复方明目口服液的制备工艺研究

目的：探究以枸杞子和菊花为主药，玫瑰花、麦芽、陈皮为辅药组建的复方明目口服液的最佳制备工艺。方法：以复合多糖提取率和浸膏得率为评价指标，采用单因素试验和正交试验对复方明目口服液的制备工艺进行探究。结果：单因素实验结果可得，复方明目口服液的最佳提取条件为料液比1∶14、提取温度80℃、提取时间2.5 h。正交实验方差分析表明液料比在提取工艺中为显著性因素，其次为提取温度，而提取时间对提取结果影响最小。结论：复方明目口服液的最佳制备工艺为料液比1∶14、提取温度100℃、提取时间2 h，且多糖提取率和浸膏得率分别高达15.28%±0.012%、33.89%±0.024%。     

正交试验优化水提复方慢肝宁工艺研究

目的优化复方慢肝宁中药材的水提工艺,并建立其提取及含量测定方法。方法采用L_9(3~4)正交试验,以复方提取液中人参皂苷Rb1含量、三七皂苷R1含量、丹参酮I含量、柴胡皂苷A含量的综合评分为评价指标,考察加水量、煎煮时间和煎煮次数对水提工艺的影响,并进行验证试验。结果优化的水提工艺为第1次提取加9倍水,提取90 min;第2次提取加7倍水,提取60 min;第3次提取加7倍水,提取60 min。结论优化的水提工艺稳定、可行,可用于复方慢肝宁中各药材的提取,为复方慢肝宁缓释片的制剂开发提供实验依据。     

宁虫素对蔬菜、水稻害虫的毒力及药效特性研究

本研究了复方生物农药宁虫素对蔬菜、水稻害虫的毒力及药效特性,结果表明,该药集速效性、特效性于一体,对蚜虫、褐飞虱的毒力分别为：0.63ppm和2.00ppm。田间试验表明,宁虫素可有效地控制蔬菜蚜虫、小菜蛾、菜青虫、稻飞虱等的危害,防效显。田间使用宁虫素后,平均蛛虱比明显优于噻嗪酮和吡虫啉,生态安全性好。     

絮凝酵母颗粒悬浮床反应器中试及酒精连续发酵工艺的研究

在小型悬浮床反应器研究絮凝颗粒酵母酒精连续发酵工艺的基础上,建立了单釜反应器有效容积0．5m³、二级串联的中试装置。以淀粉糖化液为底物,考察了悬浮床反应器由小试到中试的放大效应,提出了该反应器工业放大时对危险截面表观气速应加以限制的原则,以保证絮凝酵母颗粒不被剪切破坏。进而在平均稀释率为0．10h^-1的条件下．中试装置稳定运转一个月,获得终点发酵液中酒精浓度80～85g／L,残糖低于5.0g／L,设备平均生产酒精8．4g／L·h的结果。     

猴头菌颗粒治疗慢性萎缩性胃炎作用机制研究

目的：观察猴头菌提取物颗粒对大鼠慢性萎缩性胃炎的治疗效果。方法：SD大鼠分为5组,即正常对照组、模型对照组、维酶素治疗组、猴头菌颗粒小剂量组、猴头菌颗粒大剂量组;除正常对照组外,其余大鼠用热盐水灌胃方法制作慢性萎缩性胃炎模型：模型制成后连续给药28天;处死动物后通过制备病理切片和利用试剂盒测定胃粘膜氨基己糖、前列腺素E2（PGE2）含量,由此观察猴头菌提取物颗粒对慢性萎缩性胃炎的治疗作用。结果：猴头菌颗粒治疗组大鼠胃粘膜G、D细胞数量和氨基己糖、PGE2含量高于模型对照组,尤以猴头菌颗粒大剂量组,最为显著（P〈0．01）。结论：猴头菌颗粒对大鼠慢性萎缩性胃炎有明显的治疗作用。     

梅花鹿胎盘中药复方对机体运动能力的影响

梅花鹿胎盘中药复方对机体运动能力的影响谭建华，崔青山，罗基花，王绍，程远国，甄英凯，张万忠（长春农牧大学兽医研究所长春１３００６２）梅花鹿胎盘是大补之品，可补血生精，增强体质。我们制备了以梅花鹿胎盘为主药，辅以适量人参、大枣的中药复方，对其提高机体运...     

