本文選題：利多卡因 + 普魯卡因�。� 參考：《山西醫(yī)科大學(xué)》2007年碩士論文

【摘要】： 目的 1建立生物樣品中利多卡因、普魯卡因、布比卡因原體及分解產(chǎn)物的氣相色譜(GC)及氣相色譜-質(zhì)譜聯(lián)用(GC/MS)定性定量分析方法; 2建立生物樣品中利多卡因、普魯卡因和布比卡因的分解動(dòng)力學(xué)研究方法; 3研究不同存貯條件下生物樣品中利多卡因、普魯卡因和布比卡因的濃度變化規(guī)律,指導(dǎo)選擇最佳檢材存放條件,為局麻藥硬膜外麻醉意外死亡和中毒案件的法醫(yī)學(xué)鑒定提供實(shí)驗(yàn)依據(jù)。 方法 1氣相色譜/氣質(zhì)聯(lián)用分析方法: 1.1樣品處理:腦組織(勻漿)、血液、尿液樣品中加入內(nèi)標(biāo)物,經(jīng)酸、堿化處理后,乙醚萃取。 1.2氣相色譜分析條件:DB-5毛細(xì)管柱(30m×0.25mm×0.25μm),程序升溫模式:150℃(1min)→10℃·min~(-1)→280℃(2min);檢測(cè)器為NPD,溫度300℃;進(jìn)樣口空氣60KPa,氫氣2.3 KPa;載氣:N_2 1.5 ml·min~(-1)。 1.3氣質(zhì)聯(lián)用分析條件:DB-5毛細(xì)管柱(30m×0.25mm×0.25μm),電子能量70ev,質(zhì)量范圍:50～650,柱溫:150℃(1min)→10℃·min~(-1)→280℃(2min);離子源溫度:200℃,傳輸線(xiàn)溫度:250℃,載氣:He:1.5 ml·min~(-1)。 1.4定性定量方法:利用保留時(shí)間結(jié)合特征離子峰(氣-質(zhì)聯(lián)用)定性,內(nèi)標(biāo)法和工作曲線(xiàn)法(氣相色譜)定量。 2分解動(dòng)力學(xué)研究: 2.1分組:利多卡因、普魯卡因和布比卡因?qū)嶒?yàn)組犬各4只,分別按2倍極量30mg·kg~(-1)、60mg·kg~(-1)和10mg·kg~(-1)于5分鐘內(nèi)勻速注入犬蛛網(wǎng)膜下腔。 2.2分解動(dòng)力學(xué)研究:注入藥液后立即處死動(dòng)物,分別采取腦組織、心血和尿液,每種樣品分三等份分別置于20℃、4℃、-20℃環(huán)境中保存,于死后不同時(shí)間點(diǎn)檢測(cè)各保存條件下利多卡因、普魯卡因和布比卡因的含量,winolin軟件擬合分解動(dòng)力學(xué)方程,計(jì)算利多卡因、普魯卡因和布比卡因在不同保存條件下(20℃、4℃、-20℃)不同樣品(腦組織、血液和尿液)中的分解半衰期。 2.3 SAS9.0重復(fù)測(cè)量設(shè)計(jì)資料方差分析法統(tǒng)計(jì)數(shù)據(jù),對(duì)各藥的溫度之間、時(shí)間之間進(jìn)行配對(duì)t檢驗(yàn),比較不同溫度對(duì)各藥的分解動(dòng)力學(xué)過(guò)程有無(wú)影響。 結(jié)果 1氣相色譜/氣質(zhì)聯(lián)用分析:利多卡因、普魯卡因、布比卡因、SKF_(525A)和各藥分解產(chǎn)物的保留時(shí)間分別為8.20、9.70、12.00、12.10和7.80、8.62、11.58min;特征離子片段分別為86、58、30;86、99、120;140、84、98;86、91、99和68、121、165;120、137、92;98、70、42。各峰形均對(duì)稱(chēng),可完全分離。