HPLC法测定百草枯急性中毒大鼠的体内分布

目的 应用高效液相色谱法对口服百草枯急性中毒大鼠体内分布进行测定。方法以200mg／kg剂量百草枯给予Wister大鼠灌胃,4h后脱臼处死,解剖取脑、心、肝、脾、肺、肾、胃、盲肠、肌肉等组织,应用固相萃取法提取,液相色谱法测定各器官组织中百草枯含量。结果各组织经检验,均检出百草枯;组织间百草枯含量（μg／g）相差明显,其中最高为胃（231．47±129．10）,其次为盲肠（87．08±39．86）、肺（22．73±10．20）,最低为心（2．01±0．36）。结论百草枯口服给药后组织分布较为广泛,除胃、肠外各脏器中以肺浓度最高。     

离子对SPE—HPLC法检测生物检材中的百草枯

目的建立生物检材中百草枯的简便、快速、灵敏、可靠的高效液相色谱分析方法。方法生物检材酶解后,用以十二烷基三甲基溴化铵和十二烷基硫酸钠预处理过的C18固相柱萃取,HPLC/DAD进行分析。结果回收率81%～94%,检出限为1ng·mL-1,线性范围50ng·mL-1～1mg·mL-1,结论此方法适用于中毒生物检材中百草枯的检测。     

A fatal case of parenteral paraquat poisoning.

The chief clinical and analytical aspects of the suicide of a 21-year-old male with psychiatric problems by parenteral administration of a 200 g/l aqueous solution of paraquat are described. Paraquat levels were determined in plasma, urine, kidney, liver and lung after autopsy. Tissue damage was studied by electron microscopy. The death ensued from pulmonary dysfunction 15 days after hospital admission.     

Determination of paraquat from formalin-fixed tissue

In this paper, a sulfuric acid digestion method and a clean-up technique by using cation exchange resin followed by XAD-2 resin has been developed for the determination of paraquat from formalin-fixed tissue at the submicrograms per gram level. Formalin-fixed tissue is dissolved by hot sulfuric acid, then paraquat is isolated and purified with cation exchange chromatography. The eluted paraquat forms an ion-pair with sodium dodecyl sulfate, it is then adsorbed on XAD-2 resin. Paraquat is eluted, extracted and reduced with solvent mixtures, NaCl solution and dithionite reagent, respectively. The calibration graphs of zero-order and second-derivative spectroscopy are linear in the range of 0.01-5.0 mg/kg. The relative standard deviation was less than 5% and the detection limit was 0.02 mg/kg based on 0.5-g samples. The sensitivity of the proposed method could be increased by using larger sample sizes. The method was precise and gave a quantitative recovery of paraquat spiked into formalin-fixed liver homogenates (78%). The proposed method has been satisfactorily applied to the determination of paraquat in the formalin-fixed tissues of suspected poisoned cases. It has been shown to be of great value in the field of forensic toxicology especially when formalin-fixed tissue only is available.     

大鼠百草枯中毒后体内的分布研究

目的建立生物检材中百草枯的超高效液相色谱-质谱联用检测方法,研究百草枯灌胃染毒致死的大鼠动物模型。方法大鼠以1/2 LD_(50)剂量灌胃染毒,分别于染毒后0.5h、2h、4h、8h、12h、24h、48h、72h处死解剖,采集心、肝、脾、肺、肾、脑、肌肉、膀胱和胃组织,UPLC-MS/MS法定量检测各组织中百草枯。结果试验中大鼠灌胃后,4h以内胃是主要分布器官,胃中含量最多,其他器官中含量相对较低。4h内除胃以外的脏器含量变化不大,4h后胃内百草枯含量有所下降,除胃以外的脏器含量均升高。各组织与脑组织比较具有显著性差异(P0.05)。结论百草枯在大鼠体内死后分布不均匀并且各组织含量随着时间变化有所改变。百草枯UPLC-MS/MS方法、口服染毒致死的动物模型、各组织分布规律可为甲百草枯中毒死亡案件提供检测依据。     

