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The Simultaneous Analysis of 14 Antioxidant Compounds Using HPLC with UV Detection and Their Application to Edible Plants from Asia


Antioxidants in edible herbs and spices have many benefits and are essential to human health. Therefore, to investigate each antioxidant compound present in edible herbs and spices, a new and simple high-performance liquid chromatography method was developed to simultaneously analyze 14 antioxidant compounds, namely, gallic acid, epicatechin gallate, caffeine, vanillic acid, epigallocatechin gallate, coumaric acid, epigallocatechin, rosmarinic acid, gingerol, thymoquinone, carvacrol, thymol, carnosol, and carnosic acid. This method consisted of a C18 column, where gradient elution was conducted using 1% acetic acid and acetonitrile as the mobile phase at 1.0 mL/min and subsequent UV detection at 284 nm. Results showed that the 14 antioxidant compounds were separated during the 70 min cycle with good specificity, linearity, range, intra- and inter-day precision, and recovery test results. The limit of the quantitation range was between 0.07 and 5.85 μg/mL. Furthermore, the relative standard deviation values of the intra- and inter-day precision were 0.040–0.064% and 0.003–0.016%, respectively, whereas those of the recovery test using Japanese tea and coriander were between 90.68–115.91% and 80.92–95.01%, respectively. The overall test results were satisfactory as well. Therefore, we propose that this method would effectively identify and evaluate antioxidants, including caffeine in Japanese tea, as this advanced method showed sensitive, quantitative, and reliable results suited for analysis of herbs, spices, and other plant samples.

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All data generated or analyzed during this study are included in this published article.


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The authors would like to thank Enago ( for the English language review.


This study was funded by JSPS KAKENHI Grant Number 21K05785.

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Nichawee Jongsawatsataporn: conceptualization, methodology, investigation, data curation, and writing (original draft preparation).

Ryusuke Tanaka: conceptualization, methodology, investigation, data curation, writing (original draft preparation), and supervision.

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Correspondence to Ryusuke Tanaka.

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Nichawee Jongsawatsataporn declares that he has no conflict of interest. Ryusuke Tanaka declares that he has no conflict of interest.

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Jongsawatsataporn, N., Tanaka, R. The Simultaneous Analysis of 14 Antioxidant Compounds Using HPLC with UV Detection and Their Application to Edible Plants from Asia. Food Anal. Methods 15, 1331–1340 (2022).

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  • Antioxidant
  • Catechin
  • HPLC
  • Herbs
  • Spices