Lipid analysis in Food Industry by HPLC-ELSD Brochure Lpd-1
The Best Column and Detector for Superior Lipid Analysis Introduction The characterization of the lipid fraction present in fats and oils became very important in the food industry. The composition of the lipid fraction has not only an influence on the nutritional value of the food but is also important for health related issues (digestion, prevention of heart diseases, etc). Triglycerides are a major component of the non-polar lipid fraction. The composition of this fraction is usually very complex due to various chain lengths and different degrees of unsaturation. Phospho- and sphingolipids are part of the polar lipid fraction. These compounds have not only a positive influence on biological activities in the human body (e.g. reducing blood cholesterol levels) but are also used for their functionality and emulsifying qualities in food systems. Gas chromatography has long been a preferred technique for triglyceride analysis however special care is needed for both column choice and injection system. Since evaporative light scattering detectors became available together with high efficient HPLC columns, HPLC analysis of triglycerides gained popularity. HPLC is also the preferred technique for analysis of the polar lipid fraction. The ELSD2000ES, in combination with Alltima HP High Load or Prevail Silica columns, allows high performance HPLC gradient analysis and detection of both non-polar and 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. 88:482-488. 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. The ELSD 2000 ES Superior Sensitivity and Stability vs. RI and Low-Wavelength UV Outperforms RI and Low-Wavelength UV Offers Outstanding Sensitivity Delivers Rock-Solid Gradient Baselines Independent of the degree of saturation and chain lenght of an acyl chain Minimal Sample Preparation The Alltech ELSD 2000ES Evaporative Light Scattering Detector is a universal HPLC detector offering distinct advantages over RI and low-wavelength UV for lipid analyses. 5 10X higher sensitivity, total gradient compatibility for improved resolution and faster separations, no injection events to interfere with early eluting sample components, and exceptionally quiet and stable baselines unaffected by fluctuations in solvent composition or temperature. Because the mobile phase evaporates before the light scattering measurement occurs, the ELSD is blind to the spectroscopic properties of and compositional changes in the mobile phase. This is a significant advantage over RI and low-wavelength UV detectors which place stringent requirements on the spectroscopic nature of the solvent. With the ELSD, solvent fronts are eliminated and rock-solid gradient baselines are the norm. The laser has no decline in light-intensity, a long lifetime and high intensity that leads to an excellent reproducibility, unique sensitivity and stability over longer periods of analysis. The signal of the ELSD detector is independent of the degree of saturation and chain lenght of an acyl chain unlike a low-wavelength UV. How does it work? ELSDs detect any compound less volatile than the mobile phase using a simple three-step process: 1. Nebulization Column effluent passes through a needle and mixes with nitrogen gas to form a dispersion of droplets. 2. Evaporation Droplets pass through a heated drift tube where the mobile phase evaporates, leaving a fine mist of dried sample particles in solvent vapor. 3. Detection The sample particles pass through a cell and scatter light from a laser beam. The scattered light is detected, generating a signal.
Columns for Triglycerides and Polar lipids Alltima HP HiLoad Columns Long column life - exceptional column stability minimizes downtime and reduces cost Better Peak Symmetry - high purity silica eliminates peak tailing problems Ideal for critical analysis - low to no detectable column bleed Superior retantion and loadability High cabon load (24%) The Alltima HP HiLoad columns are based on a high purity, low metal containing silica. This superior silica quality will give symmetrical sharp peaks without tailing. The material also shows exceptional stability from ph 1 to 10. Due to the extreme low bleed behaviour the material is very well suited for LC/MS and critical applications. The high carbon load of the HiLoad columns (24% C) creates a stronger retention allowing use of higher organic fractions in the mobile phase which enhances performance when coupled with MS or ELSD detectors. The high carbon load also allows to load more sample onto the columns. The good retention behaviour makes Alltima HP HiLoad a good material to analyse complex hydrophobic mixtures like triglycerides. Prevail Silica Columns Small pore silica High surface area ph stability Long column life Prevail Silica is a small pore (110 A) high quality silica with a relative high surface area (350 m²/g). This silica has good ph stability (ph 1 to 8) and will give long column life time. The high surface area will give good retention behaviour and symmetrical peaks. The material is available in 3µm particles for high efficiencies and can be packed in various column formats like narrow bore (3.0mm ID) for enhanced compatibility with ELSD detection. The good material stability makes Prevail Silica a very well suited material for demanding applications like the analysis of polar lipids in dairy products. Olive Oil Lpd101 Lecitine, 0.5 mg/ml Lpd102 3
