摘要
目的 :为基础研究和临床应用研究建立快速、准确、经济的测定大鼠血浆及小肠、肝和肌肉组织内谷氨酰胺水平的方法。 方法 :采用反相高效液相色谱柱前衍生荧光法 (RP- HPL C) ,衍生剂为邻苯二甲醛和 3-巯基丙酸 ,流动相为磷酸缓冲液 -乙腈 (V/ V,94∶ 6 ) ,L ichrosorb C18柱 ,样品与衍生剂按 4∶ 1进行衍生反应 ,激发波长 =2 30nm ,荧光检测波长 =389nm ,流速为 2 .0 ml/ min。 结果 :谷氨酰胺的保留时间为 3.15 8min,检测限为 2 5μm ol/ L(S/ N =3.5 )。线性范围 5 0～ 32 0 0μm ol/ L ,r=0 .9996。日间精密度 RSD值均不大于 6 .97%。 结论 :应用 RP-HPL C能准确、快速、灵敏地测定正常及辐射性肠粘膜损伤的大鼠血浆及组织的谷氨酰胺水平 。
Objectives:To establish a rapid,sensitive and economical method for determination of glutamine in rat plasma and tissue using reversephase high performance liquid chromatography with fluorescence detection. Methods:The glutamine was pre column derivatized with o phthaladehyde and 3 mercaptopropionic acid under an alkaline condition and separated by HPLC on a Lichrosorb C18 column.The mobile phase was consisted of phosphate buffer acetonitrile(V/V,94/6) with a flow rate of 2.0 ml/min.The volume ratio of sample and derivatization reagent solution was 4∶1(V/V).The excited and emitted wavelength were selected at 230 nm and 389 nm respectively. Results:The retention time of glutamine was 3.158 min and the detection limit of glutamine was 25 μmol/L(S/N=3.5),and the regression equation was A=16.940 5C+ 179.933 9, r =0.999 6 at the linear range of 50~3 200 μmol/L.The day to day deviation(RSD)<6.97%. Conclusions:This method is sufficiently sensitive,accurate and precise and can support pre clinical or clinical studies and application.
出处
《肠外与肠内营养》
CAS
2000年第2期93-96,共4页
Parenteral & Enteral Nutrition
关键词
反相高产液相色谱
柱前衍生
谷氨酰胺
组织
血浆
Pre volume derivatization
Reversed phase high performance liquid chromatography
Glutamine
Tissue
Plasma
