高效液相色谱/串联质谱法测定肿节风分散片中异嗪皮啶含量

目的: 建立高效液相色谱/串联质谱法测定肿节风分散片中异嗪皮啶。 方法: 采用Agilent SB-C₁₈色谱柱(2.1 mm×50 mm,1.8 μm),流动相甲醇-水(35:65),流速0.2 mL·min^-1。三重四级杆质谱检测器,正离子多反应监测模式(MRM),监测离子(m/z)222.9/189.7,222.9/161.9。 结果: 异嗪皮啶在0.48~2.40 mg·L^-1与峰面积具有良好的线性关系(r=0.999 2),平均加样回收率为99.3%,RSD 0.55% (n=9)。 结论: 方法简便易行,结果准确,重复性好,可用于肿节风分散片中异嗪皮啶的含量测定。     

阿扎他定血药浓度的测定及其药动学的研究

目的:应用LC-MS-MS法研究抗组胺药物马来酸阿扎他定片的人体药动学特征.方法:10名健康受试者分别口服马来酸阿扎他定片1mg和2mg,采用LC-MS-MS法测定给药后0～48h内人体血浆中阿扎他定的浓度,并计算其药动学参数.结果:阿扎他定质量浓度的线性范围为0.05～5.00ng/mL,最低定量下限为0.05ng/mL,主要药动学参数分别为tmax为(3.90±0.994)h,(3.70±1.059)h;Cmax为(0.795±0.187)ng/mL,(1.444±0.245)ng/mL;t1/2为(11.716±2.005)h,(10.62±2.06)h;AUC0～48为(11.213±4.042)ng·h/mL,(18.80±6.98)ng·h/mL。结论:本法操作简便、灵敏度高、干扰少、结果准确的优点,适用于阿扎他定药动学的研究。     

中药附子的化学成分

目的：体外评价黄连解毒汤主要有效成分对人肝细胞色素P450（CYP1A2,CYP2A6,CYP2B6,CYP2C9,CYP2D6,CYP3A4）活性的影响。方法：利用液质联用仪（LC-MS-MS）建立同时测定6种探针药物的分析方法,选择合适的孵育体系、孵育时间、蛋白浓度建立Cocktail模型,并通过工具药进行验证。利用Cocktail模型筛选黄芩苷、汉黄芩苷、黄芩素、汉黄芩素、小檗碱、药根碱、黄连碱、表小檗碱、巴马汀、兰花碱、栀子苷对6种酶活性抑制的体外评价。结果：黄芩素、汉黄芩素及黄芩苷对CYP1A2的IC₅₀分别为37.7,46.3,49.9 μmol·L^-1,小檗碱、药根碱、黄连碱、表小檗碱对CYP2D6抑制作用的IC₅₀分别为39.7,46.5,7.3,11.5 μmol·L^-1。结论：黄连解毒汤中黄芩及黄连的主要有效成分对细胞色素P450酶有不同程度的抑制,在临床联合用药时要注意代谢层面的相互作用,减少不良反应的发生。     

牛奶和奶粉中八种镇定剂残留量的液相色谱-串联质谱测定

采用液相色谱-串联质谱仪,建立了牛奶和奶粉中乙酰丙嗪cetopromaizine、氯丙嗪chlor-promazin、氟哌啶醇haloperidol、丙酰二甲氨基丙吩噻嗪propionylpromazine、甲苯噻嗪xylazine、阿扎哌隆azaperone及其代谢物阿扎哌醇azaperol和咔唑心安carazolol残留量的液相色谱-串联质谱测定方法。结果表明八种镇定剂在牛奶中添加水平0.10～0.50μg/kg到0.80～4.00μg/kg,其回收率在86.29%～92.49%之间,相对标准偏差在8.99%～20.56%之间;在奶粉中添加水平0.80～2.00μg/kg到2.0～8.00μg/kg,其回收率在81.59%～97.49%之间,相对标准偏差在1.76%～16.70%之间。八种镇定剂在确定的添加范围内线性关系良好。八种镇定剂在牛奶中的方法检出限为0.1～0.5μg/kg,在奶粉中的方法检出限为0.8～4.0μg/kg。     

