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Quantitation of Four Tryptophan-Related Impurities in Compound Amino Acid Injection-18 AA by HPLC–PDA

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Abstract

Compound amino acid injection-18 AA (CAAI-18 AA) is widely applied in medical care as a part of total parenteral nutrition systems. Thus, suitable quality control of CAAI-18 AA is of crucial importance for safety of the patients. Tryptophan (TRP) is one of the main components in CAAI-18 AA and may be degraded to 5-hydroxy-tryptophan (5-HTP), 5-hydroxytryptamine (5-HT), kynurenine (KYN) and indole (IN) under certain conditions, such as oxidation environments, ultraviolet light radiation and nonsterile conditions. The existence of these harmful impurities seriously affects the quality of CAAI-18 AA. Here, a simple HPLC approach with a PDA detector was developed and validated for simultaneous determination of the concentrations of 5-HTP, 5-HT, KYN and IN impurities in CAAI-18 AA. Separation of these four impurities in CAAI-18 AA was achieved under a gradient condition using acetonitrile and 10 mM phosphate buffer (containing 10 mM sodium 1-octanesulfonate, pH adjusted to 1.8 with phosphoric acid). To detect the four analytes with high sensitivity, multi-wavelength detection at 257, 270 and 276 nm was carried out during the separation. The method was validated in accordance with ICH guidelines and the results were compared to that obtained by UHPLC–MS/MS, showing that the two approaches did not provide significantly different results. Our results reveal that the simple and less expensive HPLC method holds great potential for quantitation of the four impurities in CAAI-18 AA.

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Acknowledgements

We wish to appreciate Huan-Tsung Chang at National Taiwan University for revision of the manuscript. We would also like to thank Cisen Pharmaceutical Co., Ltd, Shandong Qi Du Pharmaceutical Co., Ltd and Guangdong Litai Pharmaceutical Co., Ltd for providing us with pharmaceutical samples.

Funding

This study was funded by the Natural Science Foundation for Colleges and Universities of Jiangsu Province (17KJD150005), the Jiangsu Overseas Research & Training Program for University Prominent Young & Middle-aged Teachers and Presidents Fund, the Qing Lan Project Fund of Jiangsu Province, Science and Technology Project Fund of Lianyungang (SH1627), The Fifth Phase of 521 Project Fund of Lianyungang (KD2017LYG521002), Innovation and Entrepreneurship Training Program for College Students of Jiangsu Province (201813980006X); and Science and Technology Funds of Kangda College of Nanjing Medical University (KD2016GCCRCYJ01, KD2016GCCRCYJ02, KD2018KYJJYB001 and KD2018KYJJYB010).

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Authors and Affiliations

  1. Kangda College, Nanjing Medical University, Lianyungang, 222000, China

    Li Li, Puzhong Gu, Honglei Li, Mengqing Li, Ziyue Zhang, Ruiling Chen & Ming Zhang

  2. Lianyungang Inspection and Testing Center for Food and Drug, Lianyungang, 222000, China

    Naijiang Chen

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  1. Li Li
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Li, L., Chen, N., Gu, P. et al. Quantitation of Four Tryptophan-Related Impurities in Compound Amino Acid Injection-18 AA by HPLC–PDA. Chromatographia 83, 205–217 (2020). https://doi.org/10.1007/s10337-019-03847-1

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