Extraction and determination of trace amounts of chlorpromazine in biological fluids using magnetic solid phase extraction followed by HPLC

A simple, rapid and sensitive method termed as magnetic solid phase extraction (MSPE) combined with high-performance liquid chromatography-ultraviolet detector (HPLC-UV) has been proposed for the determination of trace amounts of chlorpromazine (CPZ) in water, urine and plasma samples. The separation and determination was performed on a C18 column under the optimal chromatographic conditions. Several factors influencing the extraction efficiency of CPZ, such as pH, surfactant and adsorbent amounts, ionic strength, extraction time, sample volume and desorption conditions, were studied and optimized. Under the optimal MSPE conditions, the extraction percentage of CPZ was 74%, 27% and 16% in water, urine and plasma samples, respectively. The limits of detection (LODs) of the proposed approach were 0.1, 5.0 and 10 ng/mL in water, urine and plasma samples, respectively. The relative standard deviations (RSDs) based on five replicate determinations at 10 ng/mL level of CPZ was 1.2%. Good linear behaviors over the investigated concentration ranges (0.25–300 ng/mL) with good coefficient of determination, R²>0.9998, were obtained. Good spike recoveries with relative errors less than 9.0% were obtained when applying the proposed method to water, urine and plasma samples.     

尿液中12种全氟化合物的固相萃取-超高效液相色谱串联四极杆线性离子阱质谱分析方法研究

  目的  建立超高效液相色谱串联四极杆线性离子阱质谱同时测定尿液中12种全氟化合物（PFCs）的方法。  方法  尿样经2%甲酸调节pH后,用WAX柱萃取净化浓缩,收集洗脱液氮吹,10 mmol/L乙酸铵-甲醇（V_水∶V_甲=70∶30）复溶,离心取上清液进样分析。超高效液相色谱采用甲醇和10 mmol/L乙酸铵水溶液在C₁₈柱上梯度洗脱进行分离,质谱采用多反应监测,内标法定量。  结果  方法在线性范围内线性良好,检出限和定量限分别为0.032～6.5 ng/L和0.10～21 ng/L。12种PFCs的加标回收率为91.5%～114%,日内精密度和日间精密度分别为0.57%～16.0%和1.88%～20.1%。将建立的方法应用于某地小学生尿液的测定,可检出9种PFCs,其中全氟丁烷磺酸和全氟辛酸为当地学生尿样的主要组成部分。  结论  建立的方法可同时测定尿液中12种全氟化合物,方法结合固相萃取和同位素内标校正,得到了良好的灵敏度和准确度。     

Chemiluminescence determination of melamine with Luminol-K3Fe(CN)6 system

A sensitive chemiluminescence (CL) method was developed for determining melamine in urine and plasma samples based on the fact that melamine can remarkably enhance the chemiluminescence of Luminol-K₃ Fe(CN)₆ system in alkaline medium. The determination conditions were optimized. Under optimum conditions, the chemiluminescence intensity had a good linear relationship with melamine in the range of 9.0 × 10^?9 – 7.0 × 10^?6 g/mL with a correlation coefficient of 0.9992. The detection limits (3σ) were 3.54 ng/mL for urine sample and 6.58 ng/mL for plasma sample. The average recoveries of melamine were 102.6% for urine sample and 95.1% for plasma sample. Melamine in samples was extracted with liquid-liquid extraction procedures and the assay results coincided very well with that determined with flow injection chemiluminescence method. The method provides a reproducible and stable approach for sensitive detection and quantification of melamine in urine and plasma samples.     

A study on bisphenol A, nonylphenol, and octylphenol in human urine amples detected by SPE-UPLC-MS

Objective To establish a comprehensive analytical method based on SPE‐UPLC‐MS for the simultaneous determination of bisphenol A (BPA), nonylphenol (NP), and octylphenol (OP) in urine samples. Methods Sixty urine samples collected from healthy subjects were analyzed for BPA, NP, and OP concentrations. The samples were de‐conjugated by adding β‐glucuronidase and sulfatase. After the enzymatic treatment, the samples were subjected to the OASIS HLB column solid phase extraction cartridges so as to be cleaned an...     

