有机铬螯合物的合成及性能研究
文献类型：学位论文
| 作者 | 唐海燕 |
| 学位类别 | 博士 |
| 答辩日期 | 2014-06 |
| 授予单位 | 中国科学院研究生院 |
| 导师 | 张懿 |
| 关键词 | 有机铬 合成 抗氧化性 稳定性 降糖活性 急性毒性 |
| 其他题名 | Study on the Synthesis and Bioactivity of Organic Chromium Complexes |
| 学位专业 | 化学工艺 |
| 中文摘要 | 铬是一种重要的战略资源，铬盐系列产品用途广泛，涉及国民经济约10%的商品品种，具有不可替代性。但是目前我国铬盐行业存在产品品种少、附加值低等缺陷。高附加值铬产品的研发已成为我国铬盐工业迫切的技术需求和市场定位导向。 三价铬是人体维持正常糖代谢和脂代谢必需的微量元素。有机铬产品与无机铬相比，具有更高的吸收率、生物活性和安全性，已作为降糖药的辅助成分、营养补充剂和肌肉塑型剂得到广泛的应用。以天然生物活性物质为配体合成有机铬产品已成为研究热点和产品开发方向。本文以氨基酸铬作为切入点，针对其合成和使用过程中存在的操作繁琐、无法实现工业化和分析检查方法不完善等问题，建立了氨基酸铬螯合物产品质量的检测方法，提出并优化了醇相体系中分别采用三价铬盐和六价铬盐为铬源制备氨基酸铬螯合物的工艺，获得了醇相体系中以三价铬盐和蛋氨酸为原料合成蛋氨酸铬的表观动力学，揭示了氨基酸铬螯合物的抗氧化活性和体外稳定性，研究了蛋氨酸铬的降糖活性和急性毒性。论文主要取得如下创新性进展： (1) 建立了蛋氨酸铬、甘氨酸铬、苏氨酸铬和苯丙氨酸铬等四种氨基酸铬产品质量的检测方法。采用ICP-OES以及硫酸亚铁铵滴定法测定了四种氨基酸铬中铬含量。提出、优化并验证了四种氨基酸铬螯合物中氨基酸含量的高效液相色谱（HPLC）测定方法。在以ZORBAX Extend-C18柱为液相色谱柱（150×4.6mm，5μm），KH2PO4溶液为流动相，流速1.0mL/min，柱温 25 ~ 35℃，波长 200 ~ 220 nm，进样量10 ~ 20 μL的色谱条件下，HPLC法能快速准确地测得上述四种氨基酸铬中氨基酸的含量。该HPLC方法具有样品前处理过程简单，操作简便等优点。方法适用性良好，通过改变检测波长等条件能够快速检测出不同氨基酸铬中氨基酸的含量。采用有机溶剂萃取法，以无水乙醇为溶剂，测定了四种氨基酸铬螯合物的螯合率。通过建立的分析方法，蛋氨酸铬、甘氨酸铬、苏氨酸铬和苯丙氨酸铬中铬的含量分别为：10.44%，17.80%，12.79%和9.24%，其中氨基酸的含量分别为89.6%，76.1%，87.2%以及87.8%，与理论含量基本一致。四种氨基酸铬的螯合率均在90%以上，产品质量优良。 (2) 提出了在醇相体系中制备氨基酸铬螯合物的方法。以三价铬盐或六价铬盐为铬源，以蛋氨酸、甘氨酸、苏氨酸及苯丙氨酸为配体，制备了蛋氨酸铬、甘氨酸铬、苏氨酸铬及苯丙氨酸铬螯合物。通过紫外/可见/近红外光谱、红外光谱、扫描电镜、热重和元素分析等方法对四种氨基酸铬产品结构进行了表征，推测它们的分子式分别为Cr(C5H10NO2S)3、Cr(C2H4NO2)3?H2O、Cr(C4H8NO3)3和Cr(C9H10NO2)3?H2O。制得的氨基酸铬产品具有良好的外观与形貌。 (3) 优化了醇相体系中以三价铬盐及六价铬盐为铬源制备氨基酸铬螯合物的工艺，分别得到了四种氨基酸铬螯合物的最佳工艺条件。以三价铬盐为原料，氨基酸铬的合成最佳工艺为：乙醇体积分数48.5% ~ 75%，c(Cr) = 0.25 mol/L，氢氧化钠与氨基酸等摩尔，氨基酸与三价铬摩尔比为3 ~ 4.31 : 1, 80 ~ 81.1℃，反应2.1 ~ 3 h。以六价铬盐为铬源，氨基酸铬合成的最优工艺条件（以0.01 mol Cr6+计）：c(Cr6+) = 0.5或1.0 mol/L，HCl 3.5 ~ 4.0 mL，乙醇20 ~ 30mL，H2O 10 ~ 20 mL，回流还原反应35 ~ 40 min，氢氧化钠与氨基酸等摩尔，氨基酸与六价铬摩尔比为4 ~ 4.5 : 1，80 ~ 81℃，螯合反应4 ~ 5 h。在优化的工艺条件下，蛋氨酸铬、甘氨酸铬、苏氨酸铬和苯丙氨酸铬的收率均在95%以上。 (4) 获得了以三价铬盐为铬源制备蛋氨酸铬螯合物的宏观动力学数据。实验表明，蛋氨酸和三价铬的反应级数分别为2和1，蛋氨酸铬的合成反应为总体是一个3级反应，反应的表观活化能、指前因子和宏观动力学方程分别为37.42 kJ/mol，3.49×104 L2?mol-2?min-1，和 。 (5) 探讨了氨基酸铬螯合物的抗氧化活性和体外稳定性。结果表明测试的氨基酸铬螯合物均具有一定程度的抗氧化性，而且其活性大于游离的氨基酸及无机氯化铬，氨基酸铬螯合物的抗氧化性能随着浓度的升高而增强。蛋氨酸铬、甘氨酸铬、苏氨酸铬和苯丙氨酸铬在不同的条件下具有良好的稳定性，工业上对于四种氨基酸铬的保存和运输比较易于实现。 (6) 采用四氧嘧啶诱导糖尿病小鼠为对象，研究了蛋氨酸铬的降糖活性，考察了蛋氨酸铬对糖尿病小鼠的糖代谢、脂代谢、肝功能和肾功能的影响，并与无机氯化铬进行了对比。结果表明，蛋氨酸铬能够促进糖尿病小鼠的糖代谢和脂代谢，显著降低糖尿病小鼠的血糖、血脂水平。蛋氨酸铬的效果与其剂量成正比关系，且优于氯化铬。蛋氨酸铬能够显著降低糖尿病小鼠谷丙转氨酶和谷草转氨酶的活性，对糖尿病造成的肝损伤有一定的保护作用，具有保肝效果。对糖尿病小鼠补充蛋氨酸铬有利于减轻糖尿病对小鼠肾脏造成的损伤。采用一次最大限量给药法研究了蛋氨酸铬的经口急性毒性，结果表明蛋氨酸铬的半数致死量（LD50）大于10.0 g/kg体重，属于实际无毒的产品。 |
| 英文摘要 | As an important strategic resource, chromium and its salts are widely used and related to about 10% of the goods varieties in national economy. The effect of chromium is irreplaceable. However, at present, the production of chromium products in China has the disadvantages of few product series, low output and so on. Therefore, the development of products that yield a high added value has become the urgent technical demand and market orientation of chrome salts industry. Trivalent chromium [Cr(III)] is essential for mammal and plays a vital role in proper carbohydrate and lipid metabolism. Lots of Cr(III) products including inorganic chromium and organic chromium have been synthesized and applied in the relative fields. Compared to inorganic chromium, organic chromium complexes are easier to be utilized by the mammals and have higher biological activity. A variety of organic chromium products