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Development and validation of HPTLC method for estimation of Tribulosin; an important biomarker in Indian Tribulus terrestris
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| Author:
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PRAJAKTA KULKARNI, KAKASAHEBMAHADIK, SATHIYANARAYANANLOHIDASAN
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| Abstract:
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A simple, accurate and precise HPTLC (High Performance Thin layer Chromatography) method was developed for
quantification of Tribulosin in Tribulus Terrestris L which is well known for the treatment of infertility. Tribulosin is one
of the markers which are not sensitive to UV (ultra violet) detection hence for quantification of such marker different
analytical techniques can be used. Determination of Tribulosin was carried out using pre-coated silica gel 60 F254 TLC
platesas stationary phase. Ascending development was followed in twin trough chamber using n-propanol: toluene:
water: glacial acetic acid (6:2.5:1:0.5 v/v). The developed method gave yellow coloured spots of Tribulosin (Rf 0.46). A
good linear relationship was obtained in the range of 200-1200ng. The correlation coefficient of 0.997 concluded good
dependence of peak area on concentration. The amount of Tribulosin in butanol fraction of Tribulus Terrestris Lwas
found to be 0.193mg/100mg w/w. The method was validated in terms of Linearity, LOD, LOQ, Robustness Accuracy,
Precision and Specificity as per ICH (International Conference on Harmonisation) guidelines. Recovery study showed
values between 96-98% which determines the accuracy of the method. The limit of detection and quantification were
found to be 72.50 and 219.70 ng/spot respectively. The developed method can be used to standardise extracts or
herbal formulations of Tribulus Terrestris L for Tribulosin.
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| Keyword:
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Tribulus Terrestris L, Tribulosin, HPTLC, validation
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| EOI:
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| DOI:
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https://doi.org/10.31838/ijpr/2019.11.02.090
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| Download:
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