HPLC法测定乙胺吡嗪利福异烟片中4组分含量

目的：建立高效液相色谱法，测定乙胺吡嗪利福异烟片中利福平、异烟肼、吡嗪酰胺和盐酸乙胺丁醇含量。方法：利福平、异烟肼和吡嗪酰胺用C18色谱柱（150mm×4．6mm，5μm），以-0．01M磷酸二氢钾溶液-乙腈为流动相进行梯度洗脱，流速1．0ml·min^-1，检测波长254nm。盐酸乙胺丁醇选用C18色谱柱（150mm×4．6mm，5μm），四氢呋喃-0．4％庚烷磺酸钠（含有0．016％硫酸铜，用磷酸调pH至4．5）（25：75）为流动相，流速1．0ml·min^-1，检测波长258nm。结果：利福平、异烟肼、吡嗪酰胺及盐酸乙胺丁醇线性范围分别为16～160μg·ml^-1（r=0．9999），16—160μg·ml^-1（r=0．9999），53—532μg·ml^-1（r=0．9998），80～320μg·ml^-1（r=0．9998），平均回收率为99．3％～99．5％。结论：方法简便，准确，可用于乙胺吡嗪利福异烟片中各组分的质量控制。     

HPLC法同时测定复方川芎胶囊中阿魏酸和川芎嗪含量

目的：建立用高效液相色谱同时测定复方川芎胶囊中阿魏酸和川芎嗪含量的方法。方法：用Inertsil ODS2 C18柱（150mm×4．6mm，5μm），流动相：甲醇-0．5％冰醋酸溶液（25：75），检测波长：280nm，流速：1．0m1·min^-1。结果：阿魏酸和川芎嗪的线性范围分别为4．4-44μg·ml^-1（r=0．9999）和2．0—20．2μg·ml^-1（r=0．9998），平均回收率分别为100．6％（RSD2．1％）和100．9％（RSD 2．2％）（n=6）。结论：该法简单、快速、重复性好，可用于样品的含量测定及质量控制。     

HPLC法同时测定三七中7种皂苷含量

目的：建立HPLC法同时测定三七中7种皂苷含量的方法。方法：Hypersil BDS C18色谱柱（250mm×4．6mm，5μm）；流动相：乙腈（A）-水（B）二元梯度洗脱；流速：1．0ml·min^-1；检测波长203nm；柱温：室温。结果：三七皂苷R1，人参皂苷Rb1，Rb2，Rc，Rd，Rg1，Re的线性范围分别为5．6～140μg·ml^-1（r=0．9992）5．8～144μg·ml^-1（r=0．9991）、5．9-148μg·ml^-1（r=0．9997）、6．1～152μg·ml^-1（r=0．9992）、5．8～144μg·ml^-1（r=0．9992）、5．8～146μg·ml^-1（r=0．9990）、5．8～144μg·ml^-1（，=0．9994）；回收率98．5％～100．1％。结论：HPLC法可同时并且简单快速分离三七中7种皂苷。     

HPLC法同时测定消炎利胆片中穿心莲内酯和脱水穿心莲内酯的含量

目的：建立消炎利胆片中穿心莲内酯和脱水穿心莲内酯的含量测定方法。方法：以XTerra RPC18色谱柱为分离柱，以甲醇-水（45：55）为流动相，采用HPLC法测定。结果：本品中穿心莲内酯含量测定线性范围为16—160μg·ml^-1（r=0．9999），平均回收率为98．87％（RSD=1．90％，n=6）；脱水穿心莲内酯含量测定线性范围为32～320μg·ml^-1（r=0．9999），平均回收率为99．54％（RSD=1．93％，n=6）。结论：此法操作简单、结果准确，可用于消炎利胆片的质量控制。     