正交试验法优选复方四参颗粒剂的提取工艺

本文研究复方四参颗粒剂的提取工艺,采用正交试验法,以丹酚酸B提取率和干膏得率为指标,考察加水量、煎煮时间和煎煮次数对提取效果的影响,确立了复方四参颗粒剂的最佳提取工艺为:加8倍量水,提取2次,第一次时间为2 h,第二次为1 h。在最佳提取工艺条件下,干膏得率为16.58%,丹酚酸B的平均量为22.42mg。与正交试验的结果相符,说明该提取方法合理、稳定、可行。     

复方五味子提取物对力竭游泳大鼠的抗疲劳作用

五味子、黄芪、丹参、枸杞和马齿苋均具有较好的抗氧化活性和抗疲劳功效。然而,尚不清楚该5种中药材组合的应用效果。本研究将这5种植物药材进行混合提取,得到了复方五味子提取物。然后考察了不同剂量的复方五味子提取物对力竭游泳大鼠的运动能力和生化指标的影响。研究显示,复方五味子提取物以剂量依赖性方式提高大鼠的力竭游泳时间(p<0.05)。复方五味子提取物以剂量依赖性方式降低血清Lac和BUN水平并升高NEFA水平(p<0.05)。复方五味子提取物显著升高大鼠体内SOD和CAT水平,并显著降低MDA水平(p<0.05)。复方五味子提取物显著降低大鼠血清CK、AST和ALT水平(p<0.05),并减弱骨骼肌组织病变程度。本研究表明,复方五味子提取物具有较好的抗疲劳作用,可通过减少乳酸堆积、增加脂肪动员、减弱氧化应激损伤来提高机体运动能力。     

中药复方研究的思路和方法

本文阐述了近年来中药复方的药效物质基础（有效成分）,复方药理研究以及复方新药研制的思路和方法,对一些新观点与假设的实现途径进行了综述。     

复方浙贝母药物血清抑制L1210/CDDP细胞增殖与诱导凋亡研究

目的：观察复方浙贝药物血清抑制耐顺铂的小鼠淋巴细胞系（L1210/CDDP）增殖和诱导凋亡效应,并在此基础上探讨复方浙贝颗粒逆转白血病细胞多药耐药机制。方法：MTT法测定不同浓度组复方浙贝颗粒含药血清抑制L1210/CDDP增殖;Annex-in-V/PI双染经流式细胞仪（FCM）定量检测其含药血清诱导L1210/CDDP细胞早期凋亡率。结果：不同剂量复方浙贝颗粒含药血清组均具有抑制L1210/CDDP细胞增殖效应,对其凋亡抑制率分别为：29.27%（高剂量组）、31.78%（中剂量组）、21.20%（低剂量组）、25.31%（阿霉素组）;实验各组的A570nm与空白对照组比较明显下降（P〈0.01）。经FCM双荧光法检测其能够诱导L1210/CDDP细胞早期凋亡,实验各组凋亡率分别为：55.12%（中剂量血清组）、34.23%（低剂量组）、25.69%（高剂量组）,24.65%（空白血清组）。结论：复方浙贝颗粒含药血清能够明显抑制L1210/CDDP细胞增殖,并能诱导其凋亡。     

酸水解蚕蛹制备复合氨基酸的研究

采用硫酸水解法,以蚕蛹制取复合氨基酸产品,得到氨基酸态氮分别为９．０５％和１３．４５％的食用复合氨基粉和精制复合氨基酸粉。食用复合氨基酸粉含有１８种氨基酸,其中必需氨基酸含量为３９．２％。食用复合氨基酸粉的制备方法经工厂小批量生产证实,其工艺简单易行,适合于中小企业采用,该产品的质量优良,生产成本低廉,具有市场竞争力。     

Supercritical fluid chromatography as a method of analysis for the determination of 4-hydroxybenzylglucosinolate degradation products

