基于UHPLC-Q-Orbitrap HRMS技术的益心颗粒化学成分研究

目的 建立一种快速、准确识别益心颗粒中复杂化学成分的定性分析方法。方法 主要通过UHPLC-Q-Orbitrap HRMS捕捉化合物的精准分子量以及多级碎片离子信息,同时将其与对照品的保留时间和质谱信息进行比对,并结合相关参考文献或Chemical Book等数据库信息最终实现对未知化合物的快速定性。结果 从益心颗粒中共鉴定出43种化学成分,主要包括有机酸类、内酯类、木脂素类、甾体皂苷类和其他类。结论 该方法可快速、准确、系统地识别益心颗粒中多种化学成分,并为其质量控制、药效物质基础研究及进一步的临床应用提供扎实的理论依据和科学的研究思路。     

基于UPLC-Q-TOF/MS的渴络欣胶囊化学成分研究

目的 采用超高效液相色谱-四级杆-飞行时间质谱（UPLC-Q-TOF-MS）技术对中药复方渴络欣胶囊中的化学成分进行研究。方法 采用ACQUITY UPLC HSS T3 C₁₈色谱柱（100 mm×2.1 mm,1.8 μm）,以甲醇-0.1%甲酸水溶液为流动相梯度洗脱,体积流量为0.3 mL/min;采用ESI源在正、负离子模式下分别采集数据。根据化合物精确相对分子质量及二级碎片离子信息,结合参考文献数据,鉴定渴络欣胶囊的主要化学成分。结果 从渴络欣胶囊中分离和鉴定出54个化学成分,包括蒽醌类化合物10个、黄酮类化合物10个、萜类化合物7个、酚类化合物16个等。结论 首次采用UPLC-Q-TOF/MS联用技术研究渴络欣胶囊中化学成分,进一步明确了其物质基础。     

基于UPLC-Q-TOF/HRMSE的活血解毒方化学成分定性分析研究

目的 采用超高效液相色谱-四级杆-飞行时间质谱（UPLC-Q-TOF/HRMS^E）技术对活血解毒方中的化学成分进行研究。方法 采用ACQUITY UPLC BEH C₁₈色谱柱（100 mm×2.1 mm,1.8 μm）,以0.1%甲酸溶液（A）和0.1%甲酸乙腈（B）作为流动相进行梯度洗脱,体积流量为0.3 mL/min,采用ESI源在正、负离子模式下采集数据。根据化合物精确相对分子质量及二级特征碎片离子信息,结合参考文献数据与UNIFI数据库,鉴定活血解毒方的主要化学成分。结果 从活血解毒方中鉴定出66个化学成分,其中黄酮类化合物8个、生物碱类化合物7个、酚类化合物6个、皂苷类化合物30个、木质素类化合物6个、萜类化合物4个和其他类化合物5个。结论 建立的UPLC-Q-TOF/HRMS^E联用技术与UNIFI结合法能够系统、快速、准确地鉴定多种化学成分,为其质量评价指标选择及药效物质基础深入研究提供了参考。     

LC-Q-TOF-MS和LC-IT-MS分析桂枝麻黄各半汤中化学成分

目的 对桂枝麻黄各半汤中化学成分进行研究。方法 采用液相色谱-四级杆飞行时间质谱(LC-Q-TOF-MS)和液相色谱-离子阱质谱(LC-IT-MS)对桂枝麻黄各半汤提取物进行分析,色谱柱为Zorbax SB-C₁₈ Rapid Resolution HT,流动相为0.05%甲酸水(A)-乙腈(B),梯度洗脱,流速为0.6 mL·min^-1,柱温30 ℃。结合LC-Q-TOF-MS提供的化合物准确相对分子质量和LC-IT-MS分析获得的化合物多级质谱信息,推测鉴定桂枝麻黄各半汤化学成分结构。结果 共推测鉴定了桂枝麻黄各半汤中119个成分,包括26个黄酮、47个三萜皂苷、11个单萜苷、7个生物碱以及28个其他化合物,其中有25个成分经过与对照品比对确认其结构。结论 基本明确了桂枝麻黄各半汤化学成分,为其质量评价及药效物质研究提供了重要参考。     

