SPDS SET 2: ASH-01

3.0] Materials and Method:

3.1) Solvents:

Solvent Name

Grade HPLC

Product No.

Batch No. R014F06 R246A07 R189A07 R180M06

Supplier Rankem

Acetonitrile

A0755

Methanol

HPLC

M0275

Rankem

Water

Ultra pure Grade

From Sartorius water purification system.

3.2) Chemicals and their details

3.2.1) Reference Standard: Withanoside IV, Withanoside V, Withaferin A, 12-Deoxy Withastramonolide,

Withanolide A, Withanolide B Reference standard with % purity >90 %.

Reference standard

Lot No. T6B107

% Purity 90% w/w

Supplier

Withanoside IV

Withanoside V

T5A096

90% w/w

Withaferin A

T7A047

99% w/w

Natural Remedies Private Limited, Bangalore

12- Deoxy withastramonolide

T7A099

95% w/w

Withanolide A

T6K051

99% w/w

Withanolide B

T6A088

99% w/w

3.2.2) Other sources of reference standard: M/s. Chromadex and M/s. Extrasynthese

3.2.3) Sample and their details: Sample

Standardized to

Batch No RD/1162

Raw material

Total withanolides >0.2% by HPLC

Raw material

Total withanolides >0.2% by HPLC

ERH-046

Water extract

Total withanolides >0.15% by HPLC

WS–05Lot20

Hydro-alcoholic extract

Total withanolides >1.0% by HPLC

WS–05Lot21

Methanolic extract

Total withanolides >1.5% by HPLC

RD–1045

Methanolic extract

Total withanolides >2.5% by HPLC

WS–06Lot08

Methanolic extract

Total withanolides >2.5% by HPLC

WS–06Lot10

Methanolic extract

Total withanolides >2.5% by HPLC

RD–1170

QC Check sample: The QC check sample is the reference extract of Withania somnifera extract.

It is prepared out of botanically authenticated Withania somnifera and confirmed for the presence

of Withanoside IV, Withanoside V, Withaferin A, 12-Deoxy withastramonolide, Withanolide A and

Withanolide B using the respective reference standards.

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

3DJH RI