Quantitative analysis of structural and signaling lipid networks
Lipid analysis in Food Industry - · PDF fileThe Best Column and Detector for Superior Lipid...
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Brochure Lpd-1 Lipid analysis in Food Industry by HPLC-ELSD
Transcript of Lipid analysis in Food Industry - · PDF fileThe Best Column and Detector for Superior Lipid...
Brochure Lpd-1
Lipid analysis in Food Industryby HPLC-ELSD
The Best Column and Detector for Superior Lipid Analysis
Introduction
The characterization of the lipid frac-tion present in fats and oils became very important in the food industry.
The composition of the lipid frac-tion has not only an infl uence 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 com-position 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 infl uence 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 injec-tion system.Since evaporative light scattering detectors became available together with high effi cient HPLC columns, HPLC analysis of triglycerides gained popular-ity. 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 detec-tion of both non-polar and polar lipid fractions.
The ELSD 2000 ESSuperior Sensitivity and Stability vs. RI and Low-Wavelength UV
• Outperforms RI and Low-Wave-length 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-wave-length UV for lipid analyses.
5–10X higher sensitivity, total gradient compatibility for improved reso-lution and faster separations, no ‘injection events’ to interfere with early eluting sample components, and exceptionally quiet and stable baselines unaffected by fl uctuations in solvent composition or temperature. Because the mobile phase evaporates before the light scattering measurement oc-curs, the ELSD is blind to the spectroscopic properties of and compositional changes in the mobile phase. This is a signifi cant 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 elimi-nated 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 satura-tion and chain lenght of an acyl chain unlike a low-wavelength UV.
How does it work ?
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.
ELSDs detect any compound less volatile than the mobile phase using a simple three-step process:
3. Detection
The sample particles pass through a cell and scatter light from a laser beam. The scattered light is detected, generating a signal.
2. Evaporation
Droplets pass through a heated “drift tube“ where the mobile phase evaporates, leaving a fi ne mist of dried sample particles in solvent vapor.
1. Nebulization
Column effl uent passes through a needle and mixes with nitrogen gas to form a dispersion of droplets.
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Chromatograms with the courtesey of Prof. Dr. Ir. K. Dewettinck and Ir. R. Rombaut - Ghent University (Belgium) Dept of Food Technology & Nutrition.
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 effi ciencies 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.
Column: Alltima HP C18 HL, 3µm, 150 x 3.0mm (P/N 87610)Mobile Phase: Dichloromethane:Acetonitrile, gradient Flowrate: 0.72 ml/minColumn Temp: 20°CDetector: ELSD
Olive Oil Lecitine, 0.5 mg/ml
Column: Prevail Silica, 3µm, 150 x 3.0mm (P/N 99341)Mobile Phase: Chloroform:Methanol:Triethylamine (pH3), gradientFlowrate: 0.5 ml/min Column Temp: 40°CDetector: ELSD
Lpd101 Lpd102
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Chromatograms with the courtesey of Prof. Dr. Ir. K. Dewettinck and Ir. R. Rombaut - Ghent University (Belgium) Dept of Food Technology & Nutrition.
Column: Alltima HP C18 HL, 3µm, 150 x 3.0mm (P/N 87610)Mobile Phase: Dichloromethane:Acetonitrile, gradient Flowrate: 0.72 ml/minColumn Temp: 20°CDetector: ELSD
Butter Coconut Oil
Applications
Corn OilCocoa butter
Triglycerides
Column: Alltima HP C18 HL, 3µm, 150 x 3.0mm (P/N 87610)Mobile Phase: Dichloromethane:Acetonitrile, gradient Flowrate: 0.72 ml/minColumn Temp: 20°CDetector: ELSD
Column: Alltima HP C18 HL, 3µm, 150 x 3.0mm (P/N 87610)Mobile Phase: Dichloromethane:Acetonitrile, gradient Flowrate: 0.72 ml/minColumn Temp: 20°CDetector: ELSD
Column: Alltima HP C18 HL, 3µm, 150 x 3.0mm (P/N 87610)Mobile Phase: Dichloromethane:Acetonitrile, gradientFlowrate: 0.72 ml/minColumn Temp: 20°CDetector: ELSD
Lpd103Lpd103 Lpd104
Lpd106Lpd105
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Chromatograms with the courtesey of Prof. Dr. Ir. K. Dewettinck and Ir. R. Rombaut - Ghent University (Belgium) Dept of Food Technology & Nutrition.
