本文關(guān)鍵詞： 去痛片 代謝物 GC-MS/MS 分布　出處：《山西醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:1.建立SLE固相支撐液液萃取、GC-MS/MS同時(shí)檢測去痛片及其代謝物的方法;2.觀察去痛片及其代謝物在大鼠體內(nèi)的分布;3.觀察去痛片及其代謝物在中毒死亡者體內(nèi)的死后分布。方法:1.方法建立利用去痛片及其代謝物標(biāo)準(zhǔn)品,建立SLE固相支撐液液萃取,GC-MS/MS法同時(shí)檢測去痛片及其代謝物。2.標(biāo)本采集16只雄性SD大鼠,分為兩組,每組8只。禁食12h后,經(jīng)口灌服去痛片950mg/kg,A組于1.5h脫臼處死,取心血肝素抗凝、心肌、肺、肝臟、脾、腎、上肢肌肉、下肢肌肉、腦組織置于-20℃待檢。B組于24h脫臼處死,取心血肝素抗凝,并收集24h尿液,置于-20℃待檢。中毒案例樣品2例,來源于山西醫(yī)科大學(xué)司法鑒定中心。3.提取檢測待檢生物樣品中添加烯丙基異丙基巴比妥(50μg/m L),氨基比林-13C2(25μg/m L)混合內(nèi)標(biāo)工作液80μL,經(jīng)SLE固相支撐液液萃取,GC-MS/MS檢測,MRM模式掃描,利用保留時(shí)間、定量離子與定性離子對比例定性,內(nèi)標(biāo)法和工作曲線法定量。4.數(shù)據(jù)處理數(shù)據(jù)用SPSS 20.0進(jìn)行正態(tài)性檢驗(yàn)和方差分析。各臟器間含量用Graphpad 6.0進(jìn)行Kruskal-Wallis H檢驗(yàn),各組間用Dunn's檢驗(yàn)多重比較得出統(tǒng)計(jì)學(xué)結(jié)果,P0.05時(shí),認(rèn)為各組間有統(tǒng)計(jì)學(xué)差異。結(jié)果:1.空白血液加入去痛片及其代謝物標(biāo)準(zhǔn)品,經(jīng)SLE分別用正己烷、二氯甲烷、乙醚、甲基叔丁基醚、乙酸乙酯2m L淋洗,結(jié)果乙酸乙酯同時(shí)提取去痛片及其代謝物的提取回收率MAA為37.57%±0.62%,AAA為41.99%±1.21%,其余化合物均在58.19%±1.41%~95.87%±1.79%之間,相較其他溶劑有較高的提取回收率。2.GC-MS/MS法可同時(shí)檢測去痛片及其代謝物,血液、尿液、肝臟加標(biāo)樣品在0.125μg/m L~16.00μg/m L呈線性關(guān)系,血液樣品工作曲線相關(guān)系數(shù)r為0.9896~0.9997,最低檢測限為0.23ng/m L~14.48ng/m L;尿液樣品工作曲線相關(guān)系數(shù)r為0.995~0.9997,最低檢測限為0.08ng/m L~4.71ng/m L;肝臟樣品工作曲線相關(guān)系數(shù)r為0.9953~0.9997,最低檢測限為0.16ng/m L~11.18ng/m L。方法準(zhǔn)確度為79.64%~122.91%,日內(nèi)精密度為0.99%~7.43%,日間精密度為2.19%~10.60%,3.經(jīng)口灌服950mg/kg去痛片,1.5小時(shí)后去痛片及其代謝物在大鼠體內(nèi)的分布:氨基比林血液含量上、下肢肌肉含量,P0.05。MAA血液、肝臟含量下肢肌肉含量,P0.05。AA血液、上、下肢肌肉、肝臟、腎臟、心肌含量胃壁含量,P0.05。FAA和AAA各臟器間含量無統(tǒng)計(jì)學(xué)差異。非那西丁血液、肺臟、脾臟含量肝臟含量,P0.05。APAP血液、下肢肌肉、心肌、腎臟、肝臟、脾臟含量胃壁含量,P0.05�？Х纫蜓�、肝臟含量上、下肢肌肉、腦組織含量,P0.05。17X、37X、13X各臟器間含量無統(tǒng)計(jì)學(xué)差異。苯巴比妥肝臟含量脾臟、腦、上、下肢肌肉含量,P0.05。經(jīng)口灌服去痛片24h后,血液中MAA、AA含量高于FAA,P0.05。非那西丁和APAP,咖啡因和17X、37X、13X間含量均無統(tǒng)計(jì)學(xué)差異。24小時(shí)尿液中AAA含量高于AM、MAA、FAA,APAP含量高于非那西丁,P0.05�？Х纫蚝推浯x物含量無統(tǒng)計(jì)學(xué)差異。結(jié)論:1.經(jīng)SLE固相支撐液液萃取,同時(shí)提取生物樣品中去痛片及其代謝產(chǎn)物,乙酸乙酯較正己烷、二氯甲烷、乙醚和甲基叔丁基醚有較高的提取回收率。2.GC-MS/MS法可同時(shí)檢測生物樣品中去痛片及其代謝產(chǎn)物,利用保留時(shí)間和二級質(zhì)譜圖定量離子和定性離子對比例定性,利用內(nèi)標(biāo)物與目標(biāo)物峰面積比值,內(nèi)標(biāo)法和工作曲線法定量。方法回收率高、專屬性強(qiáng)、特異性高、內(nèi)源性物質(zhì)對目標(biāo)物檢測干擾小,方法靈敏度高、準(zhǔn)確度高,可滿足去痛片中毒案例定性、定量的要求,可應(yīng)用于去痛片相關(guān)案例的法醫(yī)學(xué)鑒定實(shí)踐中。3.去痛片及其代謝物在大鼠體內(nèi)的分布不均勻,口服去痛片中毒案例中優(yōu)先選擇胃內(nèi)容物和胃壁作為檢材,還可提取肝、腎、心血作為檢材。針對氨基比林、非那西丁、咖啡因和苯巴比妥單一藥物或復(fù)方制劑中毒的案例,應(yīng)根據(jù)藥物的分布特點(diǎn),科學(xué)全面地選取檢材,且應(yīng)該考慮到藥物所處吸收時(shí)相或消除時(shí)相時(shí)對各臟器分布的影響,以便為中毒案件提供科學(xué)的依據(jù)。
[Abstract]:Objective: to establish 1. SLE supported by solid liquid liquid extraction, and detection method of Compound Aminopyrine Phenacetin Tablets and its metabolite GC-MS/MS; 2. were Compound Aminopyrine Phenacetin Tablets and its metabolite distribution in rat; 3. were Compound Aminopyrine Phenacetin Tablets and its metabolites in poisoning deaths postportem distribution. Methods: 1. methods to set up by Compound Aminopyrine Phenacetin Tablets and its