固相萃取-反相高效液相色谱法同时测定人血浆中五种苯二氮卓类药物浓度及其临床应用药理专业论文.docxVIP

汕头大学医学院硕士学位论文 汕头大学医学院硕士学位论文 汕头大学医学院硕士学位论文 汕头大学医学院硕士学位论文 III III IV IV 二、 建立固相萃取法预处理血浆样品 方法：采用 C1 cartridges（100 mg，1 mL，Agilent 公司），对血浆样本进行预处 理，以提取、分离、富集待测药物。 柱子依次用 5mL 甲醇、5mL 去离子水，进行活化、平衡后，用 pH=7 的磷酸缓 冲液稀释血样上柱；依次用 1mL 水、1mL 水/甲醇混合液（80:20 v/v）洗去杂质；用 2mL 甲醇洗脱、收集被测物；洗脱液在 45℃氮气下吹干，用流动相定容至 125uL； 离心，5℃，13000 rpm x 5min，取上清液以备进样。 结果：六种药物（包括内标氯丙咪嗪）的提取回收率为 81 - 101%，RSD 为 2.1 - 13 %。地西泮的最低检测限为 3.2ng/mL，奥沙西泮的最低检测限为 2.2ng/mL，氯 硝西泮的最低检测限为 1.1ng/mL，阿普唑仑的最低检测限为 1.5ng/mL，三唑仑的最 低检测限为 1.4ng/mL。 结论：建立了固相萃取法对血浆样本进行预处理，可以有效提取样本中五种苯二 氮卓类药物及内标氯丙咪嗪，回收率较高；选择性较高，有效去除内源性干扰；将待 测药物的浓度提高为原来的 4 倍，提高了分析方法的灵敏度。本方法操作简便、精密 度好、结果准确可靠，适于临床对血浆样本中此类药物含量的分析测定。 关键词：高效液相色谱法，固相萃取法，苯二氮卓类药物，地西泮，奥沙西泮， 氯硝西泮，阿普唑仑，三唑仑。 Simultaneous Determination of Five Benzodiazepines in Human Plasma by Solid Phase Extraction RP-HPLC ABSTRACT Benzodiazepines (BZDs) possess sedative, hypnotic, anxiolytic, anticonvulsant and muscle relaxant actions, which are the first choice for clinical use in a variety of indications such as insomnia, anxiety, seizures, and so on. In general, BZDs are safe and effective in short term, while long-term usage prone to cause tolerance, physical dependence, and a withdrawal syndrome upon discontinuation; large dosage can cause cognitive or memory impairments, and the potential for toxicity increases when combined with other central nervous system depressants such as alcohol, opiates or tricyclic antidepressants. BZDs are commonly misused and taken in combination with other drugs of abuse. Thus, it is important to develop feasible and widely applicable analytical methods for the analysis of BZDs, which should be able to discriminate between the different drugs even if they are co-administered and/or taken as part of poly-pharmacy regimen such as during antidepressant therapy, finally supporting safe and effective therapy for patients. Several BZDs including diazepam, oxazepam, clonazepam, triazolam and alprazolam are commonly used in clinic. When given in therapeutical doses, plasma levels of diff