Quantitative Analysis of Apisin, a Major Protein Unique to Royal Jelly

Abstract EN

Apisin, a protein that is unique to royal jelly (RJ), is known to compose the greater part of the RJ proteins and to exist as a heterooligomer containing major royal jelly protein 1 and apisimin.

However, few reports on the methods for quantifying apisin have been published.

Thus, we attempted to quantify apisin using HPLC, a widely used analytical technique, as described below.

Isoelectric precipitation and size-exclusion chromatography were used to obtain the purified protein, which was identified as apisin by SDS-PAGE and LC-MS analyses.

The purified apisin was lyophilized and then used to generate a calibration curve to quantify apisin in RJ.

The apisin content was fairly constant (i.e., 3.93 to 4.67 w/w%) in natural RJ.

This study is the first to describe a simple, standardized method for quantifying apisin using HPLC and suggests that apisin can be used as a benchmark for future evaluations of RJ quality.