血指印检测研究进展

犯罪现场血指印的发现和提取在案件侦破中起着非常重要的作用,是最有效的个人身份鉴定手段之一。血指印检测方法依据反应原理的不同分为化学方法、物理方法、生物染色和光学方法等4类。本文重点综述了血指印的检测技术、常用试剂及其优缺点,并展望了血指印检测方法的方向和前景。     

基于金属有机骨架材料Tb-BTC显现潜指印的研究

为探究金属有机骨架材料在指印显现领域的应用,本研究采用预混合迅速滴显的方法,使金属有机骨架材料Tb-BTC直接络合在指印纹线上.使用该材料对渗透性客体中A4纸、黑色卡纸、牛皮纸;非渗透性客体中透明玻璃片、瓷砖上的皮脂指印进行显现,探究了不同配比、不同滴加顺序、不同显现时间的预混合溶液对不同类型、不同遗留时间、不同遗留物...     

双重对象同时检测的新型荧光纳米粒子传感器的制备

报道了一种基于荧光共振能量转移理论合成的新型纳米粒子. 新型纳米粒子A中掺杂了由亲和素-生物素桥联并可发生荧光共振能量转移(FRET)的供体染料和受体染料; 新型纳米粒子B中只掺杂了供体染料. 在使用供体染料的特征激发波长同时激发纳米粒子A和B时, 纳米粒子A由于荧光共振能量转移而发射出受体染料的特征波长, 而纳米粒子B则只发射供体染料的特征波长. 这样, 在单一波长激发下, 可很容易地实现双重对象同时检测.     

荧光可调铜纳米簇对潜指印的可视化识别

本文以支化聚乙烯亚胺(BPEI)为模板,抗坏血酸(AA)为还原剂,谷胱甘肽(GSH)为稳定剂,硫酸铜(Cu SO4)为原料,通过温度控制,制备了一种荧光可调的新型铜纳米簇(Cu NCs).在激发光为365、415、450 nm条件下, Cu NCs分别产生蓝色、绿色、黄色荧光.将其应用于潜指印的可视化识别,成功解决了目前潜指印显示试剂的同色背景干扰问题.采用RGB色彩分析与灰度差分统计,实现了指印图像的数据化分析,提供了一种客观评价潜指印可视化效果的新方法,为该领域建立数据化的评价体系提供了参考.     

MALDI-TOF质谱和电子光谱技术研究人血清铁蛋白释放铁的动力学

采用MALDI-TOF质谱技术研究血清混合蛋白质的组成,用电子光谱技术分析人血清铁蛋白(Serumferritin,SF)释放铁的动力学过程和规律.结果表明,在人血清中,以Na₂S₂O₄为电子供体时,SF释放铁的速率极快,无法采用常规分光光度法进行检测;以抗坏血酸为电子供体时,SF以一级反应动力学方式释放铁,认为人血清中其它蛋白质协助或参与调控铁蛋白进行快速释放铁反应,从而满足人体对铁的需求.     

中空纤维支载离子液体液液微萃取法检测环境水体中的三氯生

三氯生(5 Chloro-2-(2,4-dichlorophenoxy) phenol,TCS)是一种新型环境水体污染物,具有潜在的生态与健康风险,因此发展合适的分析方法来检测水环境中这类化合物极其必要.本研究以1-辛基-3-甲基咪唑六氟磷酸离子液体( [C8 MIM][PF6])为萃取剂,基于中空纤维的离子液体液液微萃取方法,结合HPLC/UV用于环境水样中TCS的分析测定;通过对各参数(萃取剂、供体相的体积、供体相pH值、离子强度、萃取时间等)的优化在最优条件下(样品相体积为50 mL,pH值2,盐浓度为0.2 mol/L,200 r/min振荡萃取8 h),获得了较高的富集倍数(907倍)、较低的检出限(0.05 μg/L,RSD=7.4％,n=6)和较好的线性范围(0.1～100 μg/L);以4种环境水样加标实验对方法的准确性进行评估,其回收率可达94.2％～108.5％(RSD=5.5％～8.0％,n=6);本方法可广泛应用于环境水体中痕量TCS的分析检测.     

