Acid In Royal Jelly Research Articles

Effects of 10-Hydroxy-2-decenoic Acid and 10-Hydroxydecanoic Acid in Royal Jelly on Bone Metabolism in Ovariectomized Rats: A Pilot Study.

Although previous studies have demonstrated that royal jelly (RJ) may have estrogenic properties and prevent postmenopausal bone loss, the underlying mechanisms are not fully understood. This animal study aimed to investigate the effects of specific fatty acids of RJ, 10-hydroxy-2-decenoic acid (10H2DA) and 10-hydroxydecanoic acid (10HDAA), in ovariectomized rats. Ten-week-old female Wistar rats were divided into the Baseline, Sham, Ovx, Ovx + 10H2DA, and Ovx + 10HDAA groups. Rats in the Baseline group were sacrificed immediately, whereas those in the other groups were subjected to either a sham operation or bilateral ovariectomy. The animals in the Ovx + 10H2DA and Ovx + 10HDAA groups were fed diets containing 10H2DA and 10HDAA, respectively. Twelve weeks after surgery, the rats were sacrificed, and indices of bone mass and bone mechanics were analyzed. Femoral bone mineral density was significantly lower in the Ovx group than in the Sham group (p < 0.01). Administration of 10H2DA or 10HDAA did not ameliorate bone loss after ovariectomy. In addition, administration of these fatty acids diminished femur bone stