超临界流体萃取技术在中药生物碱提取中的应用

目的对超临界二氧化碳流体萃取中药中生物碱的应用进行综述。方法通过与传统的生物碱提取方法相比较 ,总结了超临界二氧化碳流体萃取生物碱的工艺特点及应用。结果与结论超临界流体萃取技术可以有效地提取中药中的生物碱 ,并具有产率高、方便快捷、后处理简单、无污染等优点     

HPLC-MS/MS测定延胡索中的12种农药残留

目的 建立液相色谱-串联质谱法测定延胡索药材中12种农药残留的方法。方法 延胡索样品用乙腈提取,过N-丙基乙二胺（PSA）柱净化,用ZORBAX Eclipse plus C18色谱柱分离,以电喷雾电离串联质谱在正离子多反应监测（MRM）模式下进行测定,内标法定量。结果 12种农药成分在相应的测定范围内线性关系良好,相关系数在0.997 7~0.999 9内,加样回收率范围为65.5%~117.8%,RSD范围为0.4%~4.5%,各农药的检出限为0.1~2.5 μg·kg-1。结论 本方法简便、灵敏、重现性好,可用于延胡索药材中12种农药残留的同时检测。延胡索药材农药残留量甚微,符合安全性要求。     

Simultaneous determination of desloratadine and its active metabolite 3-hydroxydesloratadine in human plasma by LC/MS/MS and its application to pharmacokinetics and bioequivalence

A rapid and simple liquid chromatographic-tandem mass spectrometric (LC/MS/MS) method was developed and validated for the simultaneous determination of desloratadine and its active metabolite 3-hydroxydesloratadine concentrations in human plasma. After liquid-liquid extraction with ethyl ether for sample preparation, the chromatographic separation was achieved on a CAPCELL PAK C18 column (50 mm x 2.0mm, 5 microm, Shiseido). [(2)H(4)]desloratadine and [(2)H(4)]3-OH desloratadine were used as internal standards. A mobile phase consisted of 5mM ammonium formate in water, methanol and acetonitrile (50:30:20). Detection was by positive ion electrospray tandem mass spectrometry on a Sciex API3000. A quadratic regression (weighted 1/concentration) gave the best fit for calibration curves over the concentration range 0.05-10 ng/mL for both desloratadine and 3-OH desloratadine. The method was shown to be accurate, rapid and sufficiently sensitive to be successfully applied to a pharmacokinetic and bioequivalent study.     

Antemortem and postmortem rodenticide analysis in forensic toxicology as a part of an LC-MS/MS-based multi-target screening strategy

Since rodenticides represent a substance group relevant in toxicological analyses, the aim of this work was the development of a complex multi-target screening strategy for the identification with liquid chromatography–tandem mass spectrometry. A simple protein precipitation was used as the sample preparation strategy. Further, a Luna 5 μm C18 (2) 100 Å, 150 × 2 mm analytical column was applied for the separation of relevant analytes with a Shimadzu HPLC. Signal detection was performed with a SCIEX API 5500 QTrap MS/MS system. The rodenticides investigated (α-chloralose, brodifacoum, bromadiolone, coumatetralyl, difenacoum, and warfarin) could be incorporated effectively into a multi-target screening strategy covering about 250 substances representing different groups with a limit of detection appropriate for substance identification. The strategy can easily be modified to perform semi-quantitative measurements for this substance group and could be supplemented by quantification based on standard addition.     

HPLC同时测定不同产地延胡索中的6种生物碱

目的 采用HPLC法同时测定延胡索药材中原阿片碱、盐酸黄连碱、盐酸巴马汀、盐酸小檗碱、脱氢紫堇碱和延胡索乙素等6种生物碱的含量,并比较不同产地、不同商品等级中6种生物碱含量的差异.方法 采用Durashell C18色谱柱(250mm ×4.6 mm,5μm),流动相为乙腈(A)-0.2％冰乙酸溶液(B,三乙胺调pH5.05),梯度洗脱,检测波长280 nm,流速1.0 mL·min-,柱温35℃;用上述方法对48批延胡索药材进行测定,并对结果进行聚类分析.结果 测定了延胡索药材中6种生物碱类成分的含量,在线性范围内的线性关系良好(r2＞ 0.9993),平均回收率为96.19％～97.59％,RSD为1.19％ ～2.81％;48批延胡索样品中6种生物碱均可检测到,不同产地的延胡索中6种生物碱的含量有明显的差异,部分达到显著性差异(P＜0.05).总体来说,不同商品等级延胡索中生物碱的含量无明显差异,二级延胡索质量优于一级;采用聚类分析能将不同产地的48批延胡索药材分为3类.结论 所用方法操作简单、准确、稳定、分离效果好,适用于同时测定延胡索药材中6种生物碱的含量;生物碱的含量受产地、环境因素影响较大,延胡索的品质表现出一定的地域及生境依赖性,商品等级对生物碱的含量无显著影响.     

