高效液相色谱法测定人血浆及尿液中头孢呋辛和舒巴坦

建立反相HPLC-UV法同时测定人血浆及尿中头孢呋辛(CXM)和舒巴坦(SUL)的方法.采用Welch Materials XB-C18分析柱(150 mm×4.6 mm,5 μm ) ,流动相为乙腈-0.01 mol/L KH2PO4 (血浆测定15∶ 85(V/V),尿样测定10∶ 90(V/V),含0.04%三乙胺,以H3PO4调节至pH 3.2),流速1.0 mL/min,紫外检测波长SUL为220 nm、CXM及内标咖啡酸为274 nm.血浆样品在酸性条件下用乙醚提取浓集后进样,以内标法进行定量分析.尿样稀释后直接进样,以外标法进行定量分析.血浆测定中SUL和CXM浓度分别在0.25～200 mg/L和0.5～400 mg/L范围内线性良好; 萃取回收率分别为75.6%～88.1%和68.6%～77.4%; 批内与批间RSD均小于9%,方法回收率分别为94.6%～113.6%和91.9%～101.8%.尿样中SUL和CXM浓度分别在5～5120 mg/L和10～10240 mg/L范围内线性良好; 批内与批间RSD均小于3%; 方法回收率分别为92.9%～111.1%和98.0%～105.3%.本方法快速简便, 灵敏准确,可用于同时测定血浆及尿中SUL和CXM浓度,亦可用于药代动力学、生物利用度研究.     

化学发光酶免疫方法检测克伦特罗残留

建立了间接竞争化学发光酶免疫法(competitive indirect chemilumine scent enzyme immunoassy,ic-CLEIA)检测猪尿中残留克伦特罗(clenbuterol,CLB)的方法,并优化了竞争反应时间,Tween-20含量,PBS离子强度与pH4个参数。优化后的ic-CLEIA检出限可达0．01μg/L,检测范围为0．04～25．8μg/L,ED50为0．54μg/L;尿样检测的平均回收率为98％～120％,批内与批间相对标准偏差(variation of coefficent,CV)均小于10％。这些参数均优于目前广泛使用的ELISA方法,说明本CLEIA方法可替代ELISA法,用于实际尿样的检测。     

大鼠尿液与粪便中氟噻草胺的UPLC-MS/MS法测定

建立了一种测定大鼠尿液和粪便中氟噻草胺含量的超高效液相色谱-串联质谱(UPLC-MS/MS)分析方法。粪便与尿液样品采用乙腈提取;液相色谱分离采用Phenomenex反向C18色谱柱(50 mm×4. 6 mm,5μm),以0. 1%甲酸水和乙腈为流动相,流速0. 6 m L/min;质谱检测采用电喷雾离子源,正离子模式和多反应监测(MRM)方式进行扫描。结果表明:氟噻草胺在尿液(0. 10～10. 0 mg/L)和粪便(0. 25～50. 0 mg/L)中线性关系良好,相关系数r 0. 99,尿液和粪便中氟噻草胺的定量下限分别为0. 10 mg/L和0. 25 mg/L;质控样品的日内与日间相对标准偏差不大于9. 9%。样品稳定性为93. 7%～108%,尿中平均提取回收率为97. 0%～98. 8%,基质效应为98. 8%～107%,均符合生物分析方法验证的要求。考察了大鼠单次灌胃给予氟噻草胺400 mg/kg后的排泄动力学,144 h内尿液与粪便的总累积排泄率为12. 62%,其中尿中的累积排泄率为1. 12%,粪便中的累积排泄率为10. 13%,表明氟噻草胺主要经粪便排泄。该法灵敏、专属、准确,可用于大鼠尿液、粪便中氟噻草胺浓度的测定。     

纳米磁流体用于三相中空纤维液相微萃取/HPLC法对尿液中尼古丁的快速测定

首次将磁流体粉末代替磁子,在磁场作用下,以三相中空纤维液相微萃取(three-phase hollow fiberbased liquid phase microextraction,TP-HF-LPME)动态模式进行样品前处理,高效液相色谱(HPLC)为检测手段,建立了一种快速、准确、环境友好的尿中尼古丁含量的测定方法.系统优化了TP-HF-LPME技术的有机溶剂、磁流体粉末加入量、搅拌速率和萃取时间等条件.方法的线性范围0.05～50 mg/L,相关系数0.999 8,检出限为3μg/L(S/N=3),相对标准偏差小于3%.该方法用于测定人体尿样中尼古丁,主动和 被动吸烟者尿中尼古丁含量分别为5.39 mg/L和2.08 mg/L,相对回收率95%～100%,富集倍数17.9倍.     

