用GC、GC/MS方法检测人尿中Selegiline及其代谢物

本文采用 GC、GC/MS技术对口服 Selegiline Hydrochloride,一种单胺氧化酶抑制剂的尿中代谢物进行分析测定 ,确认检出 1 0种代谢产物 ,其中三种为新检测到的羟基化代谢物。     

吲满速尿的GC／MS测定方法及其在尿中代谢的研究

本文研究了吲满速尿的ＧＣ／ＭＳ检测方法，比较了不同的衍生化方法和提取方法，优化了条件，并对吲满速尿的阳性尿的代谢进行探讨，建立了人尿中测定吲满速尿的实用方法。     

人尿中Carphedone的气相色谱-质谱分析

建立检测人尿中兴奋剂 Carphedone分析方法。 5 0 m g尿样流经 XAD-2填装小柱淋洗 ,用甲醇洗脱后收集洗脱液 ,经过缓冲和酶解后振荡、离心 ,加入衍生化试剂 N-甲基 -N-三甲硅烷基三氟乙酰胺 ( MSTFA)和三甲硅烷基咪唑 ( TMSI)进行 TMS硅烷基化。气相色谱 -质谱 ( GC/MS)分析结果表明 :可以检测到 5 0 h内的代谢产物 ,同时可以检测 Carphedone衍生化后的脱水反应产物。此方法适用于兴奋剂检测尿中Carphedone代谢物的分析     

高效液相色谱法制备大鼠体内5-甲氧色胺衍生物（5-MT）的代谢产物

建立了大鼠尿中5-甲氧色胺衍生物(5-MT)一硫酸化代谢产物的高效液相色谱分离与制备方法.大鼠单剂量按120mg/kg灌胃给药,收集0～8h尿样,将尿样品经过SPE(固相萃取)柱预处理后,用半制备型HPLC对预处理后样品中的代谢产物进行分离与制备.结果表明该方法操作简单,分离效果好,可用于5-MT硫酸化代谢产物的制备.     

尿中7-甲基异炔诺酮(Tibolone)代谢物的气相色谱-质谱研究

应用气相色谱-质谱(GC/MS)选择离子监测(SIM)方式探讨甾体类药物7-甲基异炔诺酮(Tibolone)的检测方法。根据Tibolone的代谢规律,对受试者的阳性尿和空白尿检测结果进行比较。研究发现:在阳性尿中检出△4双键异构体、3α羟基和3β羟基等三种代谢物,6位、16位的羟基化代谢物没有检出。测定了3α羟基代谢物的浓度-时间关系曲线,表明Tibolone口服后吸收迅速,9小时后即可达到峰值浓度。可为该类药物的检测提供科学依据。     

人体尿液中兴奋剂美散痛代谢物的GC／MS分析研究

本文用GC/MS方法,对人尿中美散痛代谢物进行研究,鉴定了美散痛的几个主要代谢物和它们的代谢模式。     

气相色谱-火焰离子化检测器检测血清中乙二醇及其代谢产物羟乙酸

建立了气相色谱-火焰离子化检测器（FID）检测血清中乙二醇（ethylene glycol,EG）及其代谢产物羟乙酸（glycolic acid,GA）的方法。通过优化样本预处理和衍生化方法,用气相色谱-火焰离子化检测器检测标准工作液和小鼠中毒模型血清中乙二醇及其代谢产物羟乙酸。结果表明,乙二醇与羟乙酸浓度分别在0.01～10μg/mL,0.01～5μg/mL范围内线性良好,其标准曲线的相关系数分别为0.9999与0.9988,相对标准偏差均小于5%,绝对回收率分别为91～106%和89～95%,最低检出限分别为0.005μg/mL和0.01μg/mL。本方法灵敏度高,简单快速,结果准确可靠,适用于法医学鉴定和临床检验中乙二醇与羟乙酸的检测。     