A Study on Bisphenol A, Nonylphenol, and Octylphenol in Human Urine Smples Detected by SPE-UPLC-MS

Objective To establish a comprehensive analytical method based on SPE-UPLC-MS for the simultaneous determination of bisphenol A (BPA), nonylphenol (NP), and octylphenol (OP) in urine samples.Methods Sixty urine samples collected from healthy subjects were analyzed for BPA, NP, and OP concentrations. The samples were de-conjugated by adding β-glucuronidase and sulfatase. After the enzymatic treatment, the samples were subjected to the OASIS HLB column solid phase extraction cartridges so as to be cleaned and concentrated. The UPLC separation was performed on a Acquity UPLCTM BEH C18 column (2.1x100 mm, 1.7 μm) with a gradient elution system of methanol-water as the mobile phase. Triple-quadrupole mass spectrometry analyzer was used for the qualitative and quantitative analysis of UPLC-MS/MS system.Results The limit of detection of BPA, NP, and OP was 0.10, 0.10, and 0.15 ng/mL, respectively. The recoveries of BPA, NP and OP were 80.1%-108%, 81.3%-109%, and 81.5%-98.7%, respectively. Among the 60 urine samples, BPA was detected in 8 samples at the level of 0.297-32.7ng/mL, NP was detected in 29 samples at the level of 1.69-27.8 ng/mL, and OP was detected in 17 samples at the level of 0.407-11.1 ng/mL.Conclusion The method is simple with high sensitivity and selectivity, and is suitable for the determination of BPA, NP, and OP in urine. As shown by our analysis, BPA, NP, and OP appear to be prevalent in human urine. This is particularly true for NP. The results from our study is therefore valuable for future studies to assess the exposure to BPA, NP, and OP in the general population.     

毛细管气相色谱法测定头发中尼古丁及其代谢产物可的宁

目的建立毛细管气相色谱同时测定头发中尼古丁及其主要代谢产物可的宁的方法。方法头发样品经1.5mol/L NaOH溶液消解,二氯甲烷/甲醇(V/V,3∶1)溶剂萃取,氮气吹干,甲醇定容后直接进样。经SPB-5毛细管柱分离,火焰离子化检测器(FID)检测。结果在优化发样提取和色谱分析条件下,尼古丁和可的宁在4.3ng/mL~1mg/mL和10ng/mL~2mg/mL浓度范围内线性良好(r均为0.9996),方法检出限(S/N=3)分别为4.3ng/mL和10ng/mL。发样中尼古丁和可的宁的加标回收率分别为90.33%~113.1%和92.92%~117.4%,相对标准偏差为2.1%~7.0%和4.4%~8.9%。结论本方法灵敏度高,选择性好,操作简便、快速,易于推广,可用于控烟流行病调查研究批样的检测。     

固相萃取-高效液相色谱串联质谱测定人体尿液中芳香化合物代谢物的方法研究

  目的  建立固相萃取-高效液相色谱串联质谱（high performance liquid chromatographic tandem mass spectrometric, HPLC-MS/MS）同时测定尿液中8种芳香化合物代谢物（苯巯基尿酸、甲基马尿酸、N-乙酰基-S-苯基-L-半胱氨酸、苯乙醛酸、苯乙醇酸、对氨基酚、对硝基酚和1-羟基芘）的方法。  方法  取1 mL尿液,加入20 μL β-葡萄糖醛酸酶和1 mL乙酸铵缓冲溶液,在37 ℃孵箱中酶解20 h。酶解后混匀,取适量酶解液注入PrimeHLB 固相萃取柱净化,4 mL乙腈洗脱,洗脱液氮吹至干,用0.20 mL甲醇复溶后进样HPLC-MS/MS系统分析。高效液相色谱分离采用反相C₁₈色谱柱（2.1 mm×150 mm,3.5 μm）,流动相A为含体积分数0.1%甲酸的水,流动相B为甲醇,采用梯度洗脱,流速为0.2 mL/min。质谱采用正、负离子交替扫描方式,多反应监测（multiple reaction monitoring, MRM）模式检测,内标标准曲线法定量。  结果  8种代谢物在 1~100 ng/mL范围内线性良好,相关系数均大于0.995,测定的精密度（relative standard deviation, RSD）在0.05%～9.95%之间,8种代谢物的检出限在0.041~0.12 ng/mL范围内。将所建立的方法用于尿样分析,加标回收率为80.1%～114.0%。  结论  所建立的方法灵敏、快速、准确,适用于普通人群和职业人群的芳香化合物生物监测评估。     

