An enzyme-pHBH method for specific quantification of porphyran

Abstract

Porphyran, the major polysaccharide extracted from Porphyra, exhibits tremendous potential for development as functional food or pharmaceutical due to its multiple biological activities. The quantitative analysis of porphyran is important for the quality control in product development. However, the specific quantitative method of porphyran has not been established, and the lack of reference substance makes the quantification more challenging. Here, a common component of porphyran, with high purity, similar molecular weight distribution, sourced from different Porphyra producing areas in China was first prepared by a series of isolation and purification steps, and utilized as the reference substance for porphyran quantification. Subsequently, the porphyran was fully degraded into oligosaccharides by using a β-porphyranase, followed by employing para-hydroxybenzoic acid hydrazide (pHBH) method to detect the content of the generated reducing sugar. The enzyme-pHBH method for porphyran specific quantification was established. Results showed that this method was validated with good linearity, high accuracy and precision, and reliability. Addtionally, NaCl with a concentration below 0.5 %, alcohol under 8 % and other polysaccharide including chitosan, agarose, chondrotin sulfate, alginate, hyaluronic acid and κ-carrageenan did not interfere with this method. This approach is promising for quality control of the porphyran products and offers a feasible strategy for the specific quantification of other polysaccharides.