各类硫酸软骨素制备方法概述

扼要介绍硫酸软骨素、多硫酸软骨素、低分子硫酸软骨素、硫酸软骨素金属盐和酯,以及硫酸软骨素蛋白多糖的各种制备方法。     

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。     

不同工艺制备的硫酸软骨素的理化性质差异

对硫酸软骨素按目前国内普遍采用的稀碱和浓碱两种工艺制备所得产品的理化性质的研究,表明两者存在差异。     

比色法测定复方硫酸软骨素片中硫酸软骨素的含量

硫酸软骨素的测定方法有很多,各有其优点和缺点。本文参照卫生部药品标准,用比色法测定复方硫酸软骨片(chondroitin sulfate,CS)中硫酸软骨素含量。     

猪软骨中硫酸软骨素的分离纯化

目的从猪鼻软骨中提纯硫酸软骨素。方法猪软骨经稀碱 酶解法提取 ,SephadexG 1 0凝胶过滤法、D0 6 1离子交换色谱法纯化。结果提纯的硫酸软骨素经电泳、色谱检查不含蛋白质、肽类、氨基酸及其他酸性黏多糖 ,经红外光谱检查与Sigma公司硫酸软骨素A一致 ,经高效液相离子色谱鉴定纯度已达 99.7%。结论纯化的硫酸软骨素已达色谱纯     

不同工艺硫酸软骨素的理化性质和生物活性

依据部颁质量标准,对硫酸软骨素的工艺进行探讨,筛选出浓碱、稀碱和稀碱-稀盐三种工艺,并对上述工艺产品的理化性质和生物活性进行比较。结果表明浓碱工艺产品口服用于实验性大鼠高脂血症防治,效果较好。     

硫酸软骨素及其衍生物研究进展

硫酸软骨素是一种糖胺聚糖,以动物组织为原料,或者微生物细胞发酵法而制得,在临床上主要用于治疗和预防骨关节炎、心脑血管疾病及眼科疾病。近年来研究表明,硫酸软骨素及其衍生物具有多种潜在的生物活性,如抗炎、抗肿瘤、抗凝、抗血栓等,在多种生理和病理过程中发挥至关重要的作用,具有新药开发的前景。该文从硫酸软骨素及其衍生物现有的药理活性、临床研究方面进行综述,以期为后续研究提供参考。     

硫酸软骨素及其衍生物的研究进展

综述近年来硫酸软骨素及其衍生物的研究进展,主要包括制备、药理作用及临床应用等。     

硫酸软骨素制备工艺的优化

目的建立提取硫酸软骨素优化工艺。方法通过正交实验对生产工艺中4个关键因素进行优化,并对产品进行纯度鉴定和理化性质分析。结果在此优化条件下,硫酸软骨素产率为33.8%,纯度用间苯三酚分光光度法测定为93.7%,供试品经红外光谱鉴定含有较多的硫酸软骨素A,所有指标均符合部颁要求。结论此工艺可有效提高产品纯度和产率,达到了优化目的。     

硫酸软骨素生产的优化条件

硫酸软骨素的生产工艺较多,无论是稀碱或浓碱提取法,还是稀碱稀盐的综合提取法,都要经过酶解和活性炭脱色等处理,才能达到药用口服标准。近年,魏章祥报道采用稀碱浓盐法工艺生产硫酸软骨素,我们通过实验研究,证明了此工艺具优越性,有生产实际价值,不仅可以省去酶解脱色处理步骤,还可提高产品收率和纯度。我们为摸索最佳生产条件,又进行了一些试验,作了改进。     

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%。     

RP—HPLC法测定鲨芪康胶囊中硫酸软骨素含量

目的建立RP．HPLC法测定硫酸软骨素的含量。方法采用RP-HPLC法测定,使用C18柱（4．6×250mm,5μm）,流动相为乙腈．2．28mmol·L^-1四甲基氯化铵水溶液（5：95）,流速为0．6mL·min^-1,检测波长195nm。结果硫酸软骨素总量在0．08-0．4mg·mL^-1浓度范围内线性关系良好（r=0．9996）,方法平均回收率95．52％,RSD％=1．43％。结论本方法准确、快捷、重复性。可作为该制剂中硫酸软骨素的含量测定方法。     

HPLC法同时测定复方硫酸软骨素片中硫酸软骨素和甘草酸含量的研究

目的:建立高效液相色谱梯度洗脱-波长切换法同时测定复方硫酸软骨素片中硫酸软骨素和甘草酸含量的方法。方法:采用InertSustain C₁₈色谱柱(4.6 mm×250 mm, 5μm),以5 mmol·L^-1庚烷磺酸钠溶液为流动相A,乙腈为流动相B,梯度洗脱，柱温为35℃,检测波长为199 nm(0～8 min时，检测硫酸软骨素)、251 nm(8～30 min时，检测甘草酸)。结果:硫酸软骨素钠和甘草酸分别在0.150 2～3.004 9 mg·mL^-1和0.012 67～0.316 8 mg·mL^-1范围内与峰面积呈良好的线性关系，且该检测方法专属性、精密度、回收率等良好。4批样品中，硫酸软骨素和甘草酸的含量分别为72.196 3～73.801 4和1.498 8～1.548 0 mg·片^-1。结论:该方法操作简单，结果准确，可用于复方硫酸软骨素片的质量控制。     

