Chromatographic profiles of tryptophan and kynurenine enantiomers derivatized with (S)-4-(3-isothiocyanatopyrrolidin-1-yl)-7-(N,N-dimethylaminosulfonyl)-2,1,3-benzoxadiazole using LC-MS/MS on a triazole-bonded column
Chromatographic profiles of tryptophan and kynurenine enantiomers derivatized with (S)-4-(3-isothiocyanatopyrrolidin-1-yl)-7-(N,N-dimethylaminosulfonyl)-2,1,3-benzoxadiazole using LC-MS/MS on a triazole-bonded column
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DOI：
10.1002/chir.22726
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发表时间：
2017-10-01
期刊：
影响因子：
2
通讯作者：
Fukushima, Takeshi
中科院分区：
文献类型：
--
作者：
Iizuka, Hideaki;Harashima, Takahiro;Fukushima, Takeshi
用手性试剂（S）-4-（3-异硫氰酸基吡咯烷-1-基）-7-（N，N-二甲氨基磺酰基）-2，1，3-苯并恶二唑（DBD-PyNCS）衍生化d-和l-色氨酸（Trp）和d-和l-犬尿氨酸（KYN），并通过配备三唑键合柱（Cosmosil HILIC）的高效液相色谱（HPLC）使用串联质谱（MS/MS）检测器进行对映体分离。考察了柱温、盐（HCO 2 NH 4）浓度和移动的相pH对（S）-DBD-PyNCS-d，l-Trp和-d，l-KYN对映体分离的影响，并进行了质谱检测。采用乙腈/10 mM甲酸铵水溶液（90/10）为移动的流动相，柱温50-60 ℃，分离度和质谱检测均良好。d，l-Trp和d，l-KYN的对映体分离分别产生2.22和2.13的Rs值，以及Trp和KYN对映体的1.08和1.08的分离因子（）。所提出的LC-MS/MS方法提供了Trp和KYN的两种对映体的优异检测灵敏度（5.1- 19 nM）。
d- and l-Tryptophan (Trp) and d- and l-kynurenine (KYN) were derivatized with a chiral reagent, (S)-4-(3-isothiocyanatopyrrolidin-1-yl)-7-(N,N-dimethylaminosulfonyl)-2,1,3-benzoxadiazole (DBD-PyNCS), and were separated enantiomerically by high-performance liquid chromatography (HPLC) equipped with a triazole-bonded column (Cosmosil HILIC) using tandem mass spectrometric (MS/MS) detection. Effects of column temperature, salt (HCO2NH4) concentration, and pH of the mobile phase in the enantiomeric separation, followed by MS detection of (S)-DBD-PyNCS-d,l-Trp and -d,l-KYN, were investigated. The mobile phase consisting of CH3CN/10mM ammonium formate in H2O (pH5.0) (90/10) with a column temperature of 50-60 degrees C gave satisfactory resolution (Rs) and mass-spectrometric detection. The enantiomeric separation of d,l-Trp and d,l-KYN produced Rs values of 2.22 and 2.13, and separation factors () of 1.08 and 1.08, for the Trp and KYN enantiomers, respectively. The proposed LC-MS/MS method provided excellent detection sensitivity of both enantiomers of Trp and KYN (5.1-19nM).