浊点萃取-高效液相色谱法同时测定人尿中的4种抗凝血鼠药

建立了基于浊点萃取同时测定人尿中杀鼠灵、杀鼠醚、溴敌隆和溴鼠灵的反相高效液相色谱法。以非离子表面活性剂TritonX-114为萃取剂,考察了色谱测定条件,优化了浊点萃取参数。5mL尿样中加入0.030gNaCl,30μL甲酸和5%(w/v)TritonX-114400μL,50℃萃取15min,4种目标物的萃取效率能达到85%以上。在选择的色谱条件下,以V(甲醇)∶V(1%甲酸)=90∶10为流动相,待测组分经反相C18色谱柱分离,于306nm波长处用二极管阵列检测器检测,10min可完成分析。4种鼠药中杀鼠醚在0.01～5.0mg/L,其余3种在0.02～5.0mg/L范围内呈良好的线性关系(r≥0.997);尿样的标准加入回收率为71.2%～114.5%;相对标准偏差小于7.0%。对于5mL尿液,富集相用50μL甲醇稀释,萃取浓缩因子可达到28倍。定量检出浓度分别为:杀鼠灵0.016mg/L,杀鼠醚0.001mg/L,溴敌隆0.005mg/L,溴鼠灵0.003mg/L。本法简便快速、灵敏可靠、绿色环保,能满足鼠药中毒应急检测的要求。     

毛细管电泳与电致化学发光结合用于人尿中甲磺酸培氟沙星的测定研究

基于甲磺酸培氟沙星(PM)对三联吡啶钌在铂电极上的电致化学发光信号有增敏作用,与毛细管电泳结合,建立了一种研究PM药代动力学的新方法.在最佳条件下,该药物在0.05 ～10 mg/L范围内峰高与PM浓度呈良好的线性关系,方法检出限为0.006 mg/L,峰高相对标准偏差为2.4%(n=6),尿样回收率在 93% ～112%之间.该方法用于人尿中PM的排泄速率测定,1.5 ～2 h内平均排泄速率最大,48 h内原型药物排泄率为13.5%.     

高效薄层法检测尿中氟硝西泮的代谢物7-氨基氟硝西泮

 报道了尿中氟硝西泮代谢物 7 氨基氟硝西泮 (7AFLZ)的高效薄层 (HPTLC)定性分析和半定量分析方法 ,分析物斑点用荧光胺进行荧光显现 ,灵敏度高 ,检测限 (LOD)为 5 μg/L ,测量限 (LOQ)为 15 μg/L。该方法可以检测人口服 1mg氟硝西泮后 48h内排泄尿中的 7AFLZ ,适于麻醉抢劫犯罪案件中的药物检测。     

灿烂绿共振瑞利散射法测定药物及尿样中利福平的含量

建立了测定利福平的共振瑞利散射法。在弱碱性Tris-盐酸缓冲介质中,灿烂绿与利福平反应生成的络合物使共振瑞利散射急剧增强并产生新的RRS光谱,在291 nm处产生最大散射峰,利福平的质量浓度在0.16～0.99 mg/L范围内与共振散射强度(△IRRS)呈线性关系,线性回归方程为△IRRS=-12.61+2620ρ,相关系数为0.9997,检出限(3Sb/K)为0.014 mg/L。方法可用于市售利福平药物及尿样中利福平含量的测定,结果满意。     

