杨树花高效液相色谱指纹图谱研究

目的 建立杨树花HPLC指纹图谱以控制杨树花质量。方法 采用HPLC对10批不同产地的杨树花进行指纹图谱构建和方法学考察,运用指纹图谱参数共有峰和相似度进行分析,并对图谱进行聚类分析和主成分分析。结果 建立了杨树花HPLC指纹图谱评价方法,确定了Unitary C₁₈（4.6 mm×250 mm,5µm）色谱柱和乙腈-0.1%甲酸为流动相洗脱系统,在290 nm检测波长下优化了梯度洗脱程序,得到了峰形、分离度较理想的色谱图。通过杨树花HPLC指纹图谱的构建,得到了12个色谱峰,10批杨树花的相似度均在0.9~1.0之间,聚成2类。结论 经方法学考察和统计分析,建立的杨树花HPLC指纹图谱方法稳定、可行。     

宁心宝胶囊指纹图谱研究及4种核苷类成分的含量测定

摘 要 目的： 研究宁心宝胶囊的HPLC指纹图谱,并建立同时测定宁心宝胶囊中尿嘧啶、尿苷、腺嘌呤、腺苷含量的方法。方法: 采用Agilent Zorbax SB Aq C₁₈色谱柱（250 mm×4.6 mm,5 μm）,甲醇 0.05 mol·L^-1磷酸二氢钾水溶液为流动相,梯度洗脱,流速为1.0 ml·min^-1,检测波长为212 nm（指纹图谱）、260 nm（含量测定）,柱温为30 ℃。对10 批宁心宝胶囊进行了指纹图谱及含量测定研究,采用中药色谱指纹图谱相似度评价软件进行分析。结果: 建立了宁心宝胶囊的HPLC 指纹图谱,标定了10个共有色谱峰。4种核苷类成分的分离度良好。结论:此方法简便、可靠,指纹图谱结合含量测定可以更好地控制宁心宝胶囊的内在质量。     

温经汤高效液相色谱指纹图谱的建立

目的 建立古代经典名方温经汤HPLC-DAD指纹图谱检测方法。方法 采用RP-HPLC-DAD法,利用Waters Xselect HSS T₃色谱柱（4.6 mm×250 mm,5 μm）,以乙腈-0.1%甲酸水溶液作为流动相进行梯度洗脱,柱温25℃,流速0.8 ml/min,波长254 nm,选取甘草酸铵为参照峰,分析10批次温经汤水煎液的HPLC指纹图谱,并使用中药色谱指纹图谱相似度评价系统对指纹图谱相似度进行评价。结果 在温经汤水煎液指纹图谱中标定了39个共有峰,指认了6个共有峰,10批样品指纹图谱相似度均>0.98,不同批次间差异较小。结论 该方法灵敏度高、稳定性强,数据准确可靠,基本体现了温经汤的整体化学成分特征,可用于温经汤开发的质量控制。     

酸枣叶HPLC指纹图谱研究

目的 建立酸枣叶药材的HPLC指纹图谱,为其质量标准的研究提供依据。方法 采用Kinetex C₁₈色谱柱（100 mm×2.1 mm,2.6 μm）;流动相为乙腈-0.1%磷酸酸水溶液,梯度洗脱;体积流量为300 μL/min;柱温30℃,检测波长225 nm,进样量10 μL,对12批酸枣叶药材进行了指纹图谱研究,采用中药色谱指纹图谱相似度评价系统（2012版）软件进行分析。结果 12批酸枣叶的HPLC指纹图谱有13个共有峰,其中1个共有峰得到确认,相似度均＞0.85。结论 该方法准确可靠,重复性好,为更好地控制酸枣叶内在质量提供科学依据。     

牛黄清感胶囊HPLC特征指纹图谱研究及多成分含量测定

目的 建立牛黄清感胶囊HPLC指纹图谱，并测定其中7种有效成分的含量。方法 采用HPLC，色谱柱为Agilent SB-C₁₈（250 mm×4.6 mm，5 μm），以乙腈（A）-0.1%磷酸水溶液（B）为流动相进行梯度洗脱；检测波长210 nm；柱温30℃。采用"中药色谱指纹图谱相似度评价系统（2004 A版）"进行相似度分析，并对指认的7个指标成分进行定量测定研究。结果 在特征图谱研究中，共确定牛黄清感胶囊HPLC指纹图谱22个共有峰，通过与对照品比较指认其中7个共有峰分别为绿原酸、咖啡酸、木犀草苷、黄芩苷、黄芩素、汉黄芩苷和汉黄芩素，利用相似度软件对12批样品指纹图谱进行分析，各批样品相似度均>0.90。定量分析条件通过方法学验证，平均加样回收率为99.1%~104.8%，RSD为1.30%~1.88%。结论 所建立的HPLC指纹图谱和含量测定分析方法可用于牛黄清感胶囊质量控制。     