RP-HPLC法测定人血浆和尿液中美罗培南浓度

目的 建立测定人血浆和尿液中美罗培南含量的RP-HPLC法.方法 血浆样品经乙腈沉淀蛋白,二氯甲烷2次提取去杂质,取水相进样;尿液稀释后直接进样.以甲硝唑为内标,色谱柱为Capcell Pak MG C18(4.6 mm× 150 mm.5μm)柱,流动相为乙腈-0.02 mol/L NaH2 PO4溶液(体积比1...     

分散液相微萃取联合气相色谱质谱检测水中11种有机磷农药和阿特拉津方法的建立

目的 建立分散液相微萃取气相色谱质谱测定水样中11种有机磷农药和阿特拉津的方法。方法 分别对气相色谱条件、气相质谱条件进行优化,以提高萃取效率,优选条件建立方法后检测自来水和河水水样中有机磷农药。取10 mL水样,将200 μL二甲苯和800μL甲醇分别作为萃取剂和分散剂并混合后,注入水样中乳化分散,离心后取1 μL上层有机相进样分析。气相色谱采用HP5毛细管色谱柱（30 m×0.25 mm,0.25 μm）程序升温分离,质谱采用EI离子源,选择离子监测模式定量。结果 待测农药和阿特拉津在2.0~50 ng/mL范围内线性良好。方法检出限为0.12～1.38 ng/mL,测定精密度为5.57%～9.85%。将该方法应用于自来水和河水的测定,加标回收率为75.5%~107%。结论 方法简单快速,萃取和浓缩同时进行,轻质萃取剂在萃取后更易取出,适用于水样中有机磷农药的分析。     

巯基化衍生结合分散液相微萃取-气相色谱/串联质谱-选择反应监测技术测定尿样中超痕量氯乙烯基胂酸

目的：建立路易氏剂染毒尿样中超痕量氯乙烯基胂酸（CVAOA）的分析检测方法。方法采用正交实验,优化分散液相微萃取（DLPME）条件,优化结果为：将250μl甲醇（分散溶剂）、250μl乙酸乙酯（萃取溶剂）、3,4-二巯基甲苯（DMT）（衍生试剂,用量满足摩尔比CVAOA∶DMT=1∶100）混合液,加入到pH=1的1 ml尿样中,90℃衍生并萃取60 min,使用气相色谱/串联质谱-选择反应监测模式〔GC/MS/MS（SRM）〕对衍生产物进行定性定量分析。结果本法在50 pg/ml~1μg/ml浓度范围内呈现良好的线性（r2=0.9999）,相对标准偏差（RSD）＜10%,检出限可达18 pg/ml,定量限为56 pg/ml,远低于已报道的DLPME分析方法。对低、中、高3种浓度（0.5、5和50 ng/ml）的路易氏剂染毒尿样进行检测,准确度为98.2%~104%,RSD为6.9%~8.9%。结论本文所建方法灵敏度高,专属性好,准确度和精密度良好,且操作简单,适用于真实染毒样品分析。     

Cathodic adsorptive stripping voltammetry of an anti-emetic agent Granisetron in pharmaceutical formulation and biological matrix

Granisetron showed one well-defined reduction peak at Hanging Mercury Drop Electrode (HMDE) in the potential range from ?1.3 to ?1.5 V due to reduction of C=N bond. Solid-phase extraction technique was employed for extraction of Granisetron from spiked human plasma. Granisetron showed peak current enhancement of 4.45% at square-wave voltammetry and 5.33% at cyclic voltammetry as compared with the non stripping techniques. The proposed voltammetric method allowed quantification of Granisetron in pharmaceutical formulation over the target concentration range of 50–200 ng/mL with detection limit 13.63 ng/mL, whereas in human plasma 50–225 ng/mL with detection limit 11.75 ng/mL.     

