蒙药材蓝盆花指纹图谱及一测多评含量测定方法研究

目的： 建立蒙药材蓝盆花的HPLC指纹图谱及一测多评法同时测定绿原酸、木犀草苷、异绿原酸A及异绿原酸C 4个成分的含量,评价药材质量。方法： 采用Kromasil 100-5-C₁₈色谱柱（250 mm×4.6 mm,5 μm）,以乙腈-0.2%磷酸溶液为流动相,梯度洗脱,流速1.0 mL·min^-1,检测波长分别为326 nm（含量测定）和350 nm（指纹图谱）,柱温30℃。以17批样品建立指纹图谱共有模式。以异绿原酸A为内参物,建立绿原酸、木犀草苷、异绿原酸C与内参物的相对校正因子,计算含量,并与外标法计算结果进行比较。结果： 建立了蓝盆花药材的HPLC指纹图谱,标示了10个共有峰,并指认其中的7个色谱峰,分别为绿原酸、木犀草苷、异绿原酸A、大波斯菊苷、异绿原酸C、木犀草素、芹菜素。相对校正因子重现性良好,以异绿原酸A为内参物采用校正因子计算的含量值与实测值之间无显著差异。结论： 采用指纹图谱与多指标检测模式可为评价蒙药材蓝盆花的质量提供有效方法。     

壮药毛冬青配方颗粒一测多评的定量研究

目的 建立一测多评法同时测定毛冬青配方颗粒中6种酚酸类成分的含量。方法 以绿原酸为内参物,分别建立其与隐绿原酸、咖啡酸、异绿原酸B、异绿原酸A、异绿原酸C的相对校正因子,并利用该相对校正因子计算含量;同时利用外标法测定该6种酚酸的含量,比较2种测定方法的差异,验证一测多评法的准确性和可行性。结果 相对校正因子的重现性良好,8批毛冬青配方颗粒的计算值与实测值无显著性差异。结论 该方法可用于毛冬青配方颗粒中多指标成分质量评价。     

一测多评法在冬菊利咽合剂质量控制中的应用研究

目的 建立一测多评法同时测定冬菊利咽合剂中绿原酸、木犀草苷和3，5-O-二咖啡酰基奎宁酸3种成分的含量。方法 采用HPLC：Agilent TC-C₁₈（2）色谱柱（4.6 mm×250 mm，5 μm），以乙腈-0.1%磷酸水溶液为流动相进行梯度洗脱，流速1.0 mL·min^-1，柱温30℃，检测波长334 nm；以绿原酸为内参物，建立其与木犀草苷、3，5-O-二咖啡酰基奎宁酸之间的相对校正因子（f_s∕i）；采用外标法测定冬菊利咽合剂中绿原酸的含量，通过f_s∕i计算冬菊利咽合剂中其他2种成分的含量，并将一测多评法与外标法测得的实验结果进行t检验，验证一测多评法的准确性。结果 各成分的f_s/i适用性和重复性良好，采用一测多评法和外标法的测定值无显著差异。结论 本实验所建立的一测多评法可用于冬菊利咽合剂的定量分析和质量评价。     

