HPLC法同时测定复方三维右旋泛酸钙糖浆中4中维生素的含量

目的 建立高效液相色谱(HPLC)法同时测定复方三维右旋泛酸钙糖浆中维生素B₁、维生素B₂、维生素B₆和烟酰胺的含量。方法 采用外标法进行测定,色谱柱为Thermo Betasil C₁₈ Analytical(4.6 mm×250 mm, 5 μm),流动相为乙腈-5 mmol·L^-1十二烷基硫酸钠（含0.05%甲酸）水溶液梯度洗脱,流速1.0 mL·min^-1,检测波长260 nm,柱温30 ℃。分别测定10批复方三维右旋泛酸钙糖浆。结果 维生素B₁、维生素B₂、维生素B₆、烟酰胺4种成分的线性范围分别为5.76~115.2 μg·mL^-1(r=0.999 9)、1.16~23.20 μg·mL^-1(r=0.999 9)、1.72~34.4 μg·mL^-1(r=0.999 6)和5.76~115.2 μg·mL^-1(r=0.999 9),平均加样回收率为96.2%~98.4%(RSD为2.14%~3.42%)。不同批次复方三维右旋泛酸钙糖浆中维生素B₁、维生素B₂、维生素B₆、烟酰胺含量范围分别为0.133 7~0.155 9、0.027 86~0.030 71、0.039 05~0.047 7、0.138 7~0.148 2 mg·g^-1。结论 该方法为完善复方三维右旋泛酸钙糖浆的质量标准和加强质量控制提供了新的方法和依据。     

HPLC法测定复方吡拉西坦脑蛋白水解物片中维生素B1、B2和B6含量

摘 要 目的:建立HPLC法测定复方吡拉西坦脑蛋白水解物片中维生素B₁、维生素B₂和维生素B₆含量的方法。方法: 采用Insteril ODS-3色谱柱（250 mm×4.6 mm,5 μm）,流动相：0.01 mol·L^-1庚烷磺酸钠（含0.25%三乙胺,用冰醋酸调节pH至3.8)-甲醇（75∶25）,柱温30℃,检测波长为280 nm,流速：1.0 ml·min^-1。结果: 维生素B₁、维生素B₂、维生素B₆分别在3.98～99.40 μg·ml^-1（r=0.999 7）、4.08～101.91μg·ml^-1（r=0.999 9）、2.08～52.00 μg·ml^-1（r=0.999 9）范围内线性关系良好,平均回收率分别为99.18%、99.53%、99.27%,RSD分别为0.60%、0.67%、0.71%(n=9)。结论:本法简便、快速、准确,可用于复方吡拉西坦脑蛋白水解物片中维生素B₁、维生素B₂和维生素B₆的含量测定     

联苯双酯对大鼠黄曲霉毒素B1代谢及肝毒性的影响

目的研究抗肝炎药联苯双酯对大鼠黄曲霉毒素B₁代谢和肝毒性的影响。方法大鼠po联苯双酯300 mg·kg^-1·d^-1, 连服3 d后ip黄曲霉毒素B₁1.5 mg·kg^-1。给黄曲霉毒素B₁16 h后测定血清ALT和AST水平,观察联苯双酯对黄曲霉毒素B₁引起肝损伤的保护作用以及对体外代谢的影响。结果联苯双酯(300 mg·kg^-1·d^-1,连服3 d)可明显降低黄曲霉毒素B₁引起的大鼠血清转氨酶升高，增加低毒代谢产物AFM₁的生成。联苯双酯还可增加大鼠肝脏细胞色素P450总量和胞浆GSH含量，诱导P450 2B1介导的PROD和GST的活性。此外，联苯双酯对P450 3A介导的红霉素脱甲基酶和P450 1A介导的EROD也有一定的诱导作用。结论联苯双酯可通过增加大鼠肝脏对AFB₁代谢的解毒功能起到肝保护作用。     