宁泌泰胶囊联合坦索罗辛治疗良性前列腺增生症的临床研究

目的 系统研究藤黄健骨胶囊的主要化学成分,并探讨其发挥药效的主要作用机制,为其药效物质研究提供一定的参考依据。方法 采用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱（UHPLC-Q-Orbitrap HRMS）对藤黄健骨胶囊的主要化学成分进行全面分析,根据化合物的一级、二级质谱信息,并与对照品或参考文献进行比对,以实现对药物的化学信息进行快速识别。在此基础上,采用BAT-MAN-TCM数据库对鉴定得到的化学成分进行靶点预测,进一步通过DAVID数据库进行KEGG通路注释分析和GO富集分析,初步筛选出药物的主要药效物质,并探讨其作用机制。结果 从藤黄健骨胶囊中共鉴定出34种化学成分,“成分-靶点”网络分析表明药物中的重要成分山柰酚、大豆苷元、熊果酸、刺芒柄花素和豆甾醇等可作用于Bcl-2、BAX、AKt、PPARG、PTGS1、PTGS2、TNF、IL6、F7及IL1B等关键靶点,结果分析表明破骨细胞分化信号通路、NF-κB信号通路、PI3K-Akt信号通路、肾细胞信号通路以及血小板激活等可能是其发挥壮骨健骨、补肾活血和解痉止痛治疗作用的主要途径。结论 采用UHPLC-Q-Orbitrap HRMS高分辨质谱分析结合网络药理学的方法,初步明确了藤黄健骨胶囊的化学组成及潜在作用机制,为筛选其药效成分及深入阐明作用机制提供了科学的理论依据。     

UPLC-Q-TOF-MS/MS鉴定小儿清咽颗粒化学成分

目的 基于超高效液相色谱-四极杆-飞行时间串联质谱(UPLC-Q-TOF-MS/MS)技术鉴定小儿清咽颗粒中的化学成分。方法 采用Phenomenex Kinetex XB C₁₈色谱柱(2.1 mm×150 mm，1.7 μm)，以0.1%甲酸水-乙腈为流动相体系，梯度洗脱，流速0.25 mL·min^–1，质谱采用电喷雾(ESI)离子源，以正、负离子模式采集多级质谱碎片信息。结果 通过高分辨质谱数据分析，结合参考文献数据以及对照品确认，共鉴别出59个化学成分，包括5个环烯醚萜类，15个有机酸类，5个色原酮类，14个黄酮类，4个木脂素类，9个苯乙醇苷类，2个核苷类，2个酚类及3个其他类，并对化合物的药材来源进行了归属。结论 本研究建立的分析方法灵敏、高效，可应用于小儿清咽颗粒各类化学成分的鉴定分析，为研究小儿清咽颗粒药效物质基础和作用机制提供重要依据。     

基于UHPLC-Q-TOF/MS技术快速分析消痰通腑方化学成分

目的 探明消痰通腑方的主要化学成分,为方剂药效物质基础研究和临床治疗提供科学依据。方法 采用超高效液相色谱四极杆飞行时间质谱法（UHPLC-Q-TOF/MS）对消痰通腑方化学成分进行鉴定,色谱条件为Waters ACQUITY UPLC BEH C₁₈色谱柱（2.1mm×100 mm,1.7 μm）,以 0.1%甲酸水溶液-乙腈为流动相进行梯度洗脱,柱温30 ℃,流速0.3 ml/min,进样量1 μl,检测波长254 nm;质谱条件为电喷雾离子源（ESI）,正、负离子全扫描模式采集质谱数据,根据对照品、自建质谱数据库和在线获得的化合物质谱数据库对色谱峰进行物质鉴定。结果 初步鉴定出55个化合物,分别来自黄连、重楼、大黄、天南星、法半夏。结论 本文所建立的UHPLC-Q-TOF/MS方法能系统、准确地鉴别消痰通腑方的化学成分,可为该方剂的质量标志物筛选和有效成分研究提供参考。     