LC-MS/MS法检测血液和尿液中东莨菪碱和阿托品

  目的  建立曼陀罗中毒人体血液和尿液中东莨菪碱和阿托品的LC-MS/MS检测方法。  方法  采用Shim-pack GISR-HP C18色谱柱,以乙腈-甲酸（0.1%）水溶液为流动相进行梯度洗脱,电喷雾离子源正离子多反应监测模式质谱分析,采用2个特征离子对和外标标准曲线法对曼陀罗中毒人体血液和尿液中东莨菪碱和阿托品进行定性定量分析。  结果  东莨菪碱和阿托品的定量离子对分别为m/z 304.25 [M+H]+→138.15和m/z 290.2 [M+H]+→124.15。东莨菪碱和阿托品在质量浓度1～100 ng/mL范围内线性良好（R≥0.999）,检出限（LOD）分别为:血液0.45 ng/mL和0.19 ng/mL,尿液0.38 ng/mL和0.16 ng/mL;定量限（LOQ）分别为:血液1.01 ng/mL和0.63 ng/mL,尿液1.07 ng/mL和0.38 ng/mL;提取回收率分别为:血液94.38%和94.49%,尿液94.96%和96.52%;基质效应为-3.51%～0.84%。2种目标物的准确度为89.38%～102.86%,日内和日间精密度均小于6%。  结论  建立的LC-MS/MS检测方法能快速、准确的测定生物检材中东莨菪碱和阿托品的含量。     

HPLC-MS/MS同时检测大鼠血浆中阿托伐他汀和伏立康唑的浓度

目的建立并应用同时测定大鼠血浆中阿托伐他汀和伏立康唑浓度的液相质谱联用方法（HPLC-MS/MS）,研究阿托伐他 汀单独给药及与伏立康唑联合给药后,大鼠体内阿托伐他汀的药动学变化以及伏立康唑的浓度变化。方法采用醋酸钠酸化, 甲基叔丁基醚液-液萃取法处理血浆样品,经Thermo Hypersil Gold C1（8 2.1×100 mm, 1.9 μm）色谱柱分离,流动相为乙腈-0.1%甲 酸水溶液,梯度洗脱,分析时间6 min;质谱检测采用加热电喷雾离子源（H-ESI）正离子扫描方式,选择反应监测（SRM）模式检 测,选用m/z559.2→440.2（阿托伐他汀）、m/z350.2→281（伏立康唑）、m/z370.2→252（内标兰索拉唑）作为定量分析的离子。结 果阿托伐他汀在0.01~100 ng/mL（r=0.9957）,伏立康唑在0.025~100 ng/mL（r=0.9966）范围内线性关系良好,阿托伐他汀和伏 立康唑的日间和日内精密度均小于13%,提取回收率66.50%~82.67%,血浆样品稳定性符合测定要求。单独给药和联合给药后 大鼠血浆中阿托伐他汀的AUC0-24 h分别为（438.78±139.61）和（927.43±204.12)）h·μg·L-1,CLz/F分别为（23.89±8.14）和（10.43± 2.58）L·h-1·kg-1,Cmax分别为（149.62±131.10）和（159.37±36.83）μg·L-1,t1/2分别为（5.08±1.63）和（5.58±2.11）h,Tmax分别为（0.37± 0.14）和（3.60±1.52）h,其中AUC0-24 h和CLz/F,Tmax具有显著性差异（P<0.05）。同时可测定合并给药组中伏立康唑的血药浓度。 结论本方法简单快速,灵敏度高,可用于同时检测大鼠血浆中阿托伐他汀和伏立康唑的浓度。伏立康唑和阿托伐他汀联用,使 阿托伐他汀的部分药代动力学参数发生了改变。     

