绿藻多糖CH1-1的可控降解及其寡糖的制备研究

目的 研究绿藻多糖CH1-1的可控降解及其寡糖的制备。方法 采用不同浓度的三氟乙酸（TFA）或盐酸（HCl）对绿藻多糖CH1-1进行水解,通过薄层色谱和凝胶渗透色谱法研究了水解过程中不同的反应温度和反应时间对多糖降解产物的影响;通过控制酸浓度、反应温度和反应时间,通过Bio-Gel P4凝胶色谱柱分离得到寡糖产物;采用质谱法对得到的寡糖产物进行分析。结果 采用0.1 mol?L-1 和0.05 mol?L-1 TFA或HCl,随着反应温度的升高和反应时间的延长,多糖的分子量降低,其中0.1 mol?L-1 TFA、80 ℃及0.1 mol?L-1 HCl、80 ℃水解条件对多糖的降解作用较好;最终选取0.1 mol?L-1 HCl、80 ℃、3 h的水解条件对多糖进行降解制备寡糖,并采用Bio-Gel P4凝胶色谱柱对寡糖混合物进行分离纯化,电喷雾质谱分析表明,这些寡糖组分为聚合度为1～8的硫酸阿拉伯糖。结论 建立了绿藻多糖CH1-1可控降解方法,制备得到新颖的海洋硫酸寡糖。     

海洋硫酸鼠李寡糖的制备研究

目的 探究海洋硫酸鼠李寡糖的制备方法并对所得寡糖的特征进行了研究。方法 采用三氟乙酸(TFA)和盐酸(HCl)对海洋硫酸鼠李多糖进行降解,通过薄层色谱法和凝胶渗透色谱法对降解条件进行分析,确定寡糖的制备方法;制备得到一系列硫酸鼠李寡糖,采用质谱法对其特征进行研究。结果 0.1 mol.L-1 TFA 80℃及0.1 mol.L-1 HCl 80℃水解条件对多糖的降解作用较好,且通过Bio-Gel P4凝胶色谱柱对寡糖能进行有效分离;最终选取0.1 mol.L-1 HCl 80℃10 h的水解条件对硫酸鼠李多糖进行降解,采用Bio-Gel P4凝胶色谱柱对寡糖混合物进行分离纯化,负离子模式下的电喷雾质谱分析表明,这些寡糖为聚合度为1–11的硫酸化鼠李寡糖。结论 首次建立了海洋新型硫酸鼠李寡糖的制备方法,这些硫酸鼠李寡糖为新型海洋特征寡糖。     

不同降解方法制备褐藻胶甘露糖醛酸寡糖的结构特点

综述了以不同降解方法制备的甘露糖醛酸寡糖的结构特点,及其甘露糖醛酸寡糖在质谱、核磁共振氢谱及碳谱等谱图上的特征。     

LC-ESI-MSMS表征瑞戈非尼的强制降解产物

摘 要 目的：采用LC ESI MSMS对瑞戈非尼原料药中的杂质进行表征。方法: 对瑞戈非尼原料药HPLC色谱系统进行优化,液相色谱条件为ACE Aq C18色谱柱（250 mm×4.6 mm,5 μm）,以乙腈 0.1%磷酸水为流动相,梯度洗脱,流速为1.0 ml·min-1,检测波长为254 nm,柱温为30℃。液质联用色谱条件为BEH C18 (100 mm×2.1 mm,1.7 μm)色谱柱,流动相A为25 mmol·L-1甲酸溶液,流动相B为乙腈,梯度洗脱,流速为0.3 ml·min-1,电喷雾电离源（ESI）,扫描模式为全扫（SCAN）与碰撞诱导解离（CID）,正离子模式,离子源温度300℃,气体流速12 L· min-1,毛细管电压：0.8 kV。结果: 瑞戈非尼强制降解杂质分离良好,通过一级质谱与二级质谱碎片离子推测出瑞戈非尼5个强制降解产物的结构。     

黄芪活性多糖APS-Ⅱ的酸降解及其降解寡糖的结构和免疫活性研究

课题组前期研究发现黄芪免疫活性多糖APS-Ⅱ具有较强的免疫活性,鉴于其分子量较大且结构复杂,本文旨在建立APS-Ⅱ酸降解的最优方法,对降解产物进行结构初探与分离制备以后,通过体外免疫细胞实验筛选免疫活性最强的寡糖活性中心.本研究以APS-Ⅱ为研究对象,采用自上而下的研究策略,首先通过单因素试验和正交试验优化了APS...     

