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柱前衍生化-HPLC法测定市售3种胶类药材中L-羟脯氨酸和胶原蛋白的含量
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| 篇名： | 柱前衍生化-HPLC法测定市售3种胶类药材中L-羟脯氨酸和胶原蛋白的含量 |
| TITLE： | |
| 摘要： | 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（pH 6.5，7 ∶ 93，V/V）]-[乙腈-水（4 ∶ 1，V/V）]（梯度洗脱），流速为1.0 mL/min，检测波长为254 nm，柱温为43 ℃，进样量为20 μL；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2.5～40 μg/mL（r＝0.999 9）；定量限为0.20 μg/mL，检测限为0.05 μg/mL；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。 |
| ABSTRACT： | OBJECTIVE： To establish the method for the content determination of L-Hyp and collagen in gelatinous medicinal material， and to compare the contents of two components in reference medicinal material and commercially available medicinal material. METHODS： Pre-column derivatization was adopted for pretreatment. The content of L-Hyp was determined by HPLC. The determination was performed on Kromasil C18 column with mobile phase consisted of acetonitrile-0.1 mol/L sodium acetate buffer （pH 6.5，7 ∶ 93，V/V）-acetonitrile-water （4 ∶ 1，V/V）（gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 254 nm， and column temperature was set at 43 ℃. The sample size was 20 μL. The content of collagen was calculated by using convert coefficient. RESULTS： The linear range of L-Hyp were 2.5-40 μg/mL（r＝0.999 9）. LOQ was 0.20 μg/mL， and LOD was 0.05 μg/mL. RSDs of precision， stability and reproducibility tests were all lower than 4.0％. The recoveries were 96.03％-102.07％（RSD＝2.20％，n＝9）. There was difference in the contents of L-Hyp and collagen among 28 batches of commercially available medicinal material. The contents of two components in 13 batches of commercially available Colla Corii Asini were relative close to reference medicinal material； 5 batches of Colla Carapacis et Plastri Testudinis and 7 batches of Cervi Cornus Colla were much higher than those of reference medicinal material. CONCLUSIONS： The method is accurate， reliable and suitable for the content determination of L-Hyp and collagen in gelatinous medicinal material. |
| 期刊： | 2017年第28卷第27期 |
| 作者： | 王旭波，徐利丽，郑洁，李楠，沈玉萍，王智，韦波，杨欢 |
| AUTHORS： | WANG Xubo，XU Lili，ZHENG Jie，LI Nan，SHEN Yuping，WANG Zhi，WEI Bo，YANG Huan |
| 关键字： | 胶类药材；L-羟脯氨酸；胶原蛋白；含量测定；柱前衍生化；高效液相色谱法 |
| KEYWORDS： | Gelatinous medicinal material； L-hydroxy proline； Collagen； Content determination； Pre-column derivation； HPLC |
| 阅读数： | 228 次 |
| 本月下载数： | 23 次 |
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