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题名: 高液相色谱与石墨炉原子吸收光谱联用技术(HPLC-GFAAS)分离测定金属硫蛋白及其金属含量
作者: 孙朋
学位类别: 硕士
答辩日期: 1991
授予单位: 中国科学院研究生院
授予地点: 北京
导师: 单效全
中文摘要:
      本论文共由四章组成。第1章对近期关于金属硫蛋白(MTs)的存在部位、诱导、结构、生理功能、分离测定及其金属含量等方面的研究工作进行了综述,以各种分离测定MTs的方法的原理及其实际应用作了较为细致的探讨。其中包括:金属饱和法(如Hg/TCA, Cd/hem, Ag/hem等方法),示差脉冲极谱法(DPP)、放射免疫测定法(RIA)、阴离子交换高效液相色谱与原子吸收光谱联用法,反相高效液相色谱法等。同时对各种测定MTs中金属含量的阴离子交换高效液相色谱与石墨炉原子吸收光谱联用方法(anion-exchange HPLC-GFAAS)。实验中采用DEAE-5PW阴离子交换柱,以Tris-HCl缓冲液淋洗(0.01-0.25M, pH8.6)分离兔肾中MT异构体(MT-I和MT-II),同时进行紫外检测(254nm)。由紫外吸收   峰高定量MT浓度。结果表明,以饮食方式摄入一定量金属镉可使肾脏中MTs含量增高而肝脏中MTs含量没有明显变化。MT-I与MT-II对金属镉有不同的结合能力。以饮食方式进入体内的镉可引起肾脏中锌含量的增高。第3章研究了反相高效液相色谱与石墨炉原子吸收光谱联用法分离测定MTs及MTs中金属镉含量测定。各种MT异构体在C18反相柱上用中性NaH_2PO_4缓冲液进行洗脱同时用紫外检测(254nm)。实验中发现兔肝MTs由七种异构体组成。其中五种为MT-I的亚形态。兔肾MTs只有两种异构体(MT-I和MT-II)。通过饮食摄入一定量镉可引起MTs在肾中的诱导和聚集,肝脏中MTs没有明显变化。肾脏中总镉量的约10%与MT结合。第4章利用阴离子交换高效液相色谱与石墨炉原子吸收光谱联用技术分离测定了重金属污染水貂肝脏中MTs含量及MTs中镉含量。当用从污染水中捕捞的鱼喂养水貂时,水貂肝脏中MT-I含量增高,MT-II含量没有明显变化。随着重金属污染程度的增高,MT-I中镉含量也有一定程度增高。
英文摘要:       This thesis consists of four Chapters. In Chapter 1 recent work on the occurance, induction, structure, biological roles, separation and qutitation of metallothioneins (MTs) and heavy metal content determination in MTs have been reviewed. The principles and practical application of various methods for MTs separation and determination, such as metal saturation (Hg/TCA, Cd/hem, Ag/hem), differential pulse polarography (DPP), radioimmunoassay (RIA), anion-exchange HPLC-AAS and reversed-phase HPLC, have been discussed in detail. Determination of heavy metal content in MTs by various methods has also been reviewed in comprehensive form. Chapter 2 presents a rapid method for the determination of dietary cadmium induced MTs and cadmium content in MTs in rabbit kidney by anion-exchange high-performance liquid chromatography (HPLC) coupled with graphite furnace atomic absorption spectrometry (GFAAS). Rabbit kidney MT isoforms (MT-I and MT-II) were eluted respectively from a DEAE-5PW anion-exchange column with Tris-HCl buffer (0.01-0.25 M, pH 8.6) and detected by UV absorbance at 254 nm. MT concentrations were determined based on UV absorbance peak height. The results suggested that feeding a dose of cadmium for some days resulted in the increase in MT concentration in rabbit kidneys, but not in the livers. MT-I and MT-II showed some differences associated with the oral intake cadmium. Dietary cadmium also caused zinc to accumulate in kidney to some extent. In chapter 3 the feasibility of using reversed-phase HPLC for the separation and subsequent determination cadmium in MTs by GFAAS has been studied. The MT isoforms were eluted from a radially compressed C18 column with a neutral sodium phosphate buffer and detected by UV absorbance at 254 nm. Rabbit liver MTs was found to be comprised of three distinct isoforms. with two of which were found to be subspecies of MT-I isoform. Rabbit kidney MTs exhibited only two predominant isoforms. Feeding stable cadmium in the diet resulted in the deposition of MT in the kidney rather than in the liver. About 10% of total cadmium in kidney was associated with MTs. MT in livers of minks feeding heavy metal contaminated fish and cadmium in liver MT were determined by anion-exchange HPLC coupled with GFAAS in chapter 4. Obvious increase in liver MT-I concentration rather than liver MT-II was found when the minks were fed contaminated fish captured from the heavy metal polluted water. The cadmium concentration in mink liver MT-I also increased to some extent as the contaminated level increased.
内容类型: 学位论文
URI标识: http://ir.rcees.ac.cn/handle/311016/34901
Appears in Collections:环境化学与生态毒理学国家重点实验室_学位论文

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Recommended Citation:
孙朋. 高液相色谱与石墨炉原子吸收光谱联用技术(HPLC-GFAAS)分离测定金属硫蛋白及其金属含量[D]. 北京. 中国科学院研究生院. 1991.
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