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目的:比较柱前衍生高效液相色谱法(HPLC法)和柱后衍生阳离子交换色谱法(AARO法)测定人胎盘组织液中18种氨基酸含量的差异。方法:HPLC法采用C18柱和2,4-二硝基氯苯(DNCB)进行柱前衍生化,加入0.1 mol獉L~(-1)硼砂缓冲液(p H=9.1),进行测定;AARO法采用强酸性阳离子交换色谱柱和柱后衍生化,直接测定。结果:柱前衍生高效液相色谱法和柱后衍生阳离子交换色谱法的重复性考察,AARO法各组分RSD为0.91%~1.84%,HPLC法RSD为1.04%~1.87%。结论:柱后衍生阳离子交换色谱法与柱前衍生高效液相色谱法测定人胎盘组织液种18种氨基酸,均可采用,两种方法对测定上述各氨基酸结果一致性较好,但柱前衍生HPLC法可能易造成衍生不彻底和衍生物不稳定。
OBJECTIVE: To compare the differences of 18 amino acids in human placenta tissue fluid between pre-column derivatization HPLC and post column derivatization cation exchange chromatography (AARO). Methods: The pre-column derivatization with C18 column and 2,4-dinitrochlorobenzene (DNCB) was carried out by HPLC, and 0.1 mol·L -1 borax buffer (p H = 9.1) was added for determination. AARO Method using strong acid cation exchange column and column derivatization, direct determination. Results: The repeatability of HPLC by HPLC and column-by-column derivatization with cation exchange chromatography showed that the RSD of each component of AARO method was 0.91% ~ 1.84% and the RSD of HPLC method was 1.04% ~ 1.87%. Conclusion: Post-column derivatization cation exchange chromatography and pre-column derivatization high performance liquid chromatography determination of 18 kinds of amino acids in human placental tissue liquid species can be used, the two methods for the determination of the above amino acid consistency is good, but pre-column derivatization HPLC Law may easily lead to incomplete derivation and derivatives instability.