苦碟子注射液与常用溶媒配伍的稳定性研究

目的:通过考察苦碟子注射液与常用溶媒的配伍变化,指导临床合理使用苦碟子注射液。方法:将苦碟子注射液与常用溶媒(灭菌注射用水、5%葡萄糖注射液、10%葡萄糖注射液、0.9%氯化钠注射液、复方氯化钠注射液)配伍,定时考察配伍后溶液的外观、pH、不溶性微粒及质量浓度变化情况。结果:室温4 h内,配伍后混合液外观及pH均无明显改变;光照对溶液颜色无明显影响;灭菌注射用水、5%葡萄糖注射液、10%葡萄糖注射液、0.9%氯化钠注射液作为溶媒的不溶性微粒数目均符合2010年版《中国药典》规定;与常用溶媒配伍后,只有与10%葡萄糖注射液配伍时,总黄酮质量浓度未见明显变化。结论:10%葡萄糖注射液为苦碟子注射液最佳溶媒。     

苦杏仁中苦杏仁酶的闪式提取工艺研究

目的 建立苦杏仁中苦杏仁酶的闪式提取工艺,并对其工艺条件进行优化.方法 以苦杏仁酶总活力为指标,考察了氨水浓度、提取液用量、提取时间、中和终点pH值和沉淀时乙醇体积分数对闪式提取工艺的影响:采用二硝基水杨酸(DNS)法测定苦杏仁酶的比活力,并计算其总活力.结果 苦杏仁酶的最佳提取工艺条件为:取50.0 g脱脂苦杏仁粉,...     

经前平口服液纯化工艺研究

目的:优选经前平口服液的纯化工艺。方法:以芍药苷含量和澄清度为指标,分别用60%醇沉、ZTC-Ⅲ型天然澄清剂、壳聚糖和高速离心纯化方法处理经前平提取浓缩液,筛选出醇沉为最佳纯化方法;再采用正交设计法考察乙醇浓度、醇沉时间、醇沉液pH值对芍药苷提取效果的影响,进一步优化醇沉工艺条件。结果:醇浓度60%、醇沉12h、醇沉液pH值7.0为最佳醇沉工艺。结论:该纯化工艺简便、合理、可行,符合经前平口服液工业化大生产的要求。     

HPLC内标法测定鲜蒜中蒜氨酸

目的:建立准确、方便的 HPLC 法测定鲜蒜中蒜氨酸。方法:采用微波灭酶,以利巴韦林为内标物,HPLC 紫外检测,测定鲜蒜中蒜氨酸含量。与美国药典蒜氨酸衍生化 HPLC 测定进行比较。结果:同一鲜蒜样品测定蒜氨酸含量,以微波法灭酶大蒜的含量最高,为1.291%(RSD=1.4%),回收率为98.80(±2.0)%;美国药典方法测定结果为1.287%(RSD=3.6%),回收率为96.03(±2.6)%。结论:HPLC 内标法与美国药典方法测定结果一致,操作步骤简化。     

鸦胆子油乳注射液正交试验优化调配技术及其在不同溶媒中稳定性考察

摘要：目的:优化鸦胆子油乳注射液调配工艺,考察其与0.9%氯化钠注射液、5%葡萄糖注射液、10%葡萄糖注射液及葡萄糖氯化钠注射液配伍的稳定性。方法:通过正交试验优化鸦胆子油乳注射液调配工艺,测定成品输液的pH、不溶性微粒及主要成分油酸、亚油酸的含量变化,考察鸦胆子油乳注射液在不同溶媒中配伍的稳定性。结果:鸦胆子油乳注射液用250 ml0.9%氯化钠注射液稀释效果最佳。4种溶媒配制的成品输液在存放12 h内pH以及不溶性微粒较稳定。油酸、亚油酸的含量在不同溶媒中随着时间的增加均逐渐减少,0～2 h含量减少幅度最小,9～12 h含量减少幅度最大,其中10%葡萄糖注射液中油酸、亚油酸含量均出现10%以上的降解。结论:鸦胆子油乳注射液最佳调配工艺为0.9%氯化钠注射液250 ml作为溶媒稀释10 ml药品溶液;0.9%氯化钠注射液、葡萄糖氯化钠注射液及5%葡萄糖注射液调配的成品输液12 h内稳定,10%葡萄糖注射液调配的成品输液建议现用现配。     

