Quantitative Analysis of Astragalin Contents in Ethanolic Extract of Moringa oleifera Lam. (leave) by High-Performance Thin-Layer Chromatography (HPTLC) -
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Abstract
Introduction and objective: Astragalin, a flavonoid found in drumstick tree leaves (Moringa oleifera Lam., Moringaceae family), exhibits various biological activities, including anti-inflammatory and antioxidative effects and the ability to lower blood sugar levels. This herb has traditionally been used for medicinal purposes and in developing health products. However, there is currently no specification for the active marker content in this plant. This study aimed to develop an analytical method to quantify astragalin content in ethanolic extracts of drumstick tree leaves using high-performance thin-layer chromatography (HPTLC).
Methods: The development and validation were performed by weighing 100 mg of drumstick leave-ethanolic extract and dissolved in 25.0 mL of methanol. Subsequently, 1 µL of the solution was applied to the HPTLC plate coated with Silica gel GF254 utilizing an autosampler. The mobile phase consisted of ethyl acetate, glacial acetic acid, formic acid, and water in a ratio of 20:1:1:2 (v/v/v/v). The developed plate was examined under UV366 nm after being sprayed with NP/PEG solutions, and astragalin content was quantitatively analyzed using a densitometer.
Results: This method was validated by establishing linearity for astragalin concentrations ranging from 1.96 to 11.76 µg/mL, yielding a determination correlation coefficient (r) of 0.9996. The accuracy of astragalin ranged from 97.75% to 99.49%, while the precision (%RSD) ranged from 0.22% to 0.45%. The limits of detection and quantification were determined to be 0.313 and 1.045 µg/mL, respectively. Quantitative analysis of astragalin in 12 leave-ethanolic extracts revealed an astragalin content of 0.06 ± % (w/w).
Discussion: The recently developed analytical method offers several advantages, including rapidity achieved through the simultaneous observation of multiple samples. It is also cost-effective, as the HPTLC plate employed as the stationary phase is more affordable than traditional high-performance liquid chromatography (HPLC) columns. Furthermore, this method is environmentally sustainable and safe, utilizing a reduced volume of mobile phase. Notably, the newly established method incorporates a lower acid concentration in the mobile phase than previously reported techniques. Consequently, HPTLC has been utilized to establish a method for quantification of astragalin in the ethanolic extract of drumstick tree leaves.
Conclusion and recommendation: This newly developed method is suitable and cost-effective for analyzing astragalin content in drumstick leave ethanolic extracts. The validation of the astragalin determination method in drumstick leaves using HPTLC has been successfully established and could be used to analyze astragalin content in drumstick tree leave extracts. Appropriate specifications for astragalin content in drumstick tree leave extract could be established.
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