液相色谱-串联质谱法测定健康人体血浆中米格列醇浓度及其药代动力学研究

目的建立测定血浆中米格列醇片(降血糖药)的液相色谱-串联质谱法,考察米格列醇在中国健康志愿者体内的药代动力学。方法血浆样品经液-液提取后,进行色谱分离,在三重四极杆串联质谱仪上,以多反应离子监测(MRM)方式进行定量分析,用于监测的离子为m/z 208．3→m／z 146．1 (米格列醇)和m/z 268．5→m/z 250．4(内标,伏格列波糖)。结果米格列醇的最低定量浓度为5．0 μg·L-1,线性范围为5～2 000 μg·L-1,精密度与准确度符合生物样品分析要求。结论该法操作简便、快速、灵敏度高,适于临床药代动力学研究。     

HPLC—MS／MS法测定人血浆中赖诺普利的浓度

目的:建立测定人血浆中赖诺普利浓度的 HPLC-MS/MS 法。方法:200μL待测血浆中定量加入内标后,直接加入蛋白沉淀剂三氟醋酸,涡旋离心后取上清液直接进样。采用 Zorbax Eclipse DB-C_8柱(5μm,150 mm×4.6 mm)分析柱;流动相为甲醇-12.5 mmol·L~(-1)醋酸铵缓冲液(60:40,pH=4.40),流速0.5 mL·min~(-1);进样量25μL。质谱检测采用 ESI 正离子模式,扫描方式为多反应监测方式,扫描离子对为 m/z406.3→84.0(赖诺普利)和 m/z 349.1→206.1(内标依那普利拉)。结果:本文所建立测定赖诺普利的线性范围为1.064～851.2 ng·mL~(-1),最低定量限可达1.064 ng·mL~(-1)。测定的方法回收率为97.52%～103.2%;日内 RSD<7%,日间 RSD<5%。结论:本文所建立的方法灵敏度良好、准确度高,可用于赖诺普利的药代动力学研究及临床药物浓度监测。     

液相色谱-串联质谱法测定人血浆中米格列奈浓度及其药动学研究

目的建立一种快速分析测定人血浆中米格列奈的液相色谱-串联质谱色谱法,用以研究米格列奈在健康人体内的药动学。方法以那格列胺为内标,血浆酸化后经液液萃取后,采用液相色谱-串联质谱法以多反应检测方式进行测定,选择监测的离子为m/z 316.2→298.2(米格列奈)和m/z 318.2→120.2 (那格列胺)。流动相以甲醇-10 mmol·L~(-1)醋酸铵水溶液(75:25,V/V),流速0.3 mL·min~(-1),色谱柱为Agilent Zorbax Eclipse Plus C_(18)(1.8μm,3 mm×150 mm);柱温:30℃。结果米格列奈在0.502 0～4 016μg·L~(-1)浓度范围内呈良好的线性关系(r=0.995),最低检测浓度为0.502 0μg·L~(-1),精密度和准确度试验均符合生物分析要求,应用此法测得5、10、20 mg剂量组不同给药方式、多个时间点的米格列奈血药浓度,结果呈线性动力学特征。结论该方法灵敏度高、专属性强、准确、简便,适用于米格列奈的人体药动学研究。     

HPLC-示差折光检测器测定米格列醇片的含量及有关物质

目的建立高效液相色谱-示差折光检测法测定米格列醇片的含量和有关物质。方法用磺酸基阳离子交换键合硅胶(250×4.6mm,5μm)为填充剂;以乙腈-三氟乙酸溶液(4.5→1000mL)(20∶80)为流动相;示差折光检测器检测;检测器温度为40℃。理论板数以米格列醇峰计算,应不低于2000。结果空白辅料不干扰测定,米格列醇主峰和杂质峰分离良好,定量限浓度约为10μg·mL^-1。米格列醇浓度在26.28~210.2μg·mL^-1范围内,与峰面积线性关系良好,线性方程A=2611.5C-2802,r=0.9988;平均回收率100.9%(RSD=1.0%;n=9);溶液在8h内稳定。结论本法专属性强、灵敏度高、准确度和稳定性好,可作为米格列醇片有关物质和含量测定的检测方法。     

液相色谱-质谱／质谱联用法测定人血浆中加兰他敏

目的:建立测定人血浆中加兰他敏浓度的液相色谱-质谱/质谱联用法,用于其人体血药浓度测定。方法:血浆样品经液-液萃取后,以甲醇-水-甲酸(65:35:1)为流动相,Zorbax SB-C_8柱(150mm×4.6mm,5μm)分离,采用大气压化学电离源,以选择反应监测方式进行正离子检测。内标为双氢吗啡酮,用于定量分析的离子反应分别为 m/z 288→m/z 213(加兰他敏)和 m/z 286→m/z 185(内标)。结果:血浆中加兰他敏最低定量限为0.5ng·mL~(-1),其线性范围为0.5～100ng·mL~(-1)。其高、中、低3个浓度的平均提取回收率为80.9%,方法回收率为100.5%,日内及日间 RSD 均<8%。结论:方法选择性强,灵敏度高,快速准确,可作为加兰他敏人体内药动学研究的手段。     

