发酵液中小组分环孢菌素D的HPLC测定法

建立了一种采用高效液相测定发酵液中小组分环孢菌素D含量的方法。发酵液经甲醇提取 ,浓缩至干 ,乙醚转移 ,蒸干 ,甲醇定容 ,正已烷脱脂后进样。色谱条件为YWG C1 8柱 (5μm ,4 6mm× 2 50mm) ;乙腈—水 (78∶2 2 ,v/v)为流动相 ;流速 1 0ml/min ;检测波长 2 2 0nm ,柱温 70℃。该方法的精密度、线性关系和回收率均良好 ,且操作方便 ,比较成功地解决了在杂质较多和含量较低情况下环孢菌素D的测定。     

复方氯喘片中三组分的HPLC测定法

用高效液相色谱法同时测定复方氯喘片中三组分的含量。采用μ-BondapakC18柱;流动相为甲醇-0.01mol.L~(-1)磷酸二氢钠(pH=3)(55:45 v/v);紫外检测波长为214 nm;以奈福泮为内标。本法简单、准确。盐酸氯喘、盐酸去氯羟嗪、盐酸溴己胺的平均回收率分别为99.89%、100.21%和99.91%.     

联合检测胃癌患者外周血sCD44v5和CD44v5含量的意义

目的联合检测胃癌患者血清中可溶性CD44变异体5(sCD44v5)含量及外周血淋巴细胞(PBL)CD44变异体5(CD44v5)含量,探讨其意义。方法采用酶联免疫吸附测定(ELISA)法检测40例胃癌患者术前血清中sCD44v5含量。同时应用流式细胞仪(FCM)检测36例胃癌患者术前PBLCD44v5含量并与健康组相对照。结果胃癌血清中sCD44v5含量(平均36.308ng/ml)明显高于正常对照组(平均30.946ng/ml)(P<0.05)。无转移组sCD44v5含量(平均30.981ng/ml)与正常对照组相比,无显著差异(P>0.05)。但是,胃周转移组sCD44v5含量(平均39.811ng/ml)明显高于无转移组(P<0.01)以及远处转移组sCD44v5含量(平均27.733ng/ml,P<0.05)。胃癌患者术前PBLCD44v5含量[(7.138±5.461)%]较正常对照组[(2.741±1.842)%]明显增高(P<0.01)。有转移患者PBLCD44v5含量[(8.130±5.834)%]明显高于无转移者[(4.167±2.159)%](P<0.01)。结论胃癌患者血清中sCD44v5含量及术前PBLCD44v5含量似乎与转移程度关系密切。联合检测其含量可望为判断胃癌患者有无转移、转移程度、制定个体化治疗方案提供参考指标。     

HPLC-UV法测定广东王不留行中槲皮素含量(英文)

本文首次建立了广东王不留行活性成分水解后槲皮素的含量测定方法,为评价广东王不留行的质量和制定其质量标准提供依据。采用HPLC-UV法,以C18反相色谱柱,甲醇–0.4%(v/v)磷酸溶液(50:50,v/v)为流动相等度洗脱,检测波长为360nm。对11批广东王不留行样品进行测定,实验证明该含量测定方法分离效果好、灵敏度高、重复性好,平均回收率为99%,可作为广东王不留行的质量控制方法。     

核磁共振法测定熊去氧胆酸中鹅去氧胆酸的含量

用一般的滴定方法测定熊去氧胆酸中鹅去氧胆酸的含量是很难进行的。NMR方法能有效地进行测定。样品(10～20毫克)溶解在CDCl_3和DMSO-d_6(8:1v/v)的混合溶剂中,马来酸做标准物,以δ=3.8ppm的鹅去氧胆酸C_7质子峰面积计算含量。方法的变异系数是0.45-1.3%,回收率是99.3-100.1%。     

HPLC法测定环孢素胶丸的含量

报道了环孢素胶丸的高效液相色谱测定法。固定相为H_5ODSC_(-18),乙醇-水(60:40,v/v)作流动相。检测波长UV210nm,线性范围10～400μg/ml(r=0.9995),回收率98.2％(n=6)。此法快速、准确的测定胶丸中环孢素含量,且辅料无干扰,测定差异小。     

高效液相色谱-电化学检测器法测定人肝微粒体中雌二醇的代谢产物(英文)

