不同生理状态下银荷方药物制备物黄酮类成分的质量表征特性研究

目的建立银荷方药物制备物及大鼠血浆中槲皮素、山奈酚、异鼠李素的含量测定方法,以研究银荷方药物制备物黄酮类成分的质量表征特性。方法制备物和血浆样品均加酸进行水解,并用乙醚-丙酮(14∶1)萃取法处理血浆样品,采用HPLC分析,以C_(18)为固定相,流动相为乙腈-0.2%磷酸水溶液梯度洗脱,在270 nm检测黄酮类苷元槲皮素、山奈酚、异鼠李素的含量。结果经方法学考证,槲皮素、山奈酚、异鼠李素的线性范围分别为1.49~29.70 mg/L(r=0.999 3),0.65~12.90mg/L(r=0.999 5),0.47~9.40 mg/L(r=0.999 0),日内、日间精密度RSD≤5.85%,血浆样品冻融稳定性良好,室温放置10 h,在-80℃条件下保存1个月的稳定性均良好,符合生物样品分析要求。槲皮素、山奈酚、异鼠李素在正常组中的血药浓度分别为7.25、1.71、3.73 mg/L,在血栓组中血药浓度分别为8.53、2.64、5.69 mg/L,在抗凝组中血药浓度分别为8.32、2.76、4.56 mg/L。结论本文所建立的检测方法适用于银荷方药物制备物黄酮类成分槲皮素、山奈酚和异鼠李素的体内含量测定,并可应用于分析黄酮类成分质量表征特性,为基于黄酮类成分的银荷方药物质量精准控制提供科学依据。     

Rapid and sensitive analysis of trace β-blockers by magnetic solid-phase extraction coupled with Fourier transform ion cyclotron resonance mass spectrometry

A rapid and sensitive method for analyzing trace β-blockers in complex biological samples, which involved magnetic solid-phase extraction (MSPE) coupled with Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS), was developed. Novel nanosilver-functionalized magnetic nanoparticles with an interlayer of poly(3,4-dihydroxyphenylalanine) (polyDOPA@Ag-MNPs) were synthesized and used as MSPE adsorbents to extract trace β-blockers from biological samples. After extraction, the analytes loaded on the polyDOPA@Ag-MNPs were desorbed using an organic solvent and analyzed by FTICR-MS. The method was rapid and sensitive, with a total detection procedure of less than 10 min as well as limits of detection and quantification in the ranges of 3.5–6.8 pg/mL and 11.7–22.8 pg/mL, respectively. The accuracy of the method was also desirable, with recoveries ranging from 80.9% to 91.0% following the detection of analytes in human blood samples. All the experimental results demonstrated that the developed MSPE-FTICR-MS method was suitable for the rapid and sensitive analysis of trace β-blockers in complex biological samples.     

同时定量检测水胺硫磷和水胺氧磷的液相色谱-串联质谱法及应用

建立同时测定大鼠全血中水胺硫磷及其活性代谢产物水胺氧磷的液相色谱-串联质谱(LC-MS/MS)分析方法,用于毒代动力学研究中血药浓度的测定。方法 采集大鼠全血样品后即刻用4倍体积冰冷乙腈沉淀蛋白。待测物用Zorbax SB C₁₈柱(2.1 mm×50 mm,1.8 μm)分离,以含有0.1%甲酸的水和甲醇为流动相系统进行梯度洗脱。采用ESI源正离子多反应监测模式分析,水胺硫磷、水胺氧磷和内标三唑磷的定量离子对分别为m/z 231→121、274→215和314→162。 结果 水胺硫磷和水胺氧磷在5 μg/L～5 mg/L浓度范围内线性良好,最低定量限为5 μg/L,回收率在63.1%～77.8%的范围内,准确度和精密度符合生物样品的检测要求。大鼠静脉注射3 mg/kg水胺硫磷后,水胺硫磷在血中很快下降,消除半衰期为22.2 min。活性代谢产物水胺氧磷的生成也较快,10～15 min达到(257.91±42.00)ng/ml的平均峰值,消除半衰期为17.6 min。水胺硫磷和水胺氧磷的AUC_0~t分别为(528.62±49.15)和(191.11±33.23)h ·μg/L。结论 本研究首次建立了同时定量测定大鼠全血中水胺硫磷和水胺氧磷的LC-MS/MS分析方法,方法的专属性好、灵敏度高,适用于水胺硫磷及水胺氧磷在大鼠体内的毒物代谢动力学研究。     