海带硫酸多糖的降解、分离纯化及理化性质分析

文章采用自由基方法降解海带硫酸多糖，然后通过琼脂糖凝胶电泳、凝胶色谱等方法进行分离纯化；分别利用硫酸酚法和明胶比浊法，测定糖的含量和硫酸根的含量；MN-纤维素膜层析和气相色谱测定单糖组成；利用超滤和凝胶过滤测定降解多糖的分子质量等。海带硫酸多糖可以被自由基有效地降解，降解之后海带硫酸多糖的多糖含量、硫酸根含量及单糖组成基本没有变化，分子质量可被降解到10000-15000左右。     

苯唑西林钠碱性降解物的分析研究

目的:应用液相色谱-三重四极杆质谱分析苯唑西林钠的碱性降解物。方法:采用C18色谱柱(250 mm×4.6 mm,5μm);流动相:0.01 mol.L-1醋酸铵溶液(醋酸调pH至5.0)-乙腈(75∶25);流速:1.0 mL.m in-1;检测波长:225 nm;柱温:30℃;液相出口分流20%进入四极杆质谱,以电喷雾电离源正离子模式(ESI+)进行质谱数据采集。结果:得到苯唑西林钠碱性降解物液相色谱图以及液相色谱峰对应的一级质谱图。对谱图进行分析归纳,推测苯唑西林钠碱性降解物中4个主要组分分别为苯唑西林的青霉噻唑酸及其异构体、苯唑西林的脱羧青霉噻唑酸以及6-APA与苯甲异唑酰氯的缔合物。结论:本方法灵敏、可靠、快速,为杂质控制和工艺改进提供了有力帮助,对苯唑西林钠质量的提高及杂质含量控制指标的制订具有重要的指导意义。     

蛋白质及多肽的液相色谱-电喷雾离子化质谱研究进展

蛋白质及多肽的液相色谱电喷雾离子化质谱研究进展徐友宣彭师奇（北京医科大学生源药物化学中德联合实验室，北京１０００８３）多肽及蛋白质药物对临床的重要性，已引起各国药物化学界的广泛重视。在多肽或蛋白质药物的结构研究中，ＭＳ研究有重要价值。例如ＦＡＢＭ...     

微探针电喷雾串联质谱与液相色谱-电喷雾串联质谱的基质效应对比研究

电喷雾电离(electrospray ionization, ESI)易受基质干扰,影响其定量的准确度、精密度和稳定性。探针电喷雾电离(probe electrospray ionization, PESI)是原位电离的代表之一,无色谱分离,无复杂样品前处理,具有简便、快速、高通量等诸多优点。微探针电喷雾串联质谱(micro pen electrospray ionization tandem mass spectrometry,μPen-ESI-MS/MS)是基于现有的PESI开发的新技术。本文旨在评价μPen-ESI-MS/MS方法用于血浆中药物定量分析的基质效应,并与液相色谱-电喷雾串联质谱(liquid chromatography coupled with electrospray ionization tandem mass spectrometry, LC-ESI-MS/MS)方法的基质效应进行比较。分别建立5种药物血浆样品的μPen-ESI-MS/MS和LC-ESI-MS/MS方法,计算两种方法的基质因子及内标归一化的基质因子。结果表明,他克莫司、氟桂利嗪和地氯雷他定的...     