目的:建立雌二醇在人肝微粒体中代谢产物的测定方法,研究低浓度雌二醇在人肝微粒体中的代谢机制。方法:有机溶剂提取、蒸发、重溶的方法处理样品,高效液相色谱法分离组分,电化学检测器测定含量。结果:流动相为醋酸缓冲液-乙腈(50:50,v/v,pH4.5,电压为 0.7V vs Ag/AgCl时,在C18柱上分离较好。最低检测量为100pg。结论:本法可用于测定底物浓度在1-100μmol/L时,人肝微粒体中的代谢产物。     

阿德福韦在HK-2亚细胞组分中的含量测定及线粒体损伤机制研究

通过建立亚细胞组分药物浓度测定方法,探讨阿德福韦(ADV, adefovir)对人肾小管上皮细胞株(HK-2)损伤的机制以及线粒体分裂相关蛋白drp1抑制剂对损伤的保护作用 方法 利用免疫磁性分离法分离亚细胞组分,利用液质联用仪(LC-MS/MS)测定亚细胞组分中的ADV的浓度,利用realtimePCR测定线粒体DNA(mtDNA)的相对含量。结果 HK-2细胞线粒体组分中ADV含量为12.5±3.8 ng/mL,显著高于细胞核组分中ADV含量2.3±1.7ng/mL, 经Mdivi-1预处理的线粒体组分中ADV含量显著降低,而细胞核中组分无影响(P<0.05)。ADV干预后mtDNA相对含量显著低于空白对照组,而Mdivi-1可逆转这一变化。结论 ADV主要分布在HK-2细胞线粒体中；ADV对mtDNA明显抑制作用；线粒体动力学蛋白drp1可逆转ADV对HK-2细胞mtDNA的抑制作用。     

高效液相色谱法测定氯强油搽剂中氯霉素和醋酸泼尼松的含量

目的:建立高效液相色谱法测定氯强油搽剂中氯霉素和醋酸泼尼松的含量。方法:测定氯霉素含量用Spherisorb C18柱(4.6mm×250mm,5μm),流动相为甲醇-0.02mol.L-1磷酸二氢钾(用磷酸调pH至3.0)(60∶40,v/v),流速为0.9mL.min-1,检测波长为242nm;测定醋酸泼尼松含量用C18色谱柱(4.6mm×250mm,5μm),以甲醇-水(60∶40,v/v)为流动相,检测波长为240nm,流速为1.0mL.min-1。结果:氯霉素在100.1~500.4mg.L-1范围内线性关系良好(r=0.999 6),平均回收率为100.2%;醋酸泼尼松在10~150mg.L-1范围内线性关系良好,峰面积积分值与浓度呈良好线性关系(r=0.999 6);平均回收率为99.4%(RSD=1.7%)。结论:高效液相色谱法可以用于该制剂得含量测定和质量控制,方法简便、灵敏、结果准确。     

高效液相色谱法测定泌尿宁颗粒中甘草次酸含量

目的建立高效液相色谱法测定泌尿宁颗粒中甘草次酸含量的方法,以控制泌尿宁颗粒的质量。方法采用高效液相色谱法,Diamonsil C18色谱柱(250mm×4.6mm,5μm),流动相为甲醇-水-冰醋酸(42∶57∶1v/v/v)。流速为lml/min。检测波长为254nm,柱温为室温。结果甘草次酸在0.996～2.998μg/mL范围内峰面积与浓度呈良好的线性关系,r=0.9999(n=5),平均加样回收率为97.9%,RSD为1.30%(n=6)。结论本法简便,准确度和精密度好,可用于泌尿宁颗粒中甘草次酸的含量测定。     

An in-vitro study on the metabolism of rokitamycin and possible interactions of the drug with rat liver microsomes