Alkaloid profiling of the traditional Chinese medicine Rhizoma corydalis using high performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry

Since alkaloids are the major active constituents of Rhizoma corydalis (RC), a convenient and accurate analytical method is needed for their identification and characterization. Here we report a method to profile the alkaloids in RC based on liquid chromatography-tandem quadrupole time-of-flight mass spectrometry (LC–Q-TOF-MS/MS). A total of 16 alkaloids belonging to four different classes were identified by comparison with authentic standards. The fragmentation pathway of each class of alkaloid was clarified and their differences were elucidated. Furthermore, based on an analysis of fragmentation pathways and alkaloid profiling, a rapid and accurate method for the identification of unknown alkaloids in RC is proposed. The method could also be useful for the quality control of RC.KEY WORDS: LC–Q-TOF-MS/MS, Alkaloid, Fragmentation pathway, Rhizoma corydalis     

代谢组学分析技术平台和数据处理的新进展

分析技术和生物计量学促进了代谢组学的飞速发展。代谢组学快速、灵敏、可定量、非侵入性以及系统性的特点,使其在新药研发、药物毒性筛选、疾病诊断等领域显示出广阔的前景。本文综述了代谢组学研究中的某些关键问题:样品处理方法,分析技术和数据处理的方法和原则,代谢组动态变化、生物标记物的鉴定和代谢途径的检索近年来的进展。评价了各种分析手段的优缺点,并展望代谢组学发展前景。     

Comparison of Effect of Fasting and of Five Different Diets on the Bioavailability of Single Oral Dose of Amoxicillin 500 mg Capsule

The objective of this crossover bioavailability study on amoxicillin was to compare the bioavailability under fasting and five different diets, in twelve healthy adult male human volunteers. A single dose of amoxicillin 500 mg capsule was administered at six occasions: after overnight fasting, after two vegetarian diets (high-fat and low-fat), two non-vegetarian diets (high-fat and low-fat) and milk. Serial blood samples were collected up to 8 h after dose. Plasma concentrations were determined using a validated LC-MS/MS method. T_max increased and AUC and C_max values decreased in the presence of food. The non-vegetarian diets affected the rate of absorption of amoxicillin more than the vegetarian diets.     

高灵敏度LC/MS/MS法同时测定人血浆中麻黄碱和氯苯那敏

麻黄碱是存在于草麻黄和木贼麻黄等植物中的生物碱,属拟肾上腺素类药物,包括互为差向异构体的麻黄碱和伪麻黄碱两种活性不同的成分;氯苯那敏为丙胺类组胺H1-受体拮抗剂,作用较强,用量少,中枢抑制作用小,为常用抗过敏药,临床上可用于缓解各种感冒症状,与麻黄碱联合应用,用于缓解支气管哮喘与慢性喘息性支气管炎所致支气管痉挛.     

A simple extraction and LC‐MS/MS approach for the screening and identification of over 100 analytes in eight different matrices

In the present study, a liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) multi‐analyte approach using one single work‐up approach in whole blood, plasma, serum, post‐mortem blood, liver tissue, gastric content, hair, and urine was developed for fast target screening and reliable identification of 130 analytes often requested in clinical and forensic toxicology. Samples (500 μL each) of whole blood, plasma, serum, post‐mortem blood, tissue (homogenized 1 + 4 with water), as well as 3 g of distilled gastric contents, 1 mL of urine, or 20 mg of pulverized hair were extracted at different pH values with an diethyl ether‐ethyl acetate mixture (1:1). Separation and identification were performed using LC‐QTRAP with electrospray ionization in positive mode. For identification 1 scheduled multi‐reaction‐mode (sMRM) method with 390 transitions was developed covering benzodiazepines, Z‐drugs, antidepressants, neuroleptics, opioids, new synthetic drugs, and phosphodiesterase type 5 inhibitors. For positive sMRM transitions with intensities exceeding 5000 cps, dependent scans (EPI scan collision energy, 35 eV, collision energy spread, 15 eV) were performed for library search using our in‐house library. The method was developed with respect to selectivity, matrix effects, recovery, process efficiency, limit of detection, and applicability. The simple work‐up procedure was suitable for all biosamples with exception of urine in respect to low concentrated analytes, which showed median recovery values of 59%. The method was selective for 130 analytes in all 8 biosamples. For 106 analytes, the limit of detection in whole blood, plasma, and serum was lower than the lowest therapeutic concentration listed in blood level lists. Copyright © 2014 John Wiley & Sons, Ltd.     

