connect Column Coupler) ea 00 *All-Guard Holder required Polar Lipids Prevail Columns Prevail All-Guard Cartridges* Phase i.d. x Le n g t h Qt y Part No. Silica. x.mm.0 x.mm. x.mm All-Guard Cartridge Holder (Includes Direct-Connect Column Coupler) ea 00 *All-Guard Holder required Prevail HPLC Columns Phase Format i.d. x Length Part No. Silica, Expedite MS. x 0mm µm Expedite MS. x 0mm LC/MS. x 0mm LC/MS. x 00mm 0 LC/MS. x 0mm Solvent-Reducer.0 x 0mm Expedite MS. x 0mm Expedite MS. x 0mm Analytical. x 0mm Analytical. x 00mm Analytical. x 0mm Rocket.0 x mm Rocket.0 x mm Silica, Analytical. x 0mm µm Solvent-Reducer.0 x 0mm Solvent-Reducer.0 x 0mm 0 Analytical. x 0mm Analytical. x 0mm Copyright 00 Alltech Associates, Inc. Alltech Associates Inc., is a wholly owned subsidiary of W. R. Grace & Co.-Conn. Grace Davison is a business unit of W. R. Grace & Co.-Conn., Global Headquarters 00 Grace Drive, Columbia, MD 0. The information contained herein is based on our testing and experience and is offered for the user's consideration, investigation and verification. Since operating and use conditions vary and since we do not control such conditions, we must DISCLAIM ANY WARRANTY, EXPRESSED OR IMPLIED, with regard to results to be obtained from the use of this product. Test methods are available on request. Alltech reserves the right to change prices and/or specifications without prior notification. ALL-GUARD, ALLTECH, ALLTIMA, DIRECT-CONNECT, EXPEDITE, GROM, JONES CHROMATOGRAPHY, MODCOL, PREVAIL, ROCKET, and VYDAC are trademarks of Alltech Associates, Inc. DAVISIL, DAVISON, GRACE, and GRACE DAVISON are registered trademarks of W. R. Grace & Co.-Conn. This trademark list has been compiled using available published information as of the publication date of this brochure and may not accurately reflect current trademark ownership. Grace Davison Discovery Sciences Headquarters 0 Waukegan Road Deerfield, Illinois 00- Tel:..00 Fax:..0 discoverysciences@grace.com Web: 0/0 #A

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