【作者】 贾薇；

【导师】 钟大放；

【作者基本信息】 沈阳药科大学 ， 药物分析学， 2001， 硕士

【摘要】 液相色谱-质谱联用法是专属性好，灵敏度高，鉴别能力强的先进分析技术。目前广泛应用的质量分析器主要有离子阱及四极杆，两种分析器各具特点，离子阱质量分析器由于具有多级质谱分析功能，在定性检测上占有优势；而四极杆由于具有选择性好，灵敏度高，及精密准确的特点，多用于定量分析工作。本文分别应用这两种类型的液质联用仪，进行生物样品中四环素类药物的限量检测及金刚乙胺的定量测定及其代谢物的定性分析。 液相色谱-电喷雾离子阱串联质谱法检查牛奶中4种四环素类药物残留量 四环素类为广谱抗菌药，由于常被用于治疗动物感染性疾病或促进动物生长，可能存在动物性食品及牛奶中四环素类药物的残留问题。本章旨在建立一种用电喷雾离子化液相色谱-串联质谱仪限量检测牛奶中四环素类药物残留量的方法，并探讨美他环素的质谱裂解途径。本法用Band Blut C₁₈小柱固相提取浓缩样品，通过Hypersil BDS C₁₈色谱柱分离，以乙腈-水-甲酸（37.5∶62.5∶1.2，体积比）为流动相，应用ESI LC／MSⁿ测定牛奶中4种四环素药物的残留量。结果表明，四环素、土霉素、美他环素的浓度在0.1～5.0 μg／mL范围内，金霉素浓度在0.2～10.0 μg／mL范围内的标准曲线均呈线性；牛奶中四环素，土霉素及美他环素浓度在0.05 μg／mL，金霉素在0.1 μg／mL时可以准确可靠地被检出，可满足牛奶中四环素限量检测的需要。实验还发现四环素类的质谱裂解特点不同，当选择[M+H]⁺作为母离子进行二级质谱分析时，四环素、土霉素及金霉素产生[M+H-H₂O]⁺，而美他环素给出特征的[M+H-NH₃]⁺子离子。 拍上 液相色各四极杆串联质谱法测定人血浆中金刚乙胺浓度 金刚乙胺是一种用于治疗和预防A型病毒引起的流感的药物。本文首次建立了电喷雾液相色谱一串联质谱法测定人血浆中金刚乙胺浓度的方法，并用该法对两种盐酸金刚乙胺片剂的生物等效性实验的样品进行了分析。取受试者血浆样品，用液一液提取法处理后经液相色谱一串联质谱测定，所用色谱柱为 Kramasil C18，流动相为甲醇一水一甲酸（75：25＊，体积比）。在本实验条件下，血浆中金刚乙胺的检测限可达 2．0 ng／mL，在金刚乙胺血浆浓度 2．00－400．0 ng／mL范围内，典型的标准曲线相关系数为 0．998；血浆最低定量浓度为 2刀0 ng／mL。在盐酸金刚乙胺片剂的生物等效性实验中应用本法，取得了满意的结果。 液相色谱一电喷雾离子阶质谱法鉴定人尿中金刚乙肢代谢物 金刚乙胺在体内经多种途径代谢。本章用C．8小柱固相革取样品，采用 Dikml CIS色谱柱分离，以甲醇一水一甲酸为流动相，应用 ESILC／MS”检测了健康受试者尿中金刚乙胺的代谢物。实验结果表明，金刚乙胺在人体内可产生多种代谢产物，包括间一羟基化代谢物、2个对一羟基代谢物的差向异构体，以及它们与葡萄糖醛酸的结合物。金刚乙胺原形及羟基代谢物的二级质谱主要碎片均为其准分子离子脱去一个氨分子，产生［M＋H－NH3f，其羟基代谢物葡萄糖醛酸的全扫描二级质谱主要碎片为准分子离子上脱去一个葡萄糖醛酸，产生［M斗H－C6HSO6f，并且还可继续脱去一个氨分子，产生［M＋H一C。HSO。－NH。］。更多还原

【Abstract】 Liquid chromatography-mass spectrometry is a sensitive, specific and powerful method for many organic compounds. The mass analyzers at present are mainly ion trap detector and quadrupole detector. Both of them have their characteristics. Triple stage quadrupole mass spectrometer has good accuracy and precision, and increased both selectivity and sensitivity. With the function of MS", ion trap mass spectrometer has confidently power in qualitative analysis of compounds. In this thesis, the quantitative and qualitative analyses were carried out with these two LC/MS instrumentss.Determination of residues of four tetracyclines in milk by high-performance liquid chromatography-electrospray mass spectrometryThe tetracycline antibiotics are widely used in modern agricultural practice. Such a wide use may lead to residue problems in milk.The aim of the experiment was to develop a precise liquid chromatographic determination with high sensitivity and a reliable ESI mass spectrometric confirmation of tetracycline antibiotics in milk, and interpret the pathways of mass fragmentation of metacycline. The tetracyclines were extracted from milk and concentrated using solid-phase extraction with a Bond Blut C18 cartridges. HPLC separation was carried out on a Hypersil BDS C18 column with a mobile phase of acetonitrile-water-formic acid (37.5:62.5:1.2, V/V), and the tetracyclines were detected using a ESI ion trap LC/MS/MS. With this method, the calibration curves were linear from 0.05 μg/mL to1.0 μg/mL for tetracycline, oxytetracycline, metacycline, and from 0.1 ug/mL to 2.0 ug/mL for chlortetracycline. The analytes were detectable at levels as low as 0.05 ug/mL for tetracycline, oxyletracycline, metacycline, and 0.1μg/mL for chlortelracycline. This method achived the required sensitivity to detect the residues of tetracyclines in milk. It also been found that tetracyclines have different pathways of mass fragmentation. Tetracycline, oxytetracycline and chlortetracycline gave [M+H-H2O]+ as major product ions while metacycline gave [M+H-NH3]+ as major product ion, when [M+H]+ was selected as the precursor.Determination of plasma concentration of rimantadine by ESI LC/MS/MSRimantadine is a drug used in the treatment and prophylaxis of influenza caused by the type A strain of virus. A method was developed using liquid chromatography-mass spectrometry to determine the concentration of rimantadine in plasma sample. The analytes were separated by Kramasil C18 column after extracted by liquid-liquid extraction from plasma samples. The mobile phase used was methanol-water-formic acid (75:25:1, V/V). With this method, the calibration curves were linear from 2.0 ng/mL to 400 ng/mL of plasma sample (r = 0.9981) and the detection limit was 2.0 ng/mL of plasma concention. This method was used to assess the bioequivalence of rimantadine preparations.Identification of metabolites of rimantadine in urine by ESI LC/MS/MSRimantadine, which has an asymmetric center and is given as the racemate, is extensively metabolized in man to hydroxylated metabolites. In this study, the urinary metabolites generated by healthy volunteers who had taken rimantadine were assayed using ESI LC/MS/MS followed by solid phase extration and C18 column separation. Analysis of the experimental data showed that rimantadine undergoes extensive metabolism in humans. The metabolites include m-hydroxyrimantadine, epimers of two p-hydroxyrimantadine and their glucuronide conjugates. By MS/MS analysis, rimantadine and its unconjugated metabolites all losed an ammonia from their quasi-molecular ions and produced[M+H-NH3]+,and conjugated hydroxyrimantadine losed a glucuroniderest from its quasi-molecular ion and produced[M+H-C6H8O6]+.更多还原