Method development and validation for quantification of six bioactive compounds (andrographolide, columbin, piperine, gallic, paracoumaric and oleanolic acids) by HPTLC.

OBJECTIVES: During the spread of pandemic diseases, immunity boosting herbal drugs are taken as a preventive medicine. Kapacurak Kutinir Curanam is a Siddha drug used for flu like viral infections, cold and fever. Developing an analytical method to estimate the content of active phytoconstituents in such antiviral immune boosting drug will be useful in the phyto pharmaceutical industry. METHODS: A precise, reliable and sensitive ordinary phase high performance thin layer chromatography (HPTLC) method has been developed and validated for identification and simultaneous estimation of six bioactive components namely like andrographolide, columbin, gallic acid, ρ-coumaric acid, piperine and oleanolic acid from any Indian traditional medicine, medicinal plant, drugs and food materials etc. The separation was achieved on silica gel 60F254 TLC plates using toluene: ethyl acetate: formic acid (7:3:0.5, v/v) as mobile phase. The gallic acid, ρ-coumaric acid, piperine markers were estimated using the densitometric scanning in absorption mode at 254 nm. The densitometric scanning was done after derivatization (vanillin-sulphuric acid reagent) at λ=520 nm for andrographolide, columbin and oleanolic acid. RESULTS: The linear regression analysis data for the calibration plots showed a correlation coefficient in the concentration range 1-5 µg per band for the bioactive markers with respect to area. The method was validated for accuracy, precision, limit of detection (LOD), and quantitation of limit (LOQ). CONCLUSIONS: Developed method was accurate, precise and fast to ensure the quality of Kapacurak Kutinir Curanam.

Proximate analysis, HPTLC finger print analysis and multi spectrometric analysis of Strychnos nux-vomica nuts.

OBJECTIVES: In alternative medicine, plants pay a major role. Some plants are known for their poisonous nature but still have some importance in the herbal drug industry for their medicinal value. Strychnos nux-vomica is one such plant. Its nuts are called as poison nut due to the presence of alkaloids. Both the nut and its minerals are having medicinal properties and hence the present study was indented to understand the nature of primary metabolites and multi elemental composition. METHODS: The nuts of S. nux-vomica were procured, authenticated, powdered and subjected to proximate analysis parameters, visualization of thin layer chromatographic separation (TLC) and finger print profiling through high performance thin layer chromatographic (HPTLC); surface morphology by scanning electron microscopy, energy dispersive X-ray analysis, X-ray fluorescence spectrometry, X-ray photoelectron spectrometry, powder X-ray diffractometry and inductively coupled plasma optical emission spectrometry. RESULTS: In HPTLC, 7 spots each under 254 nm, 366 nm, derivatization with vanillin sulphuric acid (VSR) reagent appeared and 2 spots with Dragendorff's reagent. In HPTLC, 12 peaks at 254 nm, 9 peaks at 366 nm, 7 peaks at 520 nm after derivatization with VSR reagent detected. Elements such as potassium, calcium, magnesium, chlorine, aluminium, iron, manganese, sodium, nickel, phosphorus, copper, zinc, sulphur and silicon were identified. PXRD revealed that the presence of potassium chloride, calcite and dolomite as major elemental composition. CONCLUSIONS: The presence of all the above elements has vital roles on human physiology. Potassium, calcium, chlorine, aluminium, nickel, phosphorus, sulphur and silicon are reported for the first time in this study.