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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

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