In the present study analytical and preparative supercritical fluid chromatography (SFC) were used for investigation of myrosinase catalysed degradation of 4-hydroxybenzylglucosinolate (sinalbin). Sinalbin occurs as a major glucosinolate in seeds of Sinapis alba L., in various mustards and other food products. The degradation products were identified and quantified by analysis based on a developed SFC method using a bare silica column. Determinations comprised transformation products of sinalbin, produced both during degradation of isolated sinalbin, and during autolysis of meal from S. alba seeds. The conditions in the developed SFC method were used as basis for the preparative SFC procedure applied for isolation of the components prior to their identification by nuclear magnetic resonance (NMR) spectroscopy. Myrosinase catalysed sinalbin hydrolysis resulted in the reactive 4-hydroxybenzyl isothiocyanate as an initial product at pH values from 3.5 to 7.5 whereas 4-hydroxybenzyl cyanide was one of the major products at low pH values. 4-Hydroxybenzyl isothiocyanate was found to disappear from the aqueous reaction mixtures in a few hours, as it reacted easily with available nucleophilic reagents. 4-Hydroxybenzyl alcohol was found as the product from reaction with water, and with ascorbic acid, 4-hydroxybenzylascorbigen was produced.     

High-yield and high-degree purification of human alpha-fetoprotein produced by adaptation of the human hepatoma cell line Hep G2 in a serum-free medium

The human hepatoma cell line Hep G2 secretes both albumin and alpha-fetoprotein when grown in the presence of serum. The present report describes how adaptation to growth in serum-free medium results in a progressive switch in the expression of the two proteins; i.e., alpha-fetoprotein becomes the main protein secreted while albumin production is greatly reduced. The culture supernatant obtained, being very enriched in the protein, allows the development of a purification procedure by preparative electrophoresis. By this procedure it is possible to easily obtain large amounts of alpha-fetoprotein from a constant and unlimited source. The availability of these protein preparations should improve the reproducibility and the quality of standardization in clinical immunoassays for alpha-fetoprotein and should permit a more accurate study of the structure and biological functions of the protein.     

HPLC-DAD-ELSD Combined Pharmacodynamics and Serum Medicinal Chemistry for Quality Assessment of Huangqi Granule

Objective

To more scientifically and reasonably control the quality of Huangqi Granules, preliminary studies on the pharmacodynamics and serum pharmacochemistry of this medicine were performed. DPPH and MTT experiments showed that water extracts of Huangqi Granules had good antioxidant activity and increased immunity. Timed blood samples collected 5 min, 15 min, and 30 min after oral administration of a set amount of Huangqi Granules were collected and tested using UPLC-ESI-MS/MS. As a result, calycosin-7-O-β-D-glucoside, ononin, calycosin, astragaloside IV, and formononetin were found to exist in rat blood after dosing, indicating that the five chemical compounds might have pharmacological activity, and based on this result, they were designated biomarkers for quality control of Huangqi Granules. Consequently, a simple, rapid and efficient method was developed in the present study for the simultaneous determination of the five characteristic compounds in Huangqi Granules using HPLC-DAD-ELSD.

Materials and Methods

The separation was performed using an Agilent Hypersil ODS column (4.6 × 250 mm, 5 μm) at 30 ℃. The mobile phase was composed of water (solvent A) and acetonitrile (solvent B) with a flow rate of 1 mL/min. The drift tube temperature of the ELSD system was set to 85 ℃, and the nitrogen pressure was 3.5 bar.

Results

All five characteristic compounds had good linear behavior with r2 values greater than 0.9972. The recoveries varied from 96.31% to 101.22%. Subsequently, the developed method was applied to evaluate the quality of Huangqi Granules from different batches, and hierarchical clustering analysis (HCA) was used to analyze the classification of the samples based on the values of the five compounds.

Conclusion

The established HPLC method combined with HCA proved to be effective to evaluate the quality of Huangqi Granules.     

