利多卡因、普鲁卡因和布比卡因在生物样品中的分解动力学研究

发布时间：2018-04-22 04:33

本文选题：利多卡因 + 普鲁卡因　；参考：《山西医科大学》2007年硕士论文

【摘要】： 目的 1建立生物样品中利多卡因、普鲁卡因、布比卡因原体及分解产物的气相色谱(GC)及气相色谱-质谱联用(GC/MS)定性定量分析方法; 2建立生物样品中利多卡因、普鲁卡因和布比卡因的分解动力学研究方法; 3研究不同存贮条件下生物样品中利多卡因、普鲁卡因和布比卡因的浓度变化规律,指导选择最佳检材存放条件,为局麻药硬膜外麻醉意外死亡和中毒案件的法医学鉴定提供实验依据。方法 1气相色谱/气质联用分析方法: 1.1样品处理:脑组织(匀浆)、血液、尿液样品中加入内标物,经酸、碱化处理后,乙醚萃取。 1.2气相色谱分析条件:DB-5毛细管柱(30m×0.25mm×0.25μm),程序升温模式:150℃(1min)→10℃·min~(-1)→280℃(2min);检测器为NPD,温度300℃;进样口空气60KPa,氢气2.3 KPa;载气:N_2 1.5 ml·min~(-1)。 1.3气质联用分析条件:DB-5毛细管柱(30m×0.25mm×0.25μm),电子能量70ev,质量范围:50～650,柱温:150℃(1min)→10℃·min~(-1)→280℃(2min);离子源温度:200℃,传输线温度:250℃,载气:He:1.5 ml·min~(-1)。 1.4定性定量方法:利用保留时间结合特征离子峰(气-质联用)定性,内标法和工作曲线法(气相色谱)定量。 2分解动力学研究: 2.1分组:利多卡因、普鲁卡因和布比卡因实验组犬各4只,分别按2倍极量30mg·kg~(-1)、60mg·kg~(-1)和10mg·kg~(-1)于5分钟内匀速注入犬蛛网膜下腔。 2.2分解动力学研究:注入药液后立即处死动物,分别采取脑组织、心血和尿液,每种样品分三等份分别置于20℃、4℃、-20℃环境中保存,于死后不同时间点检测各保存条件下利多卡因、普鲁卡因和布比卡因的含量,winolin软件拟合分解动力学方程,计算利多卡因、普鲁卡因和布比卡因在不同保存条件下(20℃、4℃、-20℃)不同样品(脑组织、血液和尿液)中的分解半衰期。 2.3 SAS9.0重复测量设计资料方差分析法统计数据,对各药的温度之间、时间之间进行配对t检验,比较不同温度对各药的分解动力学过程有无影响。结果 1气相色谱/气质联用分析:利多卡因、普鲁卡因、布比卡因、SKF_(525A)和各药分解产物的保留时间分别为8.20、9.70、12.00、12.10和7.80、8.62、11.58min;特征离子片段分别为86、58、30;86、99、120;140、84、98;86、91、99和68、121、165;120、137、92;98、70、42。各峰形均对称,可完全分离。脑组织、血液和尿液中利多卡因、普鲁卡因和布比卡因工作曲线线性检测范围分别为0.5～20μg·ml~(-1)、0.5～25μg·ml~(-1)和0.5～20μg·ml~(-1),最低检出限0.05μg·ml~(-1),方法回收率91±2.1%～114±4.2%,RSD均小于10%。 2利多卡因、普鲁卡因和布比卡因在生物样品中的分解动力学 2.1分解动力学方程及参数:利多卡因、普鲁卡因和布比卡因在生物样品中的分解动力学符合一级动力学过程,可用lgC=lgCo-kt/2.303表示,式中k为一级分解速率常数。脑组织、血液和尿液中利多卡因在20℃、4℃和-20℃存储条件下的分解半衰期t_(1/2)分别为17、30、34;27、35、35和13、14、47天;普鲁卡因的分解半衰期t_(1/2)分别为2、2、2;2、7、15和35、42、49天;布比卡因的分解半衰期t_(1/2)分别为33、64、72;51、71、80和17、60、81天。 2.2各药不同储存条件下的含量变化趋势本实验研究结果显示,20℃、4℃、-20℃保存时,脑组织、血液和尿液中利多卡因含量均在第8、48和8天显著下降(P＜0.05);脑组织中利多卡因含量在20℃保存80天、4℃保存120天降为0μg/g,-20℃降至初始值的4.2%;血液中利多卡因含量在20℃保存96天降为0μg/ml,4℃和-20℃保存120天分别降至初始值的5.0%和7.6%;尿液中利多卡因含量在20℃、4℃和-20℃保存120天分别降至初始值的1.1%、4.5%、13.0%。 20℃、4℃、-20℃保存时,脑组织、血液和尿液中普鲁卡因含量分别在保存第2、2、2,2、4、8和32、48、64天显著下降(P＜0.05);脑组织、血液和尿液在-20℃保存120天中普鲁卡因含量分别为初始值的1.0%、1.9%和35.6%。 20℃、4℃、-20℃保存时,脑组织、血液和尿液中布比卡因含量在保存第8、16、64,4、4、8和2、4、16天显著下降(P＜0.05);脑组织、血液和尿液在-20℃保存120天中布比卡因浓度分别为初始值的54.5%、35.5%和33.2%。结论 1建立了脑组织、血液和尿液中测定利多卡因、普鲁卡因、布比卡因的GC及GC/MS分析方法,可使药物原体和代谢产物完全分离,该法简便、灵敏、重现性好,适用于利多卡因、普鲁卡因和布比卡因麻醉意外案例的快速鉴定。 2建立了利多卡因、普鲁卡因和布比卡因在生物样品中的分解动力学研究模型,可应用于其法医毒理学研究。 3利多卡因、普鲁卡因和布比卡因在各保存条件下均有分解,20℃保存时分解较快,其次为4℃,-20℃保存时分解较慢,因而麻醉意外死亡和中毒案件的法医学鉴定中应注意冷藏或冷冻保存检材,并尽快检验。 4利多卡因、普鲁卡因、布比卡因在生物样品内的分解动力学符合一级动力学过程,可用lgC=lgCo-kt/2.303表示,利用该分解动力学方程,可以推断解剖或死亡当时体内药物含量,为麻醉意外死亡和中毒案件法医学鉴定中毒物检验结果的分析提供了实验依据。
[Abstract]:objective
1 the qualitative and quantitative analysis method of lidocaine, procaine, bupivacaine and decomposition products by gas chromatography (GC) and gas chromatography-mass spectrometry (GC/MS) in biological samples was established.
2 to establish a kinetic method for the decomposition of lidocaine, procaine and bupivacaine in biological samples.
3 the variation of concentration of lidocaine, procaine and bupivacaine in biological samples under different storage conditions was studied to guide the selection of the best storage conditions, and to provide experimental evidence for the forensic identification of accidental death and poisoning cases in epidural anesthesia with local anesthetics.