This in-vitro study was designed to identify the enzyme(s) involved in the major metabolic pathway of rokitamycin, i.e. the formation of leucomycin A7, and to assess possible interactions of the drug with rat liver microsomes. Formation of leucomycin A7 was NADPH-independent and was not appreciably inhibited by anti-rat NADPH cytochrome P-450 reductase serum or cimetidine, a nonspecific inhibitor of cytochrome P-450 isoforms. Eadie-Hofstee plots for the formation of leucomycin A7 were indicative of apparently monophasic behaviour for six rat liver microsomes tested. The mean (+/- s.d.) kinetic parameters, Km, Vmax and Vmax/Km, for the formation of leucomycin A7 from rokitamycin were 47+/-13 microM, 390+/-56 nmol min(-1) (mg protein)(-1) and 8.6+/-1.6 mL min(-1) (mg protein)(-1), respectively. Three esterase inhibitors (100 microM), bis-nitrophenylphosphate, physostigmine and metrifonate inhibited the formation of leucomycin A7 by more than 60%. Metabolism of rokitamycin was inhibited by terfenadine, but not by mequitazine, whereas chlorpheniramine and theophylline activated the formation of leucomycin A7. Rokitamycin, leucomycin A7, leucomycin V, erythromycin and clarithromycin were weak inhibitors of CYP3A-catalysed 3-hydroxylation of quinine with mean IC50 values ranging from 71 to >100 microM. It is concluded that in rat liver microsomes the formation of leucomycin A7 from rokitamycin is catalysed mainly by an esterase (possibly cholinesterase, EC3.1.1.8), but not by cytochrome P-450 enzyme(s). Although in this in-vitro animal study CYP3A activity was barely inhibited by rokitamycin, the possibility cannot be totally discounted in man when rokitamycin is co-administered with drugs metabolized by CYP3A.     

国产柱晶白霉素的临床效果探讨

选择呼吸道、尿道、口腔、耳等部位感染及钩体病非重症者70例(病原学阳性75.7％)。随机以国产柱晶白霉素治疗40例及红霉素治疗30例对照观察。柱晶白霉素组420mg一日三次。3～21日痊愈～显效率76％,无效12.5％,清除革兰氏阳性球菌为90.1％,且对耐红霉素者多敏感,而革兰氏阴性杆菌耐药,对淋球菌效果差,仅有消化道反应7.5％。红霉素组375mg,一日三次,3～30日痊愈～显效率77.4％,无效9.7％,清除革兰氏阳性球菌81.3％,对丙型链球菌和淋球菌效果差,消化道反应为23.3％。显示柱晶白霉素是一种安全、有效的抗生素,特别适于门诊应用。     

Synthesis and biological evaluation of novel leucomycin analogues modified at the C-3 position. I. Epimerization and methylation of the 3-hydroxyl group

The synthesis and biological evaluation of sixteen-membered macrolides modified at the C-3 position are described. 3-Epi-leucomycin A7 (9), 3-O-acyl-3-epi-leucomycin A7 analogues (11a-11e), 3-O-acylleucomycin A7 analogues (13b-13e) and 3-O-methylleucomycin analogues (16a, 16b and 22) were synthesized via fully protected intermediates (7, 5a, 5b and 20). After appropriate modification, subsequent deprotections were performed to furnish a variety of leucomycin analogues. Methylation of the 3-hydroxyl group was found to improve the pharmacoprofile of leucomycin antibiotics. 3-O-Methylrokitamycin (16b) showed enhanced antibacterial activity in vitro and 3,3'-di-O-methyl-4'-O-(3-methylbutyl)leucomycin V (22) exhibited improved metabolic stability in rat plasma in vitro.     

HPLC定量分析麦迪霉素

麦迪霉素是链霉菌发酵的次级代谢产物,在发酵过程中往往产生一系列结构相似、性质相仿的同系物,而当菌种或生产工艺不同时,都会使产品中各组分间的比例有明显不同,从而带来一些质量问题。为了考察国产麦迪霉素的质量,本文较系统地研究了反相高效液相色谱分离和测定麦迪霉素的条件,选用了日立胶3050柱,以甲醇—0.01mol/L磷酸盐缓冲液(pH5.8)(45:55)作流动相,较满意地分离、测定了麦迪霉素。方法快速、准确。     

Structure-activity relationships among the O-acyl derivatives of leucomycin. Correlation of minimal inhibitory concentrations with binding to Escherichia coli ribosomes.

The synthesis, antimicrobial activity, and binding to ribosomes of leucomycin and leucomycin derivatives are described. In general, the binding of the leucomycins and the leucomycin derivatives to ribosomes correlated with their antimicrobial activity. Some 2'-O-acyl derivatives apparently underwent gradual hydrolysis during antimicrobial assays, for their binding to ribosomes was poor compared to their relatively good antimicrobial activies. Correlation between antimicrobial activity and binding to ribosomes, their molecular site of action, provides some insight into the nature of the active molecular moieties.     