1例以腹泻为首发症状的输入性恶性疟感染病例报道

探讨一例以腹泻为首发症状的输入性疟疾病例的临床特征和流行病学资料,为输入性疟疾诊冶及预防提供参考资料.收集该病例实验室检测结果、临床诊治经历及流行病学等资料并进行分析.该患者疟疾发作以腹泻为首发症状,血细胞分析:白细胞计数(WBC)5.02×109/L,红细胞计数(RBC)3.91×1012/L,血红蛋白(HGB)13...     

四聚乙醛在亚急性中毒大鼠部分组织中的分布

目的 建立生物检材中四聚乙醛气相色谱/质谱以及气相色谱-FID的分析方法, 考察四聚乙醛在亚急性中毒大鼠部分组织中的分布情况.方法 SD大鼠口服给药四聚乙醛 (40 mg/kg, 140 mg/kg, 280 mg/kg) 1周后麻醉处死, 心血、肝脏、胃组织、肾脏提取后检测, 对照组大鼠以生理盐水口服给药, 各对应组织器官样品为空白对照样.采用二氯甲烷液-液萃取, 气相色谱/质谱进行定性分析、气相色谱-FID定量分析生物检材中的四聚乙醛的浓度.结果 建立了生物检材中四聚乙醛的GC/MS及GC-FID分析方法.GC/MS分析方法最低检出限为0.1 ng, GC-FID分析方法在0.010.1 mg/mL范围线性良好 (R=0.9992) , 平均回收率大于75%.高、中、低剂量染毒后四聚乙醛在胃组织中的浓度最高, 而常见药物代谢的肝脏组织中浓度最低.结论 四聚乙醛在亚急性中毒大鼠体的含量分布情况为:胃组织>心血>肾脏>肝脏, 该方法操作简便快速, 检测灵敏度高, 干扰小, 适用于体内四聚乙醛含量的测定.     

基于AFM对蝙蝠葛碱诱导NB4细胞凋亡的研究

【目的】 探讨蝙蝠葛碱对急性早幼粒细胞白血病(APL) NB4细胞增殖的影响和可能的细胞凋亡机制。【方法】 采用CCK-8法检测NB4细胞的存活率,原子力显微镜(AFM)探测蝙蝠葛碱作用NB4细胞前后细胞形态及超微结构的变化,流式细胞术检测细胞凋亡率、线粒体膜电位和活性氧的变化。【结果】 蝙蝠葛碱能显著抑制NB4细胞生长增殖,20~80 μmol/L 蝙蝠葛碱处理细胞24 h后,细胞存活率从(86.5±11.7)%下降到(4.1±0.5)%; 以10~40 μmol/L蝙蝠葛碱处理NB4细胞24 h,流式细胞术分析显示,细胞凋亡率和细胞内活性氧含量上升、线粒体膜电位下降。AFM 探测表明,正常NB4细胞呈圆形,细胞饱满,表面较光滑。蝙蝠葛碱处理组细胞皱缩,细胞表面的平均粗糙度增大。【结论】 蝙蝠葛碱能显著抑制NB4细胞增殖并诱导其凋亡。     

