去甲万古霉素HPLC法和EMIT法测定方法的比较及患者血药浓度监测结果的相关分析研究

发布时间：2018-08-25 18:58

【摘要】：去甲万古霉素属糖肽类抗生素,其化学结构与万古霉素相近,抗菌谱和抗菌作用与万古霉素相仿。糖肽类抗生素,是治疗革兰氏阳性菌尤其是耐甲氧西林金葡菌(MRSA)感染的首选药物。随着耐甲氧西林金黄色葡萄球菌(MRSA)和甲氧西林表皮葡萄球菌(MRSE)引起的感染日渐增多,去甲万古霉素的应用越来越频繁。去甲万古霉素不良反应大,治疗窗较窄。因此,在临床使用过程中,对其进行血药浓度监测十分必要。目前,去甲万古霉素的测定方法有荧光免疫偏振法(FPIA)、高效液相色谱法(HPLC),还有少数实验者用液相色谱-质谱/质谱联用法(LC-MS/MS)酶免疫放大法(EMIT)等,各具优缺点。日常监测中需要一种简便快捷准确的测定方法。目的:分别建立HPLC法和EMIT法测定血浆中去甲万古霉素浓度的方法,比较HPLC法和EMIT法测定患者血浆中去甲万古霉素浓度的方法学相关性。对监测结果进行相关性分析研究,为临床去甲万古霉素的合理使用提供理论依据。方法:不同测定方法的比较:分别采用HPLC法和EMIT法建立了血浆中去甲万古霉素浓度的测定方法,并分别验证这两种方法的精密度与准确度稳定性。收集使用过去甲万古霉素患者的血浆样本,分别用HPLC法和EMIT法对其浓度进行测定,对测定结果进行回归分析,比较相关性。患者血药浓度监测结果的相关性分析研究:用EMIT法测定105例血浆样本中去甲万古霉素的浓度,并与年龄、肾功能以及基础疾病等进行相关性比较分析。结果:HPLC法去甲万古霉素和万古霉素的保留时间分别为8.0min,11.0min。标准曲线为:Y=0.0207C-0.0003,r=0.9997,线性范围为2~100μg/ml,最低检测浓度为2μg/ml。低、中、高3种浓度(4、40、80μg/ml)的平均提取回收率(%)分别为101.6±10.09,107.2±2.23,107.7±1.53,日间、日内RSD均小于15%。EMIT法标准曲线:Y=-3.4×10-7C3-9.5×10-6C2+0.0042C+0.3034,r=0.9999,线性范围为2~50μg/ml,最低检测浓度为2μg/ml。HPLC法和EMIT法测定去甲万古霉素样品结果线性回归方程为Y=0.9395X-1.934(Y:EMIT法μg/ml,X:HPLC法μg/ml),r=0.9217。两组数据配对t检验结果:P0.05。血肌酐正常组与大于正常值组比较,P0.05。年龄60~75岁组和75岁组比较,P0.05。结论:HPLC法测定血浆去甲万古霉素和EMIT测定法具有较好的相关性,所监测患者去甲万古霉素血药浓度与年龄肾功能基础疾病联合应用抗生素等有一定相关性,进行血药浓度监测十分必要。
[Abstract]:Norvancomycin is a glycopeptide antibiotic, and its chemical structure is similar to that of vancomycin, and its antibacterial spectrum and antibacterial activity are similar to vancomycin. Glycopeptide antibiotics are the first choice for the treatment of Gram-positive bacteria, especially methicillin-resistant Staphylococcus aureus (MRSA) infection. With the increasing infection caused by (MRSA) and (MRSE) of methicillin-resistant Staphylococcus aureus, norvancomycin is used more and more frequently. The adverse reaction of norvancomycin was large and the window of treatment was narrow. Therefore, it is necessary to monitor the blood drug concentration during clinical use. At present, the methods for the determination of norvancomycin are fluorescence immunopolarization method (FPIA), high performance liquid chromatography (HPLC), and a small number of experimenters by liquid chromatography-mass spectrometry / mass spectrometry (LC-MS/MS)? The enzyme immunoamplification method (EMIT) has its own advantages and disadvantages. Need a simple method of daily monitoring? Fast? Accurate determination method. Aim: to establish a method for the determination of norvancomycin in plasma by HPLC and EMIT, and to compare the correlation between HPLC and EMIT in the determination of norvancomycin in plasma. The correlation analysis of the monitoring results provides a theoretical basis for the rational use of norvancomycin. Methods: HPLC and EMIT methods were used to determine the concentration of norvancomycin in plasma, and the precision and accuracy of the two methods were verified. Stability. Plasma samples were collected from patients with mevancomycin and their concentrations were determined by HPLC method and EMIT method respectively. The results were analyzed by regression analysis and the correlation was compared. Correlation analysis of blood drug concentration monitoring results: the concentration of norvancomycin in 105 plasma samples was determined by EMIT method and was compared with age, renal function and underlying diseases. Results the retention time of norvancomycin and vancomycin was 8.0 min and 11.0 min, respectively. The standard curve is 0. 0207C-0.0003. The linear range is 2 ~ 100 渭 g / ml and the lowest detection concentration is 2 渭 g / ml. The average recovery (%) was 101.6 卤10.09107.2 卤2.23107.7 卤1.53, respectively. Intra-day RSD was smaller than 15%.EMIT standard curve: -3.4 脳 10-7C3-9.5 脳 10-6C2 0.0042C 0.30342C. The linear range was 2 ~ 50 渭 g / ml. The lowest detection concentration was 2 渭 g/ml.HPLC and EMIT method. The linear regression equation was Y=0.9395X-1.934 (Y:EMIT method 渭 g 路ml X: HPLC 渭 g/ml). Two groups of data matched t test result: P0.05. The serum creatinine level in normal group was higher than that in normal group (P 0.05). The age of 60 to 75 years old group and 75 years old group comparison P 0.05. Conclusion there is a good correlation between the determination of plasma norvancomycin and EMIT, and the plasma concentration and age of norvancomycin in patients monitored. Kidney function? Basic diseases? Combined use of antibiotics has certain correlation, so it is necessary to monitor blood drug concentration.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R96

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