Diosgenin by HPLC-ELSD
Determination of Diosgenin by HPLC-ELSD
1) Reagents
a. Nanopure water b. Acetonitrile (HPLC)
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: Shimpack-VP-ODS C18 5μm , 4.6×150mm
Mobile Phase: A:Water B: Acetonitrile
A/B=10/90(isocratic)
Flow Rate: 1.0 ml/min
Injection: 10μl
Drift tube temperature :74.5℃
Gas flow:1.9L/min
Impactor: Off
Gain:2
4) Procedure
Samples Preparation
Accurately weigh 250mg 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 MEOH) using a commercial reference standard of Diosgenin，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 Diosgenin the on the sample chromatogram and compare to the corresponding peaks in the chromatogram for the sample solution.
1) Reagents
a. Nanopure water b. Acetonitrile (HPLC)
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: Shimpack-VP-ODS C18 5μm , 4.6×150mm
Mobile Phase: A:Water B: Acetonitrile
A/B=10/90(isocratic)
Flow Rate: 1.0 ml/min
Injection: 10μl
Drift tube temperature :74.5℃
Gas flow:1.9L/min
Impactor: Off
Gain:2
4) Procedure
Samples Preparation
Accurately weigh 250mg 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 MEOH) using a commercial reference standard of Diosgenin，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 Diosgenin the on the sample chromatogram and compare to the corresponding peaks in the chromatogram for the sample solution.