PLASMID MEDIATED DRUG RESISTANCE IN VIBRIO CHOLERAE 0139 BENGAL

Sixteen strains of Vibrio cholerae were isolated from cases of diarrhoea. Out of these, 12 (75%) were identified as Vibrio cholerae 0139 synonym Bengal and 4 (25%) as Vibrio cholerae El Tor by standard biochemical and serological tests. Modified CAMP reaction in sheep blood agar showed that 0139 produced moderate hemolysis, El Tor produced wider zone of hemolysis whereas Classical Vibrio cholerae produced no zone of hemolysis (CAMP negative). Break point minimum inhibitory concentration (MIC) by agar dilution method showed that all 0139 strains were resistant to ampicillin 8 mg/L, streptomycin 1 mg/L, chloramphenicol 8 mg/L, sulphamethoxazole 32 mg/L and trimethoprim 0.3-128 mg/L, 58.3% were sensitive to gentamicin 1 mg/L, and all were sensitive to norfloxacin 1 mg/L and cefotaxime 1 mg/L. Resistance to trimethoprim, sulphamethoxazole, ampicillin and gentamicin in 5 strains could be transferred to E coli K-12 by conjugation experiment at a rate of 5×10⁻⁶ to 4×10⁻³. Distinct plasmid bands of 35.8 mega daltons could be seen in agarose gel electrophoresis.KEY WORDS: CAMP test, Drug resistance, Plasmid, Vibrio cholerae 0139.     

RP-HPLC法测定大鼠血浆中姜黄素含量

目的 采用RP-HPLC法测定大鼠血浆中姜黄素的含量。方法血浆样品经处理后,按以下色谱条件测定姜黄素的含量：DiamonsilC18柱（4．6mm×250mm,5μm）,柱温：室温;流动相：乙腈-2％醋酸溶液（58：42）;流速：1．0ml／min;检测波长：426nm．结果在本色谱条件下血浆中的杂质峰较少,姜黄素和内标物及其他内源性干扰成分可成功分离。采用乙酸乙酯液液萃取法处理血浆样品,萃取回收率最高。在0．1～0．5mg／L浓度范围内,线性关系良好,回归方程y=0．0026x-0．0756,r=0．9984;日内、日间精密度RSD符合生物样品的分析要求;加样回收率和萃取回收率均在90％以上。结论所建立的测定大鼠血浆中姜黄素含量的方法简单可行、准确可靠,适用于姜黄素的血药浓度监测。     

氧化苦参碱对大肠埃希菌的体外药敏试验

目的：：观察氧化苦参碱对大肠埃希菌标准株的体外药敏情况。方法：采用二倍稀释法,用0.9%氯化钠注射液将苦参碱胶囊分别稀释为8000、4000、2000、1000、500、250、125、62.5 mg/L的药液,按药液：MH琼脂=1：9的比例分别配制含药培养基,每个浓度各3个,并设空白和0.9%氯化钠注射液组为对照。大肠埃希菌复苏后,配制菌液备用,接种环取菌液一环接种于各培养基内,置36℃的培养箱内,培养24 h,观察结果。结果：各个含药培养基大肠埃希菌生长良好,与对照组相比无明显差异。结论：氧化苦参碱对大肠埃希菌的生长无影响。     

高效液相色谱法测定家兔血浆中安妥沙星浓度及药代动力学参数

目的:建立测定家兔血浆中安妥沙星浓度的高效液相色谱方法,探讨安妥沙星在家兔体内的药代动力学特征。方法:取安妥沙星片,按25 mg/kg剂量,研细,分散于0.5%羧甲基纤维素钠溶液中,给家兔灌胃,采血,血样经高氯酸处理后进行高效液相色谱法分析。色谱柱条件:Phenomenex C₁₈柱(250 mm×4.6 mm,4μm);乙腈:50 mmol/L枸橼酸溶液:1 mol/L醋酸胺溶液(19:80:1,体积比)为流动相,流速1.0ml/min;检测波长为295nm,柱温40℃。结果:安妥沙星在家兔血浆中的线性范围为0.164~10.5μg/ml,最低定量限为0.164μg/ml。日内及日间变异均<10%,准确度相对误差<5%。血浆中回收率>80%。实验条件下安妥沙星在家兔体内的药代动力学参数:血药浓度时间曲线下面积为(46.17±13.99)mg·L-1·h-1,达峰时间为(0.9±0.14)h,峰浓度为(5.15±0.54)mg/L,生物半衰期为(11.98±4.02)h,清除率为(0.58±0.19)L·kg-1·h-1。结论:该方法经考察符合生物样品的测定要求,可用于测定家兔血浆安妥沙星浓度和药代动力学的研究。     

