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Determination of free amino acids in tea by a novel method of reversed-phase high performance liquid chromatography applying 6-Aminoquinolyl-N-Hydroxysuccinimidyl carbamate reagent

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Abstract

We describe a novel analytical method for quantification of free amino acids in tea using variable mobile phase pH, elution gradient and column temperature of reversed-phase high-performance liquid chromatography (RP-HPLC). The study of mobile phase pH 5.7 was chosen to simultaneous quantification of 19 free amino acids in tea, while it improved maximum resolution of glutamine, histidine and theanine. Elution gradient was adapted for enhancing the solution of free amino acids, mainly because of adjustment of mobile phase A and B. The column temperature of 40 °C was conducive to separate free amino acids in tea. The limit of detection (LOD) and limit of quantitation (LOQ) of this method were in the range of 0.097–0.228 nmol/mL and 0.323–0.761 nmol/mL, respectively. The relative standard deviation of intraday and interday ranged in 0.099–1.909% and 3.231–7.025%, respectively, indicating that the method was reproducible and precise, while recovery ranged between 81.06–112.78%, showing that the method had an acceptable accuracy. This method was applied for the quantification of free amino acids in six types of tea. Multivariate analysis identified serine, glutamine, theanine and leucine as the most influencing factor for classify among analyzed sample.

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (31170283), Anhui Provincial Natural Science Foundation (1508085MC59), Shandong Provincial Natural Science Foundation (ZR2014CP010) and the Anhui Major Demonstration Project for Leading Talent Team on Tea Chemistry and Health. We thank Tiejun Ling for critically reading of the manuscript. We appreciate Qi Chen, Yinghui Hu, Shaode Hu, Zhenguo Li and Chunyi Peng for assistance on technology.

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Author notes

  1. Min Li and Daxiang Li have contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei, Anhui, China

    Min Li, Daxiang Li, Yuling Tai, Chenchen Gu, Yasai Song, Weiting Jiao, Jingming Ning, Chaoling Wei, Xungang Gu & Xiaochun Wan

  2. State Key Laboratory of Crop Biology, College of Horticulture Science and Engineering, Shandong Agricultural University, Taian, 271018, Shandong, China

    Min Li

  3. Department of Food Science, Rutgers University, New Brunswick, NJ, USA

    Chi-Tang Ho

  4. Department of Plant Pathology, Faculty of Crop Protection, Sindh Agriculture University, Tandojam, Pakistan

    Jamal U. Ddin Hajano

Authors
  1. Min Li
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  2. Daxiang Li
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  3. Yuling Tai
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  4. Chenchen Gu
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Corresponding author

Correspondence to Xiaochun Wan.

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Supplementary material 1

Chromatograms for free amino acids standards with different column temperature. The temperature was used in 37 °C and 40 °C. Elution order as follows: 1. Asp, 2. Ser, 3. Glu, 4. Gly, 5. Gln, 6. His, 7. Arg, 8. Thr, 9. Ala, 10. Pro, 11. Thea, 12. Tyr, 13. Cys, 14. Val, 15. Met, 16. Ile, 17. Lys, 18. Leu and 19. Phe. * represents AMQ, ▽ represents NH3, and ◇ represents the solvent peak (TIFF 8347 kb)

Supplementary material 2

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Li, M., Li, D., Tai, Y. et al. Determination of free amino acids in tea by a novel method of reversed-phase high performance liquid chromatography applying 6-Aminoquinolyl-N-Hydroxysuccinimidyl carbamate reagent. J Food Sci Technol 55, 4276–4286 (2018). https://doi.org/10.1007/s13197-018-3366-9

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  • DOI: https://doi.org/10.1007/s13197-018-3366-9

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