高效液相色谱法检测癫癎患儿血清中氯硝西泮

HPLC assay for serum clonazepam in epileptic children

目的 :建立检测血清中氯硝西泮 (CZP)的高效液相色谱法 ,监测 CZP血药浓度 ,为临床应用提供依据。方法 :0 .5 m l样品用 Tris- HCl缓冲液 -乙醚系统萃取净化。色谱条件 :以 C1 8反相柱为分析柱 ,流动相为甲醇 -乙腈 - 0 .0 1%磷酸溶液 (35∶ 15∶ 5 5 V/ V) ,流速 1m l/ m in,柱温 30℃ ,检测波长 2 5 4nm。结果 :CZP在 2 0～ 140 0 ng/ m l范围内呈线性 ,血清中最低检测限 5ng/ m l,绝对回收率 91.8%～ 94.6 %。 9例使用剂量为 0 .0 78～ 0 .36 m g· kg- 1 · d- 1 的癫患儿稳态谷血药浓度为 79.8～ 2 74ng/ m l,症状得到有效控制 ,除嗜睡外未见其他不良反应。结论 :本法简便、快速、准确 ,适合于体液中 CZP的监测及药物动力学研究 ;本研究也表明儿科所用的

AIM: To develop a method for determination of the clonazepam (CZP) in serum for providing a scientific basis for clinical use of CZP. METHODS: 0 5 ml serum sample was added to the test tube packed with 0 5 ml Tris HCl buffer solution, and then was extracted by 3 ml×2 ethyl ether, C 18 reversed column was used, mobile phase: methanol acetonitrile -0 01% phosphoric acid solution (35∶15∶50 V/V ), flow rate 1 ml·min -1 , column temperature 30℃. The detection was carried out with an ultraviolet detector at 254 nm. RESULTS: The linear relation was in the range of 20- 1 400 ng·ml -1 . The lower detection limit of CZP in serum was 5 ng·ml -1 . The absolute recoveries of 4 concentrations of CZP in serum were 91 8%, 93 4%, 94 2%, 94 6% respectively. CZP (0 078-0 36 mg·kg -1 ·d -1 ) was administered orally to 9 epileptic children. The ( c ss ) min was in the range of 79 8-274 ng·ml -1 . All were up to the super limit of therapeutic ranges. Seizure was controlled, adverse drug reactions except sleepy and somnolence weren't discovered. CONCLUSION: This paper provides a rapid, convenient, sensitive and accurate method which is suitable for monitoring CZP and pharmacokinitic study. It is indicated that the reliability of currently used therapeutic serum concentration range for CZP in children will be investigated further.