Development and Method Validation of Orientin and Vitexin Contents in Garcinia cowa Roxb. Leaves Using Ultra-High Performance Liquid Chromatography
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Abstract
Introduction and Objective: Cha Muang has the scientific name Garcinia cowa Roxb. ex Choisy belonging to Family Clusiaceae. The taste of young leaves of Garcinia cowa is sour.
It has been used for laxative, antipyretic and mucolytic agents. In addition, it is also used as food. Cha Muang have been used as active ingredient of several drugs and products in the markets. However, the appropriate analytical methods for quality control of those compounds is still limited. This study aimed to develop analytical methods for quantification assessment of orientin and vitexin in Garcinia cowa leaves samples using the Ultra High Performance Liquid Chromatography (UHPLC) technique.
Methods: The study was divided into three steps: developing the UHPLC method for analyzing orientin and vitexin in Cha Muang leaves, testing the validity of the analysis method, and analyzing the quantity of orientin and vitexin in 9 Cha Muang leaf samples.
Results: This method was validated by establishing the linearity for orientin and vitexin at concentration range of 1.32-127.01 and 1.40-134.06 μg/mL, respectively, with a determination correlation coefficient (r) of 0.9999. The % recovery were in the range of 96.26 to 103.15%. The limit of detection of orientin and vitexin were 0.002 and 0.003% w/w (calculated on the dried-weight as Garcinia cowa leaves, µg/µg), respectively, while limit of quantification were 0.006 and 0.009% w/w (calculated on the dried-weight as Garcinia cowa leaves, µg/µg), respectively. The average content of orientin and vitexin in Garcinia cowa leaves were 0.10 and 0.06%w/w (calculated on the dried-weight as Garcinia cowa leaves, µg/µg), respectively.
Discussion: The method developed for analyzing orientin and vitexin in raw Cha Muang leaves using UHPLC involves the extraction of dried Cha Muang leaves with 50% ethanol as the solvent through the reflux method. The results of the validity test of the analysis method were all within acceptable limits. This developed analysis method requires less time for analysis and involves a simple, convenient, and rapid preparation of the mobile phase
Conclusion and Recommendation: This newly developed method is suitable and can be used to analyze the content of orientin and vitexin in Garcinia cowa leaves; and the appropriate specification of orientin and vitexin contents in Garcinia cowa leaves could be established.
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