Purification of Cytochrome b5 from Pig Testis Microsomes by Isoelectric Focusing in an Immobiline pH Gradient

Testicular cytochrome b5 was purified by a procedure including preparative isoelectrofocusing. The cytochrome b5 was determined to have an isoelectric point of 4.45 on analytical isoelectric focusing. The purified cytochrome b5 was found to be homogeneous and its molecular weight was estimated to be 16,000 on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The oxidized and reduced forms of the purified preparation exhibited absorption spectra of a typical cytochrome b5. A 69-fold purification was achieved with an overall yield of 6.2%. Following preparation of the microsomes, the purification is accomplished by a two-step procedure utilizing column chromatography and preparative isoelectric focusing.     

Rare sugars and sugar-based synthons by chemo-enzymatic synthesis

The unique catalytic potential of the fungal enzyme pyranose oxidase was demonstrated by preparative conversions of a variety of carbohydrates, and by extensive chemical characterization of the reaction products with NMR spectroscopy. The studies revealed that POx not only oxidizes most substrates very efficiently but also that POx possesses a glycosyl-transfer potential, producing disaccharides from beta-glycosides of higher alcohols. Although most substrates are oxidized by POx at the C-2 position, several substrates are converted into the 3-keto-derivatives. On the basis of these products, strategies are developed for the convenient production of sugar-derived synthons, rare sugars and fine chemicals by combining biotechnical and chemical methods.     

Purification of cytochrome P-450 from pig testis by aniline-sepharose 4B and isoelectric focusing

Testicular cytochrome P-450 was purified by a procedure including preparative isoelectrofocusing. The cytochrome P-450 was determined to have an isoelectric point of 6.47 on analytical isoelectric focusing. The purified cytochrome P-450 was found to be homogeneous and its molecular weight was estimated to be 52,000 on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The carbon monoxide difference spectrum with a peak at 448 nm exhibited absorption spectrum of a typical cytochrome P-450. 284-fold purification was achieved with an yield of 10.6%. Following preparation of the microsomes, the purification is accomplished by a two-step procedure utilizing Aniline-Sepharose 4B column chromatography and preparative isoelectric focusing.     

Systematic isolation and identification of membrane lipid oxidation products

This report discusses the analytical procedure by which it is possible to isolate and identify the oxidation products of cellular and subcellular membrane lipids. The key point of this procedure is the method used for the transmethylation of the lipid material isolated from the tissues. In effect, both the conversion of the glycerides into methyl esters and the reduction of the hydroperoxyl groups into the corresponding hydroxyl groups is performed in one step, without breaking any oxirane rings that may be present. The methyl esters containing functional groups introduced by oxidative processes are separated from the non-modified ones by preparative TLC and are identified by GLC and GC-MS.     

A method for characterization and fractionation of deoxyribonucleic acid by continuous turbidimetry of cetyltrimethylammonium-DNA precipitates

Cetyltrimethylammonium-DNA from solutions containing 1 m NaCl may be precipitated by continuously lowering the NaCl concentration. By monitoring the increasing turbidity of the reaction mixture at 600 nm as a function of dilution, a curve of apparent absorbance versus volume of diluting solution (or decreasing salt concentration) is obtained. Evaluation of these curves allows (a) an estimation of DNA concentration even in the presence of components absorbing at 260 nm; (b) analysis of the molecular weight distribution of DNA samples; (c) separation of native DNA, RNA and proteins on an analytical or preparative level; (d) optimization of yield and purity by simple procedures even with considerable excess of RNA; (e) characterization at the analytical level and partial, eventually complete separation of doublestranded from single-stranded DNA.The controlled fractionation of protein, DNA, and RNA is the basis of a preparation procedure for DNA from practically all biological objects, yielding products of high quality.     

Separation of neutral reducing oligosaccharides derived from glycoproteins by HPLC on a hydroxylated polymeric support

A prototype hydroxylated polymeric support, Glyco-PAK N, was employed to separate a number of neutral reducing oligosaccharides derived from various glycoproteins by the hydrazinolysis/re-N-acetylation/mild acid hydrolysis procedure. General features of the separation properties of the matrix are described. A major advantage of the packing over silica-based supports is its stability, a property which is advantageous both in repetitive analytical chromatography and in the preparative isolation and spectroscopic identification of pure compounds, free of column breakdown products.