Method
1 gas chromatograph / GC-MS analysis method:
1.1 sample treatment: brain tissue (homogenate), blood and urine samples were added with internal standard, after acid and alkalization treatment, ether was extracted.
1.2 gas chromatography analysis conditions: DB-5 capillary column (30M x 0.25mm x 0.25 m), temperature programmed mode: 150 C (1min), 10 C, min~ (-1) and 280 C (2min); detector is NPD, temperature 300, air 60KPa, hydrogen 2.3 KPa, carrier gas: N_2 1.5.
1.3 GC-MS analysis conditions: DB-5 capillary column (30M x 0.25mm x 0.25 m), electronic energy 70ev, mass range: 50~650, column temperature: 150 C (1min), 10 C, min~ (-1), 280 C (2min); ion source temperature: 200 c, transmission line temperature: 250 C, carrier gas: He: 1.5 ml min~ (min~).
1.4 qualitative and quantitative methods: using retention time combined with characteristic ion peaks (GC / MS), internal standard method and working curve method (gas chromatography) quantitative analysis.
2 decomposition kinetics study:
2.1 groups: lidocaine, procaine and bupivacaine in the experimental group of 4 dogs, 2 times the maximum amount of 30mg. Kg~ (-1), 60mg. Kg~ (-1) and 10mg. Kg~ (-1) were injected into the subarachnoid space at a uniform speed in 5 minutes.
2.2 decomposition kinetics study: immediately after injection of liquid, the animals were killed, and brain tissue, blood and urine were taken respectively. Each sample was divided into three equal parts at 20, 4, and -20, respectively. The contents of lidocaine, procaine and bupivacaine were detected at different time points after death, and the winolin software fitted the decomposition kinetics. The decomposition half-life of lidocaine, procaine and bupivacaine in different samples (20 C, 4 C, -20 C) in different samples (brain tissue, blood and urine) in different storage conditions was calculated.
2.3 SAS9.0 repeated measurement design data variance analysis method statistics, the temperature of each drug, time between the paired t test, compare the different temperature on the decomposition kinetics of the drugs have no effect.
Result
1 gas chromatography / GC-MS analysis: the retention time of lidocaine, procaine, bupivacaine, SKF_ (525A) and each product were 8.20,9.70,12.00,12.10 and 7.80,8.62,11.58min, respectively, the characteristic ion fragments were 86,58,30, 86,99120, 140,84,98, 86,91,99 and 68121165, 120137,92; the peaks of 98,70,42. were all symmetrical and can be completely divided. The linear detection range of lidocaine, procaine and bupivacaine in brain tissue, blood and urine was 0.5 to 20 mu g. Ml~ (-1), 0.5 to 25 mu ml~ (-1) and 0.5 to 20 g. Ml~ (-1), and the lowest detection limit was 0.05 mu g. Ml~ (-1), and the recovery rate was 91 + 2.1% to 114 + 4.2%.
2 decomposition kinetics of lidocaine, procaine and bupivacaine in biological samples
