【摘要】 目的建立高效液相色谱荧光法测定尿液中犬尿喹啉酸(KYNA)含量的方法,观察不同周龄自发性高血压大鼠(SHR)及其正常对照大鼠(WKY)的尿KYNA含量变化。方法高效液相色谱荧光法采用的色谱柱为ODS色谱柱,流动相为10 mmol/L醋酸钠缓冲液(pH4.5)和乙腈(体积95∶5),激发波344 nm,发射波398 nm。选择4周龄WKY大鼠和SHR各4只、16周龄WKY大鼠和SHR各6只检测24 h尿液中的KYNA含量,并进行统计学分析。结果KYNA浓度检测的线性范围0.220-33.000nmol/mL(R2=0.999 1),回收率为(90.37±8.47)%,日内差异为4.70%~9.40%,日间差异为5.20%~7.50%。4周龄WKY组24 h尿KYNA含量为(41.251±22.663)μg/24 h尿,SHR组为(35.387±10.814)μg/24 h尿,两组间无统计学差异(P<0.05);16周龄SHR尿KYNA含量为(39.511±15.985)μg/24 h尿,较WKY组(90.690±42.189)μg/24 h尿明显降低(P<0.05)。结论高效液相色谱荧光方法具有灵敏、稳定、特异性高等特点,可用于临床检测尿KYNA。成年SHR尿KYNA含量较WKY大鼠减少,提示可能与高血压的发生发展有关。更多还原

【Abstract】 Objective To develop a method of high-performance liquid chromatography(HPLC) for the measurement of urine kynurenic acid(KYNA) and to observe the urine KYNA of spontaneously hypertensive rats(SHR) and WKY rats with different ages. Methods Chromatography was accomplished with ODS column and fluorescence detection,with the mobile phase of sodium acetate buffer(10 mmol/L,pH=4.5) and acetonitrile(volume,95∶5),excitation wavelength at 344 nm and emission wavelength at 398 nm.Four SHR and four WKY of 4 weeks old and six SHR and six WKY of 16 weeks old were collected,and 24-h urine KYNA was measured and analysed. Results The linear range of KYNA was 0.220-33.000 nmol/mL(R2=0.9991).The method had a high recovery rate of(90.37± 8.47)%.The within-and between-day accuracy ranged from 4.70%-9.40% and 5.20%-7.50%,respectively.Urine KYNA was significantly reduced in 16-week-old SHR compared with the age-matched WKY,(39.511±15.985) μg/24 h urine and(90.690±42.189) μg/24 h urine,respectively(P<0.05).However,there was no significant difference between 4-week-old SHR and WKY rats,(35.387±10.814) μg/24 h urine and(41.251±22.663) μg/24 h urine,respectively. Conclusion The assay for urine KYNA by HPLC is ideal in sensitivity,accuracy and consistency.The urine KYNA is reduced in the SHR than the WKY rats,which indicates that KYNA metabolism may play a role in the pathogenesis of experimental hypertension.更多还原