高效液相色谱法测定人血浆罗哌卡因浓度

目的建立罗哌卡因血浆药物浓度的高效液相色谱测定方法。方法采用安捷龙1101色谱分析系统;色谱柱：Nova-pak C18柱（3.9 mm×150 mm）,内标为布比卡因;流动相：甲醇∶蒸馏水=2∶1,流速1 ml/min,检测波长230nm。结果罗哌卡因标准曲线线性方程为C=0.990A-0.031（n=8,r=0.995,P〈0.01）,线性浓度范围为（0.10-4.00）mg/L;最低检测浓度0.10 mg/L。平均回收率为100.16%,日内及日间变异均小于6%。结论反相高效液相色谱法简便、灵敏、准确,可以用于临床上罗哌卡因血浆药物浓度的监测和药代动力学研究。     

Caffeine modulates pharmacokinetic and pharmacodynamic profiles of pioglitazone in diabetic rats: Impact on therapeutics

Objectives:To determine the influence of caffeine on pharmacokinetics and pharmacodynamics of pioglitazone (PIO) in diabetic rats.Methods:This was a preclinical study conducted in the College of Pharmacy, Najran University, Saudi Arabia, using 5 groups of Wistar rats: normal rats, untreated diabetic rats, diabetic rats + caffeine (20 mg/kg), diabetic rats + PIO (10 mg/kg), and diabetic rats + PIO (10 mg/kg) + caffeine (20 mg/kg). The drugs were administered for 14 days, and fasting plasma glucose concentrations were determined on the first day, and thereafter at weekly intervals. On day 14, rat sacrifice was followed with assay of levels of biomarkers. To estimate the pharmacokinetic parameters, the diabetic animals were assigned to 2 groups: one group received PIO (10 mg/kg), while the other received PIO + caffeine (20 mg/kg). Blood samples were drawn from the retro-orbital plexus at different time intervals, and pharmacokinetic parameters were measured using high performance liquid chromatography.Results:Caffeine caused statistically marked increases in area under the curve, C_max, T_max, and half-life of PIO, and decreased clearance. Combination of PIO and caffeine produced a synergistic effect on percentage reduction in blood glucose, with 67.1% reductions observed on day 7 and 68.9% reductions observed on day 14. Liver and cardiac biomarkers were significantly decreased, suggesting cardioprotective and hepatoprotective effects.Conclusion:Co-administration of PIO with caffeine enhances its antidiabetic effect, probably due to enhanced bioavailability of PIO, leading to clinical benefits in diabetic patients.     

HPLC法测定秦艽中龙胆苦苷含量及药动学的研究

目的研究秦艽中龙胆苦苷在家兔体内的药代动力学规律。方法以秦艽水煎醇沉液家兔耳静脉注射后定时取血,以SHIM-PACKVP-ODS色谱柱（150mm×4.6mm,5μm）,测定血浆药物浓度,采用DAS软件计算药动学参数。结果龙胆苦苷血药浓度在0.683-136.6μg/ml内线性关系良好（r=0.9995）;日内、日间精密度均小于10%,回收率为89.8%-101.4.3%。药-时曲线符合三房室模型,主要药动学参数：Tmax=0.083h,Cmax=45.1mg/L,V=1.312L/kg,CL=0.761L/（h.kg）,t1/2=1.195h,AUC（0-∞）=30.78mg/（L.h）。结论该方法具有准确、简便、快速的特点,可用于秦艽中龙胆苦苷的检测。     

Analysis of docosanol using GC/MS: Method development,validation, and application to ex vivo human skin permeation studies

Docosanol is the only US Food and Drug Administration (FDA) approved over-the-counter topical product for treating recurrent oral-facial herpes simplex labialis. Validated analytical methods for docosanol are required to demonstrate the bioequivalence of docosanol topical products. A gas chromatography/selected ion monitoring mode mass spectrometry (GC/SIM-MS) method was developed and validated for docosanol determination in biological samples. Docosanol and isopropyl palmitate (internal standard) were separated on a high-polarity GC capillary column with (88% cyanopropy)aryl-polysiloxane employed as the stationary phase. The ions of m/z 83 and 256 were selected to monitor docosanol and isopropyl palmitate, respectively; the total run time was 20 min. The GC/SIM-MS method was validated in accordance with US FDA guidelines, and the results met the US FDA acceptance criteria. The docosanol calibration standards were linear in the 100–10000 ng/mL concentration range (R²>0.994). The recoveries for docosanol from the receptor fluid and skin homogenates were >93.2% and >95.8%, respectively. The validated method was successfully applied to analyze ex vivo human cadaver skin permeation samples. On applying Abreva® cream tube and Abreva® cream pump, the amount of docosanol that penetrated human cadaver skin at 48 h was 21.5 ± 7.01 and 24.0 ± 6.95 ng/mg, respectively. Accordingly, we concluded that the validated GC/SIM-MS was sensitive, specific, and suitable for quantifying docosanol as a quality control tool. This method can be used for routine analysis as a cost-effective alternative to other techniques.     