Determination of paraquat in tissue using ion-pair chromatography in conjunction with spectrophotometry

Homogenized tissue was deproteinized with sulfuric acid. Paraquat in the supernatant was quantificated directly with the dithionite reagent (step 1) or concentrated by the XAD-2 column chromatographic technique before paraquat determination (step 2). Tissue paraquat levels in the range of 0.01-75 mg/kg could be quantificated by second-derivative or zero-order spectroscopy using 2.5 g of tissues. The sensitivity could be increased tenfold by using 25 g of tissue samples. The coefficients of variation of within-run and day-to-day precisions of spiked paraquat in tissue homogenates were below 5% at concentrations of 10.0, 1.0 and 0.1 mg/kg, respectively. The recoveries of the spiked paraquat in tissues ranging from 0.1-10 mg/kg were 91% by step 1 and 74% by step 2. Using these simple methods, steps 1 and 2, the paraquat concentrations in the psoas muscle, liver, lung and kidneys of a swine dosed with 0.16 g/kg of paraquat were investigated. The results were in close agreement with those of the TCA deproteinization method followed by cation-resin column chromatography. The proposed method offers the advantages of simplicity, rapidity, reasonable sensitivity and a wide range of concentrations.     

Homicidal Paraquat Poisoning

Paraquat poisoning usually results from suicide, occupational, or accidental exposure. Herein, we report a rare fatal case of homicidal paraquat poisoning. A 58‐year‐old man was poisoned by taking paraquat‐mixed medicine and wearing paraquat‐soaked underwear. In the absence of a history of paraquat exposure, the patient was misdiagnosed with pulmonary infection and scrotal dermatitis and died of respiratory failure 24 days after the initial exposure to paraquat. Ultra‐performance liquid chromatography‐tandem mass spectrometry (UPLC‐MS/MS) was applied to detect and quantify paraquat in postmortem specimens. The concentration of paraquat in postmortem specimens from high to low is lung (0.49 μg/g), brain (0.32 μg/g), kidney (0.24 μg/g), liver (0.20 μg/g), cardiac blood (0.11 μg/mL), and stomach wall (<LOQ). Identification of homicidal paraquat poisoning is not easy for a clinician or a forensic pathologist, it is important to consider the possibility of paraquat poisoning when patients suffer from rapidly aggravating pneumonia of unknown origin.     

The detection of nifedipine in biological materials

The objective of the present study was to find optimal conditions for the isolation of nifedipine from biological materials by ethylacetate. It was shown that nifedipine can be purified from co-extracted substances of the biological material on a Silasorb C-18 column with the size of the particles 30 microns. The authors propose to use thin-layer chromatography, IR spectrophotometry, and reverse-phase high performance liquid chromatography for the identification and quantitative determination of nifedipine extracted from cadaveric liver samples.     

尸体材料中对硫磷(1605)代谢产物氨基对硫磷的检测

本文报道了对硫磷(1605)中毒后其代谢产物氨基对硫磷的测定方法。检材中的氨基对硫磷经有机溶剂提取,用气相色谱(NPD)测定,在肝或血中的回收率皆在70％以上。本法适用于腐败组织中以检测氨基对硫磷来确定1605中毒。     

The determination of benzodiazepines in biological objects on the Milichrome microcolumn liquid chromatograph]

1,4-benzodiazepines and benzophenols, the products of their hydrolytic cleavage, were measured by high-pressure liquid chromatography in expert material (stomach, liver, intestine, kidney samples). Milichrome microcolumn liquid chromatograph, designs 1 and 2, was employed, with a chromatography column packed with Separon C18 (5 microns) sorbent. A mixture of (NH4)2HPO4 (0.05 M) and acetonitryl in volume ratio 65:35 was used as the mobile phase in native compound analysis and that in volume ratio 45:55 for benzophenols and medazepam separation. The external standard method was used for quantitative estimation.     