一种寡糖胶囊剂的制备及升高白细胞作用的研究

摘要：目的：制备低聚甘露糖醛酸钠（DPM207）胶囊剂,对其质量进行考察,并初步评价其改善白细胞减少的效果。方法：采用湿法制粒,以辅料的种类、用量为考察因素,以吸湿性、休止角、颗粒得率等为评价指标,优化胶囊剂的制备工艺。根据《中国药典》等相关规定,对胶囊剂的水分含量、装量差异、溶出度、含量等进行测定。采用辐照所致白细胞减少的动物模型对DPM207升高白细胞的效果进行评价。结果：DPM207胶囊剂的最佳成型工艺为：低聚甘露糖醛酸钠与微晶纤维素按照1:1混合,添加70%乙醇进行湿法制粒,制备的胶囊水分、装量差异、溶出度等均符合2020版《中国药典》规定。在小鼠灌胃给药5日后,DPM207不同剂量组均显著提高血液中白细胞的数量。结论：DPM207胶囊剂成型工艺稳定,胶囊质量可控,具有作为升高白细胞药物的潜力。     

Structural features of a pectic polysaccharide from mulberry leaves

A pectic polysaccharide SDA was obtained from the boiling water extract of mulberry leaves. It consisted of rhamnose, arabinose, xylose, glucose, galactose, and galacturonic acid units in the molar ratio of 5:4:1:2:6:38. Its molecular weight was determined to be 1.5 × 10⁴ by high-performance gel filtration chromatography. A combination of linkage analysis, partial acid hydrolysis, ESIMS, ¹H-NMR, and ¹³C-NMR spectral analyses revealed its structural features. It was found that SDA possessed an α-(1 → 4)-galacturonan backbone with some insertions of (1 → 2) Rha residues, with the side chains attached to the O-3 or O-2 position of GalA residues, or the O-4 position of 1,2-linked Rha residues. The side chains contained branched (1 → 5) linked arabinan, branched (1 → 3) linked rhamman, linear (1 → 4) linked xylan, linear (1 → 4) linked glucan, and linear (1 → 2) linked galactan. SDA was a new acidic polysaccharide isolated from mulberry leaves for the first time.     

Evaluation of the prebiotic effects of citrus pectin hydrolysate

Citrus pectin enzyme hydrolysate (PEH) of different hydrolysis time intervals (6 hours, PEH-6; 12 hours, PEH-12; 24 hours, PEH-24; or 48 hours, PEH-48) or concentrations (1%, 2%, and 4%) was tested for its growth stimulation effect on two probiotics, Bifidobacterium bifidum and Lactobacillus acidophilus. Higher monosaccharide concentrations and smaller molecular weights of PEHs were obtained by prolonging the hydrolysis time. In addition, higher PEH concentrations resulted in significantly higher (p < 0.05) probiotic populations, pH reduction, and increase in total titratable acidity than the glucose-free MRS negative control. Furthermore, significantly higher populations in the low pH environment and longer survival time in nonfat milk (p < 0.05) were observed when the two probiotics were incubated in media supplemented with 2% PEH-24, than in glucose and the negative control. In comparison with other prebiotics, addition of 2% PEH-24 resulted in a more significant increase in the probiotic population (p < 0.05) than in the commercial prebiotics. This study demonstrated that PEH derived from citrus pectin could be an effective prebiotic to enhance the growth, fermentation, acid tolerance, and survival in nonfat milk for the tested probiotics.     

Racemization of free and protein-bound amino acids in strong mineral acid

A new GC/MS technique was applied to measure the racemization of amino acids in strong mineral acid. The inversion rate constants of 15 free amino acids were determined under standard hydrolysis conditions (110°, 6N HCl in evacuated vacuum-sealed tubes). The variations in the inversion rate constants of all tested amino acids with the exception of serine and threonine were related to two factors: side-chain electron-withdrawing capacity (s?*) and steric hindrance in the vicinity of the α-hydrogen atom. The rate constants for serine and threonine were much lower than expected. Hydrolysis-induced racemization of protein-bound amino acids was investigated with α-lactalbumin and β-lactoglobulin. Significant differences were observed as compared with the racemization rates of free amino acids. Such discrepancies were also observed between the two proteins. In the early stage of α-lactalbumin hydrolysis, 10% inversion of methionine was measured as compared with less than 0.5% in the case of β-lactoglobulin. We attributed the particular behaviour of methionine in α-lactalbumin to the neighbourhood of a cysteine residue on the carboxyl side.     

