Ling Sharon Sheue Nee, Yuen Kah Hay, Barker Susan A
School of Pharmaceutical Sciences, University of Science Malaysia, 11800, Penang, Malaysia.
J Chromatogr B Analyt Technol Biomed Life Sci. 2003 Jan 5;783(1):297-301. doi: 10.1016/s1570-0232(02)00657-8.
A high-performance liquid chromatographic method with ultraviolet (UV) detection was developed for measuring cefotaxime in rat and human plasma. The method used direct injection of the plasma supernatant after deproteinization with 70% perchloric acid. Degradation of cefotaxime in acidic medium was retarded by adding phosphate buffer before centrifuging the sample. The mobile phase was 0.05 M aqueous ammonium acetate-acetonitrile-tetrahydrofuran (87:11:2, v/v) adjusted to pH 5.5. Analysis was run at a flow-rate of 1.0 ml/min, and a detection wavelength of 254 nm was used. The method has a quantification limit of 0.20 microgram/ml. The within- and between-day coefficients of variation and accuracy values were less than 8% and +/-3%, respectively, while the recovery values were greater than 87% over the concentration range tested (0.20-50 microgram/ml). The speed, sensitivity, specificity and reproducibility of this method make it particularly suitable for the routine determination of cefotaxime in human plasma. Moreover, only a relatively small sample plasma volume (100 microliter) is required, allowing this method to be applied to samples taken from neonates.
建立了一种带紫外（UV）检测的高效液相色谱法，用于测定大鼠和人血浆中的头孢噻肟。该方法采用70%高氯酸脱蛋白后直接进样血浆上清液。在样品离心前加入磷酸盐缓冲液可减缓头孢噻肟在酸性介质中的降解。流动相为0.05 M醋酸铵水溶液 - 乙腈 - 四氢呋喃（87:11:2，v/v），pH值调至5.5。分析流速为1.0 ml/min，检测波长为254 nm。该方法的定量限为0.20微克/毫升。日内和日间变异系数及准确度值分别小于8%和±3%，在所测浓度范围（0.20 - 50微克/毫升）内回收率大于87%。该方法的速度、灵敏度、特异性和重现性使其特别适用于人血浆中头孢噻肟的常规测定。此外，仅需相对少量的样品血浆体积（100微升），使得该方法可应用于新生儿采集的样品。
