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Quantification of Two Marker Compounds from unsaponifiable matter of Hygrophila spinosa Seeds Using Validated TLC-Densitometric Method
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| Author:
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NIRAJ VYAS, MANAN RAVAL
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| Abstract:
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Plant Hygrophila spinosa of Acanthaceae family is known as Kokilaksha in Ayurvedic System of medicine and being used as aphrodisiac and spermatogenic since ancient time. Unsaponifiable matter of the plant was prepared and screened for aphrodisiac and spermatogenic activity using and in vivo methods. A simple, accurate, precise, rapid and reproducible TLC densitometric method was developed for simultaneous estimation of ?-sitosterol and Lupeol in seeds of Hygrophila spinosa. The TLC separation was performed on silica gel 60 F254 HPTLC plate using Toluene: Ethyl Acetate: Formic acid (15:3:0.1 v/v/v) as mobile phase. Plates were derivatized with Anisaldehyde-Sulphuric acid reagent. Densitometric Analysis was performed at 520 nm. Compact bands for ?-sitosterol and Lupeol were obtained at Rf ~0.54 and 0.64 respectively. The method was linear over concentration range 150-350 ng/spot and 300-700 ng/spot with correlation coefficient 0.9989 and 0.9991 for ? -sitosterol and Lupeol, respectively. Method was validated as per ICH guideline. The accuracy of the methods was assessed by recovery studies and was found to be 99.79-101.66% and 99.53-100.51% for ?-sitosterol and Lupeol, respectively. The amount of ?-sitosterol and Lupeol was found to be 0.040 ± 1.21 gm % and 0.133 ±0.97% in seeds of Hygrophila spinosa. This method can be employed for routine quality control analysis of ?-sitosterol and Lupeol in seeds of Hygrophila spinosa.
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| Keyword:
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HPTLC, ?-sitosterol, Lupeol, Hygrophila spinosa, Aphrodisiac, Spermatogenic
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| DOI:
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