Applications Triglycerides Butter Lpd103 Coconut Oil Lpd104 Cocoa butter Lpd105 Corn Oil Lpd106 4
Triglycerides Applications Sunflower oil - impactor OFF Lpd107 Sunflower oil - impactor ON Lpd108 Refined sunflower oil Lpd109 Soy oil Lpd110 5
Applications Triglycerides Palm oil Lpd111 Palm kernel stearin fraction Lpd112 Palm kernel olein fraction Lpd113 Hydrogenated palm kernel oil Lpd114 6
Polar Lipids Applications Polar Lipid Standards Lpd115 Acid Whey Lpd116 Gouda Cheese Lpd117 Butter Lpd118 7
Ordering Information Triglycerides ALLTIMA HP HPLC COLUMNS PACKING FORMAT I.D. X LENGTH PART NO. PRICE C18 Hi-Load, Solvent-Reducer 3.0 x 150mm 87610 3µm Analytical 4.6 x 50mm 87827 Analytical 4.6 x 100mm 87685 Analytical 4.6 x 150mm 87686 Rocket 7 x 33mm 87689 Rocket 7 x 53mm 87690 ALLTIMA HP ALL-GUARD CARTRIDGES PACKING I.D. X LENGTH QTY. PART NO. PRICE C18 Hi-Load 3.0 x 7.5mm 3 87624 4.6 x 7.5mm 3 87702 All-Guard Guard Cartridge Holder ea 80101 Polar Lipids PREVAIL HPLC COLUMNS PACKING FORMAT I.D. X LENGTH PART NO. PRICE Silica, Solvent-Reducer 3.0 x 150mm 99341 3µm Analytical 4.6 x 50mm 43842 Analytical 4.6 x 100mm 99269 Analytical 4.6 x 150mm 99271 Rocket 7 x 33mm 99284 Rocket 7 x 53mm 99283 PREVAIL ALL-GUARD CARTRIDGES PACKING I.D. X LENGTH QTY. PART NO. PRICE Silica 3.0 x 7.5mm 3 99354 4.6 x 7.5mm 3 99291 All-Guard Guard Cartridge Holder ea 80101 ELSD 2000ES* Detector Specifications Light Source: Detector Element: Temperature Range: Nebulizer Gas: Inlet Pressure: ELSD 2000ES* Detector Laser diode with collimating optics, 650nm, 30mW output, Class IIIB Silicon photodiode Ambient to 120ºC in 1ºC increments Nitrogen preferred; Calibrated to 4.0L/min 60-80psig Typical Operating Range: 1.0-3.0L/min Flow Control: Digital mass flow control Mobile Phase Flow Rate: To 5.0mL/min Analog Outputs: Communications: User Interface: Power Requirements: Dimensions: Weight: IEC/FDA Classification: Warranty: 1V or 10mV full scale Inputs: TTL/contact closure for auto zero and gas shutoff Outputs: Contact closure for pump shutdown in error conditions Graphical LCD with alphanumeric keypad and /or Windows based PC control via serial I/O 240V, 50Hz 58.4cm H x 31.8cm W x 54.8cm D 16kg Class 1 laser product 1 year parts and labor DESCRIPTION PART NO. PRICE 240V 600200 Your Online Technical Resource for Chromatography www.alltechweb.com The trademarks referred to herein are the property of their respective owners. Contact your local office or Alltech European Headquaters Belgium/European Headquarters Alltech Associates, Inc. Brandstraat 12 9160 Lokeren Tel: 09 340 65 65 Fax: 09 340 65 60 Email: info@alltech.be United Kingdom and Ireland Alltech Associates /Applied Science Ltd. 6 Kellet Road Ind. Estate Carnforth Lancashire, LA5 9XP Tel: 01524 734451 Fax: 01524 733599 Email: info@alltechuk.co.uk France Alltech France sarl Centre d Activité de l Epinoy BAT. 2, CD 952 BP 95 F-59175 Templemars Tel: 03 20 62 96 00 Fax: 03 20 62 96 09 Email: info@alltech.fr The Netherlands Alltech/Applied Science Group PB 6596 NL-4802 HN Breda Tel: 076/571 75 76 Fax: 076/571 46 56 Email: info@alltech.nl Germany Alltech GmbH In der Hollerhecke 1 D-67545 Worms Tel: 06241-403 759 Fax: 06241-403 09759 Email: info@alltechweb.de Italy Alltech Italia srl, Via Meucci 15 Loc. Cascina Serinda I-20018 Sedriano MI Tel: 02/901 10 150 ra Fax: 02/901 10 131 Email: info@alltechweb.it Copyright 2005 Alltech Associates, Inc. 8 03/05 #000 Alltech reserves the right to change prices and/or specifications without prior notification. Chromatograms with the courtesey of Prof. Dr. Ir. K. Dewettinck and Ir. R. Rombaut - Ghent University (Belgium) Dept of Food Recycled Technology Paper & Nutrition. part of the Alltech Use of Our Products: Alltech Associates, Inc., supplies chromatographic equipment reagents and accessories commitment to protect our valuable for non-medical laboratory use only. This means our products:... may NOT be used as drugs,...may NOT be ISO 9002 REGISTERED COMPANY used as cosmetics,...may NOT be used as medical devices or as in vitro diagnostic devices. natural resources 015