摘要
用固相萃取法处理样品，以扑热息痛为内标物、甲醇－醋酸钠／醋酸缓冲溶液作流动相，采用反相高效液相色谱法测定血清中头孢噻肟的浓度。头孢噻肟和内标物的平均回收率分别为９６．７％和９７．７％，头孢噻肟血清浓度在１０ｍｇ／Ｌ至１５０ｍｇ／Ｌ范围内有良好线性关系，最低检测浓度为２ｍｇ／Ｌ，日内和日间相对标准偏差分别在３．０％和４．１％以内。结果表明方法准确、简便。
A reversedphase HPLC assay has been developed for measuring cefotaxime(CTX) in human serum. A solidphase extraction column (Bond Elute C 18 ) was used for the prepurification of sample. Serum (200μL) was mixed with 100μL of internal standard (acetylamino phenol) solution and 200μL of sodium acetate buffer (0.01mol/L, pH 5.8), loaded onto the conditioned column and washed with 2mL of acetate buffer, the compounds were eluted with 0.5mL×2 of methanolacetate buffer (40∶60, V/V). The eluate (20μL) was injected directly into an Apex ODS column (250mm×4.6mm i.d., 5μm) with a mobile phase of acetate buffer (0.01mol/L, pH 5.8)methanol (80∶20, V/V),detection was achieved at 254nm. The average extraction recoveries were 96.7% and 97.7% for CTX and internal standard respectively, which were higher than that of CTX(91.0%) obtained by protein precipitation with methanol. The solidphase extraction method also provided much clearer samples for HPLC analysis. The concern of acid degradation of CTX when an acid is used as a protein precipitation agent was avoided. The calibration curve of CTX was linear between 10 and 150mg/L of serum concentration with a correlation coefficient of 0999 2, and the minimum detection limit was 2mg/L. The precision was tested with three sample concentrations, and withinday RSD below 3.0% and daytoday RSD below 4.1% were achieved. Maximum serum concentrations of 25.95 and 22.89 mg/L were measured at 45min after intramuscular injection of 1g CTX to two healthy volunteers respectively. The chromatographic behaviour of CTX was also studied.
出处
《色谱》
CAS
CSCD
北大核心
1997年第6期512-514,共3页
Chinese Journal of Chromatography
关键词
固相萃取
头孢噻肟
血清
HPLC
CTX
high performance liquid chromatography, solidphase extraction, cefotaxime, serum