塞曼原子吸收汞分析仪测定尿汞的方法学评价

目的对塞曼原子吸收汞分析仪测定尿汞的方法进行评价,为实验检测及临床诊断中选用简便、快捷的测汞方法提供参考。方法采用塞曼原子吸收光谱法(ZAAS)和氢化物发生-原子吸收光谱法(HG-AAS)进行尿汞含量测定,对两种方法的检出限、准确度、精密度及一致性进行比较。结果 ZAAS在0~1000 ng/m L范围内呈良好的线性关系(R2=1.0000),HG-AAS在0~20 ng/m L范围内呈良好的线性关系(R2=0.9990)。ZAAS和HG-AAS检出限分别为0.156 ng/m L、1.593 ng/m L,ZAAS灵敏度较高。ZAAS加标回收率在97.5%~103.2%之间,HG-AAS加标回收率在95.6%~104.5%之间;重复测量10 ng/m L、100 ng/m L汞标准溶液10次,ZAAS的相对标准偏差(RSD)分别为0.30%和0.36%,HG-AAS的RSD分别为2.82%和1.11%,两种方法准确度和精密度均满足GBZ/T 210.5-2008标准的要求,ZAAS精密度较高。用两种方法对30份尿液样本进行检测,经配对t检验,差异无显著性(P0.05),两种方法检测结果有较高一致性(R2=0.9961)。结论 ZAAS能够快捷、准确测量尿汞含量,检出限较低、精密度较高。     

柱前衍生化HPLC法测定注射用氨磷汀的含量

 目的 建立一种测定注射用氨磷汀含量的HPLC法。方法 氨磷汀经氯甲酸芴甲酯(9 fluorenylmethyl chloroformate,FMOC Cl)衍生化后用HPLC法分析。采用Agilent C18 柱(150 mm×4.6 mm,5 μm) 色谱柱,流动相A:乙腈,B:水。0~4 min,A∶B=30∶70;4.1~10 min,A∶B=98∶2,每次进样前再平衡7 min;柱温25℃,流速1 mL/min,检测波长265 nm。外标法计算含量。结果 在该色谱条件下,氨磷汀在5~100 μg/mL 浓度范围内有良好的线性关系(r>0.999,n= 6),回收率为97%～103%(n=5),RSD<2.0% (n=5)。最低检测质量浓度为100 ng/mL。样品在6 h 内稳定。测得样品含量合格,在90%～110%之间。结论 经方法学实验验证,该方法简便、快速、结果准确、可靠、重现性好。     

5—单硝酸异山梨酯血浆样品的GC—ECD测定

以硝酸异山梨酯为内标，乙酸乙酯为萃取溶剂，建立了５－单硝酸异山梨酯血浆样品经液－液萃取分离后，毛细管气相色谱电子捕获检测的灵敏、准确的定量分析方法。对血浆中５－单硝酸异山梨酯测定的最低检出量为２ｎｇ／ｍｌ，血药浓度在１０～６００ｎｇ／ｍｌ范围内色谱响应线性良好。并用于５－单硝酸异山梨酯药动学的研究。     

HPLC/MS/MS同时测定血浆肾上腺素、去甲肾上腺素、可的松和氢化可的松的方法建立

目的 建立高效液相色谱-质谱-质谱联用 （HPLC/MS/MS）同时测定血浆中的肾上腺素、去甲肾上腺素、可的松和氢化可的松的方法。方法 样品经乙腈沉淀蛋白和萃取后,15 000 r/min 离心5 min,取上清液进样分析。电喷雾离子源 (ESI), 正离子检测, 多反应监测 (MRM) 方式定量分析。 结果 肾上腺素、去甲肾上腺素、可的松和氢化可的松在0.02～200.00 ng/mL浓度范围内线性良好 （相关系数均≥0.999）,检出限分别为4.13 pg/mL、4.64 pg/mL、4.29 pg/mL和4.52 pg/mL,日内和日间精密度分别为1.19%~5.42%和2.16%~6.04%;将所建立的方法用于空白人血浆样品分析,加标回收率为80.0%~109.0%,样品测定精密度为3.93%~7.57%。结论 本研究所建立的方法快速灵敏,适合血浆样品的批量分析。     