基于SERS纳米探针的细胞内硝基还原酶检测

在缺氧的肿瘤细胞内, 硝基还原酶(NTR)通常过表达且其含量高低与缺氧程度呈正相关, 因此开发高选择性检测NTR的方法对早期肿瘤诊断至关重要. 本文通过修饰对硝基苯硫酚(p?NTP)到金纳米粒子(Au NPs)表面构建了一种表面增强拉曼散射(SERS)探针. 在缺氧条件下, 以还原型烟酰胺腺嘌呤二核苷酸(NADH)作为电子供体, NTR可催化还原芳香硝基为芳香胺, 导致纳米探针的SERS光谱发生变化, 从而实现NTR的高选择性检测, 检出限低至18 ng/mL. 该探针毒性低、 生物兼容性好, 可用于缺氧条件下A549细胞内的NTR分析, 为肿瘤细胞的缺氧现象评估提供了一种有效的策略.     

模拟过氧化物酶催化性能的研究

过氧化物酶(POD)能催化H₂O₂氧化一系列氢供体(DH₂)转化成氧化型供体(D)。氢供体可以是邻联二茴香胺、4-氨基安替比林和苯酚及某些染料隐色体等。这类反应不仅可提供检测H₂O₂的灵敏方法,还可与产生H₂O₂的其它酶反应系统联用,测定葡萄糖、半乳糖、氨基酸、尿酸及胆醇等生物物质。     

D-A型共轭低聚物的电子性质及D-A比对其影响的理论研究

以[1,2,5]噻重氮并[3,4-g]喹喔啉(TQ)为受体、 噻吩(Th)、 噻吩并[3,2-b]噻吩(TTh)和吡咯(Py)为供体, 设计了6类供体-受体(D-A)型共轭低聚物. 采用杂化的密度泛函方法(B3LYP), 研究了此6类低聚物的电子结构和性质. 电子密度拓朴分析和核独立化学位移计算表明, 随着聚合度的增加, 体系共轭程度增强, HOMO-LUMO能级差逐渐减小. 同时, 随着聚合度的增加, 低聚物电离能减小, 电子亲和势增大. 供体-受体摩尔比(D-A比)对低聚物的性质有重要影响, 提高D-A比能有效地增大分子内电荷迁移, 从而使HOMO-LUMO能级差减小. Py不仅是一个强的电子供体, 还是一个潜在的氢键供体. 在含Py结构单元的低聚物中, 由于分子内氢键的存在使其具有较大的分子内电荷迁移值. 所设计的6种基于TQ的四聚体均具有较小的HOMO-LUMO能级差(<1 eV), 使其相应的聚合物的能隙更小, 可作为潜在的性能优良的导电材料.     

基于三苯胺的双氰基二苯代乙烯型双光子荧光溶剂生色探针

以双氰基二苯代乙烯为双光子荧光母体,借助电子供体-π-电子受体(D-π-A)构-效关系,以二苯氨基作电子供体,氰基作电子受体,开发了一个新型的双光子荧光溶剂生色探针(SP1),并对其结构进行了表征.性能测试结果表明,SP1在环己烷和二甲亚砜(DMSO)中的最大发射波长分别为452和604 nm,显示出很宽的溶剂生色范围(152 nm),可用于溶剂极性与黏度的检测,亦能根据最大发射波长识别溶剂种类.SP1拥有较大的双光子吸收截面,在环己烷和DMSO中的最大双光子吸收截面分别为6670和2040 GM,并具有较大的斯托克斯位移(194 nm),其光稳定性、水溶性和细胞渗透性均比较优良.该探针可用于细胞中黏度的检测与成像.     

