206 / 227
1$785$/ 5(0(',(6
3DJH RI
4.5) Accuracy:
4.5.1) Definition:
The accuracy is expressed as the difference between the average value
obtained from large series of observed value and the true value.
4.5.2) Assessment:
Accuracy is determined by Spike recovery method. A known quantity of
standards is added to the sample and the sample is re-analyzed using the same method
which is to be validated.
4.5.3)
Four different batches of
Withania somnifera
extracts were analyzed to find out actual
content of withanolides. All markers were calculated using standard dilution–III. The same
sample were spiked with different quantities of Withanoside IV, Withanoside V, Withaferin
A, 12-Deoxy Withastramonolide, Withanolide A and Withanolide B and analysed for their
total content.
4.5.4) Acceptance Criteria:
1) For Assay of >10 and < 95% recovery should be 95.0 -110%.
2) For Assay of >7.5 and < 10% recovery should be 90-115%.
3) For the Assay > 0.5 and < 7.5% recovery should be 85-120%.
4) For the Assay > 0.1 and < 0.5% recovery should be 75-125%.
5) For the Assay of < 0.1% recovery should be 70 to 130%.
4.5.5) Experiment:
a) Standard preparation: Refer 4.2.4 – Dilution-3.
b) Sample preparation:
The sample solution was prepared by weighing the quantity of the
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-53:
Batch No.
Quantity (mg)
WS/06Lot08
1714.4
WS/06Lot10
1520.4
WS/05Lot20
5029.7
WS/05Lot21
2612.5
RD/1170
1632.9
RD/1045
1640.2
RD/1162 RM
4003.8
ERH-46 RM
4091