硫酸妥布霉素和葡萄糖酸钙混合注射液的稳定性

大多数细菌感染新生儿同时亦患低血钙症,因而临床上常合并用妥布霉素与葡萄糖酸钙混合注射液。本实验检测妥布霉素在5%葡萄糖注射液以及加有葡萄糖酸钙的5%葡萄糖注射液中的稳定性。第一批试样:取含妥布霉素40 mg/ml 的硫酸妥布霉素注射液625 μl,以5%葡萄糖注射液稀释成5 ml,试样中妥布霉素浓度为5 mg/ml;按同法配制加有浓度为16 mg/ml 的葡萄糖酸钙。第二批试样:取含妥布霉素10 mg/ml 的注射液500 μl,用5%葡萄糖注射液稀释成5 ml,使试样中最终浓度为1 mg/ml;同样配制另     

甲硝唑注射液的配伍实验

甲硝唑具有良好的抗厌氧菌的作用,其临床应用越来越广,与其它药物的配伍越来越多,为合理用药,我们对临床常用的几种配伍做如下稳定性实验: 一、实验材料和仪器甲硝唑注射液(0.5% 批号890422、0.2% 批号890413武汉滨湖制药厂);10%葡萄糖注射液、葡萄糖氯化钠注射液、含钾维持液、平衡盐注射液、氯化钠注射液、均为本院自制;15%氯化钾注射液(批号890310)、维生素C注射液(批号890119)、均为北京第三制药厂;2.6—二氯吲哚酚钠(分析纯、批号841201 北京西中化工厂);硝酸银试液     

附子多糖提取纯化工艺研究

目的:通过对附子多糖类成分的含量测定,探讨附子最佳提取、纯化工艺。方法:采用蒽酮-硫酸法测定附子多糖含量,并应用正交实验筛选最佳提取工艺,应用单因素实验筛选出最佳纯化工艺。结果:附子多糖最佳提取工艺为:附子粗颗粒200g,加10倍量水,浸泡30分钟,煎煮2次,每次2小时,滤布滤过,滤液低速离心(3000转/分,5分钟),取上清液加乙醇使含醇量为80%;纯化工艺为:取沉淀加水溶解,过滤,滤液加乙醇使含醇量为80%,放置24小时,取沉淀用丙酮洗涤三次,每次50mL,沉淀加水溶解后用三氟乙酸法除蛋白3次,挥干溶剂,50℃减压干燥得附子多糖粉末。结论:验证实验证实此种提取纯化工艺是合理的。     

注射用维生素C的质量及配伍稳定性考察

目的:考察注射用维生素C的质量及在5种常用的输液中的稳定性,为临床用药提供参考。方法:采用HPLC法测定注射用维生素C的含量以及相关物质,并考察了注射用维生素C与5%葡萄糖注射液,10%葡萄糖注射液,葡萄糖氯化钠注射液,氯化钠注射液,复方氯化钠注射液配伍后的含量及pH变化。结果:与5种注射液配伍稳定,3 h内外观、维生素C含量、pH值基本不变。结论:注射用维生素C质量合格,与5%葡萄糖注射液,10%葡萄糖注射液,葡萄糖氯化钠注射液,0.9%氯化钠注射液,复方氯化钠注射液输液配伍稳定,临床可以配伍应用。     