高效液相色谱-串联质谱法同时测定中药材中的伏马毒素B_1和B_2

建立高效液相色谱-串联质谱法(HPLC-MS/MS)同时测定中药材中的伏马毒素B1和B2。样品用甲醇-水(80∶20)溶液提取,经免疫亲和柱净化后,以0.1%甲酸乙腈溶液-0.1%甲酸水溶液为流动相,通过C18色谱柱梯度洗脱,采用电喷雾离子源,正离子模式检测,内标法定量。伏马毒素B1和B2在0.5～500 ng.mL·1内线性关系良好。在4种不同基质5个加样水平的回收率实验中,伏马毒素B1的回收率为84.0%～96.1%,RSD为1.4%～7.6%;伏马毒素B2的回收率为86.3%～99.3%,RSD为1.2%～7.6%。自市售34种中药材样品中,选取10种中药材贮存于高温潮湿的条件中使其发霉,结果在上述发霉样品中有6个呈现阳性结果,其伏马毒素的总含量为82.4～2349μg.kg·1;在24种正常样品中有5种呈现阳性结果,其伏马毒素的总含量为102～729μg.kg·1。本文建立的分析方法灵敏可靠,适用于不同基质中药材中伏马毒素B1和B2的检测。     

中药及其活性成分与氯吡格雷抗血小板聚集的相互作用

氯吡格雷是临床常用的抗血小板聚集药物,临床上常与中药制剂联合使用。加强中药及其活性成分与氯吡格雷相互作用机制的研究,对提高抗血小板聚集作用,协同改善氯吡格雷抵抗提供依据。笔者综述了国内外相关文献中药及其活性成分与氯吡格雷相互作用的研究进展。细胞色素P450代谢酶、肝羧酸酯酶、P糖蛋白是中药及其活性成分与氯吡格雷发生相互作用的主要环节,也有些中药及其活性成分在药效学方面与氯吡格雷有协同抗血小板聚集作用。     

A simple method to monitor serum concentrations of fluoxetine and its major metabolite for pharmacokinetic studies

A rapid, selective and sensitive isocratic reversed-phase HPLC assay coupled with MS/MS detection for simultaneous quantification of fluoxetine and its major active metabolite in serum samples has been developed. Analytes were extracted with a simple three step liquid–liquid procedure and chromatographic separation was achieved on a C18 column.     

补肾壮阳类中药制剂中非法添加化学药物成分的检测

目的采用液质联用法检测补肾壮阳类中药制剂中非法添加的化学成分西地那非、他达拉非。方法色谱柱为AcquityBEHC18柱（2.1mm×50 mm,1.7μm）,以0.01 mol/L的醋酸铵溶液-乙腈梯度洗脱,流速0.3 mL/min,检测波长254 nm。结果在3批补肾壮阳类中药制剂中检测出西地那非或他达拉非。结论液质联用法灵敏准确,可有于补肾壮阳类中药制剂的质量检验。     

Multi-residue analysis using liquid chromatography tandem mass spectrometry for detection of 20 coccidiostats in poultry,livestock, and aquatic tissues

In this study, we developed a novel analysis method based on liquid chromatography/tandem mass spectrometry (LC–MS/MS) to allow the simultaneous identification of 20 coccidiostats in eight matrix categories, including the muscles of chicken, swine, cow, and fish as well as chicken eggs, bovine milk, and porcine viscera. In the pretreatment procedure, acetonitrile/methanol (95:5, v/v) containing 1% formic acid, 5 g of sodium acetate, and 6.0 g of anhydrous magnesium sulfate was used for extraction, followed by a clean-up procedure using n-hexane saturated with ACN to facilitate the elimination of analytes from high lipid samples. Chromatographic separations were achieved using a Poroshell 120SB C18 column and operated with a gradient mobile phase system consisting of methanol (with 0.1% formic acid) and 5 mM ammonium formate, and the MS detection was monitored simultaneously. The method was validated in accordance with the Guidelines for the Validation of Food Chemical Methods by the Taiwan Food and Drug Administration. The limit of quantitation among 8 matrices were 0.5–2 ng g⁻¹. The proposed method proved highly effective in detecting the presence of targeted veterinary drugs, providing a high degree of precision and accuracy over a broad range of matrices.     

