Hattori A, Fukushima T, Hamamura K, Kato M, Imai K
Department of Bio-Analytical Chemistry, Graduate School of Pharmaceutical Sciences, the University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.
Biomed Chromatogr. 2001 Apr;15(2):95-9. doi: 10.1002/bmc.42.
A method for the enantiomeric determination of 2,7,8-trimethyl-2-(beta-carboxyethyl)-6-hydroxy chroman (LLU-alpha, gamma-CEHC) in rat plasma was developed using high-performance liquid chromatography (HPLC) with a fluorimetric derivatization with 4-N, N-dimethylaminosulfonyl-7-piperazino-2,1,3-benzoxadiazole (DBD-PZ) followed by O-acetylation with acetyl chloride. The proposed HPLC system used two non-chiral columns (phenyl and octadecylsilica) and a chiral column (a modified cellulose type), which were connected via two column-switching valves. A derivatized sample prepared from rat plasma was first separated on the phenyl column, and the fraction including LLU-alpha derivative was introduced to the octadecylsilica column to quantify the concentration of the mixture of S- and R-LLU-alpha. Finally, the LLU-alpha derivative was directly injected into the chiral column to obtain the ratio of the enantiomers. The proposed HPLC system was applied to the enantiomeric determination of LLU-alpha in plasma after intravenous administration of racemic LLU-alpha. S-LLU-alpha was eliminated faster than R-LLU-alpha, and its concentration in plasma decreased to one-third at 2 min after dosing.
建立了一种使用高效液相色谱（HPLC）对大鼠血浆中2,7,8-三甲基-2-（β-羧乙基）-6-羟基色满（LLU-α，γ-CEHC）进行对映体测定的方法，该方法采用4-N，N-二甲基氨基磺酰基-7-哌嗪基-2,1,3-苯并恶二唑（DBD-PZ）进行荧光衍生化，随后用乙酰氯进行O-乙酰化。所提出的HPLC系统使用两根非手性柱（苯基柱和十八烷基硅胶柱）和一根手性柱（改性纤维素类型），它们通过两个柱切换阀连接。首先将从大鼠血浆制备的衍生化样品在苯基柱上分离，然后将包含LLU-α衍生物的馏分引入十八烷基硅胶柱以定量S-和R-LLU-α混合物的浓度。最后，将LLU-α衍生物直接注入手性柱以获得对映体比例。所提出的HPLC系统应用于外消旋LLU-α静脉给药后血浆中LLU-α的对映体测定。S-LLU-α的消除速度比R-LLU-α快，给药后2分钟其血浆浓度降至三分之一。
