Attaining in vivo selectivity of positive modulation of α3βγ2 GABAA receptors in rats: A hard task!

It is unclear whether GABA_A receptors (GABA_ARs) that contain the α3-subunit are substantially involved in the anxiolytic effects of benzodiazepines (BDZs). In the present study, we tested YT-III-31, a newer BDZ ligand with functional preference for α3βγ2 GABA_ARs, in two paradigms of unconditioned anxiety, the open field and elevated plus maze in rats. The effective dose of YT-III-31 (2?mg/kg) displayed a clear anxiolytic-like profile, unhampered by sedative action, in both tests. At a higher dose (10?mg/kg), YT-III-31 induced ataxia in the rotarod and sedation in spontaneous locomotor activity test. The latter effect was preventable by flumazenil and βCCt, the non-selective and α1βγ2 GABA_AR affinity-selective antagonist, respectively, demonstrating that sedative properties of YT-III-31, when attained, are mediated by the α1γ2 site. To elucidate the receptor substrate of subtle behavioral differences between YT-III-31 and diazepam, we approximated in vivo receptor potentiation for both ligands, based on estimated unbound concentrations in rat brains. Far different from diazepam, YT-III-31 has significantly lower affinity for the α1γ2 over other BDZ-sensitive sites, and at lower doses (1–2?mg/kg) was devoid of potentiation at α1βγ2 GABA_ARs. The approximation approach revealed a modest selectivity of YT-III-31 for α3γ2- in comparison to α2γ2 and α5γ2 binding sites, suggesting that its anxiolytic-like activity may not necessarily or predominantly reflect potentiation at α3βγ2 GABA_ARs. Nonetheless, as the anxiolytic effects are achievable at a dose devoid of any sedative potential, and having favorable safety (cytotoxicity) and metabolic stability profile, YT-III-31 represents a valuable candidate for further translational research.     

UPLC-MS/MS同时检测消癌平注射液中通关藤苷A、I、H的含量

目的 建立UPLC-MS/MS同时测定消癌平注射液中通关藤苷A、通关藤苷I、通关藤苷H的含量。方法 采用Phenomenex Kinetex XB-C₁₈色谱柱（2.1 mm×50 mm,2.6 μm）,以0.1%甲酸水-乙腈为流动相,梯度洗脱分离,流速为0.2 mL·min^-1,通过电喷雾离子源,多反应监测（MRM）,正离子模式。结果 通关藤苷A、I、H分别在0.05~10 ng·mL^-1,0.025~10 ng·mL^-1,0.025~10 ng·mL^-1浓度内呈良好的线性关系,r>0.998,检测限为0.012 5~0.025 ng·mL^-1,平均回收率分别为97.9%,95.7%,96.1%,RSD<3.4%。结论 方法简单快速,准确灵敏,可用于消癌平注射液中通关藤苷A、I、H的定量测定。     

UPLC-MS/MS检测大鼠体内达卢生坦的血药浓度及其药动学研究

目的 建立一种快速、高灵敏度和高选择性的UPLC-MS/MS用于大鼠血浆中达卢生坦的测定。方法 通过蛋白沉淀法完成血浆样品处理。通过Acquity UPLC BEH C₁₈柱（2.1 mm×50 mm,1.7 μm）色谱柱,流动相为0.1%甲酸水-乙腈,流速为0.4 mL·min^-1。采用ESI⁺方式检测,扫描方式为多反应离子监测方式检测。结果 达卢生坦血药浓度在0.01~2.5 μg·mL^-1内线性关系良好（R²>0.999 6）,定量下限（LLOQ）为0.01 μg·mL^-1。血浆中的达卢生坦平均回收率为87.613%~97.115%,日内及日间精密度RSD<7.71%,基质效应RSD<9.34%。结论 该方法灵敏高,快速高效,能成功应用于大鼠口服10.0 mg·kg^-1达卢生坦后的药动学研究。     

Quantitative Analysis of Cereulide Toxin from Bacillus cereus in Rice and Pasta Using Synthetic Cereulide Standard and 13C6-Cereulide Standard—A Short Validation Study