LC-MS/MS同时测定朱砂中掺假染料猩红808和玫瑰红B的含量

目的 建立液相色谱-串联质谱(LC-MS/MS)法同时测定朱砂中的掺假染料猩红808和玫瑰红B的含量。方法 采用Agilent ZORBAX SB-C₁₈(2.1 mm×50 mm,1.8 μm)色谱柱,流动相为乙腈-0.1%甲酸溶液,流速为0.3 mL·min^-1,柱温为30 ℃,离子化方式为ESI,检测模式为MRM,选择离子对：①猩红808 m/z 368.1→275.1,m/z 368.1→219.1,m/z 368.1→77.1;②玫瑰红B m/z 443.2→399.2,m/z 443.2→355.0。结果 该方法猩红808和玫瑰红B的线性范围为10~200 ng·mL^-1和9.69~193.8 ng·mL^-1;检测限分别为2 ng·mL^-1和1.9 ng·mL^-1;回收率(n=6)分别为97.86%和97.34%,RSD为0.70%和0.73%;精密度、稳定性和重复性的RSD均<4%。结论 该方法提取简单,测定方法定量准确,确证性极高、灵敏高,适用于朱砂中红色染料猩红808和玫瑰红B的含量测定,还可为其他红色中药的安全检测提供了借鉴依据。     

A simple and rapid ESI-LC-MS/MS method for simultaneous screening of doping agents in urine samples

Objective:

The use of performance enhancing substances is banned in sports by the World Anti-Doping Agency (WADA). Though most prohibited substances can be detected by GC/MS, inclusion of corticosteroids and designer drugs has made it essential to detect these critical doping agents on LC/MS/MS due to their better separation and detection.

Materials and Methods:

A common extraction procedure for the isolation of acidic, basic and neutral drugs from urine samples was developed. A total of 28 doping drugs were analyzed on API 3200 Triple quadrupole mass spectrometer using C18 column in atmospheric pressure electrospray ionization. The mobile phase composition was a mixture of 1% formic acid and acetonitrile with gradient time period.

Results:

The method developed was very sensitive for detection of 28 doping agents. The linearity was performed for each drug and the total recovery percentage ranged from 57 to 114. Limit of detection is found to be 0.5 ng/ml for carboxy finasteride and 1-5 ng/ml for other drugs. The method was successfully used to detect positive urine samples of 3-OH-stanozolol, methyl phenidate, mesocarb, clomiphene metabolite and carboxy finasteride.

Conclusion:

The method developed based on controlled pH extraction method and HPLC-mass spectrometry analysis allowed better identification and confirmation of glucocorticosteroids and a few other drugs in different categories. The validated method has been used successfully for testing of 1000 In-competition samples. The method helped in detection of chemically and pharmacologically different banned drugs in urine in a single short run at a minimum required performance limit set by WADA.     

Simultaneous determination of lithospermic acid B and its three metabolites by liquid chromatography/tandem mass spectrometry

A rapid and sensitive method based on liquid chromatography/tandem mass spectrometry (LC/MS/MS) was developed for the simultaneous determination of lithospermic acid B and its three main O-methylated metabolites in rat serum with silibinin as the internal standard. The calibration curves for LSB, and the three metabolites were linear over the ranges of 16–4096, and 8–2048 ng/ml, respectively, with coefficients of correlation >0.998. For LSB, the intra-assay coefficient of variance (CV) was less than 9.3% and the inter-assay CV was less than 8.9%. The inter-assay mean accuracy was between 92.8% and 104.7%. For the three metabolites, the intra-assay CV was less than 8.7% and the inter-assay CV was less than 9.9%. The inter-assay mean accuracy was between 92.5% and 107.9%. This quantitation method was successfully applied to a pharmacokinetic study of LSB in rats. Also, a total recovery of 5.2%was found in bile after oral administration.