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3DJH RI
2.0] Introduction
2.1) Objective:
To ensure the method [(NR/QCD/APM04 WI(17) - estimation of Withanoside IV,
Withanoside V, Withaferin A, 12-Deoxy withastramonolide, Withanolide A and Withanolide-
B in
Withania somnifera
)] developed by Natural Remedies Private Limited provides high
degree of assurance that it will consistently produce result which is accurate, precise,
reproducible and robust when performed at Natural Remedies Private Limited or at any
other laboratory by competent analyst using appropriate equipment, chemicals and
reference standards.
2.2) Scope:
This method of estimation of Withanoside IV, Withanoside V, Withaferin A, 12-
Deoxy withastramonolide, Withanolide A and Withanolide B, and total Withanolides using
Withanoside IV, Withanoside V, Withaferin A, 12-Deoxy withastramonolide, Withanolide A
and Withanolide B as reference standards applicable to
Withania somnifera
roots (raw
material) and its extract. These compounds are ranging from 0.01 to 10%. It is applicable
to all material having matrix similar to that of
Withania somnifera
roots
(raw material) and
extracts / products containing
Withania somnifera
extracts blended with excipients which
are insoluble in methanol and acetonitrile.
2.3) Purpose and application:
Withania somnifera
extract is prepared by extracting
Withania
somnifera
roots with solvents like methanol, water etc. It is standardized to 1.5%, 2.5%,
3%, 5% Total withanolides. In general the content of Withanoside IV, Withanoside V,
Withaferin A, 12-Deoxy withastramonolide, Withanolide A and Withanolide B would be 20 to
30% of Total withanolides. It is important to estimate the Withanoside IV, Withanoside V,
Withaferin A, 12-Deoxy withastramonolide, Withanolide A and Withanolide- B contents to
ensure the product meets the specification before its release to the market. Also the raw
material used for extraction to be estimated for withanolides content as the product quality
depends on raw material used for extraction. Withanolides (Withanoside IV, Withanoside
V, Withaferin A, 12-Deoxy withastramonolide, Withanolide A and Withanolide- B) can be
estimated only after clearly separating the various secondary metabolites present in the raw
material or in the extract along with the withanolides. This could be achieved only by
employing chromatographic techniques. Hplc is more suitable for this purpose.
The purpose of this method validation is to establish documental evidence which provides
adequate assurance that the method comply with the requirements of validation
characteristics suggested in the protocol which was prepared based on international
guidelines. The purpose is also to demonstrate that the method performs in accordance to
the claims made.