高效液相色谱-光化学衍生法测定湖南产莲子中黄曲霉毒素G1、G2、B1、B2

目的对不同来源的湖南产莲子中黄曲霉毒素G_1、G_2、B_2、B_1进行高效液相色谱-光化学衍生法测定。方法采用高效液相色谱–光化学衍生法,采用岛津GL Inertsil ODS-3色谱柱(250 mm×4.6 mm,5μm),流动相为甲醇–乙腈–水(35∶13∶52),柱温35℃;光化学衍生器(254 nm)激发波长λ_(ex)=360 nm,发射波长λ_(ex)=450 nm。结果黄曲霉毒素G_1、G_2、B_2、B_1分别在6.7～33.3、11.4～56.8、10.3～51.5、4.1～20.3 pg线性关系良好;平均回收率分别为98.07%、97.72%、96.11%、99.52%,RSD值分别为1.69%、1.40%、2.72%、1.34%(n=6)。9批莲子样品中,有6批未检出黄曲霉毒素,来自于农贸市场农户自存的2批次检出黄曲霉毒素B_1,实验室塑料袋包装储存1年的1批检出黄曲霉毒素B_1、B_2,但质量分数均低于法定标准限量。结论不同来源湖南产莲子中黄曲霉毒素质量分数均低于《中国药典》2020年版规定限量。该法测定结果准确、重复性良好,可为完善莲子安全性控制和质量标准提升提供实验依据。     

UHILIC-MS/MS法同时测定复方三维右旋泛酸钙糖浆中5种维生素的含量

目的 建立一种超高效亲水作用色谱串联三重四极杆质谱法（UHILIC-MS/MS）,同时测定复方三维右旋泛酸钙糖浆中维生素B₁、维生素B₂、维生素B₆、烟酰胺和泛酸钙的含量。方法 采用超高效液相色谱仪,Waters ACQUITY BEHHILIC Amide色谱柱（2.1 mm×100 mm,1.7 μm）,以90%乙腈（含0.5%甲酸）-10 mmol/L甲酸铵水（含0.5%甲酸）为流动相,进行梯度洗脱,流速0.30 ml/min;在电喷雾（ESI）正离子模式下,用多反应监测（MRM）模式进行含量测定。结果 在5 min内,样品中5种维生素分别在各自考察的浓度范围内呈良好的线性关系,相关系数（r）均大于0.998 4;整体加样回收率在93.27%~100.39%之间,RSD为1.41%~4.96%;10批样品中维生素B₁、维生素B₂、维生素B₆、烟酰胺、泛酸钙的含量测定结果分别为32.40~38.91、7.002~8.462、9.677~11.17、33.64~39.58、3.276~3.771 mg/250 g。结论 本研究建立的UHILIC-MS/MS方法可快速实现对复方三维右旋泛酸钙糖浆中5种维生素类成分定性鉴定或定量检测,为复方三维右旋泛酸钙糖浆的开发利用和质量评价提供了可靠的技术检测方法。     

高效液相色谱法与荧光光度法检测中药材中黄曲霉毒素的比较

目的比较研究了免疫亲合柱（IAC）净化-HPLC柱后溴化衍生荧光法检测黄曲霉毒素B₁，B₂，G₁和G₂的含量与溴化荧光光度法（SFB法）检测黄曲霉毒素总量的方法。方法在IAC-HPLC柱后溴衍生荧光法中，药材经甲醇-水（70∶30）溶剂系统提取后，采用免疫亲合柱净化、富集黄曲霉毒素，净化后的样品溶液通过高效液相色谱柱分离后，进入柱后衍生管中与过溴化溴化吡啶溶液发生反应，最后进入荧光检测器分别检测黄曲霉毒素B₁，B₂，G₁和G₂含量。在SFB法中，样品经甲醇-水（70∶30）提取后，再经免疫亲合柱净化、富集，在处理后的样品溶液中添加一定量一定浓度的溴水溶液后，迅速置于荧光光度计中读数。结果IAC-HPLC柱后溴衍生荧光法中，考察了3种不同药材中添加2个浓度水平的混合对照品的回收率实验，回收率平均值在93%-97%之间。黄曲霉毒素B₂和G₂的最低检出限为0.06 μg·kg^-1，黄曲霉毒素B₁和G₁的最低检出限为0.20 μg·kg^-1。精密度实验的RSD值在0.8%-1.4%之间。运用此高效液相色谱法检测了39种中药中黄曲霉毒素的含量。同时运用SFB法检测了上述39种中药中黄曲霉毒素的总量。结论SFB法不适合用来检测中药中黄曲霉毒素的总量。     