Exploratory analysis of chromatographic fingerprints to distinguish rhizoma Chuanxiong and rhizoma Ligustici

Identification and quality control of products of natural origin, used for preventive and therapeutical goals, is required by regulating authorities, as the World Health Organization. This study focuses on the identification and distinction of the rhizomes from two Chinese herbs, rhizoma Chuanxiong (from Ligusticum chuanxiong Hort.) and rhizoma Ligustici (from Ligusticum jeholense Nakai et Kitag), by chromatographic fingerprints. A second goal is using the fingerprints to assay ferulic acid, as its concentration provides an additional differentiation feature. Several extraction methods were tested, to obtain the highest number of peaks in the fingerprints. The best results were found using 76:19:5 (v/v/v) methanol/water/formic acid as solvent and extracting the pulverized material on a shaking bath for 15 min. Then fingerprint optimization was done. Most information about the herbs, i.e. the highest number of peaks, was observed on a Hypersil ODS column (250 mm × 4.6 mm ID, 5 μm), 1.0% acetic acid in the mobile phase and employing within 50 min linear gradient elution from 5:95 (v/v) to 95:5 (v/v) acetonitrile/water. The final fingerprints were able to distinguish rhizoma Chuanxiong and Ligustici, based on correlation coefficients combined with exploratory data analysis. The distinction was visualized using Principal Component Analysis, Projection Pursuit and Hierarchical Clustering Analysis techniques. Quantification of ferulic acid was possible in the fingerprints of both rhizomes. The time-different intermediate precisions of the fingerprints and of the ferulic acid quantification were shown to be acceptable.     

高效液相色谱-质谱联用法检测免疫抑制剂及其合并用药

建立高效液相色谱-质谱联用法同时测定人血浆中免疫抑制剂及合并用药12种药物浓度的方法.选用Kromasil-C18色谱柱(50 mm× 4.6 mm×5 μm),以甲醇-乙腈-1mmol/L乙酸铵溶液为流动相,采用梯度洗脱进行分离,样本用甲醇沉淀蛋白后进样,流速:1.1 mL/min;柱温:35℃;进样量:20μL.选用3200QTrap型液相色谱-串联质谱仪的多反应监测(MRM)扫描方式进行检测.12种药物的线性范围为0.2～1000μg/L;定量下限为0.2 μg/L.准确度与精密度结果显示方法日间、日内RSD均小于15％;相对偏差-13％～9.33％,稳定性较好.本方法快速、灵敏,专属性强、重现性好,可用于人体血浆中免疫抑制剂及其常用合并用药共12种药物浓度的测定.     

超高效液相色谱-串联质谱法测定人血浆中盐酸氨溴索及其制剂的生物等效性评价

采用超高效液相色谱-串联质谱（UHPLC-MS/MS）技术,建立了快速、简单、灵敏的测定人血浆中盐酸氨溴索含量的方法,并用于盐酸氨溴索人体生物等效性预试验研究。取50 μL血浆样品,采用蛋白沉淀法处理,以盐酸氨溴索-d5为内标。采用Waters XBridge BEH C18色谱柱（50 mm×2.1 mm,2.5 μm）,以0.1%（v/v）甲酸水-含0.1%（v/v）甲酸的甲醇为流动相,在0.4 mL/min流速下进行梯度洗脱。采用电喷雾电离（ESI）源以正离子模式进行MRM检测。结果显示,盐酸氨溴索在2~400 ng/mL范围内线性关系良好,相关系数（r）为0.998,准确度为97.1%~108.7%,精密度为1.0%~5.6%。将该方法用于6名健康受试者口服盐酸氨溴索受试制剂和参比制剂30 mg后血药浓度的测定,结果显示二者相对生物利用度为（102.3±14.8）%,血药浓度-时间曲线下面积（AUC_0-t、AUC_0-∞）和最大血药浓度（C_max）的90%置信区间均在80.0%~125.0%范围内,两种制剂生物等效。     

Determination of heterocyclic aromatic amines by capillary electrophoresis coupled to mass spectrometry using in-line preconcentration

