Pre-column derivatization HPLC trace detection Enzymatic γ DL glutamyl tryptophan reaction.doc

 PAGE \* MERGEFORMAT 10 Pre-column derivatization HPLC trace detection Enzymatic γ DL glutamyl tryptophan reaction On: Tu Chunyan Wu Yan Jiao Zhong Yao Yuan Yanjuan U.S. Huawei Ping Tang [Abstract] γ D glutamyl L tryptophan (SCV 07) is a novel broad-spectrum immunomodulatory compounds. For the enzymatic reaction system for synthesis of the characteristics of SCV 07, o-nitrobenzene sulfonyl chloride as a pre-column derivatization , RP HPLC for the separation mode, the choice of Lichrospher C18 column, 20 mmol / L phosphate buffer (pH 3.0) and acetonitrile as mobile phase, gradient elution, detection wavelength 230 nm, determined to establish a simultaneous analysis of multiple amino acids and small peptides method, all components in the elution within 15 min. Amino Acids and Dipeptides in 0.96 ~ 260 μmol / L a good linear range, correlation coefficient greater than 0.9991; recoveries in between 97.8% ~ 101.6%; the relative standard deviation of 1.2% -2.9%. This method is suitable for SCV 07 purity analysis, and enzymatic preparation of SCV 07 peptide transfer reaction process tracking and detection, analytical speed, high accuracy and easy operation. [Keywords:] tryptophan, o-nitrobenzene sulfonyl chloride, derivatization, HPLC 1 Introduction γ D glutamyl L tryptophan (γ D Glutamyl L tryptophan, SCV 07) is a novel broad-spectrum immunomodulatory compounds, similar to the role of thymosin @ 1, carried out experiments in mice have confirmed its fight against tuberculosis biological activity [1]. Suzuki et al [2] was first established in 2004, Enzymatic synthesis of SCV 07 technology, in recent years the country will also have the relevant reports [3,4]. This method is the use of γ-glutamyl GGT ( γ Glutamylt ranspeptidase, γ GT, EC), γ glutamyl acylation small peptide, but in the reaction when the L glutamine (L Glutamine, L Gln) as a glutamyl donor, there is peptide response to a certain degree of rotation [5], while, γ GT can SCV 07 for the glutamy