大鼠血清中人参皂甙Rb1的免疫检测

目的建立大鼠血清中人参皂甙Rb1（G-Rb1）的免疫检识方法。方法利用杂交瘤细胞技术制备抗G-Rb1单克隆抗体（mAb）。含G-Rb1的大鼠血清用甲醇沉淀蛋白，制备供试样品．点样于聚醚砜膜上．以乙腈：水：冰醋酸按25：75：1的比例展开；NaIO4处理，与BSA结合，使G-Rbl固定于膜上。将PES膜上的G-Rb1与抗G-Rb1mAb反应后,加过氧化物酶标记的羊抗小鼠kG，用含0．03％H2O2的lmg／ml 4-氯-1-萘酚磷酸缓冲液显色。结果在聚醚砜膜上，在标准品和供试样品的相同位置可清晰地观察到一个青蓝色G-Rb1斑点，该方法检测限为0．25μg。结论这种G．Rb1的免疫检识方法较薄层层析具有更好的专一性和可靠性，灵敏度接近高效液相色谱。且不需要昂贵的仪器．便于操作。

Immunodetection of ginsenoside Rb1 in rat serum

OBJECTIVE: To establish a novel immunoassay for qualitative detection of ginsenoside Rb1 in rat serum. METHODS: Anti-G-Rb1 monoclonal antibody (mAb) was through a hybridoma approach. Rat serum containing G-Rb1 was deproteinized with methanol to prepare the sample for testing, which was loaded onto polyethersulfone (PES) membrane and developed in the mixture of acetonitrile, water and acetic acid (25:75:1). After treatment with NaIO(4), the membrane was transferred to 1% BSA solution for immobilization of G-Rb1. The membrane was subsequently treated with anti-G-Rb1 mAb solution, followed by addition of peroxidase-labeled goat anti-mouse IgG and color development using 4-chloro-1-naphthol-0.03% H(2)O(2). RESULTS: On the PES membrane, a clear blue spot representing G-Rb1 occurred where the rat serum for testing and the standard G-Rb1 samples were blotted. The limit of this immunodetection was 0.25 microg. CONCLUSION: This immunoassay has greater specificity and reliability than thin-layer chromatography with a sensitivity similar to that of high-performance liquid chromatography, and does not require sophisticated equipment for convenient G-Rb1 detection in rat serum.