SPDS SET 2: ASH-01

Reproducibility of Results:

4.4)

4.4.1) Experiment

Eight different samples containing various percentages were analysed in triplicate and

Withanolide content of each injection was calculated, the mean of three values and the RSD was determined.

a) Standard preparation: 2.18mg of Withanoside IV (Purity 90%), 0.91mg of Withanoside V (Purity90%), 2.04mg of Withaferin A (Purity 99%), 1.86mg of 12-Deoxy withastramonolide (Purity 95%), 1.92mg of Withanolide A (Purity 99%), 1.58mg of Withanolide B (Purity 99%)

was dissolved in 5ml of methanol.

The sample solution was prepared by weighing the quantity of the

b)

Sample preparation:

sample shown in the Table in a 250 ml beaker, extracted with 100ml of methanol, boiling on water bath for 10-15 minutes and repeated the procedure 3-4 times till the material is completely extracted or till the extracts turn colourless. Combined all the fractions, concentrated and made up the volume to 50ml with methanol. Filtered through 0.45 microns membrane filter paper.

Table-42

Name of the sample

Sample weight

WS/06Lot08

1551.9

WS/06Lot10

1535.8

WS/05Lot20

5047.6

WS/05Lot21

1527.0

RD/1170

1503.3

RD/1045

1653.2

RD/1162

4335.7

ERH-46

4228.6

c) Procedure: 20 µ l of standard solutions and sample solutions were injected in triplicate and

the chromatograms were recorded. From the mean peak area of standard and samples Withanolide content is calculated.

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3DJH RI