SPDS SET 2: ASH-01

Dilution-6: 5ml of Dilution-5 solution was pipetted to10ml volumetric flask and made up to 10ml

with acetonitrile.

Table-10 Sl. No.

Reference standard

Concentrations (mcg/ml)

1.

20.64

Withanoside IV

2.

18.76

Withanoside V

3.

23.05

Withaferin A

4.

22.21

12-Deoxy withastramonolide

5.

21.59

Withanolide A

6.

21.09

Withanolide B

b) Procedure: All the above 1, 2, 3, 4, 5 and 6 dilutions were injected five times each and chromatograms were recorded. A graph was plotted using concentrations on X-axis and the respective peak areas on Y-axis. Correlation co-efficient r 2 was calculated.

4.2.5) Observation: Correlation co-efficient r 2 for

r 2

Compound

Concentration range(mcg/ml)

Withanoside IV

0.999

20.0 to 1321

Withanoside V

0.999

18.7 to 1200

Withaferin A

0.996

23.0 to 737

12-Deoxy Withastramonolide

0.990

22.0 to 355

Withanolide A

0.992

21.0 to 345

Withanolide B

0.984

21.0 to 337

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3DJH RI