盐酸阿夫唑嗪缓释片的人体生物等效性研究

目的 考察自制盐酸阿夫唑嗪缓释片及其市售制剂在健康人体的相对生物利用度及生物等效性.方法 采用双周期自身随机交叉试验设计.20名健康男性志愿者口服受试制剂或参比制剂,血药浓度采用液相色谱一串联质谱测定.结果 受试制剂及参比制剂的主要药动学参数为:Cmax分别为(16.8±5.9)和(17.5±5.7)ng/ml;Tma...     

LC-MS/MS同时测定犬干血浆样品中栀子苷、山栀苷及京尼平龙胆双糖苷的浓度

目的：建立LC-MS/MS法同时测定犬干血浆样品中栀子苷、山栀苷及京尼平龙胆双糖苷浓度的分析方法,并应用于犬药代动力学研究中。方法：采用Ecosil C18 （4.6 mm×150 mm,5 μm）色谱柱,流动相为甲醇-20 mmol/L 甲酸铵（0.1%甲酸）,梯度洗脱;使用电喷雾离子源（ESI）,正离子多反应监测模式检测。应用干血浆采样技术收集犬静脉输注栀子提取液后的血浆样品,超声提取后进行LC-MS/MS分析。结果： 3种环烯醚萜苷类成分在线性范围内的相关性良好（r〉0.99）,日内、日间变异均〈15%,绝对回收率在 79.9%~91.4%之间,4 ℃ 放置和室温放置 24 h、75 d 稳定性良好。药代动力学参数显示栀子苷、山栀苷、京尼平龙胆双糖苷的t1/2分别为 6.86、0.687、2.11 h,AUC0→t分别为158、9.16、34.0 μg·mL^-1·h。结论：建立的同时检测干血浆样品中栀子苷、山栀苷、京尼平龙胆双糖苷浓度的液相色谱-串联质谱分析法,灵敏度好、专属性强。干血浆样品室温放置稳定性好,易于储存运输,适用于药代动力学研究中。     

Identification of Pancreatic Cancer-Associated Tumor Antigen from HSP-Enriched Tumor Lysate-Pulsed Human Dendritic Cells

Purpose

Vaccine strategies utilizing dendritic cells (DCs) to elicit anti-tumor immunity are the subject of intense research. Although we have shown that DCs pulsed with heat-treated tumor lysate (HTL) induced more potent anti-tumor immunity than DCs pulsed with conventional tumor lysate (TL), the underlying molecular mechanism is unclear. In order to explore the molecular basis of this approach and to identify potential antigenic peptides from pancreatic cancer, we analyzed and compared the major histocompatibility complex (MHC) ligands derived from TL- and HTL-pulsed dendritic cells by mass spectrophotometry.

Materials and Methods

Human monocyte-derived dendritic cells were pulsed with TL or HTL prior to maturation induction. To delineate differences of MHC-bound peptide repertoire eluted from DCs pulsed with TL or HTL, nanoflow liquid chromatography-electrospray ionization-tandem mass spectrometry (nLC-ESI-MS-MS) was employed.

Results

HTL, but not TL, significantly induced DC function, assessed by phenotypic maturation, allostimulation capacity and IFN-γ secretion by stimulated allogeneic T cells. DCs pulsed with TL or HTL displayed pancreas or pancreatic cancer-related peptides in context of MHC class I and II molecules. Some of the identified peptides had not been previously reported as expressed in pancreatic cancer or cancer of other tissue types.

Conclusion

Our partial lists of MHC-associated peptides revealed the differences between peptide profiles eluted from HTL-and TL-loaded DCs, implying that induced heat shock proteins in HTL chaperone tumor-derived peptides enhanced their delivery to DCs and promoted cross-presentation by DC. These findings may aid in identifying novel tumor antigens or biomarkers and in designing future vaccination strategies.     

