Identification of furosemide in hair in a post‐mortem case by UHPLC‐MS/MS with guidance on interpretation

Testing for drugs in hair raises several difficulties. Among them is the interpretation of the final concentration(s). In a post‐mortem case, analyses revealed the presence of furosemide (12 ng/mL) in femoral blood, although it was not part of the victim's treatment. The prosecutor requested our laboratory to undertake an additional analysis in hair to obtain information about the use of furosemide. A specific method was therefore developed and validated to identify and quantify furosemide in hair by UHPLC‐MS/MS. After decontamination of 30 mg of hair, incubation in acidic condition, extraction with ethyl acetate, the samples were analyzed by UHPLC‐MS/MS. Furosemide was found in the victim's hair at 225 pg/mg. However, it was not possible to interpret this concentration due to the absence of data in the literature. Therefore, the authors performed a controlled study in two parts. In order to establish the basis of interpretation, several volunteers were tested (four after a single 20 mg administration and twenty‐four under daily treatment). The first part indicated that a single dose is not detectable in hair using our method. The second part demonstrated concentrations ranging from 5 to 1110 pg/mg with no correlation between dosage and hair concentrations. The decedent's hair result was interpreted as repeated exposures. In the case of furosemide analysis, hair can provide information about its presence but cannot give information about dosage or frequency of use.     

人体毛发中硝甲西泮的HPLC-MS/MS检测

目的建立测定涉毒人员毛发样本中硝甲西泮的液相色谱-质谱/质谱(HPLC-MS/MS)分析方法。方法对质谱主要参数进行优化,采用电喷雾多反应监测模式(MRM)测定不同浓度标准样品并建立标准曲线,通过准确度、灵敏度和实际案例检材的检验对方法进行评价。结果标准溶液最低检出限0.1ng/mL,线性范围宽,R²>0.999,空白毛发外添加标样回收率>80%。结论优化后的毛发样本中硝甲西泮的HPLC-MS/MS操作简便快捷,灵敏度高,重现性好,具备定性定量的适用性和可靠性。     

基于相差显微镜无损检验头发染色问题研究

为了有效识别犯罪现场提取的黑色头发是否是染色头发,利用OLYMPUSBX51相差显微镜无损观察未染色及几种情况染色的黑色头发。结果表明,黑色头发中染色与未染色、黑色再染黑与白色染黑、染色两周与刚刚染色头发之间在相差显微镜下都存在显著差别。因此,文中基于相差显微镜无损检验方法为法医学鉴定头发染色问题提供了有效的检验方法。     

Characterization of Mitochondrial DNA Sequence Heteroplasmy in Blood Tissue and Hair as a Function of Hair Morphology*,†,‡

Abstract: This study characterizes mitochondrial DNA (mtDNA) sequence heteroplasmy in blood tissue and hair as a function of hair morphology. Bloodstains (127 individuals) and head hairs (128 individuals) were typed using the mtDNA LINEAR ARRAY? assay. A total of 1589 hairs were interpreted: 1478 (93%) were homoplasmic and 111 (7%) exhibited heteroplasmy at one or more positions. Seventy‐one percent (82/116) of individuals were homoplasmic, whereas 29% (34/116) exhibited heteroplasmy in at least one hair. The results demonstrate intra‐ and inter‐tissue differences in heteroplasmy within individuals. Sequence heteroplasmy among hairs from each individual varied from 0 to 90%; the frequency does not differ significantly with population group, cosmetic treatment, age, gender, medulla morphology, region of the scalp, hair growth phase, or, when comparing living and deceased donors. However, the results support a correlation between heteroplasmy and hair pigmentation; typically, lighter‐pigmented hairs exhibit a higher incidence of sequence heteroplasmy compared to darker hairs.     

微波消解ICP-MS法测定人头发中24种元素的含量

目的建立人头发中24种无机元素的电感耦合等离子体质谱（inductively coupled plasma-mass spec-trometry,ICP-MS）分析方法。方法采用微波消解法处理样品,以铟（115In）作内标,用ICP-MS分析人头发中的24种元素含量。同时检测56例健康志愿者和10例海洛因滥用者头发中24种元素含量。结果 24种元素的方法检出限范围为0.0003~10.14μg/g,标准物质的测得值与标准值基本相符。海洛因滥用者经戒毒治疗后头发中镁、镓、钡含量下降。结论该方法灵敏度、准确度高,适用于头发中24种元素的测定。     

