摘要
目的建立检测酒精依赖标记物糖缺陷转铁蛋白(CDT)的高效液相紫外(HPLC-UV)色谱分析方法。方法抽取人体血样3 mL,分离准备血清,加碳酸氢钠(500 mmol/L)和氯化铁(10 mmol/L),使血清中的铁离子饱和;再用硫酸葡聚糖(100 g/L)和氯化钙(1 mmol/L)沉淀血清中的脂蛋白,该样品放置在4℃冰箱中30~60 min,取出样品2 000 r/min离心5 min,将离心后的样品上清液置于冻存管中,用超纯水稀释10倍,将稀释样品经超声波排除气泡,用0.45 mm微孔滤膜过滤,送样品用高效液相色谱仪—紫外检测器进行测定。采用质控样品和空白样品对照法,计算糖缺陷转铁蛋白(CDT)在所有的唾液酸转铁蛋白中的百分含量。结果 HPLC-UV检测CDT灵敏度为95.7%,特异性为92.3%,并且与金标准毛细管电泳法(CZE)间具有良好的线性关系,回归方程为Y=0.896 8X+0.691 3,R^2=0.935 1。结论 HPLC-UV检测CDT有较高的灵敏度和特异性,对于法医司法和临床酒精依赖的诊断和治疗监控有较高的应用价值。
Objective To establish high-performance liquid chromatography ultraviolet(HPLC-UV)methods to detect carbohydrate-deficient transferrin(CDT)in serum.Methods Blood samples were taken from drinkers and 3 ml was prepared.Serum 0f alcohol-dependent drinkers and nondrinkers was saturated with iron by adding NaHCO-3(500 mmol/L)and FeCl-3(10 mmol/L).After mixed and stored for 30-60 min at 4℃,the lipoproteins were precipitated by adding dextran sulfate(100 g/L)and CaCl-2(1 mol/L).After the mixture was centrifuged,the supernatent was diluted tenfold with water.The prepared samples were filtered by ultrasound and 0.45 m microporous membranes and determined by HPLC-UV detector.The percentage of CDT in all sialic acid transferrins was calculated by quality control and blank sample control.Results HPLC-UV method had good sensitivity(95.7%)and specificity(92.3%),and the gold standard method of capillary electrophoresis(CZE)had a good linear relationship with the regression equation:Y=0.896 8X+0.691 3,R2=0.935 1.Conclusion HPLC-UV has high sensitivity and specificity in the detection of CDT.It has higher application value in forensic identification and clinical alcohol dependence diagnosis and treatment monitoring.
作者
黄培培
谢辉
米尔孜合买提.买买提明
王世雯
艾克拜尔.热合曼
HUANG Peipei;XIE Hui;Mirzahmat Mamtimin;WANG Shiwen;Akbar Rahman(Department of Forensic Medicine,School of Preclinical Medicine,Xinjiang Medicial University,Urumqi 830011,China;Department of Alcohol Abstinence,The Fourth People′s Hospital of Urumqi,Urumqi 830000,China)
出处
《新疆医科大学学报》
CAS
2018年第4期472-477,共6页
Journal of Xinjiang Medical University
基金
国家自然科学基金(81360466)
关键词
糖缺陷转铁蛋白
高效液相
酒精依赖
carbohydrate-deficient transferrin(CDT)
high-performance liquid chromatography(HPLC)
alcohol dependence
