一测多评法测定小鼠脑内单胺类递质及其相关物质的含量  

Simultaneous determination of monoamine neurotransmitters and related substances in mouse brain tissue by QAMS

作　　者：朱旦[1] 韦芳芳[1] 曾常青[1] 

机构地区：[1]广东药科大学中药学院/国家中医药管理局中药数字化质量评价技术重点研究室,广东广州510006

出　　处：《广东药学院学报》2016年第2期181-186,共6页Academic Journal of Guangdong College of Pharmacy

基　　金：贵州省剑河县中药材产业科技合作专项计划项目[剑科合字(2014)4号]

摘　　要：目的建立并优化一种简单、灵敏、高选择性的高效液相色谱荧光检测法,通过一测多评法测定小鼠脑内单胺类递质及其相关物质。方法采用Thermo Scientific Syncronis a Q C18色谱柱,流动相为0.1mol/L Na Ac溶液(内含0.05 mol/L EDTA,用HAc调p H值4.70)∶甲醇(体积比95∶5),流速为1.0 m L/min,激发波长为280 nm,发射波长为330 nm。以多巴胺盐酸盐(DA)对照品为内参物,建立其与去甲肾上腺素(NE)、盐酸肾上腺素(E)、酪氨酸(Tyr)、3,4-二羟苯乙酸(DOPAC)、3-甲氧基-4-羟基苯乙二醇半哌嗪盐(MHPG)、5-羟色胺盐酸盐(5-HT)、色氨酸(Trp)、5-羟吲哚乙酸(5-HIAA)、高香草酸(HVA)的相对校正因子,并进行了方法学研究。结果采用校正因子法和外标法分别计算小鼠脑内该10种成分的质量浓度,两法计算所得各成分平均质量浓度之间差异均无统计学意义(P>0.01)。结论本方法操作简便、快速,测定结果准确度较高,可用于小鼠脑内10种单胺类递质及其相关物质的测定。Objective To establish a quantitative analysis of multi-components by single maker（ QAMS）method for simultaneous determination of monoamine neurotransmitters in mice brain and its related substances by simple,sensitive and selective high-performance liquid chromatography with fluorescence detection（ HPLC-FLD）. Methods The analytes were separated on a thermo scientific syncronis a Q C18 column.The mobile phase was consisted of 0.1 mol / L sodium acetate buffer（ 0.05 mol / L EDTA,pH4.70）and methanol（ 95 ∶5,φ） at a flow rate of 1.0 m L / min. The FLD wavelength was set at 280 nm and 330 nm for excitation and emission,respectively. Dopaminehydroehloride（ DA） was used as the internal reference substance. The relative correlation factors（ RCF） of norepinephrine（ NE）,L-epinephrine hydrochloride（ E）,tyrosine（ Tyr）,3,4-dihydroxy-phenylacetic acid（ DOPAC）,4-Hydroxy-3-methoxyphenylglycol hemipiperazinium saltl（ MHPG）,5-hydroxytryptamine（ 5-HT）,tryptophan（ Trp）,5-hydroxyindole-3-acetic acid（ 5-HIAA） and homovanillic acid（ HVA） were determined by HPLC-FLD. The methodology validation was carried out. Results The contents of the ten components were calculated by both QAMS and external standard method. No significant differences were observed in the quantitative results of ten monoamine neurotransmitters（ P〉 0.01） and its related substances determined by external standard method and QAMS. Conclusion The QAMS method is feasible and accurate,and can be used for the quality control of monoamine neurotransmitters and related substances in mouse brain tissue.

关 键 词：一测多评法 单胺类神经递质 高效液相-荧光检测器 

分 类 号：R-33[医药卫生]