Simultaneous Determination of Gallic Acid, Ellagic Acid, and Eugenol in Syzygium aromaticum and Verification of Chemical Antagonistic Effect by the Combination with Curcuma aromatica Using Regression Analysis
This study was designed to perform simultaneous determination of three reference compounds in Syzygium aromaticum (SA), gallic acid, ellagic acid, and eugenol, and to investigate the chemical antagonistic effect when combining Curcuma aromatica (CA) with SA, based on chromatographic analysis. The values of LODs and LOQs were 0.01–0.11？μg/mL and 0.03–0.36？μg/mL, respectively. The intraday and interday precisions were <3.0 of RSD values, and the recovery was in the range of 92.19–103.24%, with RSD values <3.0%. Repeatability and stability were 0.38–0.73% and 0.49–2.24%, respectively. Compared with the content of reference and relative peaks in SA and SA combined with CA (SAC), the amounts of gallic acid and eugenol were increased, while that of ellagic acid was decreased in SAC (compared with SA), and most of peak areas in SA were reduced in SAC. Regression analysis of the relative peak areas between SA and SAC showed values >0.87, indicating a linear relationship between SA and SAC. These results demonstrate that the components contained in CA could affect the extraction of components of SA mainly in a decreasing manner. The antagonistic effect of CA on SA was verified by chemical analysis. 1. Introduction The therapeutic efficacy of herbal formula originates from the combination of two or more herbal medicines and is exerted by the interaction between them. Several types of herbal combinations have been used for the purpose of increasing or enhancing the therapeutic effect, reducing toxicity or the adverse effect, weakening the action of another combined herbal medicine, or causing toxicity or an adverse effect, expressed as “seven types of herbal combination.” Their combinations are expressed as mutual reinforcement, mutual assistance, mutual restraint, mutual suppression, mutual inhibition, and single effect [1]. Besides single effect, mutual reinforcement and mutual assistance have been considered as desirable (and positive) for exhibiting the therapeutic effect (synergistic), while mutual suppression and mutual inhibition have been used due to their aggravating or negative effect (antagonistic). Most reports in the literature have dealt with synergic interactions of herbal medicine such as Astragali radix with Rehmanniae radix to promote diabetic wound healing [2] and Glycyrrhiza glabra with Solanum xanthocarpum and Adhatoda vasica to stabilize mast cells [3]. There are no reports of mutual inhibition or mutual suppression. The bud of Syzygium aromaticum (SA, syn. Eugenia caryophyllata) is a representative herbal medicine of mutual suppression when
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