应用表面增强拉曼光谱技术快速检测尿样中的β-兴奋剂

应用表面增强拉曼光谱技术与化学计量法相结合分析克伦特罗、沙丁胺醇和莱克多巴胺3种β-兴奋剂的标准溶液.在取自10头猪的尿样中,分别添加5个不同浓度的莱克多巴胺(1～20 mg/L),采用快速的液液萃取法对样品进行前处理,再进行表面增强拉曼测试.结果表明,克伦特罗和沙丁胺醇标准溶液的最低检测浓度为2 μg/L,莱克多巴胺标准溶液的最低检测浓度为0.1 mg/L;通过偏最小二乘法建立模型进行定量分析,3种药物的实际值与预测值的相关系数(R2)为0.9134～0.9368;本方法可检测尿样中1 mg/L莱克多巴胺,经外部验证后模型的实际值与预测值的相关系数(R2)为0 881,相对分析误差(RPD)为2.83;分析尿液中的莱克多巴胺含量所需时间小于30 min,为快速检测莱克多巴胺提供新途径.     

异丙醇-硫酸铵-水体系浮选硫氰酸亚铜法间接测定硫普罗宁

研究表明:控制溶液pH4.0,在SCN-存在下,Cu(Ⅱ)被硫普罗宁分子中的巯基(SH)还原生成的Cu(Ⅰ)与SCN-形成CuSCN白色乳状沉淀。该沉淀能被浮选在异丙醇/水两相界面之间。通过测定溶液中剩余Cu(Ⅱ)的量,从而间接测定硫普罗宁的量。硫普罗宁测量的线性范围0.30～12.0mg/L;检出限为0.20mg/L。本方法可直接用于片剂、血清、尿样中硫普罗宁含量的测定,结果满意。     

重庆地区儿童尿碘重复测量数据混合线性模型研究

为研究8～10岁儿童白天时段尿碘含量变化规律,给统一尿样采集时间提供理论依据,采集重庆市区一所住宿小学的60名8～10岁儿童1 d的晨尿、10:00 am、12:30、4:00 pm时段的尿样,采用砷铈催化分光光度法检测了尿碘含量,用混合线性模型(mixed linear models)分析了尿碘数据。结果表明,不同年龄、不同时间点的尿碘含量存在差异,10:00 am尿与晨尿尿碘值最为接近。提示应用10:00 am尿样通过乘以校正系数1.094,0.997和1.072的修正值代替晨尿可以使尿碘水平具有代表性和可行性,混合线性模型能够有效地处理重复测量数据。     

离子色谱安培法测定尿液中的胱氨酸

建立离子色谱安培法检测尿液中胱氨酸的分析方法。将尿液稀释处理后,过OnGuardⅡRP前处理柱,去除尿液中的有机物,选用低容量阳离子交换柱IonPac CS17进行分离,以甲烷磺酸梯度淋洗,柱后用300 mmol/L NaOH溶液衍生,离子色谱脉冲安培法检测,外标法定量。胱氨酸的质量浓度在0.1~5.0 mg/L范围内与其色谱峰面积呈良好的线性,线性相关系数r^2=0.999 9,检出限为0.05 mg/L。测定结果的相对标准偏差为0.71%(n=6),样品加标回收率为94%~108%。该方法具有操作简单,专属性强,灵敏度高等优点,可用于临床快速检测尿液中的胱氨酸。     

Rapid determination of lysine in biological samples by isocratic liquid chromatography.

A simple, rapid, and sensitive method was developed for detection and quantitation of lysine (Lys) in various biological samples by isocratic liquid chromatography (LC). Samples containing Lys and other amino acids were derivatized with 9-fluorenylmethyl chloroformate (FMOC-CI). The mobile phase used for isocratic elution was 50 mmol/L sodium acetate buffer (pH 4.20)-acetonitrile (43 + 57, v/v). Lys was detected with a UV detector at 265 nm. The derivatized Lys eluted from a LiChrospher 100 RP-18 (150 x 4.0 mm id) column at a retention time of 5.6 min. The limit of detection was 0.73 mumol/L (signal-to-noise [S/N] ratio, 3:1), and the limit of quantitation was 2.37 mumol/L (S/N ratio, 10:1). Lys recoveries from fortified biological samples were > 97.5%. Average Lys contents found in rumen fluid samples collected before the morning feeding and at 2.0, 4.0, and 6.0 h after feeding were 4.26, 3.34, 3.58, and 3.82 mumol/L, respectively. The hydrolysate of a sample of mixed rumen microorganisms collected before the morning feeding was determined to contain 1.372 mumol/mg microbial nitrogen in the form of Lys. The Lys concentrations of human plasma, goat plasma, human urine, and goat urine were 140.0, 102.0, 58.0, and 32.0 mumol/L, respectively.     