HPLC测定维药祖卡木胶囊中大黄素、大黄酚的含量

目的：采用高效液相色谱法测定祖卡木胶囊中大黄素和大黄酚的含量。方法：采用Waters Symmetry C18（150mm×4．6mm，5μm）色谱柱，流动相：甲醇-0．1％磷酸溶液（80：20）；检测波长：254mm；流速1．0ml·min^-1；柱温35℃。结果：大黄素在0．8—6．6μg·ml^-1（r=0．9999，n=5）范围内，有良好的线性关系，平均回收率为98．8％（n=9），RSD为1．7％。大黄酚在2．0—10．2μg·ml^-1（r=0．9999，n=5）范围内，有良好的线性关系，平均回收率为101．3％（n=9），RSD为1．6％。结论：方法具有操作简单、准确的优点，能有效控制祖卡木胶囊的质量。     

RP-HPLC法同时测定茶碱麻黄碱胶囊中2组分的含量

目的：建立反相高效液相色谱法同时测定茶碱麻黄碱胶囊中茶碱和盐酸麻黄碱含量的方法。方法：色谱柱为C18柱（150mm×4．6mm，5μm），流动相为甲醇-0．06mol·L^-1磷酸二氢钾溶液（用磷酸调节pH至2．5）（13：87），流速为0．8ml·min^-1，检测波长为210nm，柱温35℃。结果：茶碱和盐酸麻黄碱浓度分别在18．4—183．8μg·ml^-1（r=0．9998）和4．1-40．8μg·ml^-1（r=0．9993）范围内与峰面积呈良好的线性关系，平均回收率分别为98．9％（RSD=1．1％）和97．6％（RSD=1．2％）。结论：方法简便、准确、重复性好，可用于该制剂的质量控制。     

HPLC法同时测定雅静胶囊中厄贝沙坦和氢氯噻嗪的含×量

目的：建立HPLC法测定雅静胶囊中厄贝沙坦和氢氯噻嗪的含量。方法：色谱柱：Dianmonsil TMC18柱（250mm×4．6mm,5μm）;流动相：乙腈-0．08mol·L^-1磷酸溶液（用三乙胺调至pH5．0）（40：60）;流速：1ml·min^-1;紫外检测波长：225nm;进样量：20μl;柱温：25℃。结果：厄贝沙坦和氢氯噻嗪分别在15～135μg·ml^-1（r=0．9999）和1．2～10．8μg·ml^-1（r=0．9999）浓度范围内线性关系良好,平均回收率分别为100．0％和100．5％。结论：本方法操作简便、快捷,重复性好,灵敏度高,结果准确可靠,适用于雅静胶囊的质量控制。     

青霉素V钾咀嚼片的药动学和生物等效性研究

目的：研究青霉素V钾咀嚼片在健康人体的药物动力学及生物等效性。方法：19名健康志愿者随机双交叉、单剂量口服受试制剂和参比制剂0．472g，用液相色谱-串联质谱法（LC—MS／MS）测定人血浆中青霉素V的浓度，利用DAS2．0药物动力学软件计算有关药物动力学参数、相对生物利用度，并评价2种制剂的生物等效性。结果：受试制剂和参比制剂的tmax分别为（0．54±0．09）h和（0．53±0．13）h；Cmax分别为（10．64±3．84）μg·ml^-1和（10．87±4．43）μg·ml^-1；t1／2分别为（0．74±0．20）h和（0．75±0．10）h。AUC0-6h分别为（11．92±4．04）μg·ml^-1·h和（12．34±4．17）μg·ml^-1·h；AUC0-∞分别为（12．00±4．04）和（12．43±4．17）μg·ml^-1·h。青霉素V钾咀嚼片的相对生物利用度为（97．9±12．8）％。结论：两种制剂具有生物等效性。     

反相高效液相色谱法测定口腔溃疡涂剂中甲硝唑和盐酸丁卡因的含量

目的：建立以反相高效液相色谱法测定口腔溃疡涂剂中甲硝唑和盐酸丁卡因含量的方法。方法：色谱柱为Thermo ODS hypersilC18（50mm×4．6mm，5μm）；流动相为甲醇-水-乙腈-三乙胺（35：50：15：0．7）（用冰醋酸调pH=6．8）；流速1．0ml·min^-1；检测波长为310nm；柱温20℃。结果：甲硝唑和盐酸丁卡因分别在20．2～161．6μg·ml^-1（r=0．9994）和6．68～53．44μg·ml^-1（r=0．9993）范围内线性关系良好，平均回收率分别为99．89％（RSD0．62％）和99．84％（RSD0．67％）。结论：本法简便、快速、重复性好，可用于口腔溃疡涂剂的质量控制。     

