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  • Validated determination of primulasaponins in primula root by a high-performance-thin-layer-chromatography densitometric approach.

Validated determination of primulasaponins in primula root by a high-performance-thin-layer-chromatography densitometric approach.

Journal of pharmaceutical and biomedical analysis (2012-07-31)
Silvia A Coran, Stefano Mulas
ABSTRACT

A novel HPTLC-densitometric method was developed for separation and quantitation of primulasaponin I and II in different matrices. HPTLC silica gel 60 F254(S), 20 cm × 10 cm, plates with ethyl acetate:water:formic acid (5:1:1 v/v) as the mobile phase were used. Densitometric determinations were performed in reflectance mode at 540 nm after derivatization with vanillin reagent. The method was validated giving rise to a dependable and high throughput procedure well suited to routine applications. Primulasaponins were quantified in the range of 150-450 ng with RSD of repeatability and intermediate precision between 0.8 and 1.4% and accuracy within the acceptance limits. The method was tested on commercial herbal medicinal preparations claiming to contain primula root extract.

MATERIALS
Product Number
Brand
Product Description

Supelco
Vanillin melting point standard, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Mettler-Toledo Calibration substance ME 51143093, Vanillin, traceable to primary standards (LGC)
Sigma-Aldrich
Vanillin, ≥97%, FCC, FG
Sigma-Aldrich
Vanillin, natural, ≥97%, FCC, FG
Sigma-Aldrich
Vanillin, tested according to Ph. Eur.
Sigma-Aldrich
Vanillin, ReagentPlus®, 99%
Vanillin, European Pharmacopoeia (EP) Reference Standard
Supelco
Vanillin, Pharmaceutical Secondary Standard; Certified Reference Material