一测多评法测定藏茴香中异绿原酸A、新绿原酸、绿原酸、隐绿原酸、异绿原酸B、异绿原酸C

目的 建立一测多评法（QAMS）测定藏茴香中6种咖啡酰奎宁酸类成分含量,并验证该方法在藏茴香质量评价中应用的可行性与适用性。方法 取藏茴香粉末（过三号筛）0.5 g,精密加入70%甲醇 20 mL ,超声处理（250 W、频率 53 kHz）30 min,制备供试品溶液;采用Phenomenex GeminiR C₁₈（250 mm×4.6 mm,5 μm）色谱柱,以乙腈-0.1%甲酸水溶液为流动相,梯度洗脱,检测波长为 330 nm,体积流量 1.0 mL·min^-1,柱温 30 ℃,进行专属性、供试品提取条件、检测波长选择、色谱条件、线性关系、精密度、重复性、稳定性、加样回收率方法学考察,建立异绿原酸A、新绿原酸、绿原酸、隐绿原酸、异绿原酸 B、异绿原酸 C成分含量检测的 HPLC法;以异绿原酸 A为内参成分,分别计算新绿原酸、绿原酸、隐绿原酸、异绿原酸B、异绿原酸C 5种成分的相对校正因子,分别采用3种不同色谱仪和3种色谱柱进行相对校正因子、相对保留时间耐用性考察,对藏茴香样品同时采用外标法与 QAMS 测定 6 种成分的质量分数,比较 2 种测定方法结果的差异。结果 建立的6种成分的HPLC检测方法的专属性、供试品提取条件、检测波长选择、色谱条件、线性关系、精密度、重复性、稳定性、加样回收率均符合要求;新绿原酸、绿原酸、隐绿原酸、异绿原酸B、异绿原酸C的相对校正因子平均值分别是 1.362、1.257、1.335、1.470、1.134,3种不同色谱仪和 3种色谱柱对相对校正因子、相对保留时间均无明显影响;QAMS与外标法2种方法测定3批藏茴香样品中6种成分得到的结果之间无显著差异。结论 建立的QAMS简便、准确、可靠,可用于藏茴香中6种咖啡酰奎宁酸类成分——异绿原酸A、新绿原酸、绿原酸、隐绿原酸、异绿原酸B、异绿原酸C的定量分析。     

一测多评法同时测定妇乐颗粒中4种成分的含量

目的 采用一测多评法同时测定妇乐颗粒中绿原酸、大黄素、大黄酚与延胡索乙素的含量。方法 采用Agilent ZORBAX Eclipse XDB C₁₈色谱柱（4.6 mm×250 mm,5 μm）,流动相为乙腈（A）-0.2%磷酸水溶液（B）,梯度洗脱：0~8.0 min,85% B;8.0~15.0 min,85%→45% B;15.0~30.0 min,45%→20% B;分段变波长测定：0~8.0 min为327 nm,8.0~15.0 min为254 nm,15.0~30.0 min为280 nm;流速为1.0 ml·min^-1;柱温为30℃。测定大黄素与绿原酸、大黄酚和延胡索乙素间的相对校正因子,考察其重复性,比较计算值与测得值的差异。结果 用一测多评法对6批妇乐颗粒中绿原酸、大黄素、大黄酚与延胡索乙素进行测定,与外标法实测值基本一致。结论 该方法可靠,结果准确,可用于妇乐颗粒的质量控制。     

一测多评法测定复方人参片中的8种苷类成分

目的 建立一测多评法（quantitative analysis single-marker,QAMS）测定复方人参片中8种苷类成分的含量。方法 采用Agilent XDB-C₁₈色谱柱（4.6 mm×250 mm,5 μm）,流动相水（A）-乙腈（B）,梯度洗脱;体积流量1.0 mL·min^-1;检测波长203 nm;柱温：25℃。以人参皂苷Rb₁为内标,计算人参皂苷Rg₁、人参皂苷Re、淫羊藿苷、人参皂苷Rc、人参皂苷Rb₂、人参皂苷Rb₃、人参皂苷Rd的相对校正因子,测定其含量。结果 8种成分在各自范围内线性关系良好（r ≥ 0.999 0）,加样回收率为95.6%~104.4%,RSD为1.22%~2.73%,QAMS测定结果与外标法测定结果无显著性差异。结论 该法准确度、灵敏度高、专属性好、操作简单、重复性好。     

一测多评法测定肉苁蓉中2种有效成分的含量

摘 要 目的： 建立同时测定肉苁蓉中松果菊苷、毛蕊花糖苷的高效液相色谱一测多评法。方法: 以松果菊苷和毛蕊花糖苷为考察指标, 建立两者间的相对校正因子,并用校正因子计算另一个成分的含量,将一测多评法的计算值与外标法实测值进行比较。结果: 建立的相对校正因子重现性良好,采用校正因子计算的含量值和外标法实测值之间没有显著性差异。结论: 同时测定肉苁蓉中2 种成分的一测多评法方法可靠,结果准确,可用于控制肉苁蓉药材及其饮片的质量。     