This paper shows the potentiality of capillary electrophoresis (CE) coupled to mass spectrometry (MS) for the analysis of heterocyclic aromatic amines obtaining good results in terms of sensitivity and precision. These compounds have a special interest since they can be carcinogenic for humans. The optimization of a CE-MS method was performed and the best conditions were obtained using a 16 mM formic acid/ammonium formate solution at pH 4.5 with 60% methanol as running electrolyte. For CE-MS coupling, a sheath liquid methanol/20 mM formic acid (75/25) solution at a flow rate of 3 microL/min and hydrodynamic injection of methanol mixtures for 10 s were used. Detection limits ranging from 18 ng/g to 360 ng/g and precisions up to 1.4% and 12% for migration time and concentration, respectively, were obtained. In order to improve sensitivity, field-amplified sample injection was applied as an in-line preconcentration method. Methanol/5 mM formic acid (50/50) as a sample solvent, 3 s hydrodynamic injection (0.5 psi) of a methanol plug, and 25 s of electrokinetic injection (10 kV) of the sample were found to be the optimum conditions. Detection limits up to 25 times lower and similar precisions than those reported for hydrodynamic injection were obtained.     

超高效液相色谱-电喷雾串联质谱法测定生活饮用水中49种药物及5种代谢物

建立了超高效液相色谱-电喷雾串联质谱(UPLC-ESI MS/MS)分析生活饮用水中54种药物的方法。采用HLB固相萃取柱对水样中的目标化合物进行富集净化,以5 mL甲醇洗脱;洗脱液用氮气吹至近干,用0.4 mL 0.1%甲酸水溶液定容,上机分析;ACQUITY UPLC^TMBEH C₁₈柱用作色谱分离,以0.1%甲酸水溶液-甲醇为流动相进行梯度洗脱;多反应监测(MRM)模式进行检测;目标药物使用基质外标法定量。54种药物在自备井水、市政末梢水和地表水中的加标回收率分别为58.7%~104.4%、53.1%~109.5%和50.7%~118.8%,相对标准偏差(n=6)分别为0.3%~12.8%、1.0%~15.5%和0.4%~19.3%;方法定量限为0.002~5.000 ng/L。将建立的方法应用于北京部分自备井水、市政末梢水和地表水样品的分析,结果在自备井水样中检出26种药物。     

免疫亲和柱净化-超高效液相色谱-三重四极杆质谱法高灵敏测定尿液和血浆中3种鹅膏毒肽

建立了超高效液相色谱-三重四极杆质谱高灵敏测定尿液和血浆中α-鹅膏毒肽、β-鹅膏毒肽和γ-鹅膏毒肽的方法。经过免疫亲和柱净化,尿液样品浓缩20倍、血浆样品浓缩10倍,以Kinetex Biphenyl色谱柱(100 mm×2.1 mm, 1.7 μm)作为分析柱,甲醇-0.005%(v/v)甲酸水溶液作为流动相进行梯度洗脱分离,电喷雾电离、负离子、多反应监测模式下检测,外标法定量。3种鹅膏毒肽的线性范围为0.1~200 ng/mL,相关系数(r)>0.999。尿液和血浆中3种鹅膏毒肽的基质效应和提取回收率分别为92%~108%和90%~103%,变异系数均小于13%。尿液中3种鹅膏毒肽的准确度为-9.4%~8.0%,重复性和中间精度分别为3.0%~14%和3.5%~18%,当取样量为2.00 mL时,方法的检出限均为0.002 ng/mL;血浆中3种鹅膏毒肽的准确度为-13%~8.0%,重复性和中间精度分别为3.9%~9.7%和5.5%~12%,当取样量为1.00 mL时,方法的检出限均为0.004 ng/mL。该法操作简单、灵敏、准确,已在中毒患者摄入野生蘑菇后138 h的尿液中检出0.0067 ng/mL α-鹅膏毒肽和0.0059 ng/mL β-鹅膏毒肽。该法已成功解决中毒患者尿液和血浆中超痕量鹅膏毒肽的检测难题,对于疑似中毒病人的早诊断、早治疗、降低死亡率都具有非常重要意义,也为今后开展此类毒素毒理作用及机体代谢规律的研究提供了可靠的技术支撑。     

Simultaneous quantitation of two diastereoisomers of lobaplatin in rat plasma by supercritical fluid chromatography with tandem mass spectrometry and its application to a pharmacokinetic study

