Publication: Bioanalytical method validation for determination of rosmarinic acid in simulated biological media using hplc
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2021-01-01
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09757058
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2-s2.0-85103288181
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Mahidol University
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SCOPUS
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International Journal of Applied Pharmaceutics. Vol.13, No.2 (2021), 110-113
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Kotchaphan Chooluck, Piyanuch Rojsanga, Chutima Phechkrajang, Montree Jaturanpinyo Bioanalytical method validation for determination of rosmarinic acid in simulated biological media using hplc. International Journal of Applied Pharmaceutics. Vol.13, No.2 (2021), 110-113. doi:10.22159/ijap.2021v13i2.40372 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/78980
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Bioanalytical method validation for determination of rosmarinic acid in simulated biological media using hplc
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Objective: This study aims to develop and validate a simple, rapid and accurate HPLC method for the determination of rosmarinic acid (RA), an active marker of Thunbergia laurifolia (TL) tea, in Hank's Balanced Salt Solution (HBSS) using HPLC. Methods: The separation was performed using a Xterra® C 18 reversed-phase column (150 × 3.9 mm, 5 µm). The mobile phase consisted of 0.5% (v/v) glacial acetic acid (A) and acetonitrile (B). The flow rate was 1 ml/min and detection was carried out at 330 nm with UV-visible spectrophotometer. The method was validated according to the US FDA guidance on bioanalytical method validation in 2018. Results: The method was successfully validated in the range of 50-500 ng/ml of RA. Intra-and inter-day precision ranged from 1.6 to 2.1% and 1.4 to 4.5%, respectively. Intra-and inter-day relative errors (% bias) were less than 7.6 and 3.6%, respectively. In addition, it was found that the stability of RA in HBSS could be dependent on the pH and temperature. Conclusion: The developed method met the validation requirements and could be further applied to the permeability study of RA using an in vitro Caco-2 cell monolayer model.