清脂胃舒佐餐茶HPLC法指纹图谱的研究及4个成分的含量测定

目的 建立清脂胃舒佐餐茶水溶性成分的高效液相色谱（HPLC）法指纹图谱,并测定没食子酸、绿原酸、柚皮苷、橙皮苷的含量。方法 采用HPLC法,色谱条件：Kromasil C₁₈柱（250 mm×4.6 mm, 5 μm）;流动相为乙腈-0.1%磷酸水溶液,梯度洗脱;流速为0.8 ml/min;检测波长为300 nm;柱温为35℃。结果 通过中药色谱指纹图谱相似度评价系统（2004A）,确定了清脂胃舒佐餐水溶性成分的HPLC指纹图谱共有模式,标定了26个共有峰,整体相识度为0.934~0.995;并测定了10批制剂中绿原酸、柚皮苷、橙皮苷的含量分别为0.672、0.194、1.247和0.532 mg/g,RSD分别为1.35%、1.89%、0.69%和0.79%。结论 清脂胃舒佐餐水溶性成分的HPLC指纹图谱及其中没食子酸、绿原酸、柚皮苷、橙皮苷的含量检测方法简便、稳定、科学,为本品的质量控制提供了科学依据。     

ASE-HPLC测定仙茅中仙茅苷的含量及其指纹图谱研究

目的 获得仙茅药材的HPLC指纹图谱,同时研究快速萃取仙茅中仙茅苷及其含量测定的方法,为其质量控制提供依据。方法 采用正交试验优选ASE 350快速溶剂萃取系统的最佳提取方法,采用HPLC测定仙茅苷的量,色谱柱为Thermo Syncronis C₁₈（100 mm×3 mm,3 μm）,流动相为乙腈-0.1%磷酸水溶液,梯度洗脱,体积流量为0.5 mL·min^-1,检测波长为285 nm,柱温为40℃。指纹图谱共有模式采用国家药典委员会中药色谱指纹图谱相似度评价系统（2.0版）进行处理分析。结果 采用萃取温度100℃、静态萃取时间5 min、循环提取2次的快速提取方法最优。在指纹图谱研究中,标定了15个共有峰,建立了对照指纹图谱,20批药材与共有模式之间相似性良好,相似度均>0.9。结论 本方法快速、准确,可用于仙茅药材的综合质量评价。     

不同产地锁阳药材HPLC指纹图谱研究及2种黄酮成分含量测定

目的 建立不同产地锁阳药材HPLC指纹图谱,并测定其中2种黄酮成分的含量。方法 采用HPLC-DAD技术,以Dikma Spursil C₁₈色谱柱（4.6 mm×250 mm,5 μm）,甲醇-0.2%甲酸溶液为流动相梯度洗脱,建立锁阳药材的指纹图谱并进行含量测定;采用中药指纹图谱相似度评价系统（2012版）对12批样品进行共有峰确认及相似度评价;通过SPSS 21.0统计软件采用聚类分析（CA）和主成分分析（PCA）对HPLC指纹图谱进行模式识别研究。结果 建立了锁阳药材指纹图谱,12批锁阳药材的相似度均>0.90;确定共有峰22个,并对其中儿茶素、根皮苷含量进行测定,其含量均值分别为0.320,0.057 mg·g^-1。CA将不同批次锁阳药材分为4类,反映了12个不同产区锁阳药材的质量特征;通过PCA筛选出累计贡献率达到89.349%的5个主成分,得到决定锁阳药材质量的4个化学成分。结论 建立的HPLC指纹图谱结合含量测定、CA、PCA方法可以客观、全面、有效地用于锁阳药材的质量评价。     

注射用五味子提取物指纹图谱研究

目的 采用高效液相色谱法建立注射用五味子提取物指纹图谱,并对2015—2016年制备的40批注射用五味子提取物进行指纹图谱相似度分析。方法 试验考察了不同流动相组成和浓度、不同检测波长、不同洗脱方法、不同品牌C₁₈色谱柱（250 mm×4.6 mm,5 μm）、不同流速、不同柱温等条件下供试品溶液的色谱行为,利用中药色谱指纹图谱相似度评价系统对试验所得色谱图进行相似度分析。结果 确定了高效液相色谱检测条件,建立了20批注射用五味子提取物的对照指纹图谱（相似度不低于0.991）,确定了12个共有峰,并对5个共有峰进行了指认,其余20批供试品与对照图谱的相似度均值为0.994。结论 本试验建立的注射用五味子提取物指纹图谱可以为其质量控制提供科学的依据,为完善注射用益气复脉（冻干）质量标准提供数据支持。     

