尿中s-羟乙基半胱氨酸的OPA自动柱前衍生高效液相色谱法测定

DETERMINATION OF S-HYDROXYETHYL CYSTEINE IN URINE BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH PRE-COLUMN DERIVATIZATION WITH O-PHTHALALDEHYDE.

目的 :建立尿中 s-羟乙基半胱氨酸的 OPA自动柱前衍生高效液相色谱法测定方法。方法 :尿液经 Sep- PakC18小柱富集和纯化 ,样液中 s-羟乙基半胱氨酸在 HP10 5 0高效液相色谱仪自动进样器中使用邻苯二甲醛 (OPA)自动衍生 ,经 L ichrospher 10 0 RP- 18e色谱柱分离 ,用 DAD检测器在 338nm波长下测定吸收值。结果 :s-羟乙基半胱氨酸浓度范围0 .0 mm ol/ L ～ 0 .5 mm ol/ L 的校准曲线相关系数为 0 .9999,方法检出限为 0 .0 0 2 m mol/ L;s-羟乙基半胱氨酸浓度 0 .0 5 mm ol/L、 0 .30 m mol/ L、 0 .5 0 mm ol/ L组内相对标准偏差 (RSD)分别为 1.6 7%、 1.0 2 %和 1.4 0 % (n=6 ) ;不同日期测定组间相对标准偏差 (RSD)分别为 4 .78%、 3.82 %和 3.74 % (n=6 )。尿中加标 0 .10 mm ol/ L、 0 .30 mm ol/ L 的回收率分别为 90 %、93%。结论 :方法线性好 ,精密度和回收率高 ,操作简便 ,自动化程度高 ,可用于测定尿中

Objective:To develop method of detecting s hydroxyethyl cysteine in urine by high performance liquid chromatography(HPLC) with pre column derivatization with o phthalaldehyde(OPA).Methods:Urine samples were purified and richen by small column (Sep Pak C18).s hydroxyethyl cysteine in purified samples reacted with OPA in autosampler of HPLC,separated by column Lichrospher 100 RP-18e and detected by DAD detector in wavelengh 338nm.Results:The value of s hydroxyethyl cysteine standard curve(concentration 0 0~0 5mmol/L) was 0 9999.Detection limit of method was 0 002mmol/L.Inner relative standard deviates (RSD) of 0 05mmol/L?0 30mmol/L?0 50mmol/L s hydroxyethyl cysteine were 1 67%?1 02% and 1 40%( n =6),respectively.Outer relative standard deviates (RSD) of 0 05mmol/L?0 30mmol/L?0 50mmol/L s hydroxyethyl cysteine were 4 78%?3 82% and 3 74%( n =6),respectively.Recoveries of 0 10mmol/L?0 30mmol/L s hydroxyethyl cysteine were 90%?93%,respectively.Conclusion:The method was linear and repetitive,and have satisfied recoveries. The program of method was also automatic and simple.Therefore,it was suitable for detecting s hydroxyethyl cysteine in urine.