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目的:建立测定胶类药材中L-羟脯氨酸(L-Hyp)和胶原蛋白含量的方法,并比较对照药材与市售药材中两种成分的含量。方法:采用柱前衍生化法进行前处理,并采用高效液相色谱法测定样品中L-Hyp的含量:色谱柱为Kromasil C18,流动相为[乙腈-0.1 mol/L醋酸钠缓冲液(p H 6.5,7∶93,V/V)]-[乙腈-水(4∶1,V/V)](梯度洗脱),流速为1.0 m L/min,检测波长为254 nm,柱温为43℃,进样量为20μL;再通过折算系数计算样品中胶原蛋白的含量。结果:L-Hyp检测质量浓度线性范围为2.5~40μg/m L(r=0.999 9);定量限为0.20μg/m L,检测限为0.05μg/m L;精密度、稳定性、重复性试验的RSD<4.0%;加样回收率为96.03%~102.07%(RSD=2.20%,n=9)。28批市售药材中L-Hyp和胶原蛋白的含量有一定差异,其中13批市售阿胶药材与阿胶对照药材中两种成分的含量较为接近,而5批龟甲胶和7批鹿角胶药材中两种成分的含量高于其对照药材。结论:该方法准确、可靠,适用于测定胶类药材中L-Hyp和胶原蛋白的含量。
OBJECTIVE: To establish a method for the determination of L-Hyp and collagen in Gums, and to compare the contents of two components in the control and commercial herbs. Methods: Pre-column derivatization was used to determine the content of L-Hyp in samples. The chromatographic column was Kromasil C18 and the mobile phase was acetonitrile-0.1 mol / L sodium acetate buffer (p H 6.5,7:93 V / V]] - [acetonitrile-water (4: 1, V / V)] with gradient elution at a flow rate of 1.0 m L / min and a detection wavelength at 254 nm 43 ℃, the injection volume of 20μL; then through the conversion factor to calculate the sample collagen content. Results: The linear range of L-Hyp was 2.5-40 μg / m L (r = 0.999 9), the limit of quantification was 0.20 μg / ml and the limit of detection was 0.05 μg / ml. The precision, stability and reproducibility The RSD of the test was <4.0%. The recoveries of samples were 96.03% ~ 102.07% (RSD = 2.20%, n = 9). The contents of L-Hyp and collagen in 28 batches of medicinal herbs were different. Among them, the contents of the two components in 13 batches of gelatin and gelatin were close, while 5 batches of turtle shell gum and 7 batches of antler gum The content of the two components is higher than that of the reference drug. Conclusion: The method is accurate and reliable, and is suitable for the determination of L-Hyp and collagen in Gums.