基于聚类分析的牛黄清感胶囊HPLC指纹图谱研究

摘 要 目的：建立牛黄清感胶囊的指纹图谱,采用相似度计算和聚类分析的方法评价牛黄清感胶囊的质量,为其真伪鉴别及质量控制提供依据。方法: 采用HPLC法。色谱柱为Waters Sunfire ODS C18（250 mm×4.6 mm,5 μm）,以乙腈（A） 0.1%磷酸水溶液（B）为流动相,梯度洗脱;检测波长：210 nm,柱温：30℃,进样量：10 μl,对15批牛黄清感胶囊样品进行分析,采用中药色谱指纹图谱相似度评价系统结合聚类分析对其质量进行评价。 结果: 建立了牛黄清感胶囊的指纹图谱,确定了19个共有峰,指认了其中5个色谱峰,15批牛黄清感胶囊与对照图谱的相似度为0.924~0.999,聚类分析可将不同生产时间段的牛黄清感胶囊很好的分为3类,且分类结果与相似度结果一致。 结论: 所建立的牛黄清感胶囊指纹图谱分析方法全面、准确、稳定,结合聚类分析研究可有助于牛黄清感胶囊的整体质量评价,同时为其质量评价提供一种有效手段。     

EFFECT OF POLICOSANOL SUCCESSIVE DOSE INCREASES ON PLATELET AGGREGATION IN HEALTHY VOLUNTEERS

Policosanol is a cholesterol-lowering drug with hypocholesterolemic effects demonstrated in experimental models, healthy volunteers and type II hypercholesterolemic patients. In addition, antiplatelet effects of policosanol have been shown in experimental models and healthy volunteers. The effect of successively increasing doses of policosanol on platelet aggregation was investigated in a randomized, placebo-controlled, double-blind study conducted in 37 healthy volunteers. The volunteers were on a placebo-baseline period (two tablets per day) for 7 days and thereafter they received randomly, under double-blind conditions, placebo or policosanol (10mgday⁻¹) for 7 days. After this period dosage was doubled to 20mgday⁻¹for the next 7 days and then again doubled to 40mgday⁻¹, while the control group received placebo tablets all the time. Platelet aggregation as well as coagulation time was measured at baseline and after each dosing step. Results showed that antiplatelet effects of policosanol were successfully enhanced throughout the study, thus suggesting a dose-dependent relationship. No significant effect was reached during the first dosing period, but significant reductions of epinephrine and ADP-induced platelet aggregation were observed after the second one. Finally, a significant inhibition of platelet aggregation induced by all the agonists was observed at the last dosing step. Coagulation time remained unchanged during the trial.     

NEURAMIDE STIMULATES ADRENOCORTICOTROPIN BUT NOT PROLACTIN RELEASE FROM RAT PITUITARY

Neuramide (NMD), a substance found in crude preparations of porcine stomach extract, is a viral inhibitor that also has putative immunostimulatory effects. The effects of NMD on stress-hormone (ACTH and prolactin—PRL) release were assessed inin vivoandin vitrostudies. In the former, blood levels of corticosterone and PRL were measured in NMD-treated male rats.In vitroexperiments were performed to evaluate the effects of NMD and three of its fractions (obtained with high performance liquid chromatography) on ACTH and PRL release from perfused rat pituitary slices. NMD increased plasma corticosterone levelsin vivoand produced dose-dependent increases inin vitropituitary release of ACTH. No effects on PRL secretion were observedin vivoorin vitro. The stimulatory effects on ACTH release were caused by the NMD fraction with a molecular weight of >5000<10000Da.     

