Agilent High Resolution Analysis of Triglycerides in Cooking Oil using HPLC with ELSD Manual
Contents
1. e o e e J e Author Stephen Ball Agilent Technologies Inc High Resolution Analysis of Triglycerides in Cooking Oil using HPLC with ELSD Application Note Introduction The composition of triglycerides in cooking oil is very complex and oil contains many chemically similar species In order to develop a high resolution HPLC separation of these components gradient elution is required normally involving solvents with very different properties For this reason RI is not viable as a detection method and UV can be difficult due to baseline disturbance arising from the choice of solvents used in the gradient PLRP S 100A columns are ideally suited to the analysis of triglycerides because the very small pore sizes have extremely high surface areas available to the solutes Evaporative light scattering detection ELSD with Agilent s ELS detectors offers significant benefits over RI and UV as it is unaffected by the optical properties of the solvents Additionally it offers high sensitivity for detection of low levels of contaminants that may be present in adulterated oils EE Agilent Technologies Instrumentation Column PLRP S 100A 5 um 150 x 4 6 mm p n PL1111 3500 Detector Agilent ELSD neb 40 C evap 40 C gas 1 6 SLM Materials and Reagents Eluent A Acetonitrile Eluent B Dichloromethane Sample Preparation Concentration 1 mg oil mL Conditions Gradient 10 B hold for 5 min 10 70 B for 30 min Flo2. es For example 100 water at a flow rate of 5 mL min can be removed at 30 C The novel design of the Agilent ELS detector provides superior performance compared to detectors from other vendors for the analysis of semi volatile compounds www agilent com chem This information is subject to change without notice Agilent Technologies Inc 2011 Published in UK May 11 2011 5990 8196EN j Agilent Technologies
3. w Rate 0 5 mL min Injection Volume 10 uL Results and Discussion The chromatogram in Figure 1 shows an HPLC separation of triglycerides in unadulterated cooking oil using a PLRP S column and an Agilent ELS detector The lack of contamination in the sample is evident LL 0 min 20 Figure 1 Agilent ELSD and a PLRP S column produce an extremely stable baseline across the gradient in the analysis of triglycerides Conclusion Using an Agilent ELS detector combined with a PLRP S 100A column successfully resolved triglyceride constituents in cooking oil because of the optimum match of column detector and sample PLRP S columns are ideally suited to the analysis of many small molecules The 100A pore size has an exceptionally high surface area that is accessible to the solutes It is more retentive for small molecules than the majority of alkyl bonded silicas PLRP S media possess a much greater surface area than alkyl bonded silicas and therefore even polar molecules such as carboxylic acids may be retained much longer resulting in greater resolution Agilent ELS detectors surpass other ELSDs for low temperature HPLC applications with semi volatile compounds Its innovative design represents the next generation of ELSD technology providing optimum performance across a diverse range of HPLC applications The Agilent ELS detector s unique gas control permits evaporation of high boiling solvents at very low temperatur
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