Determination of Bitter Melon Momordicoside by HPLC 1) Reagents a. Methanol (HPLC) b. Acetronitrile(HPLC) c. Nanopure water
2) Apparatus HPLC (Shimadzu) Analytical balance (1/10000) Ultrasonic cleaner 0.45μm PTFE filter 50ml volumetric flask Screw cap vials
3) HPLC condition Column: Kromasil ODS C18 5μm , 4.6×150mm Detector: UV208nm Mobile Phase: A=Water B= MeOH:Aceton=1:1 Flow Rate: 0.8ml/min Injection: 10μl
4)Procedure
Samples Preparation Accurately weigh 120mg 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.
Reference preparation /calibration Prepare an reference solution (approximately 120mg/50ml in MeOH) using a commercial reference of Bitter Melon Momordicoside,Sonicate for about 10 minutes and transfer to screw-cap vials and can be stored in the refrigeration.
5) Calculation Compare the peak areas of the Bitter Melon Momordicoside on the sample chromatogram and the corresponding peaks in the chromatogram for the reference solution.
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