Gold standard for method validation
- Mohamad Shazeli Che Zain
- Feb 19, 2023
- 2 min read
Updated: Oct 7, 2023
Our new manuscript is just published in an open-access journal by Molecules under special issue Method Development of Sampling Preparation Techniques. Thank you to co-authors for your contributions. The work involved the development and validation of the analytical method, UHPLC-UV/PDA for the simultaneous quantification of major compounds: luteolin and apigenin derivatives present in oil palm leaves. We adopted International Conference on Harmonization (ICH) guidelines for this validation. We tested our plant extracts derived from various processing techniques to quantify the amount of compounds of interest and combined them with partial least square analysis to see the comparison between the extracts and compounds. This approach can be employed as quality control for the development of standardized herbal preparation.

Abstract
Luteolin and apigenin derivatives present in oil palm (Elaeis guineensis) leaves (OPL) are reported to possess excellent antioxidant properties relating to numerous health benefits. To meet the global demand for flavonoids, OPL, which is plentifully generated as an agricultural by-product from oil palm plantations, can be further exploited as a new source of natural antioxidant compounds. However, to produce a standardized herbal preparation, validation of the quantification method for these compounds is required. Therefore, in this investigation, we developed and validated an improved and rapid analytical method, ultra-high-performance liquid chromatography equipped with ultraviolet/photodiode array (UHPLC-UV/PDA) for the quantification of 12 luteolin and apigenin derivatives, particularly focusing on flavonoid isomeric pairs: orientin/isoorientin and vitexin/isovitexin, present in various OPL extracts. Several validation parameters were assessed, resulting in the UHPLC-UV/PDA technique offering good specificity, linearity, accuracy, precision, and robustness, where the values were within acceptable limits. Subsequently, the validated method was employed to quantify luteolin and apigenin derivatives from OPL subjected to different drying treatments and extraction with various solvent systems, giving total luteolin (TLC) and apigenin content (TAC) in the range of 2.04–56.30 and 1.84–160.38 µg/mg extract, respectively. Additionally, partial least square (PLS) analysis disclosed the combination of freeze dry-aqueous methanol yielded OPL extracts with high TLC and TAC, which are strongly correlated with antioxidant activity. Therefore, we provide the first validation report of the UHPLC-UV/PDA method for quantification of luteolin and apigenin derivatives present in various OPL extracts, suggesting that this approach could be employed in standardized herbal preparations by adopting orientin, isoorientin, vitexin, and isovitexin as chemical markers.
Read full article: https://doi.org/10.3390/molecules26041084
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