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双壳贝类血淋巴中儿茶酚胺的检测方法初步研究
高 菲1,2, 杨红生1, 许 强1,2, 陈慕雁1,2, 张 涛1, 周 毅1
1.中国科学院 海洋研究所 海洋生态与环境科学重点实验室;2.中国科学院 研究生院
摘要:
研究的目的是探索一种检测双壳贝类血淋巴中儿茶酚胺含量的方 法。以栉孔扇贝(Chlamys farreri)和长牡蛎(Crassostrea gigas)为实验对象,研究了取样、抗氧化剂、氧化铝用量以及 Tris缓冲液用量等对实验结果的影响。结果表明,高效液相色谱电化学检测器法(HPLC-ECD)可以灵敏高效地对双壳贝类血淋巴中儿茶酚胺进行定性、 定量检测。实验表明,取样时间最长不超过1.5min;还原性谷胱甘肽作抗氧化剂效果较好;前处理各试剂与血样量的最适配比为血样量1.5mL,Tris 缓冲液(1.5mol/L,pH8.6)1mL和氧化铝25mg;水洗后离心并尽可能吸干氧化铝中的水分。去甲肾上腺素、肾上腺素和多巴胺的回收率分别为 53%~69%、47%~73%和48%~56%。
关键词:  双壳贝类  血淋巴  儿茶酚胺  检测  HPLC-ECD
DOI:
分类号:
基金项目:国家重点基础研究973规划资助项目(G1999012012)
Preliminary study of the detection of catecholamines in the hemolymph of the bivalves
Abstract:
The study is intended to establish a method for the detection of catecholamines in the hemolymph of bivalves, Chlamys farreri and Crassostrea gigas. Different procedures and reagent matching in sample preparation are examined to acquire maximum recovery rate. The results are as follows: (1) the collection time of hemolymph can’t exceed 1.5 minutes; (2) the optimum reagent matching during the extraction process is: 1.5ml hemolymph, 25mg alumina and 1ml Tris buffer; (3) glutathione(GSH) proves to be an effective antioxidant; (4) high-performance liquid chromatography with the electrochemical detector is a sensitive instrument for catecholamines detection. The recovery rates of norepinephrine, epinephrine and dopamine in the study are 53%~69%, 47%~73% and 48%~56% respectively.
Key words:  bivalve  hemolymph  catecholamines  detection  HPLC-ECD
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