高效液相色谱法测定羊水中兴奋性氨基酸、抑制性氨基酸的含量  

Determination of Excitatory Amino Acids and Inhibitory Amino Acids in Amniotic Fluid by High Performance Liquid Chromatography

作　　者：李平[1] 张建林[2] 王爱勤[1] 杜建丽[3] 

机构地区：[1]中国科学院兰州化学物理研究所,兰州730000 [2]甘肃省药品检验所,兰州730000 [3]兰州大学第二医院,兰州730030

出　　处：《分析化学》2005年第6期772-776,共5页Chinese Journal of Analytical Chemistry

摘　　要：建立了反相高效液相色谱法测定羊水中兴奋性氨基酸天门冬氨酸、谷氨酸,抑制性氨基酸甘氨酸含量的方法,色谱柱为LUNAC18柱(4.6mm×250mm,5μm),流动相为乙腈水(V/V),按低压梯度洗脱,梯度系统0～4min,乙腈40%;4～20min,乙腈40%～52%;20～30min,乙腈52%～60%;30～40min,乙腈60%～40%。流速0.8mL/min,柱温40℃,紫外检测波长260nm;进样量10μL。结果显示,天门冬氨酸、谷氨酸和甘氨酸三者的线性范围均为2.5～85mg/L;标准加入的回收率为99.6%～102.6%。本方法灵敏度高、测定结果可靠。用本方法对201例不同孕期的羊水作兴奋性和抑制性氨基酸检测,发现羊水中3种氨基酸随妊娠进展而增加的趋势,胎儿畸形或宫内缺氧时羊水中这3种氨基酸水平明显升高。A determination method of excitatory amino acids and inhibitory amino acids in amniotic fluid by reversed-phase high-performance liquid chromatography ( HPLC) was established. The chromatographic column LUNA C-18 column ( 4.6 mm x 250 mm, 5 mu m) was used, the temperature of column was 40 degrees C and detected wavelength at 260 nm. Mobile phase was acetonitrile-water, the separation according to low press gradient, the gradient system was 0 4 min,40% acetonitrile, increased to 40%similar to 52% acetonitrile at 4 similar to 20 min, and 52%similar to 60% acetonitrile at 20 similar to 30 min, and held at 60%similar to 40% acetonitrile until 30 similar to 40 min. All HPLC solvents were filtered through 0.45 mu m (pore size) filters and degassed, the flow rate was 0.8 mL/min; injection volume 10 mu L. The linear range of aspartic acid, glutamic acid and glycin were 2.5 similar to 85 mg/L, the average recovery were 99.6%similar to 102.6%. In this study, an accurate, credible and sensitive reversed-phase high-performance liquid chromatographic method has been developed for the determination of excitatory amino acids and inhibitory amino acids in amniotic fluid. The excitaotry amino acids and inhibitory amino acids in 201 cases of different pregnant weeks aminotic fluid were detected by this method, the results showed that three amino acids were increase with extensible pregnant weeks, three amino acids were obrious increase in the amniotic fluid of fetus abnormality and in trauterine asphyxia.

关 键 词：兴奋性氨基酸 抑制性 羊水 反相高效液相色谱法 天门冬氨酸 紫外检测波长 C18柱 梯度洗脱 梯度系统 线性范围 标准加入 测定结果 胎儿畸形 谷氨酸 乙腈 酸含量 6mm 色谱柱 流动相 min 进样量 甘氨酸 回收率 灵敏度 柱温 

分 类 号：R446.1[医药卫生—诊断学]