Analytical and Bioanalytical Chemistry

, Volume 403, Issue 5, pp 1451–1456 | Cite as

Combination of sugar analysis and stable isotope ratio mass spectrometry to detect the use of artificial sugars in royal jelly production

  • Marine Wytrychowski
  • Gaëlle Daniele
  • Hervé Casabianca
Technical Note


The effects of feeding bees artificial sugars and/or proteins on the sugar compositions and 13C isotopic measurements of royal jellies (RJs) were evaluated. The sugars fed to the bees were two C4 sugars (cane sugar and maize hydrolysate), two C3 sugars (sugar beet, cereal starch hydrolysate), and honey. The proteins fed to them were pollen, soybean, and yeast powder proteins. To evaluate the influence of the sugar and/or protein feeding over time, samples were collected during six consecutive harvests. 13C isotopic ratio measurements of natural RJs gave values of around −25 ‰, which were also seen for RJs obtained when the bees were fed honey or C3 sugars. However, the RJs obtained when the bees were fed cane sugar or corn hydrolysate (regardless of whether they were also fed proteins) gave values of up to −17 ‰. Sugar content analysis revealed that the composition of maltose, maltotriose, sucrose, and erlose varied significantly over time in accordance with the composition of the syrup fed to the bees. When corn and cereal starch hydrolysates were fed to the bees, the maltose and maltotriose contents of the RJs increased up to 5.0 and 1.3 %, respectively, compared to the levels seen in authentic samples (i.e., samples obtained when the bees were fed natural food: honey and pollen) that were inferior to 0.2% and not detected, respectively. The sucrose and erlose contents of natural RJs were around 0.2 %, whereas those in RJs obtained when the bees were fed cane or beet sugar were as much as 4.0 and 1.3 %, respectively. The combination of sugar analysis and 13C isotopic ratio measurements represents a very efficient analytical methodology for detecting (from early harvests onward) the use of C4 and C3 artificial sugars in the production of RJ.


Royal jelly IRMS Sugar Bee feeding Gas chromatography 



We would like to acknowledge the GPGR beekeepers who conducted the feeding experiments.

Thanks also to France Agrimer and the Ministère de l’Agriculture et de la Pêche for their financial support.


