次甲基蓝显色法定量海参硫酸软骨素

Determination of Chondroitin Sulfate from Sea Cucumber by Methylene Blue Colorimetry

提取仿刺参、海地呱、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8．00mL5mmol／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r0．18rag／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。该方法操作简便，检出限为1．6～1．8μg／mL，定量限为5．2～6．2μg／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。

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 K2HPO4/KH2PO4 buffer and 1.00 mL of methylene blue （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- tion of sea cucumber chondroitin sulfate.