208 / 227
1$785$/ 5(0(',(6
3DJH RI
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
resultant solution was
filtered through 0.45 microns membrane filter paper.