A single laboratory validation study of a rapid and sensitive quantitative method for the analysis of cereulide toxin produced by Bacillus cereus using ultra high performance liquid chromatography-electrospray-tandem mass spectrometry is presented. The analysis of this cyclic peptide toxin was validated for pasta and rice samples using a newly presented synthetic cereulide peptide standard, together with ¹³C₆-cereulide that previously have not been commercially available. The use of cereulide standard was also compared to the most frequently used surrogate standard, the antibiotic valinomycin. The performance of the method was evaluated by analyzing spiked sample pools from different types of rice and pasta, as well as 21 individual rice and pasta samples from differently prepared meals. Inoculation of samples with three cereulide toxin-producing strains of Bacillus cereus was finally used to mimic naturally contaminated foods. The quantification range of the method was 1–500 ng/g (R² = 0.999) and the limits of detection and quantification were 0.1 and 1 ng/g, respectively. The precision varied from 3% to 7% relative standard deviation and the trueness from −2% to +6% relative bias at different concentration levels in cooked rice and pasta.     

基于抵当汤减毒的钩吻生物碱体内外含量变化研究

目的 建立体内、体外钩吻生物碱成分测定方法,并测定抵当汤减毒前后钩吻生物碱成分含量,探讨抵当汤减缓钩吻毒性的可能作用机制.方法 采用UPLC-QDa体外检测钩吻与抵当汤混合前后6种生物碱成分变化,采用UPLC-MS/MS检测抵当汤联合钩吻灌胃对小鼠血浆中钩吻生物碱成分含量的影响.结果 与抵当汤混合后,钩吻中钩吻素子、钩...     

吴茱萸水提取物化学成分研究

目的:研究吴茱萸Euodia rutaecarpa果实水提取物的化学成分。方法:采用大孔吸附树脂,硅胶,小孔树脂凝胶(MCI),羟丙基葡聚糖凝胶(Sephadex LH-20),ODS柱色谱以及半制备高效液相色谱等方法进行分离纯化,根据理化性质和波谱数据鉴定化合物结构。结果:从吴茱萸果实的水提物中分离鉴定了11个化合物,包括5个酚酸及其苷,分别为绿原酸(1),新绿原酸(2),隐绿原酸(3),松柏苷(4),咖啡酸(5);4个黄酮及其苷,分别为槲皮素-3-O-α-D-吡喃阿拉伯糖苷(6),金丝桃苷(7),芦丁(8),槲皮素(9);1个柠檬苦素苷,为limonin diosphenol 17-β-D-glucopyranoside(10);1个生物碱,为去氢吴茱萸碱(11)。结论:化合物2~4,6为首次从该属植物中分离得到。通过二维核磁波谱确证,首次完整报道化合物10的氢谱数据。鉴于吴茱萸的基原有3个(吴茱萸、石虎和疏毛吴茱萸),通过对照品的超高效液相色谱-质谱联用(UPLC-MS)分析,对3种植物基原的吴茱萸药材进行成分确认,从石虎和疏毛吴茱萸中首次鉴定出绿原酸(1),新绿原酸(2)和隐绿原酸(3),从石虎中鉴定出金丝桃苷(7)。     

TranscutolP对痛宁凝胶中延胡索乙素在大鼠体内的药代动力学影响

[目的] 建立一种快速灵敏的超高效液相色谱-串联质谱（UPLC-MS/MS）方法测定大鼠血浆中延胡索乙素含量,研究含7%单乙基二甘醇油醚（TransP）的痛宁凝胶在大鼠体内的药代动力学过程,考察其透皮吸收促进效果。[方法] 采用血药浓度法,UPLC-MS/MS测定大鼠体内痛宁凝胶中延胡索乙素含量,同时与空白痛宁凝胶体内药代动力学参数进行比较。[结果] 7% Trans P组的T_max、C_max与空白对照组相比差异具有显著性（p<0.05）,药时曲线下面积AUC_{（0→24h）}和AUC_{（0→∞）}与空白对照组相比差异具有显著性（p<0.05）。[结论] 应用UPLC-MS/MS技术测定痛宁凝胶中延胡索乙素的血药浓度,药代动力学参数表明7%TransP组促渗效果明显优于空白对照组,可进一步应用于痛宁凝胶制剂中。     

