Determination of three bioactive holostane-type saponins in Apostichopus japonicus Selenkaby matrix solid-phase dispersion - high performance liquid chromatography - ultraviolet absorption detector

A method for the quantitative analysis of three holostane-type saponins (holotoxin B1, E, and A1) in Apostichopus japonicus Selenkawas developed using matrix solid-phase dispersion (MSPD) for pretreatment and high performance liquid chromatography-ultraviolet (HPLC-UV) for detection. The precision, reproducibility, stability, and recovery of the method were investigated. The conditions for MSPD were as follows: silica gel as a dispersant, 1:1 (0.5 g:0.5 g) sample/dispersant ratio, 40 mL of ethyl acetate as an impurity-eluting solvent, and 60 mL of methanol as eluent. The conditions for HPLC-UV were as follows: Tnature C18 column (5 μm, 4.6 × 250 mm), ACN:H2O (V:V) = 35.5:64.5 mobile phase, 1 mL/min flow rate, and 203 nm detection wavelength. The linear equations of the three standards were as follows: Y = 594.38105X + 26.5934 (R2=0.999) for holotoxin B1, Y = 657.94293X + 25.72468 (R2=0.999) for holotoxin E, and Y = 395.54309X + 1.59529 (R2=0.999) for holotoxin A1. The linear ranges of holotoxin B1, E, and A1were 0.24–9.55, 0.25–9.85, and 0.21–8.31 μg, respectively, and their limits of detection were 5.05, 4.56, and 7.58 ng, respectively. The spiked recovery, intraday precision, and interday precision of the method were all good. The average contents of holotoxin B1, E, and A1in A. japonicusSelenka were determined as 6.24, 9.58, and 96.2 μg/g, respectively. This experiment provides a new scheme for the determination of rare holostane-type saponins in A. japonicusSelenka and a method for the quality control of these sea cucumbers.