Gupta R N, Dauphin A
Department of Laboratory Medicine, St. Joseph's Hospital, Hamilton, Ont., Canada.
J Chromatogr B Biomed Appl. 1994 Aug 5;658(1):113-9. doi: 10.1016/0378-4347(94)00214-2.
A convenient and economical procedure for the extraction of bupivacaine and its major metabolites, desbutylbupivacaine and 4'-hydroxybupivacaine, from serum is described. A 0.25-ml aliquot of 4 micrograms/ml solution of N-pentyl-2,6-pipecoloxylidide used as the internal standard and an equal volume of the serum sample are applied to a 1-ml BondElut C18 silica extraction column which has been conditioned by subsequent washing once with 1 M HCl, twice with methanol and once with water. After passing the sample at a slow rate, the column is washed twice with water and once with acetonitrile. The desired compounds are then eluted with a 0.25-ml aliquot of a mixture of 35% perchloric acid-methanol (1:40, v/v). A 5-microliters aliquot of the eluate is injected onto a 150 x 4.6 mm I.D. column packed with 5-microns C8 silica particles, which is eluted at ambient temperature with a mobile phase of acetonitrile-10 mM KH2PO4 (25:80, v/v) at a flow-rate of 1.5 ml/min. The peaks are detected at 205 nm. The extraction recovery of all the compounds is in the range of 90%. The chromatogram is clean and the desired peaks are well separated from each other and from the extraneous peaks.
本文介绍了一种从血清中提取布比卡因及其主要代谢物去丁基布比卡因和4'-羟基布比卡因的简便经济方法。将0.25 ml浓度为4 μg/ml的N-戊基-2,6-哌啶基利多卡因溶液作为内标，与等体积的血清样品加入到1 ml BondElut C18硅胶萃取柱中，该萃取柱先用1 M盐酸洗涤一次，再用甲醇洗涤两次，最后用水洗涤一次进行预处理。样品缓慢通过柱子后，柱子先后用水洗涤两次，用乙腈洗涤一次。然后用0.25 ml 35%高氯酸-甲醇（1:40，v/v）混合溶液洗脱所需化合物。取5 μl洗脱液注入内径为150×4.6 mm、填充5 μm C8硅胶颗粒的柱子中，在室温下用乙腈-10 mM KH2PO4（25:80，v/v）作为流动相，以1.5 ml/min的流速进行洗脱。在205 nm处检测峰。所有化合物的萃取回收率在90%范围内。色谱图干净，所需峰彼此之间以及与杂质峰之间分离良好。
