新型抗结核化合物TBI-166在比格犬体内的生物利用度

目的：建立比格犬血浆中TBI-166的LC-MS/MS测定方法,研究比格犬口服TBI-166的血浆药代动力学特征和生物利用度。方法比格犬血浆样品中加入TBI-166和内标普萘洛尔后经乙腈沉淀蛋白。 HPLC分离采用Symmetry C8（2.1 mm×50 mm,3.5μm）柱,流动相为含0.1%甲酸的乙腈-水,梯度洗脱,流速为0.2 ml/min质谱检测采用电喷雾离子源,正离子选择反应监测（SRM）模式检测m/z 590→478（TBI-166）和m/z 260→183（普萘洛尔,内标）。应用上述LC-MS/MS方法测定比格犬口服TBI-166（3 mg/kg）和静脉注射（0.5 mg/kg）后的血浆药物浓度,用WinNonlin软件计算药代动力学参数和绝对生物利用度。结果比格犬血浆中TBI-166在2～1000 ng/ml浓度范围内线性关系良好,日内和日间精密度<10%、回收率>98.6%、无明显基质效应且血浆样品稳定性好。雌雄比格犬口服TBI-166（3mg/kg）后血药浓度分别于（4.4±3.5）和（1.4±0.5）h达峰,血药峰浓度分别为（122.0±34.6）和（65.4±2.3）ng/ml,AUC（0-t）为（2615.1±1524.4）和（897.2±318.6）h·μg/L。雌雄比格犬静注TBI-166（0.5mg/kg）后t1/2z为（112.9±25.3）和（69.6±35.3）h、CL为（0.3±0.1）和（0.2±0.1）L/（h·kg）、AUC（0-t）为（3222.4±1656.2）和（1798.0±729.2）h·μg/L。雌雄比格犬口服TBI-166生物利用度分别为13.5%和8.3%。结论本研究建立了比格犬血浆中TBI-166的LC-MS/MS测定方法,该方法准确、简便、灵敏。比格犬口服TBI-166后体内消除较慢,具有一定性别差异,生物利用度为8.3%～13.5%。     

液相色谱-串联质谱法测定大鼠血浆中黄体酮的浓度

目的建立测定大鼠血浆中黄体酮的液相色谱-串联质谱(LC-MS/MS)法。方法血浆样品采用液-液萃取法,经正己烷-二氯甲烷-异丙醇(100∶50∶5,V/V/V)提取后,以甲醇-水(75∶25,V/V)为流动相,以Hypersil ODS柱(50 mm×2.1 mm,5μm)为分析柱,采用ESI源以多反应监测(MRM)方式进行正离子检测。用于定量分析的离子反应为m/z 315.4→m/z(97.0+109.2)(黄体酮)和m/z 385.4→m/z 267.4(醋酸甲地孕酮,内标)。结果黄体酮血浆浓度测定方法的线性范围为0.5~200 ng/ml,日内、内间精密度(RSD)均<6.7%,准确度(RE)在±6.2%之间。结论该方法分析时间短、操作简便、灵敏度高、专属性强,适用于黄体酮的药动学研究。     

两种盐酸特拉唑嗪片的人体生物等效性研究

目的 评价单剂量口服国产和进口盐酸特拉唑嗪片的人体生物等效性。 方法 采用单中心、随机、开放、双周期交叉试验设计,21名受试者在不同周期分别空腹口服国产和进口盐酸特拉唑嗪片2 mg,于给药前0 h及给药后60 h内不同时间点采集静脉血4 ml,采用液-质联用(LC-MS/MS)法测定受试者血浆中特拉唑嗪的浓度。 结果 国产和进口盐酸特拉唑嗪片的t_1/2分别为(13.2±2.39)和(12.5±1.93) h;t_max分别为(1.01±0.83)和(1.08±0.69) h;C_max分别为(40.1±10.6)和(37.3±9.57) ng/ml;AUC_0-∞分别为(428±82.1)和(426±85.2) ng·h/ml。国产盐酸特拉唑嗪片的相对生物利用度为(101.2±14.7)%。国产与进口盐酸特拉唑嗪片AUC_0-t和C_max几何均值比的90%置信区间(CI)均落在80%~125%之间。 结论 国产和进口盐酸特拉唑嗪片具有生物等效性。     

