SPDS SET 2: ASH-01

Table-52: Assay content of withanolides in sample: WS/06Lot10 Concentration W –IV W –V WF-A 12-D

W –A

W –B

Sum

Concentration-1 (7.5mg/ml)

Trial -1

0.802

0.681

0.605

0.263

0.209

0.064

2.625

Trial -2

0.821

0.697

0.617

0.268

0.212

0.066

2.680

Trial -3

0.828

0.696

0.626

0.272

0.221

0.066

2.709

Trial -4

0.821

0.684

0.615

0.265

0.218

0.064

2.667

Concentration-2 (10.0mg/ml)

Trial -1

0.808

0.684

0.609

0.265

0.211

0.065

2.642

Trial -2

0.827

0.703

0.621

0.270

0.215

0.065

2.701

Trial -3

0.818

0.684

0.612

0.265

0.217

0.064

2.661

Trial -4

0.826

0.687

0.618

0.268

0.220

0.065

2.684

Concentration-3 (15.0mg/ml)

Trial -1

0.813

0.685

0.608

0.265

0.214

0.064

2.649

Trial -2

0.818

0.691

0.618

0.269

0.216

0.065

2.678

Trial -3

0.818

0.690

0.616

0.268

0.221

0.065

2.677

Trial -4

0.821

0.684

0.612

0.266

0.219

0.064

2.666

Concentration-4 (20.0mg/ml)

Trial -1

0.806

0.686

0.607

0.265

0.210

0.065

2.638

Trial -2

0.811

0.685

0.612

0.267

0.212

0.065

2.651

Trial -3

0.822

0.686

0.616

0.266

0.223

0.065

2.677

Trial -4

0.815

0.68

0.611

0.267

0.219

0.064

2.657

Concentration-5 (25.0mg/ml)

Trial -1

0.800

0.677

0.601

0.264

0.211

0.064

2.617

Trial -2

0.817

0.679

0.610

0.267

0.218

0.065

2.656

Trial -3

0.818

0.679

0.612

0.268

0.221

0.065

2.663

Mean

0.816

0.686

0.613

0.267

0.216

0.065

2.663

RSD

0.98

0.98

0.96

0.83

2.01

1.01

0.88

W-IV: Withanoside IV W-V: Withanoside V WF-A: Withaferin-A 12-D: 12-Deoxy Withastramonolide

W-A: Withanolide A W-B: Withanolide B

4.4.5) Conclusion: Relative standard deviation of assay of sum of withanolides is < 1.0% The above results indicate the method is precise and reproducible irrespective of the

sample concentration taken for analysis.

For standard chromatograms Enclosure – 5

Enclosure-9: Sample chromatograms (19 Pages).

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

3DJH RI