Quantitative determination of trans-10-hydroxy-2-decenoic acid as a marker and quality control of Royal jelly products by High Performance Liquid Chromatography

10-HDA is an unsaturated fatty acid found in royal jelly produced from the hypopharyngeal and mandibular gland secretions of honeybees. Royal jelly has been reported to have anti-tumor, anti-biotic, immunomodulatory, estrogenic, neurogenic, and innate immune response modulating activities [1]. 10-hydroxy-2-decenoic acid or 10-HDA is a bioactive compound found in royal jelly [2]. The amount of 10-HAD in pure royal jelly varies due to the origin of the jelly [3]. The presence of 10-HAD in royal jelly can be used as a marker to validate the quality of royal jelly from other bee products in many countries [4]. Analysis of trans-10-hydroxy-2-decenoic acid (10-HDA) in royal jelly was carried out via reversed phase high performance liquid chromatography (RP-HPLC). The 10-HAD was extracted by sonication in a mixture of methanol and phosphate buffer (pH=2.5) (45:55, v/v) in 0 ºC. 0.2 g royal jelly in 2 ml deionized water is mixed and added to 38 ml extraction solution. The mixture centrifuged 15 min in 3000 rpm. One milliliter from supernatant poured in ten milliliter volumetric balloon and diluted with mobile phase solvent. 10-HAD separated using a ZORBAX ODS-C18 column (250 mm(5 µm) × 4.6 mm ID) with a polar mobile phase. In the case of 10-HDA analysis, a mixture of methanol, water, and phosphoric acid (55:45:5, v/v/v) was used as a mobile phase. The flow rate was 1.0 mL/min and the UV detection was performed at 216 nm at 25 C. Our results show that the concentration of 10-HDA lies between 1.85 and 3.92 wt% for fresh royal jelly creams.