HPLC-MS/MS法测定人血浆中阿莫西林的质量浓度

目的建立测定人血浆中阿莫西林质量浓度的高效液相色谱-质谱(HPLC-MS/MS)联用检测方法,并用于生物等效性研究。方法以阿莫西林-d4为内标,血浆样品经甲醇沉淀后,经HPLC-MS/MS分析。色谱柱为ZORBAX SB-Aq (150 mm×4.6 mm,3.5μm);流动相为甲醇-2 mL·L~(-1)甲酸溶液(40∶60);流速为0.6 mL·min~(-1)。采用电喷雾离子源(ESI),多重反应选择离子监测(MRM)负离子模式,阿莫西林和阿莫西林-d4的定量离子对分别为m/z 364→223和m/z 368→227。结果阿莫西林血药质量浓度在0.088 57～39.370 00μg·mL~(-1)范围内线性关系良好,选择性、准确度、精密度、提取回收率、基质效应和稳定性等均符合要求。结论该方法简单、快速、灵敏、准确,可用于阿莫西林血药质量浓度的检测及生物等效性研究。     

液-质联用法测定人血浆伊潘立酮的药物浓度

目的:建立高效液相串联质谱方法测定人血浆中伊潘立酮的药物浓度。方法:方法色谱柱为Kromasil 60-5CN(100mm×2.1mm,5μm),流动相:乙腈与5mmol·L-1醋酸铵缓冲液(含0.1%甲酸)体积比为35∶65,流速为0.3mL·min-1,吡格列酮为内标,采用电喷雾离子源,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分别为m/z 427.2→m/z261.2(伊潘立酮),m/z357.2→m/z133.8(吡格列酮,内标)结果:伊潘立酮的血浆浓度在20～20 000pg·mL-1范围内线性良好,定量下限为20pg·mL-1,日内精密度<3%,日间精密度<9%,回收率为96.9%～101%。结论:该法操作简单,灵敏,准确,重现性好,适用于伊潘立酮人体药动学研究及生物等效性研究。     

液相色谱-质谱联用法测定人血浆中多潘立酮

目的:建立测定人血浆中多潘立酮的液相色谱-电喷雾串联质谱(LC/ESI-MS/MS)法。方法:待测血浆0.1 mL 用甲醇沉淀蛋白,取离心后的上清液进样10μL在 Phenomenex Gimim-C_(18)柱(2.00 mm×50 mm,5 μm)上分离,流动相为甲醇-水(40:60,v/v,含0.3%醋酸),流速0.2 mL·min~(-1),LC/ESI-MS/MS 采用多离子反应监测,正离子模式,用于定量分析的离子反应分别为 m/z426→m/z175(多潘立酮)和 m/z 379→m/z 264(氨溴索,内标)。结果:血浆中的内源性物质不干扰测定,每个样品分析时间约2 min;本法线性范围为0.3～100 ng·mL~(-1),最低定量浓度为0.3 ng·mL~(-1);日内、日间 RSD 分别小于7.1%和13.3%,相对误差小于5.4%。结论:该法操作简便、快速、准确,灵敏度高,可用于多潘立酮临床治疗剂量的药物动力学研究。     

液相色谱-串联质谱法测定人血浆中格列本脲

目的:建立测定人血浆中格列本脲的液相色谱-串联质谱法,并用于临床药代动力学研究。方法:血浆样品经液-液萃取后,以乙腈-水-甲酸(90:10:0.2)为流动相,采用 Zorbax SB-C_8 柱(150mm×4.6mm,5μm)分离,通过大气压化学电离源四极杆串联质谱,以选择反应监测(SRM)方式进行检测。用于定量分析的离子反应分别为 m/z494→369(格列本脲)和m/z 324→127(内标格列齐特)。结果:LC-MS/MS 法测定人血浆中格列本脲的线性范围为0.50～500 ng·mL~(-1),定量下限为0.50 ng·mL~(-1)。以3个浓度水平的质量控制样品求得各浓度水平日内、日间精密度(RSD)均小于5.4%,相对偏差(RE)在±2.3%以内。在临床药代动力学研究中,应用此法测试了20名受试者口服盐酸二甲双胍格列本脲胶囊后血浆中格列本脲的浓度。结论:该法灵敏、快速、准确,操作简便,线性范围宽,适用于临床药代动力学研究。     

