酶法合成中对羟基苯甘氨酸的检测

建立了酶法合成中对羟基苯甘氨酸的定量检测方法，该检测体系不仅能准确地测定酶法合成过程中产物对羟基苯甘氨酸的含量，而且对底物对羟基本海因能同时进行测定。避免培养液中各成分的干扰，通过标准加入法测定对羟基苯甘氨酸及对羟基苯海因的回收率分别为97．8％～102.1％、98.3％～101.6％。结果表明该检测体系简单、实用，测定结果准确、可靠。     

酶法制备阿莫西林的工艺优化研究

目的 酶法合成阿莫西林工艺优化和稳定性研究.方法 利用对羟基苯甘氨酸甲酯为侧链在青霉素酰化酶催化下,与底物6-氨基青霉烷酸(6-APA)合成阿莫西林;对温度、pH、侧链与底物投料比、投酶量等条件进行优化;取化学法和酶法阿莫西林进行6个月加速实验,对比含量变化,考察成品稳定性.结果 工艺优化后阿莫西林摩尔收率达84.3%;6个月加速实验后酶法阿莫西林含量平均下降值比化学法少0.78%.结论 酶法阿莫西林工艺流程简单,方法绿色环保,杂质少,产品稳定性优于化学法.     

顶空气相色谱法测定阿莫西林钠中残留溶剂

目的：建立阿莫西林钠原料药中有机溶剂残留量的分离测定方法.方法：采用顶空气相色谱法,载气为氮气,FID检测器,色谱柱为HP-1毛细管柱,程序升温,外标法测定了阿莫西林钠中乙醇、异丙醇、二氯甲烷、二乙胺、三乙胺的残留量.结果：5种有机溶剂完全分离,在所考察的浓度范围内具有良好的线性关系,r为0.999 0～0.999 5,平均回收率为92.9%～105.7%,RSD均小于10%,最低检测限为0.004～0.05μg·ml-1.结论：方法快速、灵敏,适用于阿莫西林钠中有机溶剂残留量的测定.     

GC测定注射用阿莫西林钠中残留的有机溶剂

目的 建立注射用阿莫西林钠有机溶剂残留量的测定方法。方法 采用顶空进样毛细管气相色谱法,色谱柱为DB-624毛细管柱(30 m×0.25 mm,1.4 μm),柱温：40 ℃,顶空温度85 ℃,平衡时间：30 min,进样口温度：200 ℃,FID检测器温度：250 ℃,以正丙醇为内标,高纯氮气为载气,测定了注射用阿莫西林钠中7种有机溶剂的残留量。结果 各有机溶剂均能得到有效分离,在所考察的浓度范围内线性关系良好,r为0.999 0~0.999 8,平均回收率为91.3%~105.7%。结论 本方法灵敏、简便、结果准确,可用于注射用阿莫西林钠中残留溶剂的检测。     

GC测定注射用阿莫西林钠中残留的有机溶剂

目的 建立注射用阿莫西林钠有机溶剂残留量的测定方法。方法 采用顶空进样毛细管气相色谱法,色谱柱为DB-624毛细管柱(30 m×0.25 mm,1.4 μm),柱温：40 ℃,顶空温度85 ℃,平衡时间：30 min,进样口温度：200 ℃,FID检测器温度：250 ℃,以正丙醇为内标,高纯氮气为载气,测定了注射用阿莫西林钠中7种有机溶剂的残留量。结果 各有机溶剂均能得到有效分离,在所考察的浓度范围内线性关系良好,r为0.999 0~0.999 8,平均回收率为91.3%~105.7%。结论 本方法灵敏、简便、结果准确,可用于注射用阿莫西林钠中残留溶剂的检测。     

宿主蛋白残留检测及数据分析中存在的问题

目的 探讨宿主蛋白残留检测及数据分析中存在的问题,为相关部门或生物制品生产企业对宿主蛋白残留检测提供依据.方法 结合实例分析在宿主蛋白残留检测及数据分析中存在的问题.结果与结论 在实际分析中应严格按照试剂盒说明书要求对所得数据进行多参数拟和,才能得到准确反映实验结果的曲线,从而回归出准确的结果.     