HPLC法对不同蒲地蓝消炎制剂中绿原酸和咖啡酸含量的测定

目的建立高效液相色谱(HPLC)法同时测定蒲地蓝消炎片、蒲地蓝消炎胶囊和蒲地蓝消炎口服液中绿原酸和咖啡酸的含量。方法以75%甲醇为溶剂,采用超声法提取上述3种不同蒲地蓝剂型中绿原酸和咖啡酸;使用高效液相色谱法测定其含量,色谱柱为waters SunfireTMC18(5μm,4.6×150 mm),流动相为乙腈-0.4%磷酸水溶液(9∶91),流速为1.0 mL.min-1,检测波长为327 nm,柱温为室温。结果绿原酸的检测浓度在14.7～3750.0 ng.mL-1范围内与峰面积积分值呈良好线性关系(r2=0.9974),平均回收率为101.6%,RSD=1.3%;咖啡酸在68.4～4376.0 ng.mL-1范围内与峰面积积分值呈良好线性关系(r2=0.9984),平均回收率为101.1%,RSD=1.3%。结论本法灵敏、简便、准确,可用于不同剂型蒲地蓝消炎制剂中绿原酸和咖啡酸含量测定和质量控制;不同剂型蒲地蓝消炎制剂中绿原酸和咖啡酸含量不一致,应该尽快建立其质量标准。     

人血浆中阿戈美拉汀浓度的测定及其药代动力学

建立测定人血浆中阿戈美拉汀浓度的LC-MS/MS方法,测定单剂量口服25 mg阿戈美拉汀片后其在中国健康受试者体内的血药浓度,利用Phoenix WinNonlin 6.3.0软件计算药代动力学参数。该方法中阿戈美拉汀浓度的线性范围为0.051 3~10.38 ng/mL(r=0.999 4),最低检测浓度为0.051 3 ng/mL。日内、日间精密度测定RSD均小于15%。单次口服给药后,阿戈美拉汀的c_max为(13.25±15.78)ng/mL,t_max为(0.68±0.41)h,AUC_last为(14.00±19.26)ng/mL·h,AUC_inf为(14.14±19.35)ng/mL·h,t_1/2为(0.78±0.44)h。本方法简便、准确、重现性好,可用于阿戈美拉汀血药浓度的测定及其人体药代动力学研究。     

LC-MS/MS法测定大鼠血浆中芹黄春浓度及其在药代动力学研究中的应用

目的：研究大鼠分别灌胃和静脉给药芹黄春（芹菜素-7-O-β-D吡喃葡萄糖苷）溶液后,血浆中的芹黄春药物浓度及其药代动力学性质。方法6只雌性Sprague Dawley大鼠随机分为两组,每组3只,分别经灌胃(10 mg/kg)和静脉注射(2 mg/kg)给予芹黄春溶液后,按设计的时间点从大鼠眼内眦静脉丛采集血液样品,置于肝素化的离心管中,低温离心得血浆。采用甲醇沉淀法处理血浆生物样品,利用LC-MS/MS方法对血浆生物样品进行分析。药代动力学参数由Kinetica 2000软件进行计算。结果芹黄春浓度在1~2000 ng/mL范围内线性关系良好(r=0.9974),定量下限为1 ng/mL,低(8 ng/mL)、中(100 ng/mL)、高(1200 ng/mL)3个浓度的提取回收率均大于90%,日内日间精密度和稳定性的RSD均小于12.8%,日内日间准确度在91.5%~107%,方法学考察均符合要求。大鼠经静脉注射和灌胃芹黄春后,Cmax分别为(2363±96) ng/mL和(13.3±5.8) ng/mL,AUC0~∞分别为(796±201) h· ng/mL和(28.9±9.2) h· ng/mL,大鼠经静脉注射和后t1/2为(1.20±0.17) h,芹黄春的口服生物利用度约为0.6%。结论所建立的LC-MS/MS分析方法方便、快速,灵敏度高,准确度好,可用于大鼠血浆芹黄春浓度测定及其药代动力学的研究。