metabolite standard, the establishment of SLE solid support liquid liquid extraction, and acquisition of 16 male SD rats and its metabolite.2. were Compound Aminopyrine Phenacetin Tablets GC-MS/MS, divided into two groups, 8 rats in each group. After fasting 12h, oral gavage of Compound Aminopyrine Phenacetin Tablets 950mg/kg, A in 1.5h group were sacrificed by dislocation and taken blood heparin, myocardium, lung, liver, spleen, kidney, upper limb the lower limb muscles, muscle, brain tissue at -20 DEG C to be detected in the.B group 24h were sacrificed by dislocation and taken blood heparin, and 24h urine collection, at -20 DEG C to be detected. 2 cases were poisoning, from the judicial identification center of Shanxi Medical University.3 . extraction to add isopropyl allyl barbituric detection in biological samples (50 g/m L), amidopyrin -13C2 (25 g/m L) 80 L internal standard working solution mixed by SLE, supported by solid liquid liquid extraction, GC-MS/MS detection, MRM scanning mode, the retention time, qualitative and quantitative and qualitative ion the ion ratio, internal standard method and the working curve method, quantitative.4. data were analyzed by SPSS 20 test of normality and variance. The organs between content of Graphpad with 6 Kruskal-Wallis H test between groups using Dunn's test multiple comparison statistical results, P0.05, that there were significant differences between the two groups. Results: 1. join Compound Aminopyrine Phenacetin Tablets and blank blood metabolite standard, respectively by SLE with n-hexane, dichloromethane, ethyl ether, methyl tert butyl ether, ethyl acetate and 2m L leaching, the extraction recovery of ethyl acetate extraction of Compound Aminopyrine Phenacetin Tablets and its metabolites rate MAA 37.57% + 0.62%, 41.99% + 1.21% AAA, the remaining compounds were between 58.19% + 1.41%~95.87% + 1.79%, compared with other solvent extraction recovery rate is higher in.2.GC-MS/MS method for simultaneous determination of Compound Aminopyrine Phenacetin Tablets and its metabolites, blood, urine, liver samples with a linear relationship in the 0.125 g/m L~ 16 g/m L, blood sample curve correlation coefficient r is 0.9896~0.9997, the minimum detection limit is 0.23ng/m L~14.48ng/m L; urine sample curve correlation coefficient r is 0.995~0.9997, the minimum detection limit is 0.08ng/m L~4.71ng/m L; liver samples curve correlation coefficient r is 0.9953~0.9997, the minimum detection limit is 0.16ng/m L~11.18ng/m L. the accuracy of 79.64%~122.91%, the within day precision is 0.99%~7.43%. The inter day precision is 2.19%~10.60%, 3. oral gavage of 950mg/kg 1.5 hours after Compound Aminopyrine Phenacetin Tablets, Compound Aminopyrine Phenacetin Tablets and its metabolite distribution in rat: aminopyrine The content of blood, muscle content, lower limb blood P0.05.MAA content in liver, lower limb muscle P0.05.AA content, blood, liver, kidney, muscle, myocardial