自产柱晶白霉素化学结构鉴定的研究

本文对自产的柱晶白霉素不经分离,用综合光谱解析的方法,证明所含各组分的化学结构与已知柱晶白霉素Leucomycin的化学结构相同,与日本进口柱晶白霉素相同,组分A_5含量最高。     

Studies on the metabolic fate of 14C-rokitamycin. Absorption, distribution, metabolism and excretion in infant rats

14C-Rokitamycin (RKM) at the dose of 200 mg/kg was administered orally to fasted infant rats to study the absorption, distribution, metabolism and excretion in infant animals. The mean blood level of 14C-RKM reached its peak of 20.25 micrograms/ml in 30 minutes. The mean area under the curve was 93.23 micrograms.hr/ml. In vivo plasma protein binding rates of 14C-RKM were about 30% in both infant and adult rats. 14C-RKM was distributed at high concentrations into liver, kidney, lung, spleen, pancreas, bone marrow, submaxillary gland and some other tissues. Major metabolites detected in plasma, urine and bile were 10"-OH-RKM, leucomycin A7, leucomycin V and 14-OH-leucomycin V. In excretion studies, about 97% of the administered radioactivity was recovered in urine and feces within 144 hours. After intraduodenal administration to rats with cannulated bile ducts, 7.42% and 25.66% of the radioactivity were excreted within 24 hours in the urine and bile, respectively.     

Studies on the metabolic fate of 14C-rokitamycin. I. Absorption, distribution, metabolism and excretion in rats

Blood concentrations of 14C-rokitamycin (14C-TMS-19-Q) reached their peaks at 1 hour after a single oral (200 mg/kg) administration to male and female rats, and they were 28.0 +/- 0.8 and 24.9 +/- 2.0 micrograms/ml, respectively. No significant differences were observed between male and female in AUC values or maximum blood concentrations. The distribution of TMS-19-Q was good, and concentrations of 14C were high in liver, kidney, spleen, pancreas, adrenal, pituitary gland, thyroid, trachea, exorbital lacrimal gland, submaxillary gland and bone marrow. During the 72 hours period after a single oral (200 mg/kg) administration of 14C-TMS-19-Q to male rats, 8.0 and 89.6% of the dose were excreted in urine and feces, respectively and a total recovery rate was 97.5% of the dose. During the 48 hours period after a single intraduodenal (200 mg/kg) administration of 14C-TMS-19-Q in male rats, 6.9 and 36.2% of the dose were excreted in urine and bile, respectively. Reabsorption of 14C excreted from the bile was negligible. Absorptions of TMS-19-Q from the duodenum, jejunum, ileum and colon were good, but absorption from the stomach was negligible. Major metabolic reactions of TMS-19-Q were deacylation and hydroxylation, and the major metabolites in rats of TMS-19-Q found in the plasma, urine and bile after oral and intraduodenal administration were 10"-OH-TMS-19-Q, leucomycin A7, leucomycin V and 14-OH-leucomycin V.     

Enzymatic phosphorylation of macrolide antibiotics

Five macrolide antibiotics (erythromycin A, 1; oleandomycin, 3a; tylosin, 4a; spiramycins, 5a; leucomycin A3, 6a) have been phosphorylated enzymatically using cell-free extracts derived from Streptomyces coelicolor UC 5240. The necessary cofactors and the rates of the conversion have been determined.     

枝原体和尿原体对左氧氟沙星衍生物(YH7)的体外敏感性研究

采用微量稀释法测定了(S)-9氟-2，3-二氢-3-甲基-10-（4-（2-嘧啶）-1-哌嗪基）-7-氧代-7H-吡啶并[1,2,3-de][1,4]苯并恶嗪-6-羧酸（YH7）等氟喹诺酮及大环内酯类、四环素类抗生素对人型支原体（Mh）、口腔枝原体（M.ora）、唾液枝原体（M.sal）、解脲枝原体（Uu）的体外敏感性。结果显示：YH7对Mh、M、ora、M、sal、Uu的最低抑制浓度（MIC）分别为0.5、0.25、0.125、0.125mg/L。它的抑菌活性是氧氟沙星2-16倍，与红霉素、柱晶白霉素相当。Mh、Uu对YH7及交沙霉素不易产生诱导耐药性，而对红霉素和四环素易产生诱导耐药性。而对红霉素升四环素产生耐药的Mh和Uu菌株对红霉素和四环素交叉耐药而对YH7和交沙霉素无交叉耐药。