Phytochemical studies for quantitative estimation of iridoid glycosides in Picrorhiza kurroa Royle

Background: Picrorhiza kurroa Royle commonly known as 'Kutki or Kutaki' is an important medicinal plant in Ayurvedic system of medicine and has traditionally been used to treat disorders of the liver and upper respiratory tract. The plant is the principle source of iridoid glycosides, picrosides I, II and kutkoside used in various herbal drug formulations mainly as strong hepatoprotective and immune-modulatory compound. The species has become endangered to near extinction due to the unregulated collection from the wild, slower plant growth and ecological destruction of natural habitats. There is a severe shortage of plant material, while the market demand is ever increasing. Hence, it is very important to apply a simple and precise analytical method to determine and validate the concentration of the major bioactive constituents in different populations of this plant species for development of a high yielding chemotype for large scale production and its commercial exploitation on scientific lines. Results: This study assessed and validated a fast and reliable chromatography method for the determination of picroside-I and picroside-II in different populations of this priortized medicinal plant species. Separation and resolution of picrosides was carried out on a reversed phase (C-18) column by using a mobile phase of methanol and water (40:60 v/v). The detection of picrosides was carried out at 270 nm. The average levels of these two major marker compounds in all the seven accessions showed significant quantitative variation (ANOVA, p < 0.05) between mean levels of marker compounds and their accumulation in different parts of the plant viz. roots, rhizomes and leaves. The highest content of pk-I was found in the accession from Gurez altitude (3750 masl) while the highest content of pk-II was found in accession from Keller (Shopian) altitude (3300 masl) demonstrate that picroside accusation is directly correlated with altitudinal variation. The method was validated in terms of linearity, accuracy and precision (within- and between-assay variation). Conclusion: A simple chromatographic method with the ability to separate both the major chemical constituents effectively in different herbal extracts of P. kurroa and other related species has been standardized and validated, which is more suitable for regular and normal analysis of picrosides in different herbal formulations. The paper accomplish that picroside concentration in different samples showed significant variation based on altitude and other agroclimatic factors, which can be useful in the selection and collection of superior genotypes with higher concentration of these marker compounds.