超高效液相色谱-质谱联用法测定人血浆中依非韦伦的浓度

目的：建立超高效液相色谱-质谱联用法（UPLC-MS）定量测定人血浆中依非韦伦的浓度。方法：使用蛋白沉淀法处理血浆,以依非韦伦-D5为内标,色谱柱：Phenomenex Gemini C18（2 mm × 100 mm,5 μm,110 A）;柱温：40 ℃;流动相：乙腈-10 mmol·L-1乙酸铵（80∶20）,等度洗脱;流速：0.25 mL·min-1。负离子（ESI-）扫描模式,多反应监测方式（MRM）测定该药浓度。考察该方法的专属性、准确度与精密度、提取回收率与基质效应、全血稳定性及稀释可靠性等。结果：血浆样品中依非韦伦回归方程为y=0.494x+8.32×10-6（r = 0.999 1）,表明依非韦伦在0.1～8 μg·mL-1线性关系良好,定量下限为0.1 μg·mL-1。批内、批间精密度的RSD均＜15%,待测物及内标平均回收率＞95%且总体相等标准偏差（RSD）＜15%,稳定性良好。结论：该方法简便灵敏、准确可靠,适用于人体血浆中依非韦伦浓度的测定。     

液质联用法测定保健食品中非法添加57种化学药的研究

目的:建立一种同时完成抗风湿类、止咳平喘类保健食品中非法添加糖皮质激素和非甾体抗炎药(NSAIDs)等共57种化学药物的快速准确筛查和定性分析的方法。方法:样品采用乙腈超声提取。采用UPLC-MS/MS法,以Atlantis?dC18(150 mm×2.1 mm,3μm)为色谱柱,以0.1%甲酸水溶液(A)-乙腈(B)为流动相,梯度洗脱;流速为0.3 ml·min-1,柱温为40℃,进样体积为5μl。选择ESI离子源,正离子扫描,负离子扫描;多反应监测(MRM)模式,测定57种临床常用的化学药物,通过比较MRM通道中样品峰与对照品峰的分子离子峰,二级碎片离子峰等信息确定添加的化学药。结果:在上述色谱及质谱条件下,57种化学药的质谱检测的线性范围宽,相关性好(r>0.9950);方法回收率范围在80.0%~120.0%(n=9);检测限0.0083~2.5857μg·g^-1,定量限0.0285~8.6191μg·g^-1。对160批样品进行了检测,一次检测可同时对样品中添加的57种化学成分进行筛查和定性分析,有1批次样品中检出双氯芬酸钠。结论:研究结果表明,本法专属性强,灵敏度高,可作为保健食品中非法添加57种糖皮质激素和非甾体抗炎药的快速筛查和准确定性。     

UPLC-MS/MS测定地西泮对大鼠CYP450酶活力的影响

目的 基于UPLC-MS/MS同时测定大鼠血浆中的5种探针药物（安非他酮、美托洛尔、咪达唑仑、非那西丁和甲苯磺丁脲）,5种探针药物分别是细胞色素P450同工酶CYP2B6,CYP2D6,CYP3A4,CYP1A2和CYP2C9底物,评价地西泮对大鼠细胞色素P450酶活性影响。方法 将大鼠随机分成地西泮低剂量组（1 mg·kg^-1）、高剂量组（5 mg·kg^-1）和对照组。地西泮低剂量组、高剂量组腹腔注射地西泮10 d,对照组腹腔注射生理盐水10 d。5种探针药物通过灌胃给予大鼠,用UPLC-MS/MS测定血浆药物浓度。结果 与对照组相比,地西泮组安非他酮、美托洛尔和非那西丁药动学参数均有统计差异。地西泮组安非他酮、非那西丁的AUC比对照组高,但美托洛尔AUC比对照组低。结论 地西泮可能会抑制大鼠CYP2B6酶活性,抑制CYP1A2酶活性,诱导大鼠CYP2D6酶活性,而对大鼠CYP3A4、CYP2C9酶活性无影响。