Conjugates of p-hydroxymethamphetamine and 4-hydroxy-3-methoxymethamphetamine in blood obtained from methamphetamine and 3,4-methylenedioxymethamphetamine users: analysis by LC-MS-MS

A detailed procedure for simultaneous analysis of methamphetamine (MA), 3,4-methylenedioxymethamphetamine (MDMA) and their metabolites including the conjugates in whole blood by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS-MS) has been established. The methods were well validated and applied to users’ blood specimens. The blood concentrations of MA users (n = 11) ranged from <1.0 to 131 ng/ml (from <6.1 to 794 nM) for p-hydroxymethamphetamine (p-OHMA), from <2.0 to 46.7 ng/ml (from <5.9 to 137 nM) for p-OHMA-glucuronide (p-OHMA-Glu), and from 3.8 to 495 ng/ml (from 15.5 to 2020 nM) for p-OHMA-sulfate (p-OHMA-Sul). The molar conjugation percentage [(p-OHMA-Sul + p-OHMA-Glu)/(p-OHMA + p-OHMA-Sul + p-OHMA-Glu) × 100%] for every specimen except one ranged from 64.9% to 100% [average ± standard deviation (SD): 85.0% ± 12.0%, n = 10]. The blood concentrations of MDMA users (n = 3) ranged from <1.0 to 84.2 ng/ml (from <5.1 to 432 nM) for 4-hydroxy-3-methoxymethamphetamine (HMMA), from 78.1 to 508 ng/ml (from 211 to 1370 nM) for HMMA-glucuronide (HMMA-Glu), and from 190 to 610 ng/ml (from 691 to 2220 nM) for HMMA-sulfate (HMMA-Sul). The molar conjugation percentage [(HMMA-Sul + HMMA Glu)/(HMMA + HMMA-Sul + HMMA-Glu) × 100%] ranged from 89.3% to 100% (average ± SD: 95.2% ± 5.4%, n = 3). The present study clearly showed the presence of conjugated forms of p-OHMA and HMMA in human blood; the levels of sulfate conjugates were much higher than those of glucuronide conjugates. The utility of the p-OHMA-Sul and HMMA-Sul remains to be explored.     

LC-MS-MS法测定犬体内血浆中埃索美拉唑药物浓度

目的 建立液相色谱联用质谱法(LC-MS-MS)测定犬体内血浆中埃索美拉唑(esomeprazole,EMZ)药物浓度。方法 以地西泮为内标,血浆采用蛋白沉淀处理,色谱柱为Shim-pack XR-ODS柱(2 mm×100 mm, 5 μm);流动相为乙腈:2 mmol/L乙酸铵(含0. 1%甲酸);流速0. 3 ml/min;质谱条件:电喷雾离子化电离源ESI,正离子多反应检测,检测离子分别为346.2→198.2 m/z(埃索美拉唑)和285.2→193.2 m/z(地西泮)。结果 埃索美拉唑在0.5~1 200 μg/L 检测浓度范围内呈良好线性关系(r>0.99),最低定量限(LLOQ)为0.5 μg/L,平均绝对回收率在(101.0±6.5)%,低、中、高3种浓度的日内、日间RSD≤15%。结论 该方法操作简便、灵敏、准确,适用于埃索美拉唑血药浓度测定及药动学研究。     

Facile and Rapid Determination of Contamination in Sulphur Pesticide Formulations by Liquid Chromatography–Tandem Mass Spectrometry

Up to 7% of pesticide products in the European market are illegal, counterfeit or of poor quality. As a result, regulatory authorities are required to determine a wide variety of impurities in many different pesticide formulations. A simple, rapid and exceptionally reliable LC-MS-MS method for the determination of carbendazim residual quantities in sulphur formulations, used in organic agriculture, has been developed and validated. Linearity of response was established (R ² = 0.9997) over a wide range of concentrations (0.01–2 μg/mL). Recovery ranged from 94% to 101%. LOD and LOQ were 0.003 μg/mL and 0.005 μg/mL, respectively, and lack of interference was confirmed.