免疫法制备液相色谱生物样品的研究——血中沙丁胺醇的分析

 用动物免疫法制备了免疫亲和柱纯化水溶性的沙丁胺醇血浆样品。琥珀酸酐交联沙丁胺醇和牛血清白蛋白获得抗原免疫家兔抗沙丁胺醇抗体——免疫球蛋白。琼脂糖Ｓｅｐｈａｒｏｓｅ４Ｂ与抗体交联制成免疫球蛋白亲和柱。对高效液相色谱法测定中的一般提取方法和固相小柱提取法作了比较，后者具有内源性杂质干扰少的优点，是生物样品预处理的一种有效的方法。

Immunologically prepared plasma specimen of albuterol for high performance liquid chromatographic determination]

Immune-affinity method for preparation of sample in plasma is reported in this paper. Albuterol was coupled to bovine serum albumin (BSA) to prepare the antigen and immunized in rabbit to produce the anti-albuterol serum (immune globulin). The anti-albuterol serum was treated with saturated ammonium sulfate solution to obtain the antibody (immune globulin). The antibody was coupled with Sepharose 4B to prepare the immune globulin-affinity column. The plasma containing albuterol was introduced into the column and was rinsed in turn with phosphate buffer and water, and then eluted with methanol. After being concenfrated the sample was analyzed by HPLC with a sillica column (4.6 mm x 200 mm, 5 microm), a mixture of methanol: 2 mol/L ammonium acetate (99.5 : 0.5) as mobile phase with a flow rate of 1.2 mL/min, and fluorecence detector monitoring at lambdaex = 226 nm and lambdaem = 306 nm. The results showed that the immune globulin-affinity column has the characteristic of high purity and high specificity, and it is suitable for the preparation of chromatographic plasma specimen to determine albuterol in plasma. The relationship between peak area ratios and concentration from 2 to 80 microg/L were linear (r=0.998). The extraction recoveries from plasma at concentrations of 2-80 microg/L were 98.8% +/- 7.3%. The RSD for intra-day (n=5) was 5.8% and that for inter-day assay (n=5) was 7.8%.