摘要
目的建立一种新型的氧化还原柱前衍生化荧光检测高效液相色谱法测定亮氨酸脑啡肽(leu-enkephalin,L-EK)药物。方法将肽类药物L-EK与0.012mol/L高碘酸钠、0.01mol/L儿茶酚、0.3mol/L硼酸按体积比2∶2∶1∶1混合,于120℃反应10min后,用反相柱分离,以0.25mol/L硼酸盐缓冲液(pH 7.6)和甲醇为流动相,采用梯度洗脱,荧光检测器检测。同时与高效液相色谱紫外法、邻苯二甲醛(O-phthalaldehyde,OPA)柱前衍生荧光检测高效液相色谱法比较,测定体外及血样中L-EK的含量。结果氧化还原柱前衍生化荧光检测高效液相色谱法检测含L-EK血样,线性范围0.5～50.0mg/L,线性良好(r=0.999 8),血浆中检测限为0.1mg/L,且较另外两种方法不受内源性物质的干扰。结论新型的氧化还原柱前衍生化荧光检测高效液相色谱法专属性好,灵敏度高,尤其适合生物样品中L-EK药物的测定。
Objective To establish a novel high performance liquid chromatographic method with fluorimetric detector（HPLC-FLU）,by using pre-column redox derivatization for the determination of leu-enkephalin（L-EK） drugs.Methods L-EK was mixed with 0.012 mol/L sodium periodate,0.01 mol/L catecholamine and 0.3 mol/L boric acid（2 ∶ 2 ∶ 1 ∶ 1,V/V）.The mixture was heated at 120 ℃ for 10 minutes.The chromatographic separation was performed on an ODS column with 0.25 mol/L borate buffer（pH 7.6）-methanol as mobile phases with gradient elution.The derivatized compounds were detected with fluorimetric detector.L-EK was detected by this method in vitro and in human plasma,compared with HPLC-UV and HPLC-FLU with pre-column O-phthalaldehyde（OPA） derivatization.Results The linear range of HPLC-FLU with pre-column redox derivatization was 0.5-50.0 mg/L（r=0.999 8） in human plasma and the limit of detection was 0.1 mg/L.This method was better than the other two not being disturbed by endogenous substances.Conclusions HPLC-FLU with pre-column redox derivatization has advantages in detecting L-EK in human plasma.
出处
《复旦学报（医学版）》
CAS
CSCD
北大核心
2012年第3期301-304,共4页
Fudan University Journal of Medical Sciences
基金
上海市科学技术委员会中药现代化项目(08DZ1971201)~~
