SPDS SET 2: ASH-01

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

1$785$/ 5(0(',(6

3DJH RI