The invention provides a method for determining apis cerana Fabricius MRJP1 protein content (major royal jelly protein 1) in honey by utilizing liquid chromatography tandem mass spectrum. The method comprises the following steps: selecting characteristic peptide fragment, manufacturing a standard curve, pretreating the sample, performing liquid chromatographic separation and tandem mass spectrum detection quantitation and like steps. The detection method of apis cerana Fabricius MRJP1 content in honey has high accuracy, precision and sensitivity, and strong in stability, and suitable for the accurate quantitation of the apis cerana Fabricius MRJP1. The method further can be used for the truth evaluation of the honey, thereby assisting to identify whether the honey is adulterated, and is ofgreat significance for maintaining the healthy development of the honey consumption industry and the rights and interest of the honey consumer.