牛蒡苷元新制备工艺

目的:优化筛选牛蒡苷元的酶水解条件、提取工艺和大孔树脂型号,得到适合工业生产的工艺流程。方法:以时间、温度、溶媒量为考查因素,用单因素和正交试验设计法,对牛蒡苷元水解条件进行优化;以乙醇浓度、用量、提取次数、提取时间为因素,对提取方法优化;以吸附率和解吸附率对不同型号的大孔树脂优选。结果:牛蒡苷元最佳水解条件为42℃,12倍量水,水解12 h;最佳提取工艺为10倍量40%乙醇提取3次,每次0.5 h;AB-8大孔树脂纯化效果理想。结论:通过对牛蒡苷元的水解提取纯化工艺研究,确定了适合的工业生产的最佳工艺,该工艺方法简单方便,通过该工艺和进一步的硅胶柱层析分离,可得到纯度大于98%的牛蒡苷元。     

Quality of herbal remedies from Allium sativum: differences between alliinase from garlic powder and fresh garlic

Alliinase (EC 4.4.1.4) has been isolated from commercially available garlic (Allium sativum L., Alliaceae) powder and was investigated with respect to its use as ingredient of herbal remedies. The enzyme was purified to apparent homogeneity and results were compared with those obtained from a sample of fresh A. sativum var. pekinense. The purification of the enzyme involved a gel filtration step as well as affinity chromatography on concanavalin-A agarose. Vmax using L-(+)-alliin as substrate (252 mumol min-1 mg-1) was at the lower range of data given in the literature (214-390 mumol min-1 mg-1). L-(-)-Alliin was also accepted as substrate (54 mumol min-1 mg-1). Vmax for alliinase from A. sativum var. pekinense was at 332 mumol min-1 mg-1 and 90 mumol min-1 mg-1 for L-(+)- and L-(-)-alliin, respectively. The Km values for alliinase from garlic powder were estimated to be 1.6 mM for L-(+)-alliin and 2.8 mM for L-(-)-alliin. In contrast to literature values, both temperature and pH optima were somewhat higher (36 degrees C and pH 7.0 versus 33 degrees C and pH 6.5, respectively). The enzyme was found to be active in a range from pH 5 to pH 10. Gel electrophoresis gave evidence that the alliinase obtained from garlic powder consisted of two slightly different subunits with molecular weights of 53 and 54 kDa whereas alliinase obtained from fresh garlic consists of two identical subunits. It is assumed that the alliinase gets significantly altered during the drying process of garlic powder but is still capable to convert alliin to allicin.     

Stabilization and pharmaceutical use of alliinase

In recent years, numerous clinical trials were undertaken in order to elucidate the active principle of garlic (Allium sativum L., Alliaceae). The most prominent effect of garlic preparations is a contribution to the prevention of stroke and arteriosclerosis. Allicin[(2-propenyl)-2-propenethiosulfinate] and other sulfur containing compounds were suggested as active compounds. The extremely unstable allicin itself is liberated from the more stable alliin [S-(+)-2-propenyl-L-cysteine sulfoxide] by the enzyme alliinase (EC 4.4.1.4) if fresh garlic is crunched or garlic powder is moistened. Therefore, an active enzyme is required in alliin containing remedies like those prepared from garlic powder. In order to investigate enzyme stability, alliinase was isolated from garlic powder. The partially purified enzyme could be stabilized over several months by addition of sodium chloride, sucrose, and pyridoxal-5'-phosphate. Alliinase may also be freeze-dried. This allows combinations of synthetic alliin and purified alliinase as components of an acid resistant tablet or capsule. In the intestine, the pro-drug alliin would be enzymatically converted to allicin. In clinical trials, highly dosed preparations of this kind should yield a precise information about the physiological effects of allicin. In addition, alliin-homologues substances which bear a modified alkyl side chain and do not occur in nature may be tested.     