2.1 decomposition kinetics equation and parameters: the decomposition kinetics of lidocaine, procaine and bupivacaine in the biological samples conforms to the first order kinetics, and the lgC=lgCo-kt/2.303 is used to indicate that K is the first order decomposition rate constant. The decomposed half life of lidocaine in the brain tissue, blood and urine at 20, 4 and -20 centigrade is t 1/2 is 17,30,34, 27,35,35 and 13,14,47 days, and the decomposition half life of procaine t_ (1/2) is 2,2,2, 2,7,15 and 35,42,49 days, and the decomposition half life of bupivacaine is 33,64,72;
2.2 the trend of the content of different drugs under different storage conditions.
The experimental results showed that the content of lidocaine in brain tissue, blood and urine decreased significantly at 20, 4 and -20 C (P < 0.05). The content of lidocaine in brain tissue was kept at 20, 80 days at 20, 4 C for 120 days to 0 u g/g, and 4.2% at -20 C to the initial value of 4.2%; the content of lidocaine in the blood was preserved at 20 degrees for 96 days. It reduced to 0 g/ml, 4 and -20 C for 120 days to 5% and 7.6% of the initial value, and the content of lidocaine at 20, 4 and -20 was 1.1%, 4.5%, 13.0%., respectively, in the urine for 120 days.
At 20, 4 and -20, the content of procaine in brain tissue, blood and urine decreased significantly at day 2,2,2,2,4,8 and 32,48,64 (P < 0.05). The content of procaine in brain tissue, blood and urine at -20 for 120 days was 1%, 1.9% and 35.6%. respectively.
At 20, 4 and -20, the content of bupivacaine in brain tissue, blood and urine decreased significantly on day 8,16,64,4,4,8 and 2,4,16 (P < 0.05). The concentration of bupivacaine in brain tissue, blood and urine at -20 C for 120 days was 54.5%, 35.5% and 33.2%., respectively.
conclusion
1 the GC and GC/MS methods for the determination of lidocaine, procaine and bupivacaine in the brain tissue, blood and urine can separate the drug and metabolites completely. This method is simple, sensitive and reproducible. It is suitable for the rapid identification of the cases of lidocaine, procaine and bupivacaine.
2 a kinetic model for the decomposition of lidocaine, procaine and bupivacaine in biological samples has been established, which can be applied to forensic toxicology research.
3 lidocaine, procaine and bupivacaine were decomposed under the preservation conditions. The decomposition was fast at 20 C, followed by 4 degrees C, and the decomposition was slow at -20 C. Therefore, the forensic identification of the cases of accidental death and poisoning of anesthesia should be paid attention to cold storage or cryopreservation and the inspection as soon as possible.
The decomposition kinetics of 4 lidocaine, procaine and bupivacaine in biological samples accorded with the first order kinetics, which can be expressed by lgC=lgCo-kt/2.303. Using the kinetic equation, we can infer the drug content in the body at the time of dissection or death, and the analysis of the results of the toxicological test in the accidental death of anesthesia and the forensic identification of the medium toxic cases. The experimental basis is provided.

【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：D919

【参考文献】