Instituto de Investigación en Ciencias de la Alimentación (CIAL) (CSIC-UAM) CEI (CSIC+UAM), Nicolás Cabrera, 9, Campus de la Universidad Autónoma de Madrid, 28049 Madrid, Spain.
Departamento de Ingeniería Química y Ambiental, Facultad de Ingeniería, Universidad Nacional de Colombia - Sede Bogotá, Cr 30 N°45-03, Bogotá D.C., Colombia.
Food Chem. 2018 Jun 30;252:271-276. doi: 10.1016/j.foodchem.2018.01.087. Epub 2018 Jan 12.
A high-performance size-exclusion chromatography (HPSEC) method coupled to Evaporative Light Scattering (ELS) and Refractive Index (RI) detectors were evaluated and compared for the molecular mass (Mw) estimation of pectin in a wide range (0.342-805 kDa). Instrumental parameters of the ELSD were optimised by Response Surface Methodology (RSM) being 73 °C the evaporator temperature and 0.9 mL/min the air flow rate. The linear range for the ELSD concentration response was wider (10-2250 mg/L) and better (R = 0.985) than RID (10-1500 mg/L; R = 0.875). The limits of detection (LOD) and quantitation (LOQ) for all pullulans hardly changed in ELSD (LOD: 1.22-1.99 mg/L; LOQ: 4.07-6.63 mg/L); however, RID showed huge variations (LOD: 0.49-10.41 mg/L; LOQ: 1.64-34.70 mg/L), which increased with the Mw. In general, responses of both detectors were similar for the Mw estimation, although pectin characterisation with HPSEC-ELSD exhibited better results in the lowest Mw compounds.
一种高效体积排阻色谱 (HPSEC) 方法与蒸发光散射 (ELSD) 和折射指数 (RI) 检测器相结合，用于评估和比较果胶在较宽分子量范围 (0.342-805 kDa) 内的分子量 (Mw) 估计。ELSD 的仪器参数通过响应面法 (RSM) 进行优化，蒸发器温度为 73°C，空气流速为 0.9 mL/min。ELSD 浓度响应的线性范围更宽 (10-2250 mg/L)，相关性更好 (R=0.985)，而 RI 的线性范围较窄 (10-1500 mg/L; R=0.875)。所有普鲁兰的 ELSD 的检测限 (LOD) 和定量限 (LOQ) 几乎不变 (LOD：1.22-1.99 mg/L；LOQ：4.07-6.63 mg/L)；然而，RI 显示出巨大的变化 (LOD：0.49-10.41 mg/L；LOQ：1.64-34.70 mg/L)，随着 Mw 的增加而增加。总的来说，两种检测器在 Mw 估计方面的响应相似，尽管 HPSEC-ELSD 对果胶的特征化在最低 Mw 化合物中表现出更好的结果。
