Validated Chromatographic methods for estimation of Solanine in Mahamrutyunjaya rasa

Abstract

Introduction: Mahamrutyunjaya rasa is a herbomineral ayurvedic formulation used for various indications like fever, pain, cardiac arrhythmia, etc. It contains Solanum indicum as one of the ingredients having anti-inflammatory, diuretic, and diaphoretic properties. Solanine is one of the active biomarkers for Solanum indicum. The therapeutic window of Solanine is very narrow, with the LD50 value of 3-6 mg/kg body weight in humans. Objective: Estimation of such biomarkers is very significant to avoid any adverse events due to the administration of ayurvedic preparations containing Solanine. In the present study, two simple, sensitive, reliable chromatographic techniques have been developed and validated for the estimation of Solanine in Mahamrutyunjaya rasa. Materials and Methods: High-performance liquid chromatography (HPLC) separation of Solanine was performed on an reversed phase C-18 column (250 mm 9 4.6 mm ID, 5 lm particle size), with isocratic elution using a mixture of Tris buffer (10mM, pH 6.00): Acetonitrile(60:40, v/v) at a flow rate of 1 mL/min with UV detection at 218 nm for solanine. High-performance thin-layer chromatography (HPTLC) separation was done on Silica gel 60 F254 pre-coated plates using Chloroform: Methanol: Ammonia (7:3:0.5 v/v). The densitometric scanning was performed at 500 nm. The developed methods were validated for linearity, LOD, LOQ, accuracy, precision, and specificity as per ICH guidelines. Results: Solanine was eluted at 4.43 ± 0.1 min and established a linearity range of 1 – 100 μg/ml (r2 = 0.9992). In the HPTLC method, the Rf value of Solanine was 0.05 in a linearity range of 1600-4800 ng/ml. Conclusion: Reliable, rapid, simple, and sensitive chromatographic methods were developed for the quantification of Solanine in Mahamrutyunjaya Rasa.