Synthesis and evaluation of 2,6-piperidinedione derivatives as potentially novel compounds with analgesic and other CNS activities

New 2,6-piperidinediones 2_a–g and 4_a–d were prepared by initial condensation of aromatic aldehydes or cycloalkanones with cyanoacetamide to give α-cyanocinnamides l_a–g or cycloalkylidenes 3_a,b which underwent Michae1 addition with ethyl cyanoacetate or diethylmalonate. Compounds 4_a–d were alkylated by various alkyl halides to produce the N-alkylated 2,6-piperidinedione derivatives 5_a–m. Some new selected compounds 2_a–c,f, 4_a–d & 5_e,h,j were pharmacologically evaluated for potential anticonvulsant, sedative and analgesic activities. These compounds exhibited significant anticonvulsant and analgesic effects after a single I.P. administration 100 mg/kg b.wt. . On the other hand all the investigated compounds induced hypnotic activity and prolonged the phenobarbital sodium- induced sleep as compared with the control group and the most potent compound was found to be 2_f.     

INDIRECT PARASYMPATHOMIMETIC ACTIVITY OF THE CLASS III ANTIARRHYTHMIC SUBSTANCE / -SOTALOLIN VITRO: REVERSIBLE INHIBITION OF CHOLINESTERASE ISOENZYMES FROM BLOOD AND THE HUMAN CENTRAL NERVOUS SYSTEM

Inhibitory effects of the class III antiarrhythmic compound / -sotalol on acetylcholinesterase (AChE; EC 3.1.1.7) isoenzymes of both erythrocytes and the human caudate nucleus and on serum cholinesterase (ChE; EC 3.1.1.8) were studiedin vitrousing a spectrophotometric kinetic assay with acetylthiocholine (ASCh) as substrate. Sotalol concentrations in the assays varied from 0.32 to 3.2m . All isoenzymes studied were inhibited by / -sotalol in a reversible and concentration-dependent manner. Double reciprocal plots of the reaction velocity against varying ASCh concentrations revealed that / -sotalol reduced substrate affinity (apparent Michaelis constant, KM, increased) of serum ChE, but did not change the enzyme's maximal rate of ASCh hydrolysis (V_max). Thus, / -sotalol inhibition of serum ChE was of the competitive type (rate constant for reversible competitive inhibition: K_i=0.51m ). In contrast, / sotalol reduced the maximal reaction velocity of the AChE isoenzyme from the central nervous system (caudate nucleus), but had no influence on substrate affinity of the enzyme (K_Mwith ASCh unchanged) indicating purely non-competitive inhibition kinetics (rate constant of reversible non-competitive inhibition: K_i′=0.44m ). / -sotalol inhibition of erythrocyte AChE was of mixed competitive/non-competitive type (K_i=0.31m , K_i′=0.49m ). Non-competitive / -sotalol inhibition of caudate nucleus AChE and the non-competitive component of erythrocyte AChE inhibition cannot be overcome by increased concentrations of the cholinergic transmitter acetylcholine (ACh). Peak / -sotalol plasma levels as described in the literature for both humans (15μ ) and experimental animals (dogs: 18μ ; rats: 260μ ) as well as maximal myocardial concentrations of the substance (dogs: 46μ ; rats: 478μ ) are in the range of about 2% to 100% of the sotalol inhibition rate constants determined in the present paper for cholinesterase isoenzymesin vitro. Thus, / -sotalol inhibition of ACh hydrolysisin vivomay contribute to both the well known antiarrhythmic potential and proarrhythmic side effects of the compound.     

AMELIORIATION OF CHEMICALLY-INDUCED HEPATOCYTE INJURY BY CYCLOSPORINE A

Cyclosporine A, beside its current applications, possesses potential hepatoprotective effects. This study was directed to investigate the effect of Cyclosporine A pretreatment on hepatic injury due to carbon tetrachloride (CCl₄) and -galactosamine. Rats were injected by two successive doses of Cyclosporine A (5mgkg⁻¹day⁻¹). Six hours after the second dose, 1mlkg⁻¹of CCl₄was administered i.p. Effects associated with Cyclosporine A pretreatment were examined by using isolated hepatocytes and hepatocytes that were immobilized and continuously perfused. -Galactosamine (5m ) was added directly to the perfusion medium. After isolation, hepatocytes were examined histologically by light and electron microscopy, immobilized and perfused for further metabolic functional activity evaluation. Cyclosporine A pretreatmentin vivoproduced hepatoameliorative effects of various degrees which were statistically significant as manifested by: (1) an increased trypan blue exclusion after CCl₄; (2) an improved ureagenesis after CCl₄; (3) a reduction in the lipid droplets accumulation in the cytoplasm produced by CCl₄administration; (4) well preserved cytoplasmic organelles as mitochondria, endoplasmic reticulum ER, nuclear chromatin structures that were altered by CCl₄; and (5) an increased hepatocytes survival in the agarose gel matrix, reduction of LD leakage and improvement of ureagenesis after -galactosamine addition to the perfusion medium. The beneficial effect of Cyclosporine A pretreatment in modifying hepatotoxicity of chemical insults merits further studies.     

