3种鱼鳃中糖胺聚糖的分离纯化及细微结构比较分析

目的 从狭鳕 (Alaska pollack)、褐牙鲆 (Bastard halibut) 和真鲷 (Red seabream) 三种鱼鳃中提取分离糖胺聚糖并对其细微结构进行比较分析。方法 利用酶法降解并以阴离子交换色谱法分离纯化糖胺聚糖,通过单糖组成和醋酸纤维素薄膜电泳对其种类进行鉴别,采用高效液相色谱法对其相对分子量和二糖组成进行比较分析,最后利用核磁共振波谱法 (₁H-NMR) 对其结构进行表征。结果 从3种鱼鳃组织中提取分离得到54种多糖,其中18种是电荷和分子量均一的糖胺聚糖,它们集中在0.8 mol?L_-1 NaCl洗脱部分 (P4) 和1.0 mol?L_-1 NaCl洗脱部分 (P5),占多糖总量的17.4～58.4%;P4主要为硫酸软骨素C (CSC),P5主要为硫酸软骨素A (CSA) 和杂合硫酸软骨素 (HyCS),尤其,与牛气管来源CSA比较,HyCS的4S/6S比和电荷密度均较大。结论 狭鳕、褐牙鲆和真鲷鱼鳃组织中的糖胺聚糖主要为CSA、CSC和HyCS。     

鲟鱼软骨硫酸软骨素的制备及结构分析

目的 采用酶解提取法,从鲟鱼软骨中提取硫酸软骨素,对其化学结构进行研究。方法 从鲟鱼软骨中提取硫酸软骨素,通过紫外-可见吸收光谱法（UV）,傅里叶变换红外光谱法（FTIR）,PMP柱前衍生高效液相色谱法（PMP-HPLC）、核磁共振波谱法（NMR）和液质联用分析（HPLC-MS）,对硫酸软骨素的化学结构进行分析。结果 采用酶解提取法,从鲟鱼软骨中制备了硫酸软骨素。单糖分析表明,鲟鱼硫酸软骨素主要由葡萄糖醛酸（48.23%）和氨基半乳糖（46.29%）组成。结合NMR分析和HPLC-MS分析可知,从鲟鱼软骨中提取的硫酸软骨素主要含有4-单硫取代的硫酸软骨素A二糖单元,6-单硫取代的硫酸软骨素C二糖单元,其所占比例分别为34%和63%,硫酸根含量为13.13%,属于高硫酸化的硫酸软骨素。结论 从鲟鱼软骨中提取出了具有高硫酸根含量的硫酸软骨素组分,可应用于保健食品和海洋药物的开发等领域。     

真鱿软骨硫酸软骨素的提取、鉴定及结构分析

目的 采用酶解提取法,从真鱿软骨中提取1种新型的硫酸软骨素,对其化学结构进行研究。方法 采用酶解法、氯化十六烷基吡啶（CPC）沉淀法从鱿鱼软骨中提取纯化硫酸软骨素,通过PMP柱前衍生液相色谱法、聚丙烯酰氨凝胶电泳（PAGE）、核磁共振波谱法（NMR）和液质联用分析（HPLC-MS）,对硫酸软骨素的化学结构进行分析。结果 使用木瓜蛋白酶酶解鱿鱼软骨,乙酸缓冲液在料液比为1:30（g/mL）条件下,硫酸软骨素的提取率最高,蛋白质残留最低,经过CPC纯化,得到了纯度高达99.5%的硫酸软骨素。由PAGE电泳可以看出其分子量很大且能够被硫酸软骨素ABC酶酶解。单糖分析表明,鱿鱼硫酸软骨素主要由乙酰氨基半乳糖、葡萄糖醛酸以及少量的葡萄糖组成。硫酸软骨素ABC酶将其水解为聚合度2-18的偶数糖,采用HPLC-MS分析,可以得到所有寡糖的全指纹谱图。结合1H-NMR分析可知,从真鱿软骨提取出来的硫酸软骨素主要含有6-单硫取代的硫酸软骨素C二糖单元和4,6-双硫取代硫酸软骨素E二糖单元,其所占比例分别为60.3%和39.7%,硫酸根含量为22.3%,是属于高硫的硫酸软骨素。结论 从真鱿软骨中提取出了具有高硫酸根含量的硫酸软骨素组分,可用于开发保健食品和海洋药物。     

