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Abstracto

Development and Validation of Stability Indicating Hptlc Method for Determination of Picroside-I and Picroside-II in Picrorhiza kurroa

Nilam Dalavi

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. An approach for the stress degradation was successfully applied for the picrosides I, II development of stability indicating HPTLC method for the determination of Separation was achieved on the plates pecoated with silica gel 60 F254. The mobile phase used was chloroform: methanol: formic acid (8:1.5:0.5 v/v/v) and quantification was carried out wavelength at 274 nm. Stress testing of picrosides I, II was carried out according to the International Conference of Harmonization (ICH) guideline Q1A(R2). The markers were subjected to acid, base, neutral hydrolysis, oxidation, thermal degradation and photolysis. The system showed a peak for picrosides I, II at Rf value of 0.47, 0.57 ± 0.02 resp. The method was successfully validated according to ICH guidelines Q2 (R1). The data of linear regression analysis indicated a good linear relationship over the range of 200–1000 ng/band concentrations with correlation coefficient value of 0.999. The accuracy of the method was established based on the recovery studies. The LOD of picrosides I, II were found 135,108 ng/ band. LOQ of picrosides I, II were found 410,327 ng/ band, respectively. Under various tested stressed conditions, picrosides I, II showed considerable degradation under alkali, acidic, oxidative and neutral hydrolytic condition.