Automatic analytical approach for the determination of 12 illicit drugs and nicotine metabolites in wastewater using on-line SPE-UHPLC-MS/MS

In this study, we developed a novel on-line solid phase extraction (SPE)-ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS)-based analytical method for simultaneously quantifying 12 illicit drugs and metabolites (methamphetamine, amphetamine, morphine, codeine, 6-monoacetylmorphine, benzoylecgonine, 3,4-methylenedioxymethamphetamine, 3,4-methylenedioxyamphetamine, cocaine, ketamine, norketamine, and methcathinone) and cotinine (COT) in wastewater samples. The analysis was performed by loading 2 mL of the sample onto an Oasis hydrophilic-lipophilic balance cartridge and using a cleanup step (5% methanol) to eliminate interference with a total run time of 13 min. The isotope-labeled internal standard method was used to quantify the target substances and correct for unavoidable losses and matrix effects during the on-line SPE process. Typical analytical characteristics used for method validation were sensitivity, linearity, precision, repeatability, recovery, and matrix effects. The limit of detection (LOD) and limit of quantification (LOQ) of each target were set at 0.20 ng/L and 0.50 ng/L, respectively. The linearity was between 0.5 ng/L and 250 ng/L, except for that of COT. The intra- and inter-day precisions were <10.45% and 25.64%, respectively, and the relative recovery ranged from 83.74% to 162.26%. The method was used to analyze various wastewater samples from 33 cities in China, and the results were compared with the experimental results of identical samples analyzed using off-line SPE. The difference rate was between 19.91% and −20.44%, and the error range could be considered acceptable. These findings showed that on-line SPE is a suitable alternative to off-line SPE for the analysis of illicit drugs in samples.     

超声引导下妊娠囊内注射天花粉蛋白治疗异位妊娠疗效观察

目的探讨超声引导下妊娠囊内注射天花粉蛋白治疗异位妊娠的临床价值。方法 38例异位妊娠未破裂型患者,根据患者意愿,在知情同意的情况下分为2组:天花粉蛋白介入组24例(天花粉组),其中5例血人绒毛膜促性腺激素(β-HCG)≥15 000 U/L,12例4000～15 000 U/L,7例≤4000 U/L。在超声引导下经阴道后穹窿穿刺附件区包块,注射天花粉蛋白1.2～2.4 mg;14例β-HCG>1000 U/L的异位妊娠未破裂型患者为对照组,患者知情选择手术或口服甲氨蝶呤(MTX)治疗。结果天花粉组24例均治疗成功,大部分患者天花粉介入治疗后3～5 d血β-HCG下降90%以上(治疗成功),3例辅以肌注MTX或天花粉蛋白注射液治疗成功,治疗成功率100%。对照组14例中,12例口服MTX治疗2～3周后,8例血β-HCG下降80%以上(治疗成功);4例口服MTX治疗失败转为手术治疗,另外2例β-HCG>4000 U/L的患者直接行手术治疗。结论超声引导下天花粉介入治疗成功率高,为血β-HCG值较高的未破裂型异位妊娠提供了一种新的治疗方法。     

Metabolism of dauricine and identification of its main metabolites

Dauricine ( Dau) is a kind of bisbenzylte-trahydroisoquinoline alkloid extracted from the rootof Dauricum D.C.. Both clinical trials and experi-mentshave proved itsgood antiarrhythmic effect,es-pecially its character of use- dependence in prolongingAPD in guinea myocardial[1] ,which makes its verypromising to work dau into a new type of antiar-rhythmic agent.Dau has the same characters as bis-benzyltetrahydroisoquinoline alkloids such as tetran-drine,neferine,trilobine,isotetrandrine and so …     