EFFECTS OF 2-GUANIDINE-4-METHYLQUINAZOLINE ON GASTRIC ACID SECRETION IN RATS

In this study 2-guanidine-4-methylquinazoline (2-GMQ) appeared to decrease basal and stimulated gastric acid secretion, while structurally related compounds as dimethyl- biguanide, cyanoguanidine and 2-cyanoamino-4-methylpyrymidine did not. Thus, there is an antisecretory effect when the biguanide group is associated with a lipophilic structure. The antisecretive effects exerted by 2-GMQ are associated with anti H₂-histamine activity.The anti H₂-histamine nature of the effects of 2-GMQ was confirmed by the capacity of this compound of depressing the chronotropic activity of the isolated guinea pig auricle increased by histamine, as well as relaxant activity in rat uterus contracted by histamine, since both preparations are rich in H₂-histamine receptors.     

Investigation of the prevalence of tetQ, tetX and tetX1 genes in Bacteroides strains with elevated tigecycline minimum inhibitory concentrations

In this study, the antibiotic susceptibilities to tigecycline and tetracycline of 35 selected Bacteroides fragilis group strains were determined by Etest, and the presence of tetQ, tetX, tetX1 and ermF genes was investigated by polymerase chain reaction (PCR). tetQ was detected in all 12 B. fragilis group isolates (100%) exhibiting elevated tigecycline minimum inhibitory concentrations (MICs) (≥8 μg/mL) as well as the 8 strains (100%) with a tigecycline MIC of 4 μg/mL, whilst tetX and tetX1 were present in 15% and 75% of these strains, respectively. All of these strains were fully resistant to tetracycline (MIC ≥ 16 μg/mL). On the other hand, amongst the group of strains with tigecycline MICs < 4 μg/mL (15 isolates), tetQ, tetX and tetX1 were found less frequently (73.3%, 13.3% and 46.7%, respectively). All but two strains harbouring the tetQ gene in this group were non-susceptible to tetracycline, with a MIC > 4 μg/mL. These data suggest that in most cases tigecycline overcomes the tetracycline resistance mechanisms frequently observed in Bacteroides strains. However, the presence of tetX and tetX1 genes in some of the strains exhibiting elevated MICs for tigecycline draws attention to the possible development and spread of resistance to this antibiotic agent amongst Bacteroides strains. The common occurrence of ermF, tetX, tetX1 and tetQ genes together predicted the presence of the CTnDOT-like Bacteroides conjugative transposon in this collection of Bacteroides strains.     

栝楼不同药用部位质量控制的研究进展

瓜蒌子、瓜蒌皮、瓜蒌、天花粉来源于栝楼的不同药用部位,4味药材均为常用的大宗药材,现行版《中国药典》对其制定的质量标准过于简单,无法科学合理地控制其质量。本文对瓜蒌子、瓜蒌皮、瓜蒌、天花粉安全性和有效组分的研究进行综述,明确了相关研究存在的问题并针对问题提出建议,为科学全面的药材及饮片标准的制定提供参考依据。     

不同产地藤茶中二氢杨梅素含量及其与牛蒡子配伍的药效学研究

目的 探究不同产地、部位及加工工艺藤茶中二氢杨梅素含量的差异,及优选藤茶与牛蒡子配伍药效学研究。方法 二氢杨梅素高效液相方法学验证采用Agilent ZORBAX SB-C₁₈柱,流动相为甲醇-0.05 % 磷酸（30∶70）,流速为1 ml/min,检测波长为291 nm,柱温25 ℃。配伍药效验证方法采用大鼠棉球植入致炎及小鼠耳肿胀致炎模型,对大鼠棉球植入实验肉芽肿净量及小鼠耳肿胀率进行观察。结果 方法学验证二氢杨梅素在0.019 9～0.318 mg/ml范围内线性关系良好（r=0.999）,回收率在95.04 %～100.4 %之间,样品在24 h内稳定,该方法重复性较好。配伍药效学验证高剂量优选藤茶与牛蒡子配伍可致大鼠肉芽肿净量及小鼠耳肿胀率均显著低于空白对照组。结论 方法简便准确,二氢杨梅素在不同产地、部位及加工工艺中含量差异较大,其中以贵州省江口县自然晒干的芽尖部位藤茶含量最高。藤茶与牛蒡子配伍可显著改善咽部症状,减轻致炎程度,共同协同达到清咽效果。