目的: 建立1种结合优化后的Infogest体外消化静态模型的分子排阻色谱（SEC）法,测定短肽型肠内营养剂消化前后相对分子质量分布,对3种不同的短肽型肠内营养剂质量进行考察评估,探讨产品质量特性及消化稳定性,为短肽型肠内营养剂质量控制和临床个体化应用提供指导依据。方法: SEC法采用2根ECOSIL SEC G 2000色谱柱（300 mm×7.8 mm, 5 μm）串联,以乙腈-水-三氟乙酸（15 ∶ 85 ∶ 0.1）为流动相,流速0.7 mL · min-1,柱温 30 ℃,检测波长215 nm,进样量20 μL;体外模拟消化参考并优化Infogest体外消化静态模型,分为模拟胃、肠和胃肠总消化3个阶段,并采用建立的SEC法,对比消化前后相对分子质量分布的变化来评价其体外消化稳定性。结果: 在相对分子质量为165.19~12 327,8种对照品的相对分子质量对数与保留时间呈良好的线性关系。专属性试验结果显示,空白溶剂、模拟胃肠混合消化溶液对相对分子质量分布测定基本无影响;精密度、重复性、24 h稳定性试验的RSD均<1.0%。各品种短肽型肠内营养剂的相对分子质量主要集中在150~1 000,但不同品种的相对分子质量分布占比存在较大差异。肽组分的水解主要发生在生产阶段,经体外模拟消化后,其相对分子质量分布特性基本保持不变,变化程度主要与产品的原始水解程度相关。结论: SEC法结合经优化的Infogest体外消化静态模型,适用于短肽型肠内营养剂经消化前后的相对分子质量分布及消化稳定性研究,此方法操作简便、快捷,有利于该产品质量特性和营养价值的综合性评估。
Objective: To establish a method combining size exclusion chromatography (SEC) with an optimized Infogest in vitro static digestion model for determining the relative molecular mass distribution of short-peptide-based enteral nutrition before and after digestion. To evaluate and assess the quality of three different products, to explore product quality characteristics and digestive stability, and to provide guidance for quality control and clinical individualized application of short-peptide-based enteral nutrition. Methods: The SEC method was performed using two ECOSIL SEC G 2000 columns (300 mm×7.8 mm, 5 μm) in series, with acetonitrile-water-trifluoroacetic acid (15 ∶ 85 ∶ 0.1) as the mobile phase, a flow rate of 0.7 mL · min-1, the column temperature 30 ℃, the detection wavelength 215 nm, and the sample volume 20 μL. The in vitro simulated digestion process referred to and optimized the Infogest in vitro static digestion model, divided into simulated gastric digestion, simulated intestinal digestion, and simulated gastro-intestinal total digestion stages. The established SEC method was used to compare changes in the relative molecular mass distribution before and after digestion to evaluate in vitro digestive stability. Results: Within the range of relative molecular mass from 165.19 to 12 327, the logarithms of the relative molecular mass of the 8 reference standards showed good linear relationships with retention time. Specificity tests indicated that blank solvent and the simulated gastric-intestinal digestion solution had minimal impact on the measurement of relative molecular mass distribution. The RSDs for precision, repeatability, and 24 h stability tests were all less than 1.0%. The relative molecular mass of various short peptide enteral nutrition products was mainly concentrated between 150 and 1 000, but there were significant differences in the distribution ratios of relative molecular mass among different products. The hydrolysis of peptide components primarily occurred during the production stage, and after in vitro simulated digestion, their relative molecular mass distribution characteristics remained largely unchanged, with the degree of change mainly related to the original hydrolysis level of the product. Conclusion: The SEC method combined with the optimized Infogest model is suitable for studying the molecular mass distribution and digestive stability of short-peptide-based enteral nutrition. The method is straightforward and rapid, facilitating a comprehensive assessment of the products quality characteristics and nutritional value.
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