Analytical Validation of Phytometabolites in Livogrit Herbal Medicine: Integration of Ultra‐High Performance Liquid Chromatography and High Performance Thin Layer Chromatography for Rigorous Drug Standardization and Quality Control

ABSTRACT

Liver is the second largest organ, which plays a pivotal role in the body. It is intricately involved in detoxification of harmful substances, metabolizing nutrients, synthesizing proteins, regulating blood clotting, producing variety of biochemicals required for digestion, and storing energy in the form of glycogen. Liver‐related diseases encompass a wide range of conditions from viral infections like fatty acids diseases and hepatitis (A, B, C, D, and E) to autoimmune liver diseases, characterized by excessive fat accumulation, inflammation, and fibrosis. Livogrit, a triple‐herb solution infused with botanical extracts of Makoy ( Solanum nigrum ), Bhumi amla ( Phyllanthus niruri ), and Punarnava ( Boerhavia diffusa ), is believed to possess antiviral, hepatoprotective, and antioxidative properties. The study aimed to develop simple and rapid ultra‐high performance liquid chromatography (UHPLC) and high performance thin layer chromatography (HPTLC) methods for the quantification of signature phytometabolites and also explored the uniformity of Livogrit throughout all batches. The UHPLC method was validated as per ICH Q2 (R1) for the quantification of 10 signature phytometabolites, namely, methyl gallate, catechin, gallic acid, boeravinone B, ellagic acid, caffeic acid, cinnamic acid, quercetin, corilagin, and rutin. However, by HPTLC, six phytometabolites, including cinnamic acid, methyl gallate, catechin, corilagin, gallic acid, and rutin, were assessed. Both methods demonstrated acceptable performance across all validation parameters, including limit of detection and quantification, linearity, precision, specificity, system suitability, robustness, accuracy, and ruggedness. Moreover, relative assessment discovered no statistically significant variations between analytical results generated by UHPLC and HPTLC methodologies. The extent‐validated methods were found suitable for the quality estimation of Livogrit, and the uniformity was witnessed across all batches.