Current Specifications of Bioactives in
Sensoril® brand Ashwagandha
Withanolide glycosides (combined Withanoside Iv,
With id V With lid A t ) ≥ 10 0%
anos e ,
ano e , e c. :
.
Withanolide aglycones as Withaferin-A : ≤ 0.5%
Oligosaccharides : ≥ 32.0%
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Current HPLC Method - Quantification of
Withanolide Glycosides and Withaferin-A
HPLC Condtions:
Merck-Hiber
®
Pre-Packed column RT 250-4, LiChrosorb
®
RP-18, Particle
size 5μM, 4 x 250 mm cartridge column, Ord No.1.50333.0001 with a
reverse phase guard column.
A bi t
m en
Acetonitrile:Water-1: 1 (v/v)
0.6 ml/min
20 min
Isocratic
UV 225 nm
20μl (with a loop injector)
Waters HPLC 515 PDA Detector (Waters
™
2996, Photodiode Array
Detector), evaluation with Empower Software
Pdt. No. UN-1648 Acetonitrile (Merck), Pdt No. 93956 Water for HPLC
(Merck)
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Method with external standard; evaluation of area of peaks.
Withaferin A isolated from Withania somnifera by multiple column
chromatography was used as external standard. Withaferin A, 0.5 mg was
dissolved in 1 ml of methanol and four different concentrations were
applied to HPLC. Calibration curve was plotted between area and different
concentration. The linear regression equation of the calibration curve was
obtained as follows:
Y = 2217082.726 X+ 200001.715 with a correlation coefficient of 0.9998.
Where Y is the area and X is the concentration in
g.