生物检材中34种常见毒品及代谢物的液相色谱轨道阱质谱筛查方法研究

目的 建立高效液相色谱轨道阱质谱对生物检材中34种常见毒品及代谢物的筛查方法。方法 尿样和血样经乙酸乙酯提取后,氮吹浓缩和复溶;毛发样品依次用水和丙酮振荡洗涤、晾干后剪成约1 mm,置冷冻研磨仪中粉碎,甲醇提取,过滤后,滤液供仪器分析。色谱分析采用Hypersil Gold PFP色谱柱(2.1 mm×100 mm,3μm)分离,甲醇-5 mmol/L乙酸铵溶液作为流动相,梯度洗脱,流速为400μL/min。质谱分析采用电喷雾正负离子交替模式,使用一级母离子全扫描和数据依赖的二级子离子扫描(Full MS/dd-MS²)模式,采用Xcalibur4.0软件分别对仪器和数据进行控制和采集,利用TraceFinder3.3软件进行筛查识别。结果 血样、尿样和发样的34种常见毒品及其代谢物方法检出限分别为3.30～10 700 ng/L、4.43～5 440 ng/L和0.035 0～4.21μg/kg。在5.0、10、20μg/L加标水平下,日内精密度为3.50%～6.00%,日间精密度为4.18%～9.90%。采集了尿样、血样和发样共1 125份生物检材,采用该方法...     

深圳市成年居民初级农产品中铅的暴露风险评估

目的 对深圳市18岁以上成年居民通过膳食暴露铅的风险进行评估。方法 应用电感耦合等离子体质谱法（ICP-MAS）对2018年深圳市市售的11种初级农产品中铅污染水平进行检测;应用3 d入户的调查方法对深圳市18岁以上622名居民11种初级农产品的消费量进行调查;应用@risk软件以蒙特卡罗算法计算深圳市18岁以上居民通过11种初级农产品对铅的暴露水平,应用暴露边界法（MOE）对暴露风险进行评估。结果 深圳市18岁以上居民通过膳食对铅的总暴露量的最小值为0.023 0 mg/(kg·d)、中位数为0.372 6 mg/(kg·d) 、第95百分位数为0.799 3 mg/(kg·d);从食品污染的角度来看,蔬菜和畜肉中的铅污染是引起深圳成年居民膳食铅暴露风险的主要原因。50%深圳市18岁以上居民通过膳食暴露铅的MOE值大于3.22,95%的深圳成年居民MOE值大于1.5,有0.1%的18岁以上深圳居民通过膳食暴露于铅的MOE值小于1,处于危险的铅暴露模式。结论 深圳市18岁以上居民通过膳食暴露于铅的风险较低,但对膳食铅暴露的监测和评估应持续开展。     

HPLC-MS/MS法测定大鼠血浆中紫杉醇含量及其脂质体药代动力学研究

目的:建立一种测定大鼠血浆中紫杉醇浓度的高效液相色谱串联质谱（HPLC-MS/MS）分析方法,并研究其脂质体的药代动力学。方法:对6只SD大鼠静脉注射1 mg/kg的紫杉醇脂质体注射液,分别于给药后0、0.5、1、2、4、8、12、24、48 h自眼眶取血0.2 mL,C18反相色谱柱作为固定相,水（1 mmoL/L甲酸铵-0.05%甲酸）-甲醇作为流动相,ESI+扫描模式,MRM监测模式,药代动力学参数采用DAS 2.0软件进行计算。结果:紫杉醇在1～5 000 ng/mL范围内线性较好（R2 =0.999 7）,定量下限（LLOQ）为1 ng/mL,日内和日间精密度均低于10%,紫杉醇脂质体给药后血药浓度-时间曲线满足二室模型,主要药代动力学参数t1/2、Cmax、AUC0→t分别为（4.79±0.85）h、（49 785.12±1 618.45）μg/L、（38 753.08±6 144.66）μg/L·h。结论:本方法专属性和灵敏度高,结果可靠,能满足测定大鼠血浆中紫杉醇的定量需求。