GC-MS/MS法测定人头发中的大麻酚类及其代谢物

目的 建立同时检测头发中△9-四氢大麻酚(THC)、大麻酚(CBN)、大麻二酚(CBD)和△9-四氢大麻酸(THC-COOH)的分析方法.方法头发样品加入氘代内标△9-四氢大麻酸(THC-COOH-d3),经碱水解后,以混合溶剂[V(正己烷)∶V(乙酸乙酯=9∶1]进行提取,吹干,残留物经双(三甲基硅烷基)三氟乙酰胺(BSTFA)衍生化,用GC-MS/MS方法进行分析.结果 头发中THC-COOH、THC、CBN和CBD的最低检出限分别为4、4、10和20 pg· mg-1,各化合物在0.04～5ng· mg-1呈良好的线性关系(r＞0.999),方法精密度、准确度均符合要求.结论本方法选择性强、灵敏度高,适用于头发中CBD、CBN、THC及其代谢物THC-COOH的分析,并成功应用于实际案例中.     

毛发中GHB的检测及评价

目的探索毛发中外源性GHB的检测及判断的可行性,为涉GHB的鉴定提供方法和依据。方法建立毛发中GHB的GC/MS分析方法,并通过动物实验,考察毛发中内源性GHB的质量分数范围、外源性GHB在毛发中的时间过程以及给药剂量、毛发颜色与毛发中GHB的质量分数关系。结果豚鼠和中国人黑色毛发中内源性GHB质量分数分别为(3.01±1.41)ng/mg(n=28)和(1.02±0.27)ng/mg(n=20);摄GHB后毛发中GHB质量分数明显增加且与给药剂量呈正相关性;GHB在毛干中呈窄带分布;深色毛发中GHB质量分数高于浅色毛发。结论毛发中GHB的检测适用于GHB滥用和中毒的法医毒物学鉴定;根据毛发中的GHB质量分数和毛发分段分析可判断GHB的来源。     

度冷丁滥用者毛发分段分析及其结果评价

以度冷丁滥用者为研究对象,在度冷丁滥用者毛发中检出度冷丁及代谢物去甲度冷丁、N一羟甲基度冷丁和N-乙酰度冷丁。60例度冷丁滥用者头发中度冷丁和去甲度冷丁的含量分别为103±130ng／mg和117±143ng／mg。度冷丁稳定地存在于头发中,检出时限至少为药后20个月,而去甲度冷丁则随着离头发根距离的增加而降低。头发分段分析揭示度冷丁在毛干中的分布和滥用史、剂量和含量存在相关性。     

Sensitive Method for the Confident Identification of Genetically Variant Peptides in Human Hair Keratin

Recent reports have demonstrated that genetically variant peptides derived from human hair shaft proteins can be used to differentiate individuals of different biogeographic origins. We report a method involving direct extraction of hair shaft proteins more sensitive than previously published methods regarding GVP detection. It involves one step for protein extraction and was found to provide reproducible results. A detailed proteomic analysis of this data is presented that led to the following four results: (i) A peptide spectral library was created and made available for download. It contains all identified peptides from this work, including GVPs that, when appropriately expanded with diverse hair-derived peptides, can provide a routine, reliable, and sensitive means of analyzing hair digests; (ii) an analysis of artifact peptides arising from side reactions is also made using a new method for finding unexpected modifications; (iii) detailed analysis of the gel-based method employed clearly shows the high degree of cross-linking or protein association involved in hair digestion, with major GVPs eluting over a wide range of high molecular weights while others apparently arise from distinct non-cross-linked proteins; and (v) finally, we show that some of the specific GVP identifications depend on the sample preparation method.     

Simultaneous Quantitative Determination of Amphetamines,Opiates, Ketamine,Cocaine and Metabolites in Human Hair: Application to Forensic Cases of Drug Abuse

A method using liquid chromatography–tandem mass spectrometry (LC–MS/MS) to simultaneously quantify amphetamines, opiates, ketamine, cocaine, and metabolites in human hair is described. Hair samples (50 mg) were extracted with methanol utilizing cryogenic grinding. Calibration curves for all the analytes were established in the concentration range 0.05–10 ng/mg. The recoveries were above 72%, except for AMP at the limit of quantification (LOQ), which was 48%. The accuracies were within ±20% at the LOQ (0.05 ng/mg) and between −11% and 13.3% at 0.3 and 9.5 ng/mg, respectively. The intraday and interday precisions were within 19.6% and 19.8%, respectively. A proficiency test was applied to the validated method with z-scores within ±2, demonstrating the accuracy of the method for the determination of drugs of abuse in the hair of individuals suspected of abusing drugs. The hair concentration ranges, means, and medians are summarized for abused drugs in 158 authentic cases.