超高效液相色谱-串联质谱法检测中成药中添加格列本脲、格列吡嗪

目的:建立快速、准确、高灵敏度的检测中成药降糖制剂中非法添加格列本脲、格列吡嗪的分析方法。方法:采用 C_(18)(50 mm×2.1 mm,1.7 μm)柱分离,以乙腈-0.01 mol·L~(-1)乙酸铵(0.1%乙酸)缓冲溶液梯度洗脱,流速0.3 mL·min~(-1),以多反应检测(MRM)方式进行检测。用于定量分析的二级碎片离子分别为 m/z 321(格列吡嗪)和 m/z 369(格列本脲)。结果:格列吡嗪和格列本脲的线性范围分别为4.98～498 ng·mL~(-1)和4.96～496 ng·mL~(-1)。结论:此方法选择性强,灵敏度高,可作为中成药中非法添加格列本脲、格列吡嗪的分析检测方法。     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Nachweis einiger Heilmittel in der Kniegelenksynovialflüssigkeit

Zusammenfassung Mittels Gaschromatographie und Dünschichtchromatographie wiesen die Autoren 11 Substanzen nach, welche durch Injektion oder nach Verabreichung per os in die Kniegelenksynovialflüssigkeit eindrangen. In ihrer Aufstellung konnten sie eine direkte Beziehung zwischen Struktur sowie chemischphysikalischen Eigenschaften der Substanz und ihrer Fähigkeit, aus dem Blut in die Kniegelenksynovialflüssigkeit einzudringen, nicht nachweisen, außer der Tatsache, daß Substanzen mit starker Affinität zu Eiweißstoffen erst in höheren Dosen nachweisbar waren.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.     

Gender-related symptoms and correlates of alcohol dependence among men and women with a lifetime diagnosis of alcohol use disorders

This study explored gender-related symptoms and correlates of alcohol dependence in a crosssectional study of 150 men and 150 women with a lifetime diagnosis of alcohol use disorders (AUD). Participants were recruited in equal numbers from treatment settings, correctional centres and the general community. Standardized measures were used to determine participants' use of substances, history of psychiatric disorders and psychosocial stress, their sensation seeking and family history of substance use and mental health disorders. Multivariate analyses were used to detect patterns of variables associated with gender and the lifetime severity of AUD. Men had a longer history of severe AUD than women. Women had similar levels of alcohol dependence and medical and psychological sequelae as men, despite 6 fewer years of AUD. More women than men had a history of severe psychosocial stress, severe dependence on other substances and antecedent mental health problems, especially mood and anxiety disorders. There were differences in family history of alcohol-related problems approximating same-gender aggregation. The severity of a lifetime AUD was predicted by its earlier age at onset and the occurrence of other disorders, especially anxiety, among both men and women. The limitations in the generalizability of these findings due to sample idiosyncrasies are discussed.     

Biochemical and molecular characterisation of cubozoan protein toxins

Class Cubozoa includes several species of box jellyfish that are harmful to humans. The venoms of box jellyfish are stored and discharged by nematocysts and contain a variety of bioactive proteins that are cytolytic, cytotoxic, inflammatory or lethal. Although cubozoan venoms generally share similar biological activities, the diverse range and severity of effects caused by different species indicate that their venoms vary in protein composition, activity and potency. To date, few individual venom proteins have been thoroughly characterised, however, accumulating evidence suggests that cubozoan jellyfish produce at least one group of homologous bioactive proteins that are labile, basic, haemolytic and similar in molecular mass (42-46 kDa). The novel box jellyfish toxins are also potentially lethal and the cause of cutaneous pain, inflammation and necrosis, similar to that observed in envenomed humans. Secondary structure analysis and remote protein homology predictions suggest that the box jellyfish toxins may act as α-pore-forming toxins. However, more research is required to elucidate their structures and investigate their mechanism(s) of action. The biological, biochemical and molecular characteristics of cubozoan venoms and their bioactive protein components are reviewed, with particular focus on cubozoan cytolysins and the newly emerging family of box jellyfish toxins.     

Dose tolerability of chronically inhaled voriconazole solution in rodents

Invasive pulmonary aspergillosis (IPA) is a fungal disease of the lung associated with high mortality rates in immunosuppressed patients despite treatment. Targeted drug delivery of aqueous voriconazole solutions has been shown in previous studies to produce high tissue and plasma drug concentrations as well as improved survival in a murine model of IPA. In the present study, rats were exposed to 20 min nebulizations of normal saline (control group) or aerosolized aqueous solutions of voriconazole at 15.625 mg (low dose group) or 31.25 mg (high dose group). Peak voriconazole concentrations in rat lung tissue and plasma after 3 days of twice daily dosing in the high dose group were 0.85 ± 0.63 μg/g wet lung weight and 0.58 ± 0.30 μg/mL, with low dose group lung and plasma concentrations of 0.38 ± 0.01 μg/g wet lung weight and 0.09 ± 0.06 μg/mL, respectively. Trough plasma concentrations were low but demonstrated some drug accumulation over 21 days of inhaled voriconazole administered twice daily. Following multiple inhaled doses, statistically significant but clinically irrelevant abnormalities in laboratory values were observed. Histopathology also revealed an increase in the number of alveolar macrophages but without inflammation or ulceration of the airway, interstitial changes, or edema. Inhaled voriconazole was well tolerated in a rat model of drug inhalation.