摘要
目的：建立柱前衍生高效液相色谱法（HPLC）用于河蚌糖胺聚糖在大鼠血浆中的含量测定。方法：使用异硫氰酸荧光素（FITC）对河蚌糖胺聚糖进行荧光标记，色谱柱为Shodex SB-804HQ（8．0×300mm），流动相为流速为0．5mL·min^-1的0．1mol·L^-1的NaH2PO4-Na2HPO4缓冲液（pH7．8），柱温30℃，激发波长495nm，发射波长520nm。结果：FITC对河蚌糖胺聚糖的荧光标记率为70；线性范围为5～120μg·mL^-1，批内和批间精密度均小于15．0％，提取回收率为51．5～58．6％，样品在血浆中稳定性良好。结论：血浆中内源性杂质不干扰样品的测定，建立的方法符合生物样本的分析要求。
Objective： To establish a precolumn derivatization HPLC method for the determination of anodonta glycosaminogly- cans in rat plasma. Methods： FITC was used as the flurocescent labeling reagent of anodonta glycosaminoglycans. The separation was carried on a Shodex SB-804 HQ column （8.0× 300 mm） and 0.1 mol·L^-1 sodium dihydrogen phosphate-disodimn hydrogen phosphate buffer was employed as the mobile phase. The flow rate was 0.5 mL·min^-1 and the column temperature was set on 30℃. Flurocescence detection was used with the excitation wavelength at 495 nm and the emission wavelength at 520 mn. Results： The flurocescent labeling ratio was 70. The linear range of flurocescent labeling anodonta glycosaminoglycans was 5-120 μg·mL^-1. The RSD of intra- and interbatch precision were 〈- 15.0%. The extraction recovery was 51.5-58.6% and the sample was stable in plasma. Conclusions： There was no significant interference from endogenous substances and the method can be applied to determine the anodonta glycosaminoglycans in rat plasma.
出处
《现代生物医学进展》
CAS
2015年第13期2447-2450,共4页
Progress in Modern Biomedicine
基金
上海高职高专重点专业"085工程"飞跃计划项目(FY(13)400-A5-1-01)