氧杂蒽-2-羧酸及氧杂蒽酮-2-羧酸类衍生物的合成

本文合成了一系列7位拉电子基团取代的氧杂蒽-2-羧酸及氧杂蒽酮-2-羧酸化合物。经大鼠被动皮肤过敏试验(PCA),显示化合物(Ⅷ),(Ⅻ_b)和(Ⅻ_e)有50～60%的抑制率(PO,200mg/kg),(Ⅻ_d)有54%的抑制率(ⅳ,10mg/kg)。氧杂蒽酮-2-羧酸(Ⅷ)经黄鸣龙还原反应生成(Ⅸ),但收率低,推测副产物2,2′-二羟基二苯基-甲烷(Ⅹ)的生成机理,改变反应条件,从而提高了(Ⅸ)的收率。化合物(Ⅻ_d)的元素分析和质谱均证明其结构为(Ⅻ_d),但核磁共振谱(溶剂为三氟醋酸)出现异常,而与(Ⅻ_d)在醋酐中回流所得产物(Ⅻ_d)的图谱相同。表明(Ⅻ_d)在三氟醋酸中环合、脱水产生了(Ⅻ_d)。     

紫外分光光度法测定硫酸软骨素含量

<正> 一、紫外分光光度法测定硫酸软骨素的基本原理硫酸软骨素为动物的软骨,经提取制得的一种粘多糖,当用适当过量酸水解时,水解产物在288±1nm处有吸收,其吸收值与硫酸软骨素含量有线性关系,测定时先以精制硫酸软骨素(予先用重量法测定含量)为标准品,绘制不同浓度的标准曲线,然后将待测样品以同样方法测得的吸收度,即可由标准曲线上得出待测样品的含量。二、仪器与试剂 751分光光度计(上海分析仪器厂制)波     

消旋15(R)-15-甲基PGE2甲酯的合成

d,l-15(R)-15-Methyl-PGF_2α methyl ester 11-trimethylsilyl ether(II)wasprepared from selective monosilylation of d,l-15(R)-15-methyl-PGF_2αmethyl ester(I) withtrimethylsilyldiethylamine in acetone. Oxidation of(II ) with Collin's reagent gave d,l-15(R)-15-methyl-PGE₂ methyl ester 11-trimethylsilyl ether(III)which,without purification,was converted to d,l-15(R)-15-methyl-PGE₂ methyl ester(IV)under mild acidic conditions.     

HPLC-MS法鉴定制霉素和制霉菌素中成分及其潜在毒性预测

目的 采用HPLC-MS法鉴定制霉素和制霉菌素中组分,并对其潜在毒性进行预测。方法 采用HPLC-MS法测定,Sepax Bio-C₁₈色谱柱（250 mm×4.6 mm,5 μm）;流动相：[0.38%乙酸铵溶液–乙腈（71∶29）]–[0.38%乙酸铵溶液–乙腈（40∶60）],梯度洗脱;检测波长305 nm;体积流量1.0 mL/min;柱温30℃;进样量20 μL。采用电喷雾正离子化（ESI）,扫描范围m/z 100～1 700,碎裂电压100 V,毛细管电压3 500 V,雾化气压45 psi,干燥气流速10 psi,干燥气温度325℃;无二级质谱。结果 制霉素中主要成分为制霉菌素A₃、多真菌素B、制霉菌素A₁和白诺菌素,制霉菌素A₃、多真菌素B分子中具有L-洋地黄毒糖结构,白诺菌素分子中具有2,5-二酮哌嗪结构。制霉菌素以制霉菌素A₁为主要成分,制霉菌素A₁分子中无洋地黄毒糖或2,5-二酮哌嗪结构。结论 制霉素、制霉菌素中主要成分和含量均不相同,制霉素组分中的L-洋地黄毒糖结构和2,5-二酮哌嗪结构可能分别导致心脏毒性和生殖毒性。     