一测多评法测定8个银翘解毒系列制剂中的7种酚酸类成分的含量

建立8个银翘解毒系列制剂中君药金银花所含的7种酚酸类成分(新绿原酸、绿原酸、隐绿原酸、咖啡酸、异绿原酸B、异绿原酸A、异绿原酸C)的一测多评统一方法。首先,采用HPLC以绿原酸为内参物,建立绿原酸与其他6种酚酸类成分的相对校正因子(RCF),并用不同浓度、不同色谱柱、不同HPLC验证其耐用性,利用金银花提取物作随行对照定位各待测色谱峰。然后利用RCF计算8个不同银翘解毒系列制剂样品中其他6种酚酸类成分的含量,并与外标法的测定结果进行比较,对该方法进行可行性验证。结果表明,所建立的平均相对校正因子的耐用性良好(RSD在0.80%~2.56%),一测多评法测定样品中的酚酸类成分的含量与外标法的含量测定结果一致(相对平均偏差<0.93%)。因此,一测多评法可以成功应用于8个银翘解毒系列制剂中金银花的酚酸类成分的定量控制。     

唐山市协和医院 河北唐山 063000

摘 要 目的：建立基于一测多评法的银杏叶分散片溶出度研究方法。方法: 采用小杯法进行溶出度测定,以0.1 mol·L^-1的盐酸溶液为溶出介质,转速为50 r·min^-1,采用HPLC ELSD梯度洗脱结合“一测多评”法同时测定银杏叶分散片中3个总黄酮成分（山奈素、槲皮素、异鼠李素）和4个总内酯成分（白果内酯和内酯A、B、C）的溶出度。结果: 在一定的线性范围内,山奈素与槲皮素及异鼠李素的相对校正因子分别为：1.873,0.324;白果内酯与内酯A、B、C的相对校正因子分别为：2.280,1.659,1.429。且在不同的实验条件下重现性良好。5批银杏叶分散片溶出度的“一测多评”法计算值与外标法测定值无显著差异,试验所得的校正因子可信;在0. 1 mol·L^-1的盐酸溶液中,银杏叶分散片的溶出均一性良好。结论:该方法简便、准确,重复性好,可用于银杏叶分散片的溶出度测定。     

一测多评法测定华佗豆4种生物碱

目的 建立一测多评法检测华佗豆药材中4种生物碱的量。方法 以华佗豆碱乙为内标,建立华佗豆碱甲、碱丙、碱丁的相对校正因子,在考察相对校正因子的重现性后,进一步采用外标法与一测多评法所计算出的成分含量进行比较。结果 6批华佗豆药材中,各成分采用相对因子计算的量与采用外标法计算的结果无显著差异。结论 一测多评法操作简单、结果准确、可靠,可用于华佗豆4种生物碱的测定。     

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.     

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.     

鼻渊净胶囊的HPLC指纹图谱研究

目的 建立鼻渊净胶囊的高效液相色谱（HPLC）指纹图谱。方法 采用Agilent SB-C₁₈（4.6 mm×250 mm,5 μm）色谱柱,乙腈-水为流动相、以1.0 ml/min流速行梯度洗脱,检测波长210 nm,柱温30 ℃,洗脱时间为80 min。采用中药色谱指纹图谱相似度评价系统（2004A版）对检测出色谱进行指纹图谱相似度评价。结果 建立了鼻渊净胶囊的HPLC指纹图谱,确定了20个共有峰,15个峰归属到各药材,其中5个峰确认了化学成分;10批样品的指纹图谱的整体相似度与对照图谱比较,均在90%以上。结论 所建立的鼻渊净胶囊指纹图谱有助于从整体上控制该制剂的质量。     

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.     

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.     

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.     

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。     

喙果黑面神化学成分研究

目的研究大戟科植物喙果黑面神(Breynia rostrata Merr.)的化学成分。方法利用硅胶、凝胶等色谱技术分离纯化化学成分,根据化合物的理化性质和光谱数据进行结构鉴定。结果从喙果黑面神的正丁醇萃取部分分离得到4个化合物,分别鉴定为6-O-甲基丙酰基-α-D-吡喃葡糖(6-O-methylpropanoyl-α-D-glucopyranose,1);4″-苯酚基-6-O-甲基丙酰基-β-D-吡喃葡糖苷(4″-phenolic-6-O-methylpropanoyl-β-D-glucopyranoside,2);1-O-没食子酰基-β-D-吡喃葡糖苷(1-O-galloyl-β-D-glucopyranoside,3);熊果苷(arbutin,4)。结论化合物1和2为新化合物,3和4均为首次从该种植物分离得到。     

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.