Quantification of major royal jelly proteins using ultra performance liquid chromatography tandem triple quadrupole mass spectrometry and application in honey authenticity

Major royal jelly proteins (MRJPs) have been used as endogenous biomarkers for honey authentication. However, no study on quantification of MRJPs in honey authentication was reported. In order to develop a new honey authenticity method based on quantitative analysis of MRJPs in honey, trypsin-digested peptides from MRJPs were examined by mass spectrometry (MS). Three peptides, YNGVPSSLNVISK, TLQMIAGMK, and LTVA-GESFTVK were selected as specific markers for MRJP1, MRJP 2 and MRJP 3(MRJP1 similar to 3), respectively. An ultra performance liquid chromatography-triple quadrupole MS (UPLC-TQMS) method used for quantification of MRJPs in honey was developed and validated. A database on the MRJP1 similar to 3 contents of honey established by quantification of 70 authentic samples showed a potential application in honey authenticity identification. The analysis results of 10 commercial honey samples by UPLC-TQMS with reference to the MRJP1 similar to 3 database of authentic honey were compared and confirmed by widely used isotope ratio mass spectrometry, a AOAC Official Method. In conclusion, the results showed our method, a proteomic-based UPLC-TQMS method, has a high accuracy, sensitivity and specificity for honey authenticity.