微波消解–火焰原子吸收光谱法测定红土镍矿中氧化镁含量

以盐酸、硝酸、氢氟酸溶解红土镍矿样品,加入高氯酸冒烟除去硅和氟,在稀盐酸和氯化锶溶液介质中,于原子吸收光谱仪波长285.2 nm处,使用空气–乙炔火焰,测定样品中氧化镁的含量。在最佳实验条件下,镁的质量浓度在0.20~1.00 mg/L与吸光度呈线性关系,其检出限为0.036 mg/L。该方法用于红土镍矿中氧化镁含量的测定,相对标准偏差小于1.2%,加标回收率为99.1%~100.3%。氧化镁质量分数在0.15%~5.00%范围内的重复性及再现性方程分别为r=0.023m+0.037和R=0.133m–0.028。该方法适合于测定氧化镁含量在0.15%~5.00%的红土镍矿。     

焙烧层状氢氧化镁铝对水中氟离子的吸附性能

研究了焙烧层状氢氧化镁铝(CLDH)对水中氟离子的吸附性能,考察了焙烧温度、吸附时间、吸附剂用量、溶液pH值等条件对吸附的影响.发现在较宽的pH(5.5～9.5)值范围内,CLDH对水中氟离子具有良好的吸附能力,室温下0.2gCLDH可将50mL浓度为15mg/L氟离子溶液处理为符合含氟标准的饮用水.吸附平衡符合Langmuir方程,在60min内达到饱和吸附,室温下饱和吸附量为22.64 mg/g.吸附饱和后的CLDH焙烧再生,循环使用5次后饱和吸附量为10.37 mg/g.     

Determination of fluoride in drinking water and in urine of adolescents living in three counties in Northern Chihuahua Mexico using a fluoride ion selective electrode

This study was carried out to determine fluoride in drinking water and in urine of adolescents, ages 15-20 years, living in Northern Chihuahua Mexico. Participants are from a cross-sectional study on health effects of chronic fluoride exposure from drinking water. A total of 201 participants (106 female and 95 male) in the study were recruited from three counties. Samples of drinking water of each county were collected and analyzed using the U.S. EPA Fluoride Ion-Selective Method. Statistically significant difference of fluoride content in water was found among the three counties of recruitment (Cd. Juarez; 0.3 mg/L, Samalayuca, 1.0 mg/L, and Villa Ahumada, 5.3 mg/L). Fluoride content in wells and tap water samples of Villa Ahumada ranged from 5.0 to 5.7 mg/L. Fluoride content of these samples was above the level permitted by Mexican regulations. The fluoride content in bottled water obtained from local stores in Villa Ahumada ranged from 0.3 to 3.7 mg/l.Fluoride in urine samples of each participant was also analyzed using the U.S. EPA Ion-Selective Method. The mean fluoride urine concentration (reported in mg/g creatinine) in adolescents living in these counties was 0.792±0.39, 1.33±0.67 and 2.22±1.16 (Cd. Juarez, Samalayuca and Villa Ahumada), respectively. The high fluoride urinary levels found in participants from Villa Ahumada may be associated to the high fluoride level (5.3 mg/L) in dinking water.The accuracy of measurements was assessed with reference materials in water and in urine. Mean fluoride recovery was 99.0% and 99.6% in water and in urine, respectively. The levels obtained were within the assayed 5% confidence levels.     