content of gastric content, no significant difference between P0.05.FAA and AAA in various organs. Phenacetin blood, lung, spleen in liver P0.05.APAP content, blood, lower limb muscle, myocardium the liver, spleen, kidney, stomach contents of caffeine content, P0.05. content of blood, liver, muscle, brain tissue content, P0.05.17X, 37X, 13X were no significant difference between the content of benzene. Pakistan than spleen, liver, brain with content, lower limb muscle content, P0.05. oral gavage Compound Aminopyrine Phenacetin Tablets 24h, blood in MAA, the content of AA was higher than FAA P0.05., phenacetin and caffeine and APAP, 17X, 37X, 13X were no statistically significant differences between the content of AAA in the urine.24 hours is higher than AM, MAA, FAA, APAP were higher than that of phenacetin, caffeine and its P0.05. There was no significant difference in the contents of metabolites. Conclusion: 1. by SLE supported by solid liquid liquid extraction, simultaneous extraction of Compound Aminopyrine Phenacetin Tablets and its metabolites in biological samples, ethyl acetate with hexane, dichloromethane, ethyl ether and methyl tert butyl ether extract has a higher recovery rate of.2.GC-MS/MS method for simultaneous determination of Compound Aminopyrine Phenacetin Tablets and its metabolites in biological samples, with retention time and two quantitative and qualitative ion ion mass spectra on the proportion of qualitative, the internal standard and the target using the peak area ratio, internal standard method and the working curve method. The recovery rate is high, strong specificity, high specificity, endogenous substances on the target detection of small interference, high sensitivity, high accuracy, can be meet Compound Aminopyrine Phenacetin Tablets poisoning cases qualitative and quantitative requirements, and can be applied to the Compound Aminopyrine Phenacetin Tablets case forensic practice in Compound Aminopyrine Phenacetin Tablets.3. and its metabolites in uneven distribution of rats, oral In the case of poisoning Compound Aminopyrine Phenacetin Tablets preferred gastric contents and gastric wall as samples, can be extracted from the liver, kidney, blood samples. As for aminopyrine, phenacetin, caffeine and phenobarbital single drug or compound poisoning case, according to the distribution of drugs, scientifically selected samples, and should be considered drug absorption phase or eliminate phase influence on the distribution of various organs, in order to provide scientific basis for poisoning cases.

【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：D919

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 劉繼華;吳凡;王瑩;劉屹;;HPLC法測定去痛片中4種成分的含量[J];海峽藥學(xué);2016年07期

2 李明;王曉琪;竇紅允;;薄層色譜法鑒別去痛片4種主要成分[J];中國執(zhí)業(yè)藥師;2016年04期

3 冷紅梅;萬紫旭;李建;盧思英;馬軍捷;;去痛片口服致慢性腎功能不全1例[J];河北醫(yī)學(xué);2015年08期

4 楊亮;初婷婷;朱昱;;血中非那西丁的GC/MS檢測[J];廣州化工;2014年24期

5 王晉濤;郭建軍;尉志文;萬東方;王勇;王玉瑾;，

本文編號：1496502