| 郝晓蓓,汪旭,潘娅岚,等.柱前衍生 -反相高效液相色谱法测定扶正补血食疗方胶原蛋白含量[J].安徽医药,2023,27(4):674-679. |
| 柱前衍生 -反相高效液相色谱法测定扶正补血食疗方胶原蛋白含量 |
| Precolumn derivatization-reversed-phase high-performance liquid chromatography for the determination of collagen in Fuzheng Buxue dietary supplement |
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| DOI：10.3969/j.issn.1009-6469.2023.04.009 |
| 中文关键词: 膳食疗法 扶正补血 色谱法，反相 色谱法，高压液相 羟脯氨酸 柱前衍生 胶原蛋白 正交试验 |
| 英文关键词: Dietary supplement Fuzheng Buxue Chromatography, reverse-phase Chromatography, high pressure liquid phase Hydroxyproline Precolumn derivatization Collagen Orthogonal test |
| 基金项目:江苏省研究生实践创新计划项目( SJCX21-0665)； 2021年度江苏省卫生健康委医学科研项目指导性项目( Z2021065)；江苏省研究生科研创新计划项目( KYCX21-1803)；江苏省中医药科技发展计划项目( YB2020004)；南京中医药大学“江苏高校护理学优势学科建设工程资助项目”(2019YSHL097)。 |
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| 中文摘要: |
| 目的基于 RP-HPLC法对扶正补血食疗方中胶原蛋白含量进行测定，并通过正交试验优选出扶正补血食疗方中胶原蛋白的最佳提取方法。方法于 2021年 12月至 2022年 1月将样品以 6 mol/L盐酸于 110 ℃水解 24 h，利用 AQC试剂进行衍生，采用 Diamonsil-Cl8色谱柱( 250 mm×4.6 mm，5 μm)以流动相 A:无水乙酸钠 11.48 g，三乙胺 1.72 g，EDTA 1 mg，溶于 1L水中，用磷酸调 pH为 4.95；流动相 B:乙腈水溶液(体积分数，60%)，梯度洗脱；柱温 37 ℃；检测波长 254 nm；流速 1 mL/min；进样量 10 μL。以羟脯氨酸含量为评价指标，考察煎煮时间、料水比、煎煮温度对胶原蛋白含量的影响，优化其制备工艺。结果羟脯氨酸在 10~500 mg/L浓度范围内线性关系良好( Y=0.036 5X+0.517 3，R2=0.999 1)，平均回收率 95.11%，相对标准偏差( RSD)为 |
| 英文摘要: |
| Objective To determine the collagen content in Fuzheng Buxue dietary supplement based on the RP-HPLC method and to optimize the best extraction method of collagen in Fuzheng Buxue dietary supplement by orthogonal testing.Methods Samples werehydrolyzed with 6 mol/L hydrochloric acid at 110 ℃ for 24 h from December 2021 to January 2022 and derived using AQC reagent on aDiamonsil-Cl8 column (250 mm×4.6 mm, 5 μm) with mobile phase A: anhydrous sodium acetate 11.48 g, triethylamine 1.72 g, EDTA 1mg, dissolved in 1 L of water. Mobile phase B: aqueous acetonitrile (60% v/v), gradient elution; column temperature 37℃; detectionwavelength 254 nm; flow rate 1 mL/min; injection volume 10 μL. The effects of decoction time, ratio of material to water, and decoctiontemperature on the collagen content were investigated to optimize the preparation process using the hydroxyproline content as the evaluation index. Results Hydroxyproline showed good linearity in the concentration range of 10-500 mg/L (Y=0.036 5X+0.517 3, R2=0.999 1) with an average recovery of 95.11% and RSD value of 1.12% (n=6). The best extraction process for pig trotter collagen was A3B1C3, i.e., the heating temperature was 100 ℃, the ratio of material to water was 1∶8, and the extraction time was 3 h.Conclusion The method is rapid, accurate, stable and feasible and can be used as a method for the determination of collagen content in Fuzheng Buxue dietary supplement and provides a scientific basis for product quality control and clinical application. |
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