A critical evaluation of the graphite furnace conditions for the direct determination of chromium in urine

A critical study of the conditions for the direct determination of Cr in urine using GFAAS was carried out. A chemometric approach showed that the most important parameters that influence the efficiency of atomic Cr cloud formation were the atomization from the tube wall and the proper implementation of a pyrolysis step. The established procedure does not require any sample pre-treatment thus minimizing the risk of contamination. The Cr content in urine was determined using a calibration curve prepared with Cr spiked urine, easily correcting all potential matrix interferences. The heating program proposed took 68 s for a 30 μL urine sample. The procedure was applied for the determination of Cr in urine to investigate the absorption rate of Cr picolinate. The limit of detection and the characteristic mass for a 30 μL urine sample were 0.18 μg/L and 5.4 pg, respectively.     

A high-performance liquid chromatographic-tandem mass spectrometric method for the determination of ethyl glucuronide and ethyl sulfate in urine validated according to forensic guidelines

Ethyl glucuronide (EtG) and ethyl sulfate (EtS) are powerful markers for alcohol intake and abuse. Several analytical procedures for the quantification of EtG and EtG in serum and urine have been developed so far. Many of the published methods show limits of detections (LODs) or limits of quantifications (LOQs) for EtG in urine within the range of 0.1 mg/L or higher. Since this is the actual cutoff value for proving abstinence in Germany, problems may occur if urine samples are highly diluted. In this paper, the validation of a highly sensitive, fast and simple LC-MS-MS for the determination of EtG and EtS in urine is described. The calibration curves for EtG and EtS is linear over the whole range (0.025-2.0 mg/L). Very low detection limits can be achieved (LOD: EtG 0.005 mg/L, EtS 0.005 mg/L; and LOQ: EtG 0.019 mg/L, EtS 0.015 mg/L). All data for selectivity, precision and accuracy, recovery, as well as for the processed sample and the freeze/thaw stability, comply with the guidelines of the German Society of Toxicological and Forensic Chemistry. Strong matrix-related effects can be compensated for by using an internal standard. Finally, the applicability of the procedure is proven by analysis of 87 human urine samples and by successful participation in interlaboratory comparison tests.     

Simultaneous analysis of gamma-hydroxybutyric acid and its precursors in urine using liquid chromatography-tandem mass spectrometry

We have developed a rapid method that enables the simultaneous analysis of gamma-hydroxybutyrate (GHB) and its precursors, i.e. gamma-butyrolactone (GBL) and 1,4-butanediol (1,4-BD) in urine. The method comprised a simple dilution of the urine sample, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Chromatographic separation was achieved using an Atlantis dC18 column, eluted with a mixture of formic acid and methanol. The method was linear from 1-80 mg/L for GHB and 1,4-BD and from 1-50 mg/L for GBL. The limit of quantification was 1 mg/L for all analytes. The procedure, which has a total analysis time (including sample preparation) of less than 12 min, was fully validated and applied to the analysis of 182 authentic urine samples; the results were correlated with a previously published GC-MS procedure and revealed a low prevalence of GHB-positive samples. Since no commercial immunoassay is available for the routine screening of GHB, this simple and rapid method should prove useful to meet the current increased demand for the measurement of GHB and its precursors.     

Urine 2DE proteome analysis in healthy condition and kidney disease

Urine is a source of potential markers of disease. In the context of renal disease, urine is particularly important as it may directly reflect kidney injury. Current markers of renal dysfunction lack both optimal specificity and sensitivity, and improved technologies and approaches are needed. There is no clear consensus about the best sample pretreatment procedure for 2DE analysis of the urine proteome. Sample pretreatment conditions spots resolution and detection sensitivity, critically. As a first goal, we exhaustively compared eight different sample cleaning and protein purification methodologies for 2DE analysis of urine from healthy individuals. Oasis® HLB cartridges allowed the detection of the highest number of low molecular weight proteins; while PD10 desalting columns resulted in the highest number of detected spots in the high molecular weight area. Sample pretreatment strategies were also explored in the context of proteinuria, a clinical condition often associated to renal damage. Testing of urine samples from 13 patients with hypertension or kidney disease and different levels of proteinuria identified Oasis® HLB cartridge purification in combination with albumin depletion by ProteoPrep kit as the best option for urine proteome profiling from patients with proteinuric (> 30 mg/L albumin in urine) renal disease.