Allicin release under simulated gastrointestinal conditions from garlic powder tablets employed in clinical trials on serum cholesterol

The failure of five recent clinical trials to show significant reduction in elevated serum cholesterol by a single brand of allicin-standardized garlic powder tablets is in contrast to many prior positive studies with the same brand. The hypocholesterolemic activity of garlic is mainly due to allicin, a compound that is produced by the acid-sensitive garlic enzyme, alliinase, only after tablet consumption. Therefore, the allicin-releasing ability of ten lots of these tablets--manufactured over the same years that the positive and negative clinical trials were conducted (1989-1997)--was determined under simulated gastrointestinal dissolution conditions, as defined by U.S. Pharmacopeia Method 724A. It was found that the older lots were more resistant to acid-disintegration (2.5 h vs. 1.3 h, P < 0.001) and that they released three times as much allicin (44% vs. 15 % of their potential, P < 0.001) as the newer lots. A second brand of tablets employed in a recent negative trial released no detectable amount of allicin, while a third set of tablets with high allicin release was used in a trial that gave positive effects. Hence, the persons involved in the recent negative clinical trials probably received considerably less allicin than did those in the older positive studies, possibly accounting for much of the discrepancy in the outcomes. In conclusion, clinical trials using garlic powder tablets to assess any effect of garlic that might be related to allicin, as most are, cannot be considered valid for garlic when the trial shows no effect, unless the expected allicin release from the tablets has at least been determined under standardized drug release conditions (USP 724A).     

Characterization of an Alliin Lyase Preparation from Garlic (Allium sativum)

An alliin lyase (EC 4.4.1.4) preparation from garlic, ALLIUM SATIVUM L., has been purified to apparent homogeneity. The purification procedure involved liquid chromatography steps on hydroxylapatite, on an anion exchanger, and on a chromatofocussing medium. The enzyme protein was characterized by a relative molecular mass of 108,000, and was found to consist of two equal subunits. Its isoelectric point was determined to be 4.9. The enzyme appeared rather thermolabile. Simulated gastric-intestinal passage by a modified "half change test" revealed a high acid lability of the active alliinase protein. K (m)-values for different substrates were in the mM range, and activating energies for the cleavage of different substrates could be determined. A maximal specific activity for synthetic alliin in the range of 490 micromoles per min and mg protein could be achieved at 33 degrees C. There are some significant differences in the characterization of the purified protein compared to results previously reported by others on this enzyme.     

Unusual cystine lyase activity of the enzyme alliinase: direct formation of polysulphides

Garlic ( Allium sativum L.) and related species are highly estimated as foods, spices, and herbal remedies in many parts of the world. Sulphur-containing flavour compounds like allicin (allyl 2-propenethiosulphinate) are responsible for the smell and taste of freshly crunched garlic. These substances are formed by the action of alliinase (EC 4.4.1.4) on cysteine sulphoxides, e. g., alliin [ S-(+)-allyl- L-cysteine sulphoxide]. Additionally, alliinase catalyses the C-S lysis of cystine in the manner of a cystine lyase. Ammonium, pyruvate and elementary sulphur but not cysteine could be detected as reaction products. The ratios between cystine, ammonium and pyruvate are 1 : 1.9 : 1.9 suggesting a new type of reaction mechanism. Thiocysteine and disulphine were assumed as intermediates. The pH optimum of the cystine lyase activity was found at pH 7.5 and the temperature optimum was at 44 degrees C. The KM value for the homogeneous enzyme was at 2.65 mM and Vmax was at 4.12 nkat/mg using cystine as substrate. Moreover, parallel incubation of cystine and alliin gave mainly allyl (poly)sulphides as reaction products instead of allicin. These substances had not been observed as direct enzymatic products until now. Thus, the significance of alliinase and its enzymatic products has to be newly considered in terms of ecological, pharmacological, and biochemical aspects.     

Alliin Lyase from Garlic, Allium sativum: Investigations on Enzyme/Substrate, Enzyme/Inhibitor Interactions, and on a New Coenzyme

A purified alliin lyase (EC 4.4.1.4) from garlic ( ALLIUM SATIVUM L.) has been characterized by kinetic and spectral data under the addition of different substrates, inhibitors, and reducing agents. Hydroxylamine sulfate (50 microM) inhibited the alliinase activity by nearly 90%. Rotenone revealed a similar effect at a concentration of 10 nano-molar. Examination of L-cysteine, and a series of related compounds, on a competitive inhibitory effect on the alliinase-catalyzed alliin cleavage resulted in a list of essential functional groups for substances with this property. Spectral studies on the purified, yellow appearing alliinase enzyme confirmed the existance of an unknown flavinecoenzyme.     