尿和血浆中劳拉西泮的气相色谱法研究

建立一种灵敏的检测人尿和血浆中劳拉西泮的气相色谱方法。酶水解的尿和血浆样品在pH =9用乙醚提取 ,提取物用MTBDMS衍生化 ,衍生物用HP - 5毛细柱进行色谱分离 ,用电子捕获检测器检测。尿和血浆中提取率分别为 84. 4 %和 81. 3% ,检测限分别为 4. 2ng/mL和 2. 4ng/mL。方法已用于口服 2mg劳拉西泮人的尿和血浆分析 ,分析结果表明本方法适合于麻醉抢劫案中药物分析。本方法已与非衍生化、TMS衍生化气相色谱分析方法比较 ,本方法是最灵敏的方法。     

尼克刹米及其代谢产物的气相色谱／质谱法检测

本文用气相色谱/质谱法(GC/MS)证明了尼克刹米在人尿中的主要代谢产物为烟酰胺。方法简便,用样量少。     

同位素示踪法测定~(125)I-NGF在小鼠体内的血药浓度变化

本文涉及应用Idogn法对神经生长因子(NGF)进行125I标记并利用同位素示踪与电泳分离相结合的方法测定125I-NGF在小鼠体内的血药浓度与时间变化关系。结果显示,静脉注射125I-NGF在小鼠体内的代谢规律符合二房室开放模型。分布相半衰期为0.13h,消除相半衰期为3.68h,125I-NGF在小鼠体内分布和消除均较快,清除率为0.125L/h/Kg,表观分布容积为0.697L/kg,曲线所围面积为16.01μg.h/L。     

UPLC/Q-TOFMS鉴定人参皂苷Rh2在大鼠体内的代谢产物

An ultra-high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF MS) was used for the identification of metabolites in rats after oral administration of ginsenoside Rh2. The plasma, bile, urine and feces samples were collected after single oral administration of 50 mg•kg^－1 Rh2 to rats. The samples were prepared by protein precipitation with acetonitrile. After comparison with the blank samples, identification of the metabolites and their structural elucidation were performed by investigating their accurate mass data, and product ion spectra obtained from positive and negative ion detection mode using MSE data collection function(where E represents collision energy). The results reveal that the major metabolic pathways of Rh2 include deglycosylation, oxygenation, desaturation and sulfate conjugation. Protopanaxadiol is detected in rat feces. Neither parent compound nor metabolites are found in rat urine. Glutathione adduct of Rh2 is found as one of the major metabolites in rat bile, and cysteine-adduct metabolites are detected in feces. These results are helpful for the understanding of Rh2 metabolism in rat.     

烟气暴露对大鼠代谢轮廓谱的影响

运用基于液相色谱-质谱联用技术的代谢组学方法研究了不同卷烟烟气暴露对大鼠内源小分子代谢物组的影响。分别建立了大鼠血清和尿液的代谢轮廓谱,分析了烟气暴露7天、14天和30天时对照组大鼠、普通卷烟暴露组大鼠及含有天然本草添加剂的某品牌卷烟暴露组大鼠的血浆和尿液样本,采用偏最小二乘判别分析（PLS-DA）对数据进行模式识别。结果表明,普通卷烟和天然本草添加卷烟均会影响大鼠整体代谢状态,干扰大鼠磷脂、能量代谢,并对其造成氧化损伤,但天然本草添加卷烟对大鼠的损伤程度低于普通卷烟。在烟气暴露30天时,一些重要标志物在各组相对含量的变化进一步证实了天然本草添加卷烟可降低烟气对大鼠整体代谢的影响,减轻烟气造成的损伤。因此,在烟草中加入天然本草添加剂可在一定程度上减少烟气对机体的伤害,改善体内因烟气干扰而紊乱的磷脂和能量代谢。     