5-氨基水杨酸结肠定位柱塞型脉冲胶囊的制备与体外释放

目的:制备5-氨基水杨酸结肠定位柱塞型脉冲胶囊并对其体外释药行为进行评价。方法:用灌注法制备非渗透性胶囊体,粉末直接压片法压制柱塞片,湿法制粒法制备含药速崩片,将速崩片与柱塞片密封于非渗透性胶囊体内制备脉冲胶囊,考察影响释药时滞的各种因素。结果:胶囊在体外呈明显的脉冲释放,释药时滞随柱塞片中高酯果胶-乳糖或羟丙甲纤维素-低酯果胶比例的增加而增加,具有相同处方柱塞片的脉冲胶囊在模拟结肠溶出介质中释药时滞明显缩短,当高酯果胶-乳糖为4∶6和6∶4,低酯果胶/羟丙甲纤维素为9.5∶0.5和9∶1时,可达到结肠定位所需的4~5h释药时滞。结论:调节柱塞片处方组成可获得具有适当释药时滞的脉冲胶囊,以满足结肠定位释药的目的。     

一种饱和褐藻胶寡糖的制备方法

目的报道一种饱和褐藻胶寡糖的制备方法。方法以褐藻胶为原料,用酸水解法降解,在pH2.85处分级获得均聚古罗糖醛酸片段(PG)和均聚甘露糖醛酸片段(PM),分别在稀盐酸中水解(PG:pH3.8,120℃,3h;PM:pH4,120℃,4h)。水解物经凝胶渗透色谱法分离,获得寡糖组分,反复纯化后,获得系列褐藻胶寡糖。用红外光谱(IR)、核磁共振谱(NMR)、质谱(MS)等方法分析了寡糖的结构。结果褐藻胶经酸水解法降解,分离纯化后获得古罗糖醛酸寡糖二糖到六糖和甘露糖醛酸寡糖二糖到六糖,结构分析表明寡糖保持了原褐藻胶大分子的结构特征。结论本研究为褐藻胶低分子寡糖的制备提供了一种有效、简便的方法。     

Suppression of endotoxin-induced proinflammatory responses by citrus pectin through blocking LPS signaling pathways

Pectin is composed of complex polysaccharides rich in galactoside residues, and it is most abundant in citrus fruits. Pectin and modified pectin have been found to exhibit anti-mutagenic activity and inhibit cancer metastasis and proliferation, with no evidence of toxicity or other serious side effects. In this study, we investigated the inhibitory effect of pectin at different degrees of esterification (DEs) on the expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-activated macrophages. Western blot and RT-PCR analyses demonstrated that 30% esterified pectin (DE30), DE60 pectin, and DE90 pectin significantly inhibited the protein and mRNA expressions of iNOS and COX-2 in LPS-activated macrophages, and DE90 pectin was the most-potent inhibitor. To clarify the mechanisms involved, DE90 pectin was found to inhibit the phosphorylation of MAPKs and IKK kinase activity. In addition, DE90 pectin inhibited the activation of NF-kappaB and AP-1 by electrophoretic mobility shift assay and transient transfection experiments. Finally, we found that DE90 pectin could bind with LPS, and might result in decreased binding of LPS to its receptor. These results suggest that modulation of iNOS and COX-2 expressions by dietary pectin may be important in cancer chemoprevention and anti-inflammation.     