RP-HPLC测定心脑康胶囊中阿魏酸和葛根素的含量

目的：建立心脑康胶囊中阿魏酸和葛根素的RP—HPLC含量测定方法。方法：色谱柱为DIKMA-C18（250mm×4．6mm，5μm），流动相为甲醇-1％的冰醋酸（30：70），流速为1．0ml·min^-1，检测波长为254nm，柱温为室温。结果：阿魏酸和葛根素的的线性范围分别为11．2-67．2μg·ml^-1（r=0．9994），9．0～54μg·ml^-1（r=0．9992），平均回收率分别为98．5％、98．2％，RSD为0．5％、0．8％（n=5）。结论：方法简便，结果准确，可作为心脑康胶囊的质量控制方法。     

Binding of Pyrimethamine to Human Plasma Proteins and Erythrocytes

A high-performance liquid chromatographic (HPLC) assay was developed for pyrimethamine in plasma, red blood cells (RBCs), and buffer for the purpose of studying its plasma protein binding and RBC partitioning. Pyrimethamine (1000 ng/ml) was 94% bound to plasma proteins on average, depending on the pH of plasma. A comparison of the lower and upper range of plasma concentrations that would be achieved after a malaria prophylaxis dosing regimen (25 mg/week) showed that the fraction unbound was significantly lower at 120 ng/ml than at the upper plasma concentration of 360 ng/ml, 3.5 vs 4.9%, respectively. Nonlinear regression of the effect of albumin concentration (g/L) on plasma binding yielded the equation: fraction unbound = 1/[(0.421 * albumin concentration) + 1] (R ² = 0.99). There was no binding to normal levels of alpha₁-acid glycoprotein (AAG). The mean ratio of the concentration of pyrimethamine in RBCs to that in plasma (RBC:plasma ratio) was 0.42, while the mean RBC:buffer ratio was 5.2. Binding to hemolysate did not account for all of the RBC uptake, suggesting that binding to or partitioning into RBC membranes may be important. Because pyrimethamine binding depends on both albumin concentration and pyrimethamine concentration in the plasma, these studies predict greater free fractions of pyrimethamine associated with the higher doses given for toxoplasmosis (75 mg/day) and with the hypoalbuminemia associated with AIDS and malaria.     

An Outbreak of Pyrimethamine Toxicity in Patients with Ischaemic Heart Disease in Pakistan

We investigated an outbreak of darkening of skin, bleeding from multiple sites, leucopenia and thrombocytopenia in ischaemic heart disease patients. Case patients were defined as patients who had received medicines from the pharmacy of Punjab Institute of Cardiology between 1 December 2011 and 12 January 2012 and who developed any one of the following: darkening of skin, bleeding from any site, thrombocytopenia and leucopenia. Clinical and drug‐related data were abstracted. All 664 case patients had received iso‐sorbide‐mono‐nitrate contaminated with about 50 mg of pyrimethamine, and 151 (23%) died. The median age of 117 patients admitted at Jinnah Hospital Lahore was 57 years (range, 37–100) and 92 (79%) were male. The median time from intake of medicine to presentation was 37 days (range 13–72). Symptoms and signs included bleeding (in 95% of the patients), skin hyperpigmentation (in 61%), diarrhoea (in 53%) and abdominal pain (in 48%). At presentation, the median white cell count was 2.3 × 10⁹/L (range, 0.1 × 10⁹–16.0 × 10⁹), the median hemoglobin concentration was 109 g/L (range 58–169) and the median platelet count was 18 × 10⁹/L (range, 0 × 10⁹–318 × 10⁹). Bone marrow examination revealed trileneage dysplasia and severe megaloblastosis. The predictors of mortality included presentation prior to 15 January 2012, age more than 57 years, hypotension and leukocyte count less than 1.5 × 10⁹/L. None of the patients who died received Calcium folinate because all deaths occurred prior to contaminant identification. We describe an outbreak of pyrimethamine toxicity in ischaemic heart disease patients receiving medicines from a single pharmacy due to accidental contamination of iso‐sorbide mono‐nitrate tablets at industrial level. Late recognition of illness resulted in high mortality.     

