Journal of Chromatography & Separation Techniques
Open Access

Abstract

Determination of Flavonoids in Stem Bark of Manilkara hexandra by Simultaneous and Validated HPLC Methods

Sapna Sharma, Sarojkumar Katara, Mamta B. Shah*

Manilkara hexandra Roxb. Dubard, another name is Mimusops hexandra is a tall, glabrous evergreen tree belonging to the Sapotaceae family. Ethnopharmacological uses of M. hexandra in toothache, tonic, dysentery, diarrhea, alimentary disorders, infertility and veterinary use by tribal people of different regions in India. Research articles revealed immunostimulant, inhibitory effect on the SARS-CoV-2 protease enzyme, antiulcer and antioxidant activity of stem bark. A simple, accurate and reproducible reversed-phase liquid chromatographic method was developed for the qualitative and quantitative determination of four bioactive flavonoids (quercetin, luteolin, kaempferol and apigenin) from the stem bark extract of M. hexandra. Chromatographic separation was performed on a C18 column (5 μm C18, 4.6 × 250 mm) with a mobile phase consisting of 0.5% orthophosphoric acid and 100% methanol (40:60 %v/v), at a flow rate of 1.0 mL/min. The analysis was performed using a UV detector at different wavelengths. The method was validated in terms of selectivity, linearity, accuracy, precision, robustness and recovery. Good linearity was observed over the investigated concentration range of 5-30 μg/mL for quercetin and luteolin; 2-64 μg/mL of kaempferol and 2-12 μg/mL of apigenin with correlation coefficient (r2) values greater than 0.998. The intra- and inter-day precision over the concentration range was <0.57% (relative standard deviation) and the accuracy was between 98.06% and 100.65%. The %RSD of recovery for all the analytes was 0.49%-0.81%. This method was successfully applied in the quantity assessment of bioactive flavonoids in the stem bark extract of M. hexandra.

Published Date: 2025-04-17; Received Date: 2024-04-20