卡介菌多糖-核酸穴位注射治疗慢性荨麻疹的临床疗效观察

目的观察卡介菌多糖-核酸注射液穴位注射治疗慢性荨麻疹的临床疗效和安全性。方法将125例患者分为3组,全部患者均于每晚睡前口服左西替利嗪1片(5 mg),A组(42例)于双侧曲池、足三里穴注射卡介菌多糖-核酸注射液[1次/3 d,1 mg(2支)/次];B组(42例)肌肉注射卡介菌多糖-核酸注射液[1次/2 d,1mg(2支)/次];C组(41例)不加其他药物治疗。疗程均为6周。结果治疗后,A、B、C组的总有效率分别为47.61%、42.85%、29.27%(2周),71.66%、69.02%、39.02%(4周);85.71%、83.05%、47.48%(6周)。A组疗效与B组比较,无显著性差异(P>0.05);A组总有效率与C组比较,有显著性差异(P<0.05)。3组均无明显不良反应。结论卡介菌多糖-核酸注射液穴位注射治疗慢性荨麻疹临床疗效好,其疗效随着治疗时间的延长而提高。     

用于卡介菌多糖核酸制剂效力实验的小鼠模型的建立

目的  建立用于卡介菌多糖核酸制剂效力实验的小鼠模型。方法  选用猪血清、牛血清白蛋白、卵白蛋白作为致敏原,腹腔注射BALB/c小鼠以建立Ⅰ型变态反应动物模型,并通过在卡介菌多糖核酸制剂效力实验中应用建立的小鼠模型来验证其可行性。结果  猪血清、牛血清白蛋白和卵白蛋白均可作为致敏原致敏BALB/c小鼠,但牛血清白蛋白和卵白蛋白的致敏效果优于猪血清。卡介菌多糖核酸制剂效力实验显示,牛血清白蛋白致敏小鼠建立的动物模型可更好地反映卡介菌多糖核酸制剂的效力。结论  牛血清白蛋白致敏小鼠建立的动物模型可用于卡介菌多糖核酸制剂效力实验。     

Sustainable succinic acid production from rice husks

In this study, an investigation was carried out into the production of succinic acid by Actinobacillus succinogenes using the residual biomass rice husks as a sustainable carbon source. The rice husks were submitted to acid hydrolysis in autoclave and in a pressurized polytetrafluorethylene vessel. The hydrolysis conditions were optimized with the aid of a factorial design. The best results were obtained with a pressurized reactor using HCl 2.2% (v v^?1), at a temperature of 174 °C (59 bar), 46 min reaction time, and producing 19.0 g L^?1 glucose and 3.01 g L^?1 xylose. The hydrolysate was detoxified through a combination of pH regulation and adsorption on active carbon; it was subsequently, fermented in anaerobic medium at 37 °C; the nutrient concentration and the agitation speed were also optimized by factorial design. After 54 h static fermentation of the rice husks hydrolysate, supplemented with 8.40 g L^?1 yeast extract and 1.40 g L^?1 NaHCO₃, an amount of 12.5 g succinic acid L^?1 was produced, which corresponds to a yield of 59.9%. This confirms that, rice husks can definitely be used as substrate to produce succinic acid and other priority chemicals     

Structural characterization and antitumor activity of a pectic polysaccharide from the roots of Angelica acutiloba

The polysaccharide fraction from the root of Angelica acutiloba Kitagawa showed a potent antitumor activity against ascitic form of Sarcoma-180, IMC carcinoma, and Meth A fibrosarcoma as well as the solid form of MM-46 tumor. An active polysaccharide, AR-4E-2, was purified by precipitation with cetyl-trimethylammonium bromide, anion-exchange chromatography on DEAE-Sepharose, and gel filtration on Sepharose CL-4B from the polysaccharide fraction. An active polysaccharide fraction showed a weak anti-complementary activity. AR-4E-2 was composed of arabinose, galactose, and rhamnose in the molar ratios of 3.3:1.0:0.7, and also contained 14.5% galacturonic acid and 3.2% protein. Methylation analysis and base-catalysed beta-elimination studies suggested that AR-4E-2 contained a rhamnogalacturonan moiety in which 2,4-di-substituted rhamnose residues were attached to 4-substituted galacturonic acid through position 2 of rhamnose. AR-4E-2 also contained highly branched 3,5-arabinan and (1----4)-galactan.