吴茱萸致肝毒性部位在大鼠体内的多样性分析

利用超高效液相色谱-四极杆-飞行时间质谱（UPLC-Q-TOF MS）技术对吴茱萸肝毒性部位给药后大鼠胆汁、尿液、粪便中的原型成分及其代谢产物进行分析,共检测到23个原型成分和10个代谢产物。采用正离子MSE扫描模式采集质量范围m/z 50~1 200的数据,通过与UNIFI软件数据库比对,以及各成分的保留时间、精确的一级和二级质谱信息,结合化合物的裂解规律和已有文献报道,共鉴定出29个化学成分,包括23个原型成分和6个代谢产物。进一步由代谢产物逆向追踪原型成分,推测出可能的代谢途径。这些成分在体内的主要代谢途径有葡萄糖醛酸结合、硫酸酯化、甲酰基化、羟基化、去甲基化等。基本明确了吴茱萸肝毒性部位在大鼠胆汁、尿液和粪便中的主要存在形式及代谢途径,为揭示吴茱萸肝毒性物质基础提供了参考依据。     

HPLC-ESI-LTQ-Orbitrap分析芪归银方中黄酮成分体内代谢过程

为进一步明确芪归银方的药效物质基础,采用HPLC-ESI-LTQ-Orbitrap技术对大鼠灌胃芪归银方后血清中黄酮类活性成分进行表征,并通过鉴别其尿液及粪便中的代谢物对结果进行佐证。从含药血清中鉴定出8个黄酮类化合物,其中,芹菜素-6,8-二葡萄糖苷和芹菜素以原型入血,芒柄花素、木犀草素、毛蕊异黄酮、5,7,4′-三羟基黄酮醇-7-O-β-D-葡萄糖苷、木犀草素-7-O-葡萄糖苷、山奈酚-3-O-葡萄糖苷等6个化合物以Ⅱ相代谢产物入血;从尿液中鉴定出9个黄酮类成分,包括6个入血成分的相应代谢产物;从粪便中检出3个黄酮类化合物的原型形式,未检测到与入血成分相同的成分。实验结果显示,芪归银方中的黄酮类入血成分大部分是经肾脏代谢排出,这可为阐明芪归银方延缓耐药的药效物质基础提供依据。     

Mass spectrometric based approaches in urine metabolomics and biomarker discovery

Urine metabolomics has recently emerged as a prominent field for the discovery of non‐invasive biomarkers that can detect subtle metabolic discrepancies in response to a specific disease or therapeutic intervention. Urine, compared to other biofluids, is characterized by its ease of collection, richness in metabolites and its ability to reflect imbalances of all biochemical pathways within the body. Following urine collection for metabolomic analysis, samples must be immediately frozen to quench any biogenic and/or non‐biogenic chemical reactions. According to the aim of the experiment; sample preparation can vary from simple procedures such as filtration to more specific extraction protocols such as liquid‐liquid extraction. Due to the lack of comprehensive studies on urine metabolome stability, higher storage temperatures (i.e. 4°C) and repetitive freeze‐thaw cycles should be avoided. To date, among all analytical techniques, mass spectrometry (MS) provides the best sensitivity, selectivity and identification capabilities to analyze the majority of the metabolite composition in the urine. Combined with the qualitative and quantitative capabilities of MS, and due to the continuous improvements in its related technologies (i.e. ultra high‐performance liquid chromatography [UPLC] and hydrophilic interaction liquid chromatography [HILIC]), liquid chromatography (LC)‐MS is unequivocally the most utilized and the most informative analytical tool employed in urine metabolomics. Furthermore, differential isotope tagging techniques has provided a solution to ion suppression from urine matrix thus allowing for quantitative analysis. In addition to LC‐MS, other MS‐based technologies have been utilized in urine metabolomics. These include direct injection (infusion)‐MS, capillary electrophoresis‐MS and gas chromatography‐MS. In this article, the current progresses of different MS‐based techniques in exploring the urine metabolome as well as the recent findings in providing potentially diagnostic urinary biomarkers are discussed. © 2015 Wiley Periodicals, Inc. Mass Spec Rev 36:115–134, 2017.     