Over-the-Counter “Adrenal Support” Supplements Contain Thyroid and Steroid-Based Adrenal Hormones

Objective

To assess whether dietary supplements that are herbal and/or animal-derived products, marketed for enhancing metabolism or promoting energy, “adrenal fatigue,” or “adrenal support,” contain thyroid or steroid hormones.

Structures of Phytosterols and Triterpenoids with Potential Anti-Cancer Activity in Bran of Black Non-Glutinous Rice

Structures of some bioactive phytochemicals in bran extract of the black rice cv. Riceberry that had demonstrated anti-cancer activity in leukemic cell line were investigated. After saponification with potassium hydroxide, separation of the unsaponified fraction by reversed-phase high performance liquid chromatography (HPLC) resulted in four sub-fractions that had a certain degree of anti-proliferation against a mouse leukemic cell line (WEHI-3 cell), this being IC₅₀ at 24 h ranging between 2.80–467.11 μg/mL. Further purification of the bioactive substances contained in these four sub-fractions was performed by normal-phase HPLC. Structural characterization by gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS) and nuclear magnetic resonance spectroscopy (NMR) resulted in, overall, the structures of seven phytosterols and four triterpenoids. Four phytosterols, 24-methylene-ergosta-5-en-3β-ol, 24-methylene-ergosta-7-en-3β-ol, fucosterol, and gramisterol, along with three triterpenoids, cycloeucalenol, lupenone, and lupeol, were found in the two sub-fractions that showed strong anti-leukemic cell proliferation (IC₅₀ = 2.80 and 32.89 μg/mL). The other sterols and triterpenoids were campesterol, stigmasterol, β-sitosterol and 24-methylenecycloartanol. Together with the data from in vitro biological analysis, we suggest that gramisterol is a significant anti-cancer lead compound in Riceberry bran extract.     

瑞巴派特分散片的人体药动学及生物等效性

建立了液相色谱-串联质谱法测定健康志愿者口服瑞巴派特分散片(受试制剂)和普通片(参比制剂,膜固思达(R))后的血药浓度,估算两制剂的药动学参数并评价生物等效性.采用随机、开放、双周期交叉试验设计,22名男性健康志愿者单剂量口服受试或参比制剂0.1g后,血样经乙腈直接沉淀蛋白后进行LC-MS/MS分析,利用DAS2.1.1软件计算药动学参数,并进行生物等效性评价.受试和参比制剂在人血浆中瑞巴派特的cmax为(236.9±103.4)和(233.9±105.1) ng/ml,tmax为(2.7±1.1)和(2.9±1.4)h,t1/2为(2.0±0.8)和(2.3±1.5)h,AUC0→12h为(929.0±291.9)和(970.5±353.2) ng·h·ml1.受试制剂的相对生物利用度为(102.6±29.8)％.     

僵蚕抗凝成分仿生提取液的氨基酸序列分析

目的利用高效液相色谱-质谱法（LC-MS/MS）对僵蚕胃蛋白酶提取液的有效抗凝成分进行结构分析。方法采用Hola C₁₈ (2.1mm×100 mm,2.7 μm)色谱柱, 以0.05%甲酸水溶液-0.05%甲酸乙腈溶液为流动相,梯度洗脱, LC-MS/MS正离子模式分析,初步探究抗凝活性成分的相对分子量和氨基酸组成。结果僵蚕经胃蛋白酶解得到相对分子量500~1000的活性肽,且多肽为低于10个氨基酸组成的低聚肽。结论僵蚕蛋白类组分通过酶解为活性低聚肽发挥抗凝作用,胃蛋白酶提取法更具科学性。     

