In-Needle Pre-Column Derivatization for Amino Acid Quantification (iPDAQ) Using HPLC

Yuki Soma, Yoshihiro Izumi, Takehiko Shimohira, Masatomo Takahashi, Yuri Imado, Saki Tominaga, Kanako Tokito, Kosuke Hata, Shoji Shinadama, Mana Oshiro, Yoshihiro Hayakawa, Takeshi Bamba

Research output: Contribution to journalArticlepeer-review

Abstract

Pre-column fluorescent derivatization has been used for the fast quantification of amino acids using high-performance liquid chromatography (HPLC) systems. However, it generally requires an offline in-vial derivatization process with multiple derivatization reagents. The offline derivatization requires the same number of reaction vials as the number of sample vials for use as a reaction chamber for the derivatization reaction in an autosampler. Therefore, the number of samples analyzed per batch using the pre-column derivatization method is halved. To benefit from the pre-column derivatization method, we transformed the derivatization process from an offline chamber process to an online in-needle process (in-needle Pre-column Derivatization for Amino acids Quantification; iPDAQ). Fluorescent derivatization in the injection needle obviated the need for vacant vials as reaction chambers. Consequently, the throughput per batch improved up to two times, and the consumption of derivatization reagents was reduced to less than one-tenth of that in the conventional vial method. We demonstrated to separate and quantify the amino acids in various biological samples. Herein, we presented a novel HPLC-based amino acid quantification method that enables the continuous analysis of a large number of samples. The iPDAQ facilitates accurate amino acid quantification due to the automation of derivatization and achieves improvement in the throughput and reduction of analysis labor.
Original languageEnglish
Pages (from-to)807
Number of pages1
JournalMetabolites 2022, Vol. 12, Page 807
Volume12
Issue number9
DOIs
Publication statusPublished - 2022

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