Haagsma N, Keegstra J R, Scherpenisse P
Department of the Science of Food and Animal Origin, Faculty of Veterinary Medicine, University of Utrecht, Netherlands.
J Chromatogr. 1993 Jun 2;615(2):289-95. doi: 10.1016/0378-4347(93)80343-3.
A rapid clean-up procedure based on ion-pair solid-phase extraction (SPE) for the high-performance liquid chromatographic (HPLC) determination of spectinomycin in swine, calf and chicken plasma at a limit of detection of 50 ng/ml is described. After dilution with water and adjustment of the pH to approximately 5.6, the plasma is applied to a high-hydrophobic C18 SPE column treated with sodium dioctylsulphosuccinate. Spectinomycin is eluted with methanol and derivatized with 2-naphthalene sulphonyl chloride prior to chromatography. The HPLC set-up consists of a dual-column system using two Chromspher silica columns and dichloromethane-acetonitrile-ethyl acetate-acetic acid, in different ratios, as mobile phases. Detection is performed at 250 nm. Quantification is carried out using external standards prepared in blank cleaned plasma. Mean recoveries were 83 +/- 3% (n = 5), 93 +/- 6% (n = 5) and 92 +/- 6% (n = 6) for swine, calf and chicken plasma, respectively, at the 0.1 microgram/ml level.
本文描述了一种基于离子对固相萃取（SPE）的快速净化方法，用于高效液相色谱（HPLC）测定猪、牛和鸡血浆中的壮观霉素，检测限为50 ng/ml。血浆用水稀释并将pH值调至约5.6后，应用于用二辛基磺基琥珀酸钠处理过的高疏水性C18 SPE柱。壮观霉素用甲醇洗脱，并在色谱分析前用2-萘磺酰氯衍生化。HPLC装置由一个双柱系统组成，使用两根Chromspher硅胶柱，以不同比例的二氯甲烷-乙腈-乙酸乙酯-乙酸作为流动相。检测在250 nm波长下进行。使用在空白净化血浆中制备的外标进行定量。在0.1微克/毫升水平下，猪、牛和鸡血浆的平均回收率分别为83±3%（n = 5）、93±6%（n = 5）和92±6%（n = 6）。