生物制品中宿主残留蛋白的风险分析和检测方法

新版《中华人民共和国药典》对生物制品的质量要求更高,其中对残留宿主细胞蛋白(HCP)在生物制品中含量的要求也越来越严格.目前HCP研究领域进展迅速.此文对生物制品中宿主残留蛋白的风险分析和检测方法作一综述.     

阿莫西林合成工艺改进

目的 阿莫西林合成工艺的改进。方法 以6-APA为起始原料,工艺路线为6-APA胺盐溶液与混合酸酐反应得阿莫西林。结果 目标产物的结构经元素分析、IR、1H-NMR 、13C-NMR谱数据确证,且质量符合国家2005版药典。结论     

酶法合成胸苷

综述了酶法合成胸苷的反应机理，并列举了６个应用实例。     

阿莫西林合成路线的研究

阿莫西林是广谱抗生素类药物,经光学拆分的（－）－α－苄氧甲酰氨基－对羟基苯乙酸为中间体经缩合再还原制得。     

Detection of drug effects on neurons: protein synthesis inhibition by low doses of amphetamine

The effects of in vivo administered amphetamine (4 and 15 mg/kg) on neuronal and unfractionated brain cortex protein synthesis were evaluated in albino rats. In vitro incubation of brain cortex prisms with 14C-leucine and bulk isolation of neuronal perikarya were performed 1 h after drug administration, and 14C was measured by liquid scintillation counting in hot TCA-insoluble fraction. Amphetamine 15 mg/kg significantly decreased protein synthesis in both fractions, neuronal protein synthesis being more inhibited than total cortex. A lower dose (4 mg/kg) significantly inhibited neuronal protein synthesis, even when it failed to affect total cortex protein synthesis. Results presented here show that neurons may be more sensitive than other brain cells to changes in body temperature. The usefulness of the procedure in the evaluation of drug effects is also considered.     

β-内酰胺抗生素的酶法合成研究进展

β-内酰胺抗生素的酶法合成是21世纪β-内酰胺抗生素发展的必然趋势之一,本文对近年来β-内酰胺抗生素合成研究、产品的分离纯化、酶反应器研究进行概述。     

Inhibition of protein synthesis by trimethyltin

The effects of trimethyltin chloride (TMT) on protein synthesis, measured as the incorporation of [3H]valine into trichloroacetic acid-precipitable material, were investigated in mice. One hour after intraperitoneal administration of a 3.0 mg/kg dose, TMT decreased brain protein synthesis by 47% and also caused a significant decrease (4.2 degrees C) in body temperature. When hypothermia was prevented by maintaining the animals at 35 degrees C, TMT decreased protein synthesis by 20%. Twenty-four hours following administration of TMT, protein synthesis was decreased in brain and liver; however, only a reduction of brain protein synthesis was observed at 48 hr. No hypothermia was present at either time point. A regional study in brain showed that at 24 and 48 hr after TMT administration, protein synthesis was decreased by 18-23% in cerebral cortex and hippocampus but not in cerebellum. TMT also inhibited protein synthesis in vitro in mouse brain homogenates with an IC50 of about 100 microM. Neither SnCl2, nor dimethyltin or monomethyltin had any effect on protein synthesis in vitro. These results suggest that, as for other neurotoxicants such as methyl mercury or acrylamide, inhibition of protein synthesis might be involved in TMT neurotoxicity.     

头孢氨苄酶法缩合工艺的研究进展

综述了青霉素乙酰化固定化酶和头孢氨苄酶法缩合工艺的研究进展;探讨了头孢氨苄酶法缩合工艺中酶催化缩合反应的影响因素、产品的分离与纯化以及过量原料的回收和酶反应器的选择等问题.结果认为头孢氨苄酶缩合反应的适宜条件为侧链与7-氨基脱乙酰氧基头孢烷酸(7-ADCA)的投料比率为11～21,反应底物的浓度为300～600mmol·L-,反应温度在5～35℃之间,7-ADCA的转化率可以达到93%以上.     