  1. 1.
    Sabatini AG, Marcazzan GL, Caboni MF, Bogdanov S, Almeida-Muradian LB (2009) Quality and standardisation of royal jelly. JApiProd ApiMed Sci 1(1):1–6CrossRefGoogle Scholar
  2. 2.
    Garcia-Amoedo LH, De Almeida-Muradian LB (2007) Physicochemical composition of pure and adulterated royal jelly. Quim Nova 30(2):257–259CrossRefGoogle Scholar
  3. 3.
    Lercker G, Caboni MF, Vecchi MA, Sabatini AG, Nanetti A (1992) Caratterizzazione dei principali costituenti della gelatina reale. Apicoltura 8:27–37Google Scholar
  4. 4.
    Ferioli F, Marcazzan GL, Caboni MF (2007) Determination of (E)-10-hydroxy-2-decenoic acid content in pure royal jelly: a comparison between a new CZE method and HPLC. J Sep Sci 30:1061–1069Google Scholar
  5. 5.
    Boselli E, Caboni MF, Sabatini AG, Marcazzan GL, Lercker G (2003) Determination and changes of free amino acids in royal jelly during storage. Apidologie 34:129–137CrossRefGoogle Scholar
  6. 6.
    Stocker A, Schramel P, Kettrup A, Bengsch E (2005) Trace and mineral elements in royal jelly and homeostatic effects. J Trace Elem Med Biol 19(2–3):183–189CrossRefGoogle Scholar
  7. 7.
    Presoto AEF, Rios MDG, Almeida-Muradian LB (2004) Simultaneous high performance liquid chromatographic analysis of vitamins B1, B2 and B6 in royal jelly. J Braz Chem Soc 15(1):136–139CrossRefGoogle Scholar
  8. 8.
    Howe SR, Dimick PS, Benton AW (1985) Composition of freshly harvested and commercial royal jelly. J Api Res 24:52–61Google Scholar
  9. 9.
    Stocker A, Rossman A, Kettrup A, Bengsch E (2006) Detection of royal jelly adulteration using carbon and nitrogen stable isotope ratio analysis. Rapid Commun Mass Spectrom 20:181–184CrossRefGoogle Scholar
  10. 10.
    Daniele G, Wytrychowski M, Batteau M, Guibert S, Casabianca H (2011) Stable isotope ratio measurements of royal jelly samples for controlling production procedures: impact of sugar feeding. Rapid Commun Mass Spectrom 25(14):1929–1932CrossRefGoogle Scholar
  11. 11.
    Daniele G, Casabianca H (2012) Sugar composition of French Royal Jelly for comparison with commercial and artificial sugar samples. Food Chem. doi: 10.1016/j.foodchem.2012.03.008
  12. 12.
    Doner LW, White JW (1977) 13C/12C ratio relatively uniform among honeys. Science 197:891–892CrossRefGoogle Scholar
  13. 13.
    White JW, Doner LW (1978) Mass spectrometric detection of high fructose corn syrup in honey by 13C/12C ratio. A collaborative study. J Assoc Off Anal Chem 61:746–750Google Scholar
  14. 14.
    Cotte JF, Casabianca H, Lhéritier J, Perrucchietti C, Sanglar C, Waton H, Grenier-Loustalot MF (2007) Study and validity of 13C stable carbon isotopic ratio analysis by mass spectrometry and 2H site-specific natural isotopic fractionation by nuclear magnetic resonance isotopic measurements to characterize and control the authenticity of honey. Anal Chim Acta 582:125–136CrossRefGoogle Scholar
  15. 15.
    White JW, Winters K (1989) Honey protein as internal standard for stable carbon isotope ratio detection of adulteration of honey. J Assoc Off Anal Chem 72:907–911Google Scholar
  16. 16.
    White JW (1992) Internal standard stable carbon isotope ratio method for determination of C4 plant sugars in honey: collaborative study and evaluation of improved protein preparation procedure. J Assoc Off Anal Chem 75:543–548Google Scholar
  17. 17.
    Padovan GJ, De Jong D, Rodrigues LP, Marchini JS (2003) Detection of adulteration of commercial honey samples by the 13C/12C isotopic ratio. Food Chem 82:633–636CrossRefGoogle Scholar
  18. 18.
    Cordella C, Militão JSLT, Clément MC, Drajnudel P, Cabrol-Bass D (2005) Detection and quantification of honey adulteration via direct incorporation of sugar syrups or bee-feeding: preliminary study using high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) and chemometrics. Anal Chim Acta 531:239–248CrossRefGoogle Scholar
  19. 19.
    Guler A, Bakan A, Nisbet C, Yavuz O (2007) Determination of important biochemical properties of honey to discriminate pure and adulterated honey with sucrose (Saccharum officinarum L.) syrup. Food Chem 105:1119–1125CrossRefGoogle Scholar
  20. 20.
    Morales V, Corzo N, Sanz ML (2008) HPAEC-PAD oligosaccharide analysis to detect adulterations of honey with sugar syrups. Food Chem 107:922–928CrossRefGoogle Scholar
  21. 21.
    Cotte JF, Casabianca H, Chardon S, Lheritier J, Grenier-Loustalot MF (2003) Application of carbohydrate analysis to verify honey authenticity. J Chromatogr A 1021:145–155CrossRefGoogle Scholar
  22. 22.
    Ruiz-Matute AI, Rodríguez-Sánchez S, Sanz ML, Martínez-Castro I (2010) Detection of adulterations of honey with high fructose syrups from inulin by GC analysis. J Food Compos Anal 23:273–276CrossRefGoogle Scholar
  23. 23.
    Zhu X, Li S, Shan Y, Zhang Z, Li G, Su D, Liu F (2010) Detection of adulterants such as sweeteners materials in honey using near-infrared spectroscopy and chemometrics. J Food Eng 101:92–97Google Scholar
  24. 24.
    Cotte JF, Casabianca H, Chardon S, Lheritier J, Grenier-Loustalot MF (2004) Chromatographic analysis of sugars applied to the characterization of monofloral honey. Anal Bioanal Chem 308:698–705CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Marine Wytrychowski
    • 1
  • Gaëlle Daniele
    • 1
  • Hervé Casabianca
    • 1
  1. 1.Institut des Sciences Analytiques, Département Service Central d’AnalyseUniversité de LyonVilleurbanneFrance

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