Sunfl ower oil - impactor OFF Sunfl ower oil - impactor ON
ApplicationsTriglycerides
Column: Alltima HP C18 HL, 3µm, 150 x 3.0mm (P/N 87610)Mobile Phase: Dichloromethane:Acetonitrile, gradientFlowrate: 0.72 ml/minColumn Temp: 20°CDetector: ELSD
Column: Alltima HP C18 HL, 3µm, 150 x 3.0mm (P/N 87610)Mobile Phase: Dichloromethane:Acetonitrile, gradient Flowrate: 0.72 ml/minColumn Temp: 20°CDetector: ELSD
Column: Alltima HP C18 HL, 3µm, 150 x 3.0mm (P/N 87610)Mobile Phase: Dichloromethane:Acetonitrile, gradient Flowrate: 0.72 ml/minColumn Temp: 20°CDetector: ELSD
Column: Alltima HP C18 HL, 3µm, 150 x 3.0mm (P/N 87610)Mobile Phase: Dichloromethane:Acetonitrile, gradient Flowrate: 0.72 ml/minColumn Temp: 20°CDetector: ELSD
Refi ned sunfl ower oil Soy oil
Lpd107Lpd107 Lpd108Lpd108
Lpd110Lpd109Lpd109
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Chromatograms with the courtesey of Prof. Dr. Ir. K. Dewettinck and Ir. R. Rombaut - Ghent University (Belgium) Dept of Food Technology & Nutrition.
Applications
Palm kernel stearin fractionPalm oil
Triglycerides
Column: Alltima HP C18 HL, 3µm, 150 x 3.0mm (P/N 87610)Mobile Phase: Dichloromethane:Acetonitrile, gradient Flowrate: 0.72 ml/minColumn Temp: 20°CDetector: ELSD
Column: Alltima HP C18 HL, 3µm, 150 x 3.0mm (P/N 87610)Mobile Phase: Dichloromethane:Acetonitrile, gradient Flowrate: 0.72 ml/minColumn Temp: 20°CDetector: ELSD
Column: Alltima HP C18 HL, 3µm, 150 x 3.0mm (P/N 87610)Mobile Phase: Dichloromethane:Acetonitrile, gradient Flowrate: 0.72 ml/minColumn Temp: 20°CDetector: ELSD
Column: Alltima HP C18 HL, 3µm, 150 x 3.0mm (P/N 87610)Mobile Phase: Dichloromethane:Acetonitrile, gradient Flowrate: 0.72 ml/minColumn Temp: 20°CDetector: ELSD
Palm kernel olein fraction Hydrogenated palm kernel oil
Lpd111 Lpd112
Lpd114Lpd113
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Chromatograms with the courtesey of Prof. Dr. Ir. K. Dewettinck and Ir. R. Rombaut - Ghent University (Belgium) Dept of Food Technology & Nutrition.
Applications
Polar Lipid Standards Acid Whey
Gouda Cheese Butter
Polar Lipids
Column: Prevail Silica, 3µm, 150 x 3.0mm (P/N 99341)Mobile Phase: Chloroform:Methanol:Triethylamine (pH3), gradientFlowrate: 0.5 ml/min Column Temp: 40°CDetector: ELSD
Column: Prevail Silica, 3µm, 150 x 3.0mm (P/N 99341)Mobile Phase: Chloroform:Methanol:Triethylamine (pH3), gradientFlowrate: 0.5 ml/min Column Temp: 40°CDetector: ELSD
Column: Prevail Silica, 3µm, 150 x 3.0mm (P/N 99341)Mobile Phase: Chloroform:Methanol:Triethylamine (pH3), gradientFlowrate: 0.5 ml/min Column Temp: 40°CDetector: ELSD
Column: Prevail Silica, 3µm, 150 x 3.0mm (P/N 99341)Mobile Phase: Chloroform:Methanol:Triethylamine (pH3), gradientFlowrate: 0.5 ml/min Column Temp: 40°CDetector: ELSD
Lpd117 Lpd118
Lpd116Lpd115
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Chromatograms with the courtesey of Prof. Dr. Ir. K. Dewettinck and Ir. R. Rombaut - Ghent University (Belgium) Dept of Food Technology & Nutrition.
