Determination of Five Skin Ingredients in Asini Corii Colla Foods Based on Marker Peptide
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Abstract:
To establish the method based on marker peptide for determination of the skin ingredients of horse, ox, pig, camel and deer in asini corii colla foods by HPLC MS/MS. The extraction methods and enzymolysis conditions of the samples were investigated, and the separation effects of different chromatographic conditions on the components of various skin sources were compared. The mobile phase elution system was optimized. The samples were hydrolyzed by trypsin at 37 ℃. The separation was performed with the gradient elution using the mobile phase consisting of 0.1% formic acid water solution and 0.1% formic acid acetonitrile solution at a flow rate of 0.3 mL/min. The column temperature was set at 40 ℃ and a sample size of 2 µL. Marker peptides were determined in the modes of electrospray positive ionization (ESI+) and multiple reaction monitoring (MRM) with matrix matching scaling quantification. Horse and deer marker peptide showed good linear relationships within the range of 5~500 ng/mL, others showed good linear relationships within the range of 10~500 ng/mL. The limit of detection was 0.082~1.1 mg/kg. The spike recoveries (n=6) of dimethyl sulfate were 86.8%~103.5%. Marker peptides of horse and ox were detected in 29 samples. The method is specific, sensitive, simple, rapid and accurate, and can be used for the the determination ofthe skin ingredients of horse, ox, pig, camel and deer in asini corii colla pastry and asini corii colla drinks.