中药污染黄曲霉毒素B1检测分析

目的:了解中药污染黄曲霉毒素B₁(AFB₁)情况。方法:采用间接竞争酶联免疫吸附法(ELISA)对3 7种中药材和22批中成药污染AFB₁进行检测。结果:有34种中药材污染AFB₁,占92 % ;有19批中成药污染AFB1,占86%。结论:中药材和中成药都不同程度地污染AFB₁。     

HPLC同时测定斑蝥酸钠维生素B6注射液中斑蝥酸钠和维生素B6的含量

目的 建立斑蝥酸钠维生素B₆注射液中斑蝥酸钠和维生素B₆含量的同时测定方法。方法 采用HPLC，色谱柱为Syncronis C₁₈（250 mm×4.6 mm，5 μm），流动相为0.02 mol·L^-1磷酸二氢钾溶液（磷酸调pH值至2.6）-乙腈，梯度洗脱，流速为1.0 mL·min^-1，检测波长为193，290 nm，波长切换法，柱温：30℃。结果 斑蝥酸钠回归方程为Y=1.926X+0.010（r=0.999 7），在0.064 6~0.322 8 mg·mL^-1内斑蝥酸钠与峰面积的线性关系良好，平均回收率为98.9%，RSD为1.2%（n=9）。维生素B₆回归方程为Y=24.153X+0.670（r=0.999 9），在0.535 9~2.679 7 mg·mL^-1内维生素B₆与峰面积的线性关系良好，平均回收率为100.2%，RSD为1.0%（n=9）；仪器精密度、稳定性和重复性试验的RSD均<2.0%。结论 所建立的方法结果准确、重复性好，可用于斑蝥酸钠维生素B₆注射液的质量控制。     

维生素B2和B6的同步荧光分析法及其在维生素复合制剂中的应用

本文建立了维生素B₂和B₆的同步荧光分析法。以△λ=58nm进行同步扫描所得的两个同步荧光峰(以发射波长表示,分别位于526nm和389nm)可用以同时分别定量维生素B₂和B₆。方法快速、灵敏。维生素B₂和B₆的工作曲线线性范围分别为0～1μg/ml和0～1.5μg/ml,检出限分别为0.5ng/ml和1ng/ml。方法已应用于三种复合维生素B制剂中维生素B₂和B₆的分析。     

HPLC-MS/MS测定同济2号颗粒剂中黄芪甲苷、绿原酸、人参皂苷Rg1、人参皂苷Rb1和三七皂苷R1

目的 建立HPLC-MS/MS同时测定同济2号颗粒中5个主要活性成分黄芪甲苷、绿原酸、人参皂苷Rg₁、人参皂苷Rb₁及三七皂苷R₁的方法。方法 以水(0.1%甲酸)-乙腈(0.1%甲酸)为流动相,梯度洗脱,体积流量为0.4 mL/min。YMC-Pack Pro C₈色谱柱分离,采用电喷雾离子源(ESI源),负离子模式,以选择性离子监测模式(SIM)进行测定。监测离子分别是m/z 829(黄芪甲苷)、m/z 353(绿原酸)、m/z 845(人参皂苷Rg₁)、m/z 1 108(人参皂苷Rb₁)、m/z 932(三七皂苷R₁)。结果 黄芪甲苷、绿原酸、人参皂苷Rg₁、人参皂苷Rb₁和三七皂苷R₁分别在0.075～2.4、0.95～30.3、1.71～54.72、1.12～35.92、0.45～14.28 μg/mL与峰面积呈良好线性关系。结论 该方法专属性好,灵敏度高,准确快捷,适用于同济2号颗粒的快速检测,为该药的质量标准提供依据。     

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,5μm)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...     

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.     

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.     

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。