腦組織、血液和尿液中利多卡因、普魯卡因和布比卡因工作曲線(xiàn)線(xiàn)性檢測(cè)范圍分別為0.5～20μg·ml~(-1)、0.5～25μg·ml~(-1)和0.5～20μg·ml~(-1),最低檢出限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、普魯卡因和布比卡因在生物樣品中的分解動(dòng)力學(xué) 2.1分解動(dòng)力學(xué)方程及參數(shù):利多卡因、普魯卡因和布比卡因在生物樣品中的分解動(dòng)力學(xué)符合一級(jí)動(dòng)力學(xué)過(guò)程,可用lgC=lgCo-kt/2.303表示,式中k為一級(jí)分解速率常數(shù)。腦組織、血液和尿液中利多卡因在20℃、4℃和-20℃存儲(chǔ)條件下的分解半衰期t_(1/2)分別為17、30、34;27、35、35和13、14、47天;普魯卡因的分解半衰期t_(1/2)分別為2、2、2;2、7、15和35、42、49天;布比卡因的分解半衰期t_(1/2)分別為33、64、72;51、71、80和17、60、81天。 2.2各藥不同儲(chǔ)存條件下的含量變化趨勢(shì) 本實(shí)驗(yàn)研究結(jié)果顯示,20℃、4℃、-20℃保存時(shí),腦組織、血液和尿液中利多卡因含量均在第8、48和8天顯著下降(P＜0.05);腦組織中利多卡因含量在20℃保存80天、4℃保存120天降為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天降為0μg/ml,4℃和-20℃保存120天分別降至初始值的5.0%和7.6%;尿液中利多卡因含量在20℃、4℃和-20℃保存120天分別降至初始值的1.1%、4.5%、13.0%。 20℃、4℃、-20℃保存時(shí),腦組織、血液和尿液中普魯卡因含量分別在保存第2、2、2,2、4、8和32、48、64天顯著下降(P＜0.05);腦組織、血液和尿液在-20℃保存120天中普魯卡因含量分別為初始值的1.0%、1.9%和35.6%。 20℃、4℃、-20℃保存時(shí),腦組織、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天顯著下降(P＜0.05);腦組織、血液和尿液在-20℃保存120天中布比卡因濃度分別為初始值的54.5%、35.5%和33.2%。 結(jié)論 1建立了腦組織、血液和尿液中測(cè)定利多卡因、普魯卡因、布比卡因的GC及GC/MS分析方法,可使藥物原體和代謝產(chǎn)物完全分離,該法簡(jiǎn)便、靈敏、重現(xiàn)性好,適用于利多卡因、普魯卡因和布比卡因麻醉意外案例的快速鑒定。 2建立了利多卡因、普魯卡因和布比卡因在生物樣品中的分解動(dòng)力學(xué)研究模型,可應(yīng)用于其法醫(yī)毒理學(xué)研究。 3利多卡因、普魯卡因和布比卡因在各保存條件下均有分解,20℃保存時(shí)分解較快,其次為4℃,-20℃保存時(shí)分解較慢,因而麻醉意外死亡和中毒案件的法醫(yī)學(xué)鑒定中應(yīng)注意冷藏或冷凍保存檢材,并盡快檢驗(yàn)。 4利多卡因、普魯卡因、布比卡因在生物樣品內(nèi)的分解動(dòng)力學(xué)符合一級(jí)動(dòng)力學(xué)過(guò)程,可用lgC=lgCo-kt/2.303表示,利用該分解動(dòng)力學(xué)方程,可以推斷解剖或死亡當(dāng)時(shí)體內(nèi)藥物含量,為麻醉意外死亡和中毒案件法醫(yī)學(xué)鑒定中毒物檢驗(yàn)結(jié)果的分析提供了實(shí)驗(yàn)依據(jù)。
[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.

【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類(lèi)號(hào)】：D919

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