Effects of pH and Nutrients (Nitrogen) on Growth and Toxin Profile of the Ciguatera-Causing Dinoflagellate Gambierdiscus polynesiensis (Dinophyceae)

Ciguatera poisoning is a foodborne disease caused by the consumption of seafood contaminated with ciguatoxins (CTXs) produced by dinoflagellates in the genera Gambierdiscus and Fukuyoa. Ciguatera outbreaks are expected to increase worldwide with global change, in particular as a function of its main drivers, including changes in sea surface temperature, acidification, and coastal eutrophication. In French Polynesia, G. polynesiensis is regarded as the dominant source of CTXs entering the food web. The effects of pH (8.4, 8.2, and 7.9), Nitrogen:Phosphorus ratios (24N:1P vs. 48N:1P), and nitrogen source (nitrates vs. urea) on growth rate, biomass, CTX levels, and profiles were examined in four clones of G. polynesiensis at different culture age (D10, D21, and D30). Results highlight a decrease in growth rate and cellular biomass at low pH when urea is used as a N source. No significant effect of pH, N:P ratio, and N source on the overall CTX content was observed. Up to ten distinct analogs of Pacific ciguatoxins (P-CTXs) could be detected by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in clone NHA4 grown in urea, at D21. Amounts of more oxidized P-CTX analogs also increased under the lowest pH condition. These data provide interesting leads for the custom production of CTX standards.     

金莲花酚酸类成分的大鼠肝微粒体生物转化研究

目的 研究金莲花中酚酸类化合物在大鼠肝微粒体内的转化特征。方法 建立体外大鼠肝微粒体转化模型,模拟金莲花中原金莲酸、金莲花苷、藜芦酸在大鼠肝脏内的代谢过程,采用液相色谱质谱联用（LC-MS）法对代谢成分进行检测和分析。结果 在大鼠肝微粒体的作用下,原金莲酸转化为1个产物,分子式为C₁₉H₂₃O₁₀,推测为原金莲酸的葡萄糖醛酸化反应产物;金莲花苷转化为2个产物,分别为C₂₅H₃₆O₁₄和C₁₈H₂₄O₉,推测分别为增加1糖基和脱甲基生成的转化产物;藜芦酸转化为1个产物,分子式为C₈H₈O₄,推测2种结构,各为脱掉1个甲基的产物。结论 金莲花酚酸类化合物在大鼠肝微粒体中主要发生脱甲基化、糖苷化、葡萄糖醛酸化反应,为明确金莲花酚酸类成分的体内转化规律和代谢特征提供依据。     

养胃颗粒中NF-κB靶标介导的治疗胃溃疡成分的筛选与评价研究

目的筛选养胃颗粒中影响NF-κB靶标治疗胃溃疡的化学成分,并建立基于上述成分的质量评价方法。方法采用NF-κB质粒转染的RAW264.7细胞株,给药、孵育后使用PCR进行扩增和表达,测定荧光值以评价各化学成分对NF-κB靶标的影响;再以筛选所得成分为指标,采用LC-MS法进行多指标含量测定研究,色谱柱为Agilent SB C18(100 mm×2.1 mm,2.7μm),使用0.1%甲酸-乙腈作为流动相,梯度洗脱,ESI模式同时进行正负离子扫描,离子对为毛蕊异黄酮葡萄糖苷786/425,三七皂苷R1为945/925,共测定10批养胃颗粒。结果养胃颗粒的7个成分中,毛蕊异黄酮葡萄糖苷和三七皂苷R1具有较好的NF-κB靶标激动活性,在10批颗粒中,毛蕊异黄酮葡萄糖苷的平均含量为0.834 mg/g,三七皂苷R1的平均含量为0.512 mg/g。结论养胃颗粒中,毛蕊异黄酮葡萄糖苷、三七皂苷R1是通过NF-κB靶标治疗胃溃疡的活性成分,本研究以上述成分为指标建立LC-MS质量评价方法,具有准确、高效、贴近功效主治的特点。