False positive reaction for carboxyhemoglobin in blood from liver tissue

On spectrophotometric determination of carboxyhemoglobin in blood collected from the liver of three bodies at three days post-mortem, false positive results were found (5 – 15 per cent saturation), since samples of heart blood collected a few hours after death did not contain carboxyhemoglobin.     

秋水仙碱在急性中毒豚鼠体液和组织中的检测及分布

目的建立生物检材中秋水仙碱的液相色谱/串联质谱检测方法,研究秋水仙碱急性中毒在豚鼠体内的分布,为秋水仙碱中毒的法医学鉴定提供方法和评价依据。方法以4mg/kg剂量秋水仙碱给豚鼠灌胃染毒,2～8h死亡,用LC-MS/MS测定体液和组织中秋水仙碱的含量。结果秋水仙碱在豚鼠各体液和组织中含量从高到低依次为胆汁、尿液、胃、脾、肺、肾、心、胰、肝、肾上腺、睾丸,心血中秋水仙碱的含量最低。结论所建方法准确、灵敏,适用于体内痕量秋水仙碱的检测。胆汁和尿液是体内检测秋水仙碱的较佳检材。     

Qualitative and quantitative analysis of cephradine in biological materials by high performance liquid chromatography

A reversed phase high performance liquid chromatographic method was developed for the determination of cephradine, one of the commonly used antibiotics, in biological materials. Mimic samples for stomach contents, miso soup, were applied to high performance liquid chromatography (HPLC) after centrifugation and purification by Sep-Pak C18 cartridge treatment. Serum samples deproteinized or urine samples diluted were directly injected into the HPLC. The recoveries of cephradine from these materials were 95-97% and the detection limit was 0.01 microgram/injection. This method was applied to the analysis of cephradine in stomach contents obtained by autopsy. After purification by the cartridge treatment, cephradine in the sample was identified and determined by HPLC and further confirmed by thin-layer chromatography (TLC) and mass spectrometry (MS).     

Chronological study of diazinon in putrefied viscera of rats using GC/MS, GC/EC and TLC

A qualitative and qualitative weekly study of diazinon in experimental rats after a lethal dose is described. GC/MS and TLC were used for qualitative, and GC/EC for quantitative analysis. The replicate content of diazinon in stomach and intestine (S/I) revealed a high rate of decrease during the first month. The liver (L) content fluctuates through a general trend of decrease. Immediate post-mortem content of 34.5 mg in summer and 94 in winter was found in S/L samples, while it was 0.79 and 0.63 respectively for L-samples. The respective remaining amount after 2 months was 1. 16, 4.40 (S/I), 0.61 and 0.66 mg (L). A round figure of 4% remains in S/L samples. The chronological plots indicate the possibility of detection at longer periods. Interpretation of data is outlined regarding relative contents of organs and the factors affecting persistence of diazinon in putrefied viscera.     

Blowfly Larval Tissues as a Secondary Detector for Determining Paraquat-Related Death in Rabbit Carcass

Paraquat poisoning is commonly associated with suicide or homicide in Malaysia. In a case involving advanced body decomposition, pathological analysis regarding the cause of death may become difficult or almost impossible. Insects serve as common alternative matrix for poison detection in forensic analysis. Paraquat detection via secondary bioaccumulation in fly larvae tissue has never been reported. In this study, tissues from blowfly larvae collected from a rabbit carcass with paraquat poisoning were analyzed for secondary bioaccumulation. Larvae samples were collected and analyzed using liquid–liquid extraction. The detection was performed via reduction of quaternary ammonium presence in paraquat and analyzed using gas chromatography–mass spectrometry (GC-MS) with selected ion monitoring mode (SIM mode). GC-MS showed the elution of reduced paraquat was at retention time 12.8 min. Blowfly larvae tissue has proven useful as a secondary detector in paraquat-related deaths.     

