Determination of Inositol by HPLC-ELSD 1) Reagents a. Acetonitrile (HPLC) b. Nanopure water 2) Apparatus HPLC (Shimadzu) Analytical balance (1/10000) Ultrasonic cleaner 0.45μm PTFE filter 50ml volumetric flask Screw cap vials ELSD detector(Alltech) 3) HPLC condition Column: Kromasil C18 5μm , 4.6×200mm Mobile Phase: A:Water(95%) B: Acetonitrile (5%) Flow Rate: 0.5 ml/min Injection: 10μl Drift tube temperature :110.5℃ Gas flow:3.0L/min Impactor: Off Gain:2 4) Procedure Samples Preparation Accurately weigh 20mg extraction into 50ml volumetric flask. Add Methanol to volume and sonicate to mix, filter through a 0.45μm PTFE filter into a screw cap vial and proceed with the analysis. Standard preparation /calibration Prepare a standard solution (approximately 0.2mg/ml in Methanol) using a commercial reference standard of Inositol,Sonicate for about 10 minutes and transfer to screw-cap vials and can be stored in the refrigeration.
5) Calculation Determine the peak area’s logarithm of Inositol the on the sample chromatogram and compare to the corresponding peaks in the chromatogram for the sample solution
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