Distinct intraspecific variations of garlic (Allium sativum L.) revealed by the exon–intron sequences of the alliinase gene

Garlic (Allium sativum L.) has been used worldwide as a food and for medicinal purposes since early times. Garlic cultivars exhibit considerable morphological diversity despite the fact that they are mostly sterile and are grown only by vegetative propagation of cloves. Considerable recombination occurs in garlic genomes, including the genes involved in secondary metabolites. We examined the genomic DNAs (gDNAs) from garlic, encoding alliinase, a key enzyme involved in organosulfur metabolism in Allium plants. The 1.7-kb gDNA fragments, covering three exons (2, 3, and 4) and all four introns, were amplified from total DNAs prepared from garlic samples produced in Asia and Europe, leading to 73 sequences in total: Japan (JPN), China (CHN), India (IND), Spain (ESP), and France (FRA). The exon sequences were highly conserved among all the sequences, probably reflecting the fully functional alliinase associated with the flavor quality. Distinct intraspecific variations were detected for all four intron sequences, leading to the haplotype classifications. A close relationship between JPN and CHN was observed for all four introns, whereas IND showed a more divergent distribution. ESP and FRA afforded clearly different variants compared with those from Asian sequences. The present study provides information that could be useful in the development of an additional molecular marker for garlic authentication and quality control.     

生鲜葱白中粗酶的提取研究

目的:研究从生鲜葱白中提取粗酶的方法,为下一步分离纯化和临床应用提供依据。方法:采用葱白中提取的混合底物,用丙酮酸法测定葱白中酶的活力,采用磷酸盐缓冲液提取粗酶,以小牛血清白蛋白为标准,用考马斯亮蓝G-250法测定蛋白质的浓度。结果:pH6.5的Na-K磷酸盐缓冲液+10%甘油+0.2%EDTA,预冷12 h,临用前加5’-磷酸吡哆醛为浸提液所提取粗酶的相对比活度较高,故定该组分为葱白中粗酶提取的浸提液;PEG用量为15%时上清中酶活急剧减小,随后趋于平缓趋势,该浓度下的上清液中的酶的比活度相对最小,故选择该浓度为沉淀蛋白时PEG8000的浓度;随着透析时间的延长,蛋白质溶液中蛋白质含量及酶活性均在下降,透析6 h和24 h的蛋白质溶液中酶活性和蛋白质含量均相差不大,可以说明当透析6 h后,透析基本已经达到平衡。结论:该方法操作简便可以用于生鲜葱白中粗酶的提取。     

Echinocandin B的分离纯化工艺研究

目的开发echinocandin B的分离纯化工艺。方法采用大孔吸附树脂吸附解吸-溶剂萃取-溶剂结晶的分离纯化方法。结果 HZ830树脂为最佳吸附剂,吸附流速选择为2BV/h,洗脱剂为85%乙醇,结晶溶剂为丙酮。在此实验条件下所得产品纯度≥95%,提取总收率大于60%。结论此方法简单,分离效果好,适于工业化生产。     

A high-throughput method for the quantitative determination of alliin

The quality of most garlic (Allium sativum L., Alliaceae) preparations made from garlic powder or garlic dry extract is determined by their content of alliin. Therefore, a comprehensive documentation of alliin concentration beginning with the crude material up to the final remedy is required. The newly developed analytical method described in this paper was designed in order to fulfill these demands. In contrast to conventional HPLC methods, neither a pre-column derivatization nor a chromatographic separation are involved in this analytical procedure allowing a high throughput of samples. The currently investigated technique is based on immobilized alliinase (EC 4.4.1.4) which was combined with a two-channel flow injection analyser (FIA) coupled to an ammonia detecting device. A high specificity for alliin could be demonstrated and a variety of garlic samples including garlic powders, dry extracts, and garlic preparations was analysed. The results were in good correlation with those obtained by conventional HPLC methods.