have been developed and widely used as auxiliary components of hypoglycemic drugs, nutritional supplements, muscle forming agents and so on. Therefore, the preparation of organic chromium complexes with natural ligands and less or non-toxicity is of great interest. Amino acid is a good alternative. The operation for present synthetic methods of chromium amino acid (CrAA) complexes is complicated, and none of these methods reached considerable commercial application. Moreover there is no sophisticated analytic method for the quality of CrAA complexes. In order to solve the above problems, in this dissertation, the methods to analyze the quality of CrAA complexes were proposed. The synthetic methods of CrAA complexes using trivalent and hexavalent chromium salts as raw materials in aqueous ethanol solvent were developed and optimized as well. The apparent reaction kinetics for the synthesis of chromium metnionine (CrMet) using trivalent chromium salt as raw material was also investigated. The antioxidant activity and in vitro stability of CrAA complexes were revealed. Besides, the hypoglycemic activity and acute toxicity of CrMet were studied. The following innovative achievements and progresses were obtained: (1) The methods for the determination of the quality of CrAA complexes were established. The chromium contents in CrAA complexes were assessed using ICP-OES and titration method with ferrous ammonium sulphate solution. A rapid and high-throughput method using HPLC-DAD analysis has been developed, optimized, and validated to determine the contents of amino acids in CrAA complexes. The HPLC-DAD method was rapid, sensitive and cost-effective for quantitating amino acids contents in CrAA complexes on a Zorbax Extend-C18 (150×4.6 mm, 5μm) analytical column flushed with KH2PO4 as the mobile phase pumped at a flow rate of 1.0 mL/min at 25 ~ 35 oC after injecting 10 ~ 20 μL sample with UV detection at 200 ~ 220 nm. The proposed HPLC method was a fast, accurate and robust method requiring minimal sample preparation and simple instruments. The HPLC method was also feasible and applicable in different cases. The chelating rates of the CrAA complexes were determined through organic solvent extraction method, and absolute ethanol was used as the solvent. The chromium contents in CrMet, chromium glycinate (CrGly), chromium threoninate (CrThr) and chromium phenylalaninate (CrPhe) were assessed as 10.44%，17.80%，12.79% and 9.24%, respectively. The amino acid contents in CrMet, CrGly, CrThr and CrPhe were determined as 89.6%，76.1%，87.2% and 87.8%, respectively, which were in accordance with their theoretical contents. All the chelating rates of CrMet, CrGly, CrThr and CrPhe were above 90%, which indicated the good quality of these CrAA complexes. (2) Four CrAA complexes, including CrMet, CrGly, CrThr and CrPhe were synthesized successfully in aqueous ethanol solution by the chelating reaction of chromium and amino acids. Electronic spectroscopy, infrared spectroscopy, scanning electron microscope, thermalgravimetric analysis and elem |
| 语种 | 中文 |
| 公开日期 | 2015-07-08 |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/15525]  |
| 专题 | 过程工程研究所_研究所（批量导入） |
| 推荐引用方式 GB/T 7714 | 唐海燕. 有机铬螯合物的合成及性能研究[D]. 中国科学院研究生院. 2014. |
入库方式： OAI收割
来源：过程工程研究所
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