Differences in Different Parts of Sea Cucumber Based on HPLC Fingerprinting and Holotoxin A1 Content Determination Combined with Chemical Pattern Recognition
Objective:To establish high performance liquid chromatography(HPLC)fingerprints from different parts of Apostichopus japonicus including body wall,intestine,sperm and egg with a method for determination of the content of sea cucumber saponin holotoxin A1,and to evaluate their differences using chemical pattern recognition.Methods:An Agilent ZORBAX SB-C18 column(4.6 mmx150 mm,5μm)was used for the separation,with a gradient elution using 0.1％phosphoric acid aqueous solution(A)and acetonitrile(B)as mobile phases,at a flow rate of 1.0 mL/min and a column temperature of 30 ℃.Eight batches of samples from each part were analyzed for fingerprinting and holotoxin A1 content.Chemical pattern recognition was performed using SPSS 25.0 and SIMCA 14.1 for differential evaluation of the different parts of Apostichopus japonicus.Results:The fingerprints of Apostichopus japonicus body wall showed 17 peaks,28 peaks in intestine,25 peaks in sperm,and 29 peaks in eggs,with similarity greater than 0.9.The control product showed 3 peaks of cyclic(leuco-isoleucine)dipeptide,phenylalanine,and holotoxin A1,of which the content of holotoxin A1 was found to be the highest in the body wall,followed by intestine,and the lower one in sperm and eggs.In the chemical pattern recognition analysis,the samples were clustered into three categories,from the orthogonal partial least squares-discriminant analysis(OPLS-DA)plot,it can be seen that the chemical compositions of intestine and sperm were more similar,and three differential components were screened.Conclusion:The method established stable and easy-to-follow HPLC fingerprints for different parts of Apostichopus japonicus,while simultaneously determining the content of holotoxin A1.Combined with chemical pattern recognition analysis,it can effectively evaluate the differences between the various parts of Apostichopus japonicus.
万方数据
维普