Lobaplatin, consisting of two diastereoisomers, is a third‐generation platinum antineoplastic agent that has shown encouraging anticancer activity in a variety of tumor types. To investigate any stereospecificity in the pharmacokinetics of lobaplatin, a novel, simple, rapid and sensitive supercritical fluid chromatography with tandem mass spectrometry method was developed for the simultaneous quantitation of lobaplatin diastereoisomers in rat plasma. After a simple protein precipitation with methanol, the analytes and dexpantoprazole (internal standard) were chromatographed on an Acquity UPC² system with a Chiralcel OZ‐RH column using a mobile phase consisting of carbon dioxide and methanol (65:35, v/v) at 40°C over 6 min. The assay was linear over a concentration range of 25–15 000 ng/mL for both diastereoisomers using 100 μL of rat plasma for sample preparation. The lower limit of quantification was 25 ng/mL for both compounds, which was sufficient to detect the diastereoisomers in the incurred samples within this study. Intra‐ and inter‐day precisions were below 11.8% and the accuracies were below 4.5%. The validated method was successfully applied to a pharmacokinetic study after an intravenous administration of 7.6 mg/kg lobaplatin to rats. There was no apparent stereospecificity in the pharmacokinetics between the two diastereoisomers of lobaplatin.     

高通量固相萃取-超高效液相色谱-串联质谱法测定人尿中8种环境酚类内分泌干扰物

建立了人尿中8种环境酚类化合物的96孔板固相萃取-超高效液相色谱-串联质谱(96-well SPE LC-MS/MS)检测方法,其中包括7种双酚类化合物和三氯生。尿样解冻到室温,经β-葡萄糖醛酸苷肽酶/芳基磺酸酯酶37 ℃过夜酶解。实验比较了3种96孔板固相萃取柱和不同淋洗条件对人尿样的净化效果和目标化合物的回收率。结果显示,采用Oasis HLB 96孔板(60 mg)对样品进行萃取和用30%(v/v)乙腈水溶液进行淋洗净化的纯化效果最好。纯化后目标物用甲醇溶液洗脱,经氮气吹干,用0.5 mL甲醇-水(1∶1, v/v)溶液定容,目标化合物用UPLC-MS/MS进行检测。比较了2种分析柱(C₁₈和T3分析柱)以及不同的有机流动相对分离样品中目标物的影响。结果显示,以BEH C₁₈(100 mm×2.1 mm, 1.7 μm)作为分析柱,乙腈/水作为流动相,以流速0.3 mL/min梯度洗脱时,目标物的分离效果最好。质谱条件选择串联质谱负离子电喷雾(ESI^-)多反应监测模式(MRM)进行检测。对样品的基质效应进行评估发现,双酚A、双酚F、双酚S、双酚B和双酚AF的绝对基质效应为3.47%~15.32%,不需要补偿措施;四氯双酚A、四溴双酚A和三氯生的绝对基质效应分别是49.58%(中等基质效应)、71.99%和86.93%(强基质效应),均需要补偿效应。因此,该方法采用了一一对应的同位素内标法抵消基质效应。用6份实际尿样基质评估相对基质效应,8种内标的峰面积的相对标准偏差为3.63%~9.06%,说明相对基质效应稳定。在优化条件下,双酚A和双酚AF在0.50~50 μg/L内、四氯双酚A和双酚S在0.05~50 μg/L内、双酚F和四溴双酚A在0.01~50 μg/L内、双酚B在1.00~50 μg/L内、三氯生在5.00~200 μg/L内线性关系良好,相关系数大于0.9995。方法检出限为0.002~1.09 μg/L,定量限为0.007~3.63 μg/L。3个加标水平的加标回收率为81.0%~101.9%,日内精密度为0.4%~19.4%,日间精密度为2.5%~17.8%。应用该方法对2019-2020年采集的北京地区64份尿样进行测定,结果发现8种目标分析物中,除双酚B和双酚AF未检出外,其余均有检出,其中双酚A和双酚S的检出率最高,分别为100%和96.9%。三氯生、四溴双酚A、四氯双酚A和双酚F的检出率分别为57.8%、46.9%、23.4%和21.9%。尿样中8种目标物含量的中位值以降序排列分别为1.44 μg/L(三氯生)、0.69 μg/L(双酚A)、0.086 μg/L(双酚S)、0.0032 μg/L(四溴双酚A)、0.00050 μg/L (四氯双酚A)、0.00 μg/L(双酚F、双酚B和双酚AF)。以上尿样检测结果显示,北京市居民存在普遍的环境酚类化合物暴露,值得关注。该方法操作简单,定量准确,样品需求量小,有机试剂消耗少,适合大批量样本的测定。     