Pyrimethamine pharmacokinetics and its tissue localization in mice: effect of dose size

The plasma pharmacokinetics and mass fate of [¹⁴ C]pyrimethamine were investigated in the mouse, following dosage with 12.5, 25, 50, and 75 mg kg^?1 (i.p.). Peak plasma concentrations of pyrimethamine were reached between 1 and 2 h and then declined monoexponentially. The mean values for AUC 0 → 30 h increased linearly in relation to the administered dose of pyrimethamine (r = 0.979, P < 0.001). The mean values for intraperitoneal clearance and half-life were not significantly different between dose groups, indicating that the plasma pharmacokinetics of pyrimethamine were independent of dose. The percentage of the administered dose excreted in urine as pyrimethamine (1.3?3.5%) and ¹⁴ C-radioactivity (21.7?29.1%) did not change with increasing dose. In contrast, the cumulative percentage of the dose excreted as ¹⁴ C-radioactivity in faeces (16.7?22.8%) after the three highest doses 25, 50 and 75 mg kg^?1 was significantly less than that seen with the lowest dose of 12.5 mg (50.3%). This suggests extensive biliary excretion of radioactivity, and that the capacity of this process may have been exceeded with the highest doses. Seven days after the administration of each of the three highest doses, a significantly greater percentage of [¹⁴ C]pyrimethamine was localized in the soft tissues; i.e. heart, lung and kidney (7.8?13.8%), gut (5.4?9.4%) and particularly the liver (25.0?27.9%) when compared with the lowest dose of the drug (1.2, 1.0, 0.3% respectively). Following each dose, between 85 and 97% of the administered radioactivity was accounted for. These studies indicate, that with higher doses of pyrimethamine, the parent drug and/or metabolites may accumulate in soft tissue, particularly the liver, but without appreciable effects on the plasma disposition and urinary excretion of the drug.     

Pyrimethamine Impairs Host Resistance to Infection with Listeria monocytogenes in BALB/c Mice

Increased mortality has been observed when HTV-infected patientswere treated with pyrimethamine (Pyr) as prophylaxis for toxoplasmicencephalitis, suggesting that Pyr might possess im-munosuppressiveactivity. To analyze this in an animal model, immune functionwas assessed in BALB/c mice using a battery of in vivo and exvivo assays and an in vivo model of host resistance to Listeriamonocytogenes infection. Treatment for 30 days with 60 mg/kgPyr decreased circulating white blood cell and lymphocyte countsbut not neutrophil, red blood cell, or platelet counts or hemoglobinlevels. Splenic B cell percentages and lipopolysaccha-ride-inducedB cell proliferation decreased significantly after treatmentwith 60 mg/kg Pyr, as did levels of anti-keyhole limpet hemocyanin(KLH) IgM in serum 7 days after immunization with KLH. Anti-KLHIgG levels 14 days after immunization were not affected. Percentagesof splenic T cells and macrophages and T cell proliferationin the presence of concanavalin A or allogeneic cells were notdecreased by Pyr treatment. An ex vivo assay of T-cell-mediatedcytotoxicity was also unaffected. When host resistance to L.monocytogenes infection was assessed, dramatic increases inmortality were observed in Pyr-treated compared to control mice.Increased numbers of L. monocytogenes organisms were observedin liver and spleen of Pyr-treated mice, compared to controls.The reduction in Listeria resistance, which is T cell mediated,contrasts with the fact that no significant changes in T-cell-mediatedimmunity were observed. It is possible that Pyr affects parametersof innate immunity, which were not monitored in this Study.     

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.