应用气相色谱电子捕获检测器检测生物体内拟除虫菊酯杀虫剂

本文研究了一种同时测定生物样品中二氯苯醚菊酯、氯氰菊酯、杀火菊酯、溴氰菊酯的方法,并详细介绍了试管提取净化的过程。在安装电子捕获检测器的气相色谱仪上,选择不同极性的色谱柱和优选操作参数,能使上述4种杀虫剂达到基线分离,并在20～25分钟内同时完成分析。添加生物样品肝、胃,脑、血中的回收率达到70～90％;变异系数在±0.03～0.08范围。应用本文建立的方法,对中毒兔(溴氰菊酯和杀灭菊酯)各脏器均检出了原体杀虫剂,并成功地运用于一些法庭中毒案件的鉴定。     

Fatal combined anileridine-pethidine poisoning. A gas chromatography, thin layer chromatography and mass spectrometry investigation.

Anileridine and pethidine were established by gas and thin layer chromatography and mass spectroscopy. In the mass spectrum the main peak of anileridine is found at m/e 246 and that of pethidine at m/e 71. The determination was made by gas chromatography from the blood, urine, liver, muscle and stomach contents.     

溴敌隆及其代谢物-苄叉丙酮在犬体内的死后分布

目的研究溴敌隆及其代谢物-苄叉丙酮在中毒致死犬体内死后分布规律,为溴敌隆中毒检材的采取提供实验依据。方法分别经口给予犬2倍和4倍LD_(50)溴敌隆,待其死亡后迅速解剖取材,气相色谱-质谱联用法测定心血、外周血、尿、胆汁、心、肝、脾、肺、肾、脑、左下肢肌、膀胱、胃、胃内容、胰等脏器和体液中溴敌隆和代谢物-苄叉丙酮的含量。结果犬经2倍和4倍LD_(50)溴敌隆灌胃染毒后3d开始出现出血症状,(178.40±20.94)h后死亡。溴敌隆和代谢物-苄叉丙酮在各组织脏器及体液中的死后分布为:溴敌隆2LD_(50)组溴敌隆:胆汁尿、肝、心、肾心血、外周血、脾、肺等;苄叉丙酮:胆汁、尿、心血、外周血、肺、胃内容中含量高于其他脏器。溴敌隆4LD_(50)组溴敌隆:胆汁、尿肝、外周血心血、胃内容物等脏器。苄叉丙酮:胆汁、尿、肺浓度高于其他脏器。结论溴敌隆及其代谢物-苄叉丙酮在中毒致死犬体内死后分布不均匀,溴敌隆在胆汁、尿、肝脏、心血和外周血含量较高,代谢物-苄叉丙酮在胆汁、尿、肺较高。胆汁、尿、肝脏、心血、外周血可作为疑似溴敌隆中毒毒物分析的检材。     

Chlorobutanol poisoning. Report of a fatal case.

A fatal chlorobutanol intoxication is reported and a gas chromatographic method for the determination of this drug in human fluids and tissues is described. Analytical findings for blood, urine, stomach contents, bile, liver, kidney, spleen, and brain are reported.     

Evaluation of Postmortem Bacterial Migration Using Culturing and Real‐Time Quantitative PCR

Postmortem bacteriology can be a valuable tool for evaluating deaths due to bacterial infection or for researching the involvement of bacteria in various diseases. In this study, time‐dependent postmortem bacterial migration into liver, mesenteric lymph node, pericardial fluid, portal, and peripheral vein was analyzed in 33 autopsy cases by bacterial culturing and real‐time quantitative polymerase chain reaction (RT‐qPCR). None suffered or died from bacterial infection. According to culturing, pericardial fluid and liver were the most sterile samples up to 5 days postmortem. In these samples, multigrowth and staphylococci were not or rarely detected. RT‐qPCR was more sensitive and showed higher bacterial positivity in all samples. Relative amounts of intestinal bacterial DNA (bifidobacteria, bacteroides, enterobacter, clostridia) increased with time. Sterility of blood samples was low during the studied time periods (1–7 days). The best postmortem microbiological sampling sites were pericardial fluid and liver up to 5 days after death.