Rapid determination of amoxicillin in premixes by HPLC

A rapid analytical procedure for routine identification and quantification of amoxicillin in premixes by high performance liquid chromatography was developed and tested. The ground premix samples were extracted for 10 min using 100ml extraction mixture water-methanol (800:200, v/v). The extract was analyzed by reversed-phase on Agilent Zorbax SB-C18 column (4.6 mm x 150 mm, i.d., 5 microm particle size) with water-methanol-phosphoric acid-triethylamine (842:150:4:4) containing 10 mM hexane-1-sulfonic acid sodium salt (pH 3.5) as mobile phase. UV detection was carried out at 230 nm. The method was validated for specificity, linearity, solution stability, accuracy, precision, limit of detection, and limit of determination. The detector response for amoxicillin was linear over the selected concentration range from 2.0 to 40.0 mg ml(-1) with a correlation coefficient 0.9999. The mean accuracy was 100.1% with a standard deviation of 0.6%. The limit of detection and the limit of determination are 0.1 and 0.3 mg ml(-1), respectively, which corresponds to 10 and 30 mg kg(-1), respectively, in real premix sample. The sample and standard solutions were stable for 4 h. The method is selective and can be used in routine analysis.     

阿莫西林克拉维酸钾口服干混悬剂含量测定方法研究

目的：对阿莫西林克拉维酸钾口服干混悬剂含量测定方法研究。方法：采用高效液相色谱法，在３×３ＣＲＣ１８柱（４ｍｍ×３．５ｃｍ）上，以ｐＨ４．４磷酸二氢钠溶液－甲醇（９５∶５）为流动相，流速１．０ｍｌ·ｍｉｎ－１，检测波长为２２０ｎｍ。结果：阿莫西林和克拉维酸浓度分别在２５～５００μｇ·ｍｌ－１及１０～２００μｇ·ｍｌ－１范围内有良好的线性关系，平均方法回收率分别为９９．４％±１．９％和９９．５％±２．０％，日内精密度分别在１．１％～２．３％和１．７％～２．６％之间，日间精密度分别＜３．１％和＜２．５％。结论：本法实用简便，结果可靠。     

Absolute quantitation of protein therapeutics in biological matrices by enzymatic digestion and LC-MS

The advancement of biotechnology has led to an increase in biotherapeutic drugs, especially recombinant proteins and monoclonal antibodies. Ligand-binding assays or immunoassays are the standard methods of choice in pharmacokinetic studies in support of drug discovery and development for protein therapeutics. LC-MS-based methodologies are increasingly used as alternatives to immunoassays for absolute protein quantitation in biological samples. We review recent advancements in absolute quantitation of protein therapeutics in biological matrices by enzymatic digestion and LC-MS.     

Stimulation of brain-stem protein synthesis by morphine

Rats were intoxicated with morphine as intraperitoneal (i.p.) single doses, or for 4 days (final dose 130 mg/kg b.w.) or for 13 days (final dose 340 mg/kg b.w.) using an ingestion method where intoxicated and control rats received the same amount of calories and fluid. The intoxicated groups showed different degrees of physical dependence, demonstrated by variously expressed abstinence symptoms after withdrawal of the drug or after administration of the opiate receptor antagonist naloxone. Soluble protein synthesis was measured in vivo in brain stem by double labelling with 3H and 14C valine and followed over time in the various rat groups after i.p. morphine injection in different doses. Protein synthesis in astroglial-enriched primary cultures from brain stem and secretion of labelled protein to the serum free incubation medium was also evaluated after morphine treatment. There were dose- and time-dependent effects of morphine on brain stem protein synthesis with an initial decrease and a later increase, 1-3 hr after a single dose of morphine administration. Following a morphine single dose of 25 mg/kg b.w. the stimulation was more rapid in onset and more pronounced in rats with a higher degree of physical dependence. Specific protein fractions including one with a subunit M.W. of approx. 80,000 were identified by electrophoretic separation of labelled proteins. Some similar protein fractions increased in synthesis and were released to the serum-free incubation medium when separating astroglial primary culture proteins after morphine treatment. It might be that the biphasic changes in protein synthesis after morphine administration underlie adaptive phenomena such as tolerance/physical dependence development and that some of the identified proteins including proteins synthesized in astroglial cells and secreted to the incubation media participate in these processes.