Mass spectrometric determination of insulins and their degradation products in sports drug testing

Insulins' anabolic and anti-catabolic properties have supposedly led to its misuse in sport. Hence, doping control assays were developed to allow the unequivocal identification of synthetic insulin analogs and metabolic products derived from human insulin and its artificial counterparts in urine and plasma specimens. Analyses were based on immunoaffinity purification and subsequent characterization of target analytes by top-down sequencing-based approaches, which were conducted with hybrid tandem mass spectrometers that consisted of either quadrupole-linear ion trap or linear ion trap-orbitrap analyzers. Diagnostic product ions and analytical strategies are presented and discussed in light of the need to unambiguously identify misused drugs in urine and plasma specimens for doping control.     

基于质谱技术的黄芩治疗2型糖尿病的粪便代谢组学研究

采用粪便代谢组学方法,运用超高效液相色谱-四极杆飞行时间质谱（UPLC-Q-TOF MS）技术研究了黄芩治疗2型糖尿病大鼠的作用机制。采用主成分分析（PCA）和正交偏最小二乘法判别分析方法 （OPLS-DA）对健康对照组、2型糖尿病模型组和黄芩治疗组的大鼠粪便中内源性代谢物进行分析,寻找黄芩治疗2型糖尿病大鼠的潜在生物标志物。结果表明,健康对照组、2型糖尿病模型组和黄芩治疗组的大鼠粪便代谢图谱有显著的区分;发现并鉴定了11种潜在的生物标志物。黄芩对2型糖尿病大鼠的鞘脂类代谢和脂肪酸代谢具有调节作用;对三羟基三甲基吲哚酮、白三烯E4、亮氨酰脯氨酸和雌二醇的含量具有调节作用;同时,大鼠体重和空腹血糖的变化趋势表明,黄芩具有改善糖尿病症状的作用。     

尿中双氢睾酮代谢物的质谱研究

本文采用GC/MS联用技术检测双氢睾酮阳性尿,利用选择离子方式并与空白尿对照,发现一个代谢物,经质谱推测为16-羟基双氢睾酮,此代谢物于用药后24小时内可被检测到。通过检测这个代谢物,可以有效地控制兴奋剂双氢睾酮的滥用。     

兴奋剂甾体类药物质谱检测方法的研究

本文介绍兴奋剂甾体类药物在人体内代谢的规律、质谱裂解规律及其ＧＣ／ＭＳＤ检测方法。运用ＧＣ／ＭＳＤ选择离子定量做为内源性甾体双氢睾酮和睾酮的检测方法。     

液相色谱-电喷雾离子阱质谱法分析大鼠尿样中的黄芩苷及其异构体

采用液相色谱 -电喷雾离子阱串联质谱 ( LC/ MSn)技术分析了大鼠分别灌胃和静脉注射给予黄芩苷后尿样中原形药及其异构体的化学结构以及其排泄情况。根据代谢物的多级质谱和紫外吸收光谱数据推测 ,两种给药途径均检测到原形药黄芩苷及其异构体黄芩素 6-O-葡萄糖苷酸。收集大鼠灌胃 ( 1 0 0 mg/ kg)和静脉注射 ( 1 5 mg/ kg)给予黄芩苷后 0～ 48h尿样 ,以槲皮素作为内标 ,经乙酸乙酯提取后 ,以甲醇 -水 -甲酸 ( V(甲醇 )∶ V(水 )∶ V(甲酸 ) =60∶ 40∶ 1 )混合液为流动相 ,用 Diamonsil C18柱进行分离 ,采用电喷雾离子阱质谱 ,以选择反应监测 ( SRM)方式检测尿中黄芩素的两种葡萄糖醛酸结合物。黄芩素 6-O-葡萄糖苷酸的形成 ,说明黄芩苷在胃肠道可水解成苷元形式重新与葡萄糖醛酸结合 ,然后被排出体外。大鼠灌胃给予黄芩苷后 ,黄芩苷和黄芩素 6-O-葡萄糖苷酸的尿样累积排泄量分别为 1 .49%和 0 .77% ;大鼠静脉注射给予黄芩苷后黄芩苷和黄芩素 6-O-葡萄糖苷酸的尿样累积排泄量分别为 3 0 .80 %和 0 .2 8% ;以药物原形和尿药浓度评价黄芩苷在大鼠体内的绝对生物利用度为 4.84%。