Quantification of lipid modified estrogenic derivative (ESC8) in rat plasma by LC‐MS: application to a pharmacokinetic study

A lipid‐conjugated, estrogenic derivative molecule, ESC8, compared with other estrogenic molecules, encourages cell death in both ER‐positive and ER‐negative breast cancer cells. A rapid and highly sensitive assay method has been developed and validated for the estimation of a ESC8 in rat plasma using liquid chromatography coupled with mass spectrometry under positive‐ion mode with electrospray ionization. The sample process includes using methanol for precipitation of ESC8 and dextromethorphan (internal standard, IS) from plasma. Chromatographic separation was achieved with methanol–water–formic acid (70:30:0.1% v/v/v) pumped at a flow rate of 0.3mL/min and a C₁₈ column (50 × 2.1 mm i.d., 1.7 μm particle size) with a total run time of 5 min. The m/z ions monitored were 568.5 and 272.1 for ESC8 and IS, respectively. The lower limit of quantitation achieved was 1.08 ng/mL and linearity was observed from 5 to 500 ng/mL. The intra‐ and inter‐day precisions were <4%. The proposed method was successfully applied to a preliminary pharmacokinetic study of ESC8 liposomal formulation following an intraperitoneal administration of 3.67 mg/kg in rats. The concentrations of ESC8 in plasma were quantifiable up to 36 h. The peak concentration of ESC8 was found to be 110.72 ng/mL, the area under the concentration–time curve was 1625.23ng/mL h and the half‐life was 11.72 h.     

超高效液相色谱-串联质谱法测定血浆与尿液中14种麻痹性贝类毒素北大核心CSCD

人体生物基质中麻痹性贝类毒素的检测对其引起的食物中毒诊断和救治具有重要意义。研究建立了超高效液相色谱-串联质谱法测定血浆、尿液中14种麻痹性贝类毒素的分析方法。实验比较了不同固相萃取柱的影响,优化了前处理条件和色谱条件,血浆样品采用0.2 mL水、0.4 mL甲醇、0.6 mL乙腈提取后直接上机测定,尿液样品采用0.2 mL水、0.4 mL甲醇、0.6 mL乙腈提取,聚酰胺(PA)固相萃取柱净化后上机测定。采用Poroshell 120 HILIC-Z色谱柱(100 mm×2.1 mm,2.7μm)对14种贝类毒素进行分离,流动相为含0.1%(v/v)甲酸的5 mmoL/L甲酸铵缓冲溶液和0.1%(v/v)甲酸乙腈溶液,流速为0.50 mL/min。在电喷雾模式(ESI)下进行正负离子扫描,采用多反应监测(MRM)模式检测,外标法定量。结果表明,对于血浆和尿液样品,14种贝类毒素分别在0.24~84.06 ng/mL范围内线性关系良好,相关系数均大于0.995。尿液检测的定量限为4.80~34.40 ng/mL,血浆检测的定量限为1.68~12.04 ng/mL。尿液和血浆样品在1、2和10倍定量限加标水平下平均回收率为70.4%~123.4%,日内精密度为2.3%~19.1%,日间精密度为4.0%~16.2%。应用建立的方法对腹腔注射14种贝类毒素小鼠血浆和尿液进行测定,20份血浆样本中检出含量分别为19.40~55.60μg/L和8.75~13.86μg/L。该方法操作简便,样品取样量少,方法灵敏度高,适用于血浆和尿液中麻痹性贝类毒素的快速检测。