Shafawi, Nur Atielah Nabila Mohd
(2022)
Method validation of delphinidin-3-o-glucoside
chloride in rat plasma by
High-Performance Liquid
Chromatography (HPLC).
Masters thesis, Universiti Sains Malaysia.
Abstract
Hibiscus sabdariffa (Roselle) is one of the enormously explored medicinal plants with
high anthocyanin, which is essential in treating various diseases. This study was
conducted on the high-performance liquid chromatography (HPLC) method for detecting
and quantifying delphinidin-3-o-glucoside chloride, one of the anthocyanins present in
Hibiscus sabdariffa in the rat plasma. In addition, the HPLC method was validated
following the solid-phase extraction method to demonstrate that this analytical method is
suitable and to assure its reliability of H.sabdariffa pharmacokinetics and bioavailability
study in rat plasma. Six validation parameters were assessed according to FDA guidelines:
linearity, recovery, precision, accuracy, limit of detection, and limit of quantification. The calibration curve of delphinidin-3-0-glucoside chloride had a coefficient determination
(R2) of ≥95.0%. The recovery percentage of delphinidin3-0-glucoside was found to be
80.377%. Delphinidin-3-o-glucoside chloride also showed good precision and accuracy.
The method detection and quantification limits for delphinidin-3-o-glucoside chloride
were 47ng/mL and 213ng/mL, respectively. In conclusion, the validated HPLC method
in this study can be used to determine anthocyanins in the future pharmacokinetic study
of roselle.
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