美托洛尔光学异构体在犬体内的药动学-药效学结合模型

用麻醉犬研究美托洛尔(Met)光学异构体药动学—药效学结合模型。结果表明,ig(+)-Met或(-)-Met后,其药动学符合二室模型,药动学参数V_d/F,CLs/F和AUC在两种Met之间有显著性差异。(+)-Met和(-)-Met的效应和血药浓度关系呈逆时针滞后环。引入效应室理论后,药效和效应室浓度之间的关系符合Sigmoid-E_max模型。应用CAPP软件进行模型拟合,血药浓度的预测值和药效的预测值皆与其实测值较为接近。(+)-Met抑制V_max,dp/dt_max及HR效应的Ce₅₀分别是(-)-Met的3.7,6.8,6.3倍,证实(-)-Met对犬心脏的抑制作用强于(+)-Met。     

羽叶三七叶中甙类成分的研究

从羽叶三七叶中分离到十三种甙类成分,经FAB-MS,¹³CNMR谱,双照射¹HN-MR谱,¹H-¹H COSY谱及与标准品直接对照,证明十一种为已知化合物,分别为人参皂甙F₁(Ⅰ),F₂(Ⅱ),F₃(Ⅲ),R_g2(Ⅳ),R_a(Ⅴ),R_d(Ⅵ),R_b1(Ⅷ),R_b3(Ⅷ),24(S)-假人参甙F₁₁(Ⅸ),人参黄酮(Ⅹ)和珠子参甙F₁(Ⅺ);另外两种为新的达玛烷型皂甙,命名为羽叶三七甙F₁(Ⅻ)和F₂(ⅫⅠ),并确定其化学结构。同时修正珠子参甙F₃的结构。进一步阐明人参黄酮甙结构中的两个糖的连接方式。     

肿瘤的化学治疗 ⅩⅧ.若干取代查耳酮的制备

用2,6-二羟基苯乙酮和相应的苯甲醛在60%氢氧化钾溶液存在下制备了六个2′,6′-二羟基取代查耳酮(Ⅳ_a-f),又从2-羟基取代苯乙酮和相应的苯甲醛制备了2′-羟基取代查耳酮(Ⅴ_a-t)二十个。利用相似的方法制备了另一系列的取代查耳酮(Ⅵ_a-f)六个。化合物Ⅳ_d,Ⅴ_a,Ⅴ_b,Ⅴ_c,Ⅴ_e,Ⅴ_f和Ⅴ_i在浓度为125微克/每毫升,化合物Ⅳ_a,Ⅳ_b,Ⅳ_f,Ⅴ_d,Ⅴ_k,Ⅴ_r,Ⅴ_s,Ⅳ_a,Ⅳ_b,Ⅳ_c和Ⅳ_f在浓度为250微克/每毫升体外试验对Ehrlich腹水瘤有明显抑制作用。但体内试验上述三系列32个查耳酮在口服时对小白鼠肉瘤-180或Ehrlich腹水瘤无明显抑制作用。     

Effect of orally administered chondrosine on uptake of 35S sulfate into rat cartilage

Chondroitin sulfate is widely distributed in animal tissues and possibly plays an important role in different types of metabolic reactions as well as protecting joints, the internal wall of blood vessels, skin, bone, etc. In cartilage, glycosaminoglycans have a protective function; in particular, chondroitin sulfate stabilizes fibrous and cellular elements of the connective tissue and, at the same time, lubricates and protects the membranes in joints. Recently, chondroitin sulfate has been used as a nutraceutical for the treatment of joint diseases such as osteoarthritis, although acidic and large molecules such as chondroitin sulfate might not be able to be absorbed through digestive apparatus such as the intestine. In this study, we investigated the effects of orally administered chondrosine derived from shark chondroitin sulfate on the uptake of inorganic (35)S sulfate into rat cartilage and found that chondrosine stimulates the incorporation of (35)S sulfate into cartilage compared with intact chondroitin sulfate.     

Induction of inflammatory cytokines by cartilage extracts

Shark cartilage extracts were examined for induction of cytokines and chemokines in human peripheral blood leukocytes. Primary leukocyte cultures were exposed to a variety of aqueous and organic extracts prepared from several commercial brands of shark cartilage. From all commercial sources of shark cartilage tested the acid extracts induced higher levels of TNFalpha than other extracts. Different commercial brands of shark cartilage varied significantly in cytokine-inducing activity. TNFalpha induction was seen as early as 4 h and IFNgamma at detectable levels for up to four days. Shark cartilage extracts did not induce physiologically significant levels of IL-4. Results suggest that shark cartilage, preferentially, induces Th1 type inflammatory cytokines. When compared to bovine cartilage extract, collagen, and chondroitin sulfate, shark cartilage induced significantly higher levels of TNFalpha. Treatment with digestive proteases (trypsin and chymotrypsin) reduced the cytokine induction response by 80%, suggesting that the active component(s) in cartilage extracts is proteinaceous. The induction of Th1 type cytokine response in leukocytes is a significant finding since shark cartilage, taken as a dietary supplement for a variety of chronic degenerative diseases, would be contraindicated in cases where the underlying pathology of the chronic condition is caused by inflammation.     

咔唑法测定鲨鱼硫酸软骨素含量的方法学研究

目的建立用咔唑法测定鲨鱼硫酸软骨素含量的方法。方法采用咔唑法,以鲨鱼硫酸软骨素为对照品,在浓硫酸介质中经咔唑显色反应后,于515nm波长处分别测定对照品溶液和供试品溶液的吸收度,按标准曲线法计算含量。结果硫酸软骨素量在29~145μg范围内与吸收度呈良好的线性关系,相关系数r=0.999 8,加样回收率为100.5%,RSD=1.3%(n=6)。结论此法简单、快速,结果准确,可用作鲨鱼硫酸软骨素含量的测定方法。     

Classification of chondroitin sulfate A, chondroitin sulfate C, glucosamine hydrochloride and glucosamine 6 sulfate using chemometric techniques

Chondroitin sulfate A, chondroitin sulfate C, glucosamine hydrochloride and glucosamine sulfate are natural products that are becoming increasingly popular in the treatment of arthritis. They belong to a class of compounds known as glycosaminoglycans (GAGs). They are available over the counter as nutritional supplements. However, increasing use has led to increasing scrutiny of the quality of products on the market. There is also interest in the pharmacological properties of these compounds. To facilitate this, there is a need for better qualitative and quantitative methods of analysis. This paper describes methods for achieving the qualitative identification of chondroitin sulfate A, chondroitin sulfate C, glucosamine hydrochloride or glucosamine sulfate. Fourier transform infrared spectroscopy coupled with a variety of chemometric methods successfully classified these compounds. Using soft independent modeling of class analogies (SIMCA), hierarchical cluster analysis (HCA) and principal components analysis (PCA) samples were classified as either chondroitin sulfate A, chondroitin sulfate C, glucosamine hydrochloride or glucosamine sulfate. This work also examined the discriminating ability of different sections of the spectrum. It was found that for the classification of these compounds that using the finger print region of the spectrum (below 2000 cm(-1)) gave the best discrimination.     

Metabolic fate of exogenous chondroitin sulfate in man.

Chondroitin sulfate is administered as a drug to man by intravenous, intramuscular or oral route. However, few data are available on the metabolic fate of exogenous chondroitin sulfate in man. After intravenous administration of 0.5 g of chondroitin sulfate to healthy volunteers, the plasma level decreases according to a two-compartmental open model. The half-lives of distribution and elimination are 25.5 +/- 6.6 and 281 +/- 32 min, respectively. The volumes of central and tissue compartments are 6.0 +/- 1.0 and 22.9 +/- 7.7 l, respectively. More than 50% of the administered chondroitin sulfate is excreted with urine during the first 24 h as high and low molecular weight derivatives. After oral administration of 3 g of chondroitin sulfate to 12 healthy volunteers, a main peak (11.4 +/- 3.7 micrograms/ml) preceded by a lower peak is observed after 190 +/- 21 min. The elimination half-life is 363 +/- 109 min. The absolute bioavailability following oral administration calculated from AUC of plasma concentration is 13.2%. A peak of oligo- and polysaccharides with a molecular weight lower than 5000 Daltons derived from partial digestion of exogenous chondroitin sulfate is also present in plasma. These observations indicate that the metabolic fate of exogenous chondroitin sulfate is similar in man and in experimental animals.     

养殖鲟鱼软骨中硫酸软骨素的优化提取工艺研究

目的以养殖鲟鱼软骨为原料,提取硫酸软骨素、筛选提取工艺条件。方法用稀碱-盐解-酶解法提取硫酸软骨素,并对各工艺中影响提取效果的不同因素进行正交实验。结果碱提取最佳工艺组合为:碱浓度5%,料液比1∶6,碱提取时间3h;盐解最佳工艺组合为:盐浓度4%,盐解温度90℃,盐解时间20min;酶解最佳工艺组合为:酶浓度0.4%,酶解温度52℃,酶解时间2.5h。结论采用上述组合提取硫酸软骨素的得率为31.56%,显著高于(P<0.05)传统工艺的25.86%。     

Effects of chondroitin sulfate-C on articular cartilage destruction in murine collagen-induced arthritis

The effects of chondroitin sulfate-C (CAS 25322-46-7, Chondroitin ZS Tab) on type II collagen (CII)-induced arthritis (CIA) in mice were evaluated. DBA/1J mice were immunized with bovine CII emulsified in Freund's complete adjuvant, followed by a booster injection 21 days later. Chondroitin sulfate-C at doses of 100, 300 and 1000 mg/kg was administered orally once daily beginning 14 days before initial immunization. An arthritis index and hind paw edema were examined from day 0 to day 49, when the mice were killed by ether anesthesia for histopathological examination. The delayed-type hypersensitivity (DTH) reaction, serum anti-CII antibody titer, and histopathologic characteristics of both synovitis and destruction of articular cartilage were analyzed. Both the arthritis index and the serum anti-CII antibody titer were reduced by treatment with chondroitin sulfate-C in a dose-dependent manner. Chondroitin sulfate-C (1000 mg/kg) significantly inhibited hind paw edema, synovitis and destruction of the articular cartilage, but not DTH reaction.     

Quality of different chondroitin sulfate preparations in relation to their therapeutic activity

Objectives Chondroitin sulfate is currently recommended by the European League Against Rheumatism (EULAR) as a SYSADOA (symptomatic slow acting drug for osteoarthritis) in Europe in the treatment of knee and hand osteoarthritis based on research evidence and meta‐analysis of numerous clinical studies. Furthermore, recent clinical trials demonstrated its possible structure‐modifying effects. Chondroitin sulfate, alone or in combination with glucosamine or other ingredients, is also utilized as a nutraceutical in dietary supplements in Europe and the USA. However, it is derived from animal sources by extraction and purification processes. As a consequence, source material, manufacturing processes, the presence of contaminants and many other factors contribute to the overall biological and pharmacological actions of these agents. We aim to review the quality control of chondroitin sulfate in pharmaceutical‐grade preparations and nutraceuticals. Key findings Pharmaceutical‐grade formulations of chondroitin sulfate are of high and standardized quality, purity and properties, due to the stricter regulations to which this drug is subjected by local national health institutes as regards production and characteristics. On the contrary, as several published studies available in literature indicate, the chondroitin sulfate quality of several nutraceuticals is poor. Additionally, there are no definite regulations governing the origin of the ingredients in these nutraceuticals and the origin of the ingredients in natural products is the most important factor ensuring quality, and thus safety and efficacy, in particular for chondroitin sulfate, due to its extraction from different sources. Conclusions Due to the poor chondroitin sulfate quality of some nutraceuticals, we conclude that stricter regulations regarding their quality control should be introduced to guarantee the manufacture of high quality products for nutraceutical utilization and to protect customers from low‐quality, ineffective and potentially dangerous products. There is a need for specific and accurate analytical procedures, which should be enforced to confirm purity and label claims both for raw materials and finished chondroitin sulfate products, and also to govern the origin of ingredients. Until these stricter regulations are in place, then it is strongly recommended that pharmaceutical‐grade chondroitin sulfate is used rather than food supplements.