SPDS SET 2: ASH-01

filter paper. 20 µ l of standard solution dilution-3 (Refer 4.2.4) and sample solutions were

injected and chromatograms were recorded. From the obtained chromatograms individual withanolides content were calculated.

3) Samples of RD/1170 ( 313.9 mg) was weighed in to 250 ml beaker and 0.94mg of Withanoside IV, 2.43mg of Withanoside V, 1.14mg of Withaferin A, 0.97mg of 12-Doxy withastramonolide, 0.91mg of Withanolide A and 1.02mg of Withanolide B were added to the same beaker. The mixture was 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.

20 µ l of standard solution dilution-3 (Refer 4.2.4) and sample solutions were injected and

chromatograms were recorded. From the obtained chromatograms individual withanolides content were calculated.

4) Samples of RD/1162-RM ( 755.9 mg) was weighed in to 250 ml beaker and 0.96mg of Withanoside IV, 2.43mg of Withanoside V, 1.26mg of Withaferin A, 1.22mg of 12-Doxy withastramonolide, 1.25mg of Withanolide A and 1.07mg of Withanolide B were added to the same beaker. The mixture was 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. 20 µ l of standard solution dilution-3 (Refer 4.2.4) and sample solutions were

injected and chromatograms were recorded. From the obtained chromatograms individual

withanolides content were calculated.

5) Samples of WS/06Lot08 ( 1714.4 mg), WS/05Lot20 ( 5029.7mg ), RD/1045 ( 1640.2 mg), ERH-46 ( 4071mg ) were weighed in to a separate 250 ml beaker and was 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 100ml with methanol.

1ml each of these sample solutions were mixed with 1ml of standard solution containing 0.164mg of Withanoside IV, 0.056mg of Withanoside V, 0.23mg of Withaferin A, 0.19mg of

12-Doxy withastramonolide, 0.16mg of Withanolide A and 0.156mg of Withanolide B. The

filtered through 0.45 microns membrane filter paper.

resultant solution was

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