Identification and Quantification of Four Bioactive Markers in Sanjivani Vati Using High-Performance Thin Layer Chromatography

Background

Sanjivani Vati (SV) is an extensively used multicomponent Ayurvedic medicine, recommended for fever, indigestion, dysentery, gastroenteritis and more. The pharmaceutical standardization of SV and estimation of its tannin and embelin content have been reported in previous studies. However, the identification and quantification of other bioactive constituents namely, aconitine, gallic acid and β-asarone, etc. remain unaddressed. Also, concerns arise regarding one of its ingredients i.e. Vatsanabha (Aconite species), historically recognized as one of the most poisonous plants. It is used after meticulous processing specified in classical texts. Still, it is crucial to identify the amount of aconitine being delivered in the drug to ensure optimal therapeutic benefit while minimizing adverse reactions. In addition, another issue is that Vatsanabha is being sold as a mixture of various species in Indian markets. Previous research has highlighted significant qualitative and quantitative variations in the total alkaloid content among different species of Vatsanabha.

Materials and Methods

The present communication deals with identifying and quantifying four bioactive markers viz. Gallic acid, Piperine, β-asarone and Aconitine in SV prepared with two species of Vatsanabha namely Aconitum nepellus L. (SV -1) and Aconitum balfourii Holmes ex Stapf (SV-2) using High-performance Thin Layer Chromatography.

Results

Gallic acid, piperine and β-asarone were detected in both SV groups. However, aconitine was not detected in any of the samples. Identification was confirmed by matching the R_f and spectrum overlay. The selected solvent system yielded effective separation from the matrix with an 𝑅_𝑓 value of 0.45±0.01 for gallic acid, 0.27±0.01 for piperine and 0.62±0.01 for β-asarone. The amount of gallic acid was determined to be 1266 µg and 1395 µg in 100 mg of SV1 and SV2 respectively. Piperine was 35.70 µg and 49.95 µg per 100 mg of SV1 and SV2, respectively. β-asarone was 135.3 µg and 236.1 µg in SV1 and SV2 respectively.

Conclusion

The concentration of all the detected biomarkers is higher in SV-2 than in SV-1. The presence of these marker compounds helps explain the probable mode of action of SV and offers simple, efficient methods for estimating these biomarkers.