次甲基蓝显色法定量海参硫酸软骨素-食品科学.PDF

246 2013, Vol.34, No.22 食品科学 ※分析检测 次甲基蓝显色法定量海参硫酸软骨素 续晓琪，薛长湖，张翠玉，于 龙，王彦超，常耀光* ( 中国海洋大学食品科学与工程学院，山东 青岛 266003) 摘  要：提取仿刺参、海地瓜、梅花参、美国肉参及北大西洋瓜参5种海参中的海参硫酸软骨素作为标准品，以次 甲基蓝与阴离子聚合物形成显色的超分子复合物为理论基础，建立一种海参硫酸软骨素的定量方法。将1.00mL质 量浓度适当的海参硫酸软骨素溶液与8.00mL 5mmol/L 磷酸氢二钾-磷酸二氢钾缓冲液(pH7.0)及1.00mL次甲基蓝溶液 (0.18mg/mL)混合，其后立即测定545nm波长处的吸光度，吸光度代入对应标准曲线即可计算得出样品质量浓度。 μ μ 该方法操作简便，检出限为1.6～1.8 g/mL ，定量限为5.2～6.2 g/mL ，精密度、回收率高，可实现对海参硫酸软骨 素的快速定量测定。 关键词：海参；海参硫酸软骨素；次甲基蓝；比色；测定 Determination of Chondroitin Sulfate from Sea Cucumber by Methylene Blue Colorimetry * XU Xiao-qi ，XUE Chang-hu ，ZHANG Cui-yu ，YU Long ，WANG Yan-chao ，CHANG Yao-guang (College of Food Science and Engineering, Ocean Universitity of China, Qingdao 266003, China) Abstract ：In this study, employing sea cucumber chondroitin sulfate standard samples extracted from 5 species of sea cucumber, an efficient and rapid colorimetric method for determination of chondroitin sulfate from sea cucumber was es- tablished based on the theory that methylene blue and anionic polymer can form supramolecular complex. The colorimet- ric method was determined as follows: detection at 545 nm immediately following mixing of 1.00 mL of sea cucumber chondroitin sulfate solution with 8.00 mL of 5 mmol/L pH 7.0 K HPO /KH PO buffer and 1.00 mL of methylene blue 2 4 2 4 (0.18 mg/mL) and subsequent calculation of chondroitin sulfate concentration from the absorbance using the pre-prepared calibration curve. This method proved to be simple, accurate and high-recovery and allow rapid quantitative determina-