摘要
目的 :建立一种运用高速扫描技术测定血中环孢素A浓度的HPLC法。方法 :取全血 2 .0ml,加盐酸溶液 2ml后 ,以3ml乙醚两次提取 ;分离乙醚液后用氢氧化钠液洗涤后蒸干 ;以乙腈 /水溶液 6 0 μl重组 ,再用正己烷洗涤后进样 2 0 μl。色谱条件 :Elite 2 2 0mm× 4 .6mmC18柱 ,柱温 6 5℃ ,以甲醇 水 (88∶12 )为流动相 ,流速 1.0ml·min-1,扫描波长 2 0 5～ 2 5 0nm(间隔5nm) ,积分波长 :2 10nm。结果 :CsA浓度在 5 0～ 10 0 0ng·ml-1范围内 ,具有良好线性关系。本法血样的平均萃取回收率达92 .5 %。采用随机提供的FOCUS软件中的光谱分析功能 ,可判断样品峰的纯度。结论 :本法操作较简便 ,稳定性好 ,可判断每次测定结果是否受到干扰 。
OBJECTIVE To develop a high performance liquid chromatographic method utilizing high speed scanning technology for the determination of cyclosporin A (CsA) in human whole blood. METHODS Adding HCl 2 ml of HCl solution( 0.025 mol·L -1 ) to whole blood sample, mixed the mixture well and extracted with 3 ml of ether twice. Ether layer was washed with 2 ml of NaOH solution( 0.025 mol·L -1 ) and evaporated to dryness after isolation. The residue was dissolved in 60 μl of acetonitrile water(70∶30) solution and 20 μl was injected for the determination after washed by 800 μl of n hexane. Elution was performed on Elite C 18 column(220 mm× 4.6 mm ) with methanol water(88∶12) as mobile phase. The flow rate was 1.0 ml·min -1 and column temperature was maintained at 65℃. The scanning wavelengths were from 205 nm to 250 nm with a 5 nm interval and 210 nm was elected as the integration wavelength.RESULTS A linear relationship was obtained between the peak areas and CsA blood concentration from 50 ng·ml -1 to 1000 ng·ml -1 ( γ = 0.9989 ). The extraction recoveries of CsA from whole blood were 92.03% ～ 93.13% . RSD within day and between days were less than 5.3% ( n =5) and 6.1% ( n =5) respectively. Spectra analysis function of spectra FOCUS Software could be used to analyze the peak purity. CONCLUSIONS The method is accurate,stable and simple. The analyst could find if the peak of CsA is interfered by other concomitant drugs, so the method is very suitable for routine clinical monitoring of CsA concentration.
出处
《中国医院药学杂志》
CAS
CSCD
北大核心
2001年第12期709-711,共3页
Chinese Journal of Hospital Pharmacy