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Recycled Paper — part of the Alltech Chromatograms with the courtesey of Prof. Dr. Ir. K. Dewettinck and Ir. R. Rombaut - Ghent University (Belgium) Dept of Food Technology & Nutrition.Recycled Paper — part of the Alltech Chromatograms with the courtesey of Prof. Dr. Ir. K. Dewettinck and Ir. R. Rombaut - Ghent University (Belgium) Dept of Food Technology & Nutrition.commitment to protect our valuable natural resources
© Copyright 2005 Alltech Associates, Inc.
Alltech reserves the right to change prices and/or specifi cations without prior notifi cation.Chromatograms with the courtesey of Prof. Dr. Ir. K. Dewettinck and Ir. R. Rombaut - Ghent University (Belgium) Dept of Food Technology & Nutrition.
Alltech reserves the right to change prices and/or specifi cations without prior notifi cation.Chromatograms with the courtesey of Prof. Dr. Ir. K. Dewettinck and Ir. R. Rombaut - Ghent University (Belgium) Dept of Food Technology & Nutrition.
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Chromatograms with the courtesey of Prof. Dr. Ir. K. Dewettinck and Ir. R. Rombaut - Ghent University (Belgium) Dept of Food Technology & Nutrition.
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ELSD 2000ES* DetectorDESCRIPTION PART NO. PRICE
240V 600200
ELSD 2000ES* Detector Specifi cationsLight Source: Laser diode with collimating optics, 650nm, 30mW output, Class IIIB
Detector Element: Silicon photodiode
Temperature Range: Ambient to 120ºC in 1ºC increments
Nebulizer Gas: Nitrogen preferred; Calibrated to 4.0L/min
Inlet Pressure: 60-80psig
Typical Operating Range: 1.0-3.0L/min
Flow Control: Digital mass fl ow control
Mobile Phase Flow Rate: To 5.0mL/min
Analog Outputs: 1V or 10mV full scale
Communications: Inputs: TTL/contact closure for auto zero and gas shutoff
Outputs: Contact closure for pump shutdown in error conditions
User Interface: Graphical LCD with alphanumeric keypad and /or Windows® based PC control via serial I/O
Power Requirements: 240V, 50Hz
Dimensions: 58.4cm H x 31.8cm W x 54.8cm D
Weight: 16kg
IEC/FDA Classifi cation: Class 1 laser product
Warranty: 1 year parts and labor
Ordering Information
TriglyceridesALLTIMA™ HP HPLC COLUMNS™
PACKING FORMAT I.D. X LENGTH PART NO. PRICE
C18 Hi-Load, Solvent-Reducer 3.0 x 150mm 876103µm Analytical 4.6 x 50mm 87827
Analytical 4.6 x 100mm 87685Analytical 4.6 x 150mm 87686Rocket™ 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 876244.6 x 7.5mm 3 87702
All-Guard™ Guard Cartridge Holder ea 80101
Polar LipidsPREVAIL™ HPLC COLUMNS™
PACKING FORMAT I.D. X LENGTH PART NO. PRICE
Silica, Solvent-Reducer 3.0 x 150mm 993413µm Analytical 4.6 x 50mm 43842
Analytical 4.6 x 100mm 99269Analytical 4.6 x 150mm 99271Rocket™ 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 993544.6 x 7.5mm 3 99291
All-Guard™ Guard Cartridge Holder ea 80101
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