Royal jelly, a honey bee secretion, plays a critical role in caste determination in honey bees because it serves as the source of nutrition for young larvae destined to become queens. It is also fed to adult queens. Royal jelly possesses numerous functional properties and thus has been used as a medication, health food, and cosmetic in many countries. In this paper, we first introduce a traditional method for producing royal jelly by artificial larvae grafting and a newly developed method that does not require grafting of larvae. We describe protocols for the storage and freeze-drying of royal jelly to preserve its biological properties. Routine methods for determination of two important quality criteria, water content and trans-10-hydroxy-2-decenoic acid content, are outlined. On a dry basis, protein, carbohydrate, and fatty acids were found to be the 3 most abundant components of royal jelly. Methods for their isolation, identification, and quantification are described. Because royal jelly is susceptible to contamination with veterinary drugs and acaricides, we also describe methods for detection and quantification of some veterinary drugs and acaricides in royal jelly.Mé todos estándar para la investigació n de la jalea real de Apis mellifera La jalea real, una secreció n de abejas, desempeña un papel crítico en la determinació n de castas en la abeja melífera, ya que sirve como fuente de nutrició n para larvas jó venes destinadas a convertirse en reinas. También alimenta a las reinas adultas. La jalea real posee numerosas propiedades funcionales y por lo tanto se ha utilizado como un medicamento, alimento saludable y cosmético en muchos países. En este artículo, introducimos un método tradicional para producir jalea real mediante el injerto artificial de larvas y un método recientemente desarrollado que no requiere injerto de larvas. Describimos protocolos para el almacenamiento y la liofilizació n de la jalea real para preservar sus propiedades bioló gicas. Se describen métodos rutinarios para la determinació n de dos importantes criterios de calidad, el contenido de agua y el de ácido trans-10-hidroxi-2-decenoico. En una base seca, proteínas, carbohidratos y ácidos grasos fueron los tres componentes más abundantes de la jalea real. Se describen métodos para su aislamiento, identificació n y cuantificació n. Debido a que la jalea real es susceptible a la contaminació n con medicamentos veterinarios y acaricidas, también describimos métodos para la detecció n y cuantificació n de algunos medicamentos veterinarios y acaricidas en jalea real.

Until now, the properties of honey have been defined based exclusively on the content of plant components in the nectar of given plant. We showed that apalbumin1, the major royal jelly (RJ) protein, is an authentic and regular component of honey. Apalbumin1 and other RJ proteins and peptides are responsible for the immunostimulatory properties and antibiotic activity of honey. For the quantification of apalbumin1, an enzyme-linked immunosorbent assay (ELISA) was developed using polyclonal anti-apalbumin1 antibody. The method is suitable for honey authenticity determination; moreover it is useful for detection of the honey, honeybee pollen and RJ in products of medicine, pharmacy, cosmetics, and food industry, where presences of these honeybee products are declared. Results from the analysis for presence and amount of apalbumin1 in honeys will be used for high-throughput screening of honey samples over the world. On the basis of our experiments which show that royal jelly proteins are regular and physiologically active components of honey we propose to change the definition of honey (according to the EU Honey Directive 2001/110/EC) as follows: Honey is a natural sweet substance produced by honey bees from nectar of plants or from secretions of plants, or excretions of plant sucking insects, which honey bees collect, transform by combining with major royal jelly proteins and other specific substances of their own, deposit, dehydrate, store and leave in the honey comb to ripen and mature.