毛细管电泳紫外法检测重度创伤患者血清谷氨酰胺含量的实验研究

Determination of Glutamine in Serum of the Patients with Severe Trauma by Capillary Electrophoresis with UV Detection

目的探讨重度创伤患者血清中谷氨酰胺(glutamine,Gln)含量准确、简便、快速的检测方法。方法选择我院重度创伤10例(创伤组),分别在术前1 d及术后5、10 d采集静脉血制备血清。选择同期健康体检10例为对照组,同法制备血清。以四硼酸钠(20 mmol/L,pH 9.33)、十二烷基硫酸钠(166 mmol/L)和乙腈按10∶10∶3配置缓冲液,异硫氰酸苯酯为衍生剂,在电压15 kV、紫外吸收波长254 nm下用直径75μm、长58 cm的石英毛细管检测两组血清Gln含量。结果 Gln浓度在0.03～2.00 mmol/L范围内线性关系良好,相关系数为0.9901;精密度为3.14%,平均回收率为92.99%～110.28%;最低检测限浓度为0.01 mmol/L。对照组Gln含量0.79 mmol/L,创伤组术后5 d时Gln出现相对最低点,且术前1 d及术后5、10 d血清Gln浓度均低于对照组,差异有统计学意义(P<0.05)。结论该毛细管电泳紫外法快速、准确而简便,灵敏度高,重复性好,能有效分离检测重度创伤患者血清中Gln含量。

Objective To explore an accurate, simple and rapid method for determination of glutamine in serum of severe trauma patients. Methods Severe trauma patients（trauma group） were selected, and their venous blood samples were respectively taken one day before the operation, five and ten day after trauma, then serum samples were prepared. In the corresponding period, the normal physical examination group were selected to be control group（ n = 10）, and their serum samples were prepared with the same method. Bouncy silica capillary： 75 /μm dia, 58 cm from the inlet to UV detection. Electrolyte： sodium tetraborate（ pH 9.33, 20 mmol/L）, lauryl sodium sulfate（166 mmol/L） and acetonitrile, which was prepared according to the ratio of 10： 10： 3. Phenylisotbiocyanate （PITC） was used to derivatize Gin. Running voltage was 15 kV and detection wavelength was 254 nm. Two groups serum samples were detected. Results There was a good linearity range when the concentration of Gln was between 0.03 mmol/L and 2.00 mmoL/L（ r = 0. 9901 ）. Precision was 3.14% and the average recovery rate was 92.99% -110.28%. Lowest detectable limit was 0.01 mmol/L. The Gin content in normal group was 0.79 mmol/L and the relatively low content of Gin appeared on the 5th day after operation in trauma group. Conclusion The capillary eleetrophoresis method with UV detection is rapid, accurate and simple in the effective separation of glutamine in serum of severe trauma patients. It has high sensitivity and good reproducibility.