HPLC-Q-Exactive-MS/MS快速鉴定蒙药三子散化学成分

目的 采用HPLC-Q-Exactive-MS/MS对三子散复方中化学成分进行快速定性分析。方法 SHIMADZU GIST C₁₈色谱柱(4.6 mm×150 mm，5 μm)，以0.1%甲酸水-甲醇为流动相，梯度洗脱，流速0.5 mL·min^-1，柱温30 ℃，正、负离子模式下扫描三子散的一级、二级质谱信息。以总离子流图分析的分子离子峰和碎片离子的质谱信息、分子式、保留时间，结合Chemspider数据库及参考文献查找的分子式和结构式，对三子散中各成分进行定性归属。结果 通过分析各类成分的质谱裂解规律和文献信息，初步推测出95个可能的化学成分，包括39个酚酸类成分、20个鞣质成分、9个有机酸酯类成分、5个单萜类成分、12个环烯醚萜类成分、8个三萜类成分和2个黄酮类成分。其中，57个来自诃子，30个来自栀子，10个来自川楝子，其中芦丁同时来源于诃子、栀子和川楝子。结论 HPLC-Q-Exactive-MS/MS检测方法分离度好、灵敏度高，可快速、高效地推测出三子散中各类成分，为鉴定三子散中的化学成分建立了一种快速、高效的分析方法。     

基于UPLC-Q-TOF/HRMSE小陷胸汤化学成分定性分析

目的 采用超高效液相色谱-四级杆飞行时间质谱（UPLC-Q-TOF/HRMS^E）对小陷胸汤的化学成分进行研究。方法 色谱柱使用Waters ACQUITY UPLC BEH C18色谱柱（2.1 mm×100 mm,1.8 μm）,流动相使用0.1%甲酸水-0.1%甲酸乙腈进行梯度洗脱,柱温为35℃,体积流量为0.3 mL/min,采用电喷雾电离源ESI,在正、负离子模式下分别进行质谱检测分析。结果 根据化合物精确分子量及质谱裂解二级碎片离子,结合相关文献及UNIFI 1.8软件对小陷胸汤进UPLC-Q-TOF/HRMS^E分析,共鉴定得到99个化学成分,正离子模式下55种,负离子模式下44种,初步定性63个化学成分,分别为有机酸类23个,姜辣素类（含姜酮、姜二酮、姜油酮、姜酚、姜烯酚）共9个,黄酮类共8个,生物碱类6个,木脂素类共8个,氨基酸类3个,核苷类3个,其他类型3个。结论 该方法快速、准确、便捷,为小陷胸汤药效物质基础及代谢过程的深入研究奠定了基础,也为其进行质量控制提供了参考。     

五神汤化学成分鉴定及京尼平苷酸、秦皮甲素、咖啡酸、木犀草苷、β-蜕皮甾酮和毛蕊花糖苷定量检测

目的 采用超高效液相色谱串联四极杆静电场轨道阱质谱法（UPLC-Q-Exactive Orbitrap-MS/MS）对五神汤中的化学成分进行鉴定,进而对五神汤中京尼平苷酸、秦皮甲素、咖啡酸、木犀草苷、β-蜕皮甾酮和毛蕊花糖苷 6种成分进行含量测定。方法 UPLC-Q-Exactive Orbitrap-MS/MS 法鉴定五神汤中的化学成分,采用 ACQUIFY UPLCTMBEH C₁₈ 色谱柱（100 mm×2.1 mm,1.7 μm）进行分离,以0.1%甲酸-乙腈为流动相进行梯度洗脱,检测波长为 254 nm,柱温30 ℃;电喷雾离子源（ESI）全扫模式;根据一级质谱中的分子离子,推测化合物的相对分子质量和元素组成;再利用二级质谱信息,与对照品及文献报道中化合物信息库进行比对,推测五神汤的体外化学成分。HPLC 法检测五神汤中京尼平苷酸、秦皮甲素、咖啡酸、木犀草苷、β-蜕皮甾酮和毛蕊花糖苷,Aglient Eclipse XDB-C₁₈色谱柱（150 mm×4.6 mm,5 mm）,流动相为0.1%磷酸水（A）-乙腈（B）,梯度洗脱,检测波长254 nm,体积流量1.0 mL·min^-1,柱温30 ℃,进样量10 μL。结果 共检测到五神汤中的71种化学成分,主要包括黄酮类、酚酸类、三萜皂苷类以及香豆素类;6种成分在浓度范围内线性关系良好,平均加样回收率为89.6%～93.0%。京尼平苷酸、秦皮甲素、咖啡酸、木犀草苷、β-蜕皮甾酮和毛蕊花糖苷在10批样品中的平均质量浓度依次为 2.92、2.48、1.72、1.38、2.08、1.77 mg·g^-1。结论 建立的 UPLC-Q-Exactive Orbitrap-MS/MS、HPLC法可用于五神汤化学成分的鉴定和含量测定。     

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.     

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.