Oxidative stress in ovariectomy menopause and role of chondroitin sulfate

Oxidative stress due to reactive oxygen species (ROS) can cause oxidative damage to cells. Cells have a number of defense mechanisms to protect themselves from the toxicity of ROS. Mitochondria are especially important in the oxidative stress as ROS have been found to be constantly generated as an endogen threat. Mitochondrial defense depends mainly on superoxide dismutase (SOD) and glutathione peroxidase (GPx), whereas microsomal defense depends on catalase (CAT), which is an enzyme abundant in microsomes. SOD removes superoxide anions by converting them to H2O2, which can be rapidly converted to water by CAT and GPx. Also, GPx converts hydroperoxide (ROOH) into oxidized-glutathione (GSSG). Ovariectomized (OVX) rats are used as an oxidative stress model. An ovariectomy increased the levels of MDA, one of the end-products in the lipid peroxidative process, and decreased levels of the antioxidative enzymes; SOD, CAT and GPx. However, Chondroitin sulfate (CS) decreased the levels of MDA, but increased the levels of SOD, CAT and GPx in a dose-dependent manner. Moreover, inflammation and cirrhosis of liver tissue in CS- treated rats were significantly decreased. These results suggest that CS might be a potential candidate as an antioxidative reagent.     

Simultaneous determination of chondroitin sulfate sodium,allantoin and pyridoxine hydrochloride in pharmaceutical eye drops by an ion-pair high-performance liquid chromatography

An ion-pair high-performance liquid chromatography (HPLC) method has been developed for the simultaneous determination of chondroitin sulfate sodium (CSS), allantoin and pyridoxine hydrochloride (VB₆) in a commercial eye drops dosage form. An Alltima C₁₈ column (250 mm × 4.6 mm i.d., 5 μm) was used for the separation at room temperature, with 25 mM ammonium dihydrogen phosphate (containing 0.01% heptanesulfonic acid sodium salt) and acetonitrile (95:5, v/v) as the mobile phase at the flow rate of 0.5 mL min⁻¹. The detection wavelength for CSS, allantoin and VB₆ was 195 nm, 215 nm and 291 nm, respectively. The method showed good linearity for CSS, allantoin and VB₆, with correlation coefficients greater than 0.9996, in the range of 203.96–815.84 mg L⁻¹, 371.16–1488.64 mg L⁻¹, and 23.32–93.28 mg L⁻¹, respectively. The instrumental and method precisions were adequate with all relative standard deviations lower than 2.0%. The accuracy of this method, measured by the recovery of three compounds from spiked placebo solutions, was from 99.01% to 101.92%. The three components, CSS, allantoin and VB₆ were well separated from other ingredients and degradation products. This method is fast, simple, and can be used for direct and simultaneous determination of CSS, allantoin and VB₆ in the pharmaceutical preparation.     

Bile acid-conjugated chondroitin sulfate A-based nanoparticles for tumor-targeted anticancer drug delivery

Chondroitin sulfate A-deoxycholic acid (CSA-DOCA)-based nanoparticles (NPs) were produced for tumor-targeted delivery of doxorubicin (DOX). The hydrophobic deoxycholic acid (DOCA) derivative was conjugated to the hydrophilic chondroitin sulfate A (CSA) backbone via amide bond formation, and the structure was confirmed by ¹H-nuclear magnetic resonance (NMR) analysis. Loading the DOX to the CSA-DOCA NPs resulted in NPs with an approximately 230 nm mean diameter, narrow size distribution, negative zeta potential, and relatively high drug encapsulation efficiency (up to 85%). The release of DOX from the NPs exhibited sustained and pH-dependent release profiles. The cellular uptake of DOX from the CSA-DOCA NPs in CD44 receptor-positive human breast adenocarcinoma MDA-MB-231 cells was reduced when co-treated with free CSA, indicating the interaction between CSA and the CD44 receptor. The lower IC₅₀ value of DOX from the CSA-DOCA NPs compared to the DOX solution was also probably due to this interaction. Moreover, the ability of the developed NPs to target tumors could be inferred from the in vivo and ex vivo near-infrared fluorescence (NIRF) imaging results in the MDA-MB-231 tumor-xenografted mouse model. Both passive and active strategies appear to have contributed to the in vivo tumor targetability of the CSA-DOCA NPs. Therefore, these CSA-DOCA NPs could further be developed into a theranostic nanoplatform for CD44 receptor-positive cancers.     

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。     

A sensitive and simple method for the determination of chondroitin sulfate a with crystal violet by resonance rayleigh scattering technique

Resonance Rayleigh scattering (RRS) spectra resulting from interaction between chondroitin sulfate A (CSA) and crystal violet (CV) have been investigated and applied to the determination of CSA. Though the intensity of RRS has proved very weak for CSA and CV, respectively, it can be greatly enhanced when both interact and form a supramolecular complex. A new RRS spectrum appears with a maximum scattering peak at 328 nm. In this paper, the optimum conditions of the interaction, influencing factors, and the relationship between the relative intensity of RRS (DeltaI) and the concentration of CSA have been thoroughly investigated. A new method of determination for the trace amount of CSA has been developed, which combines a simple procedure, high sensitivity, and a low detection limit of 4.8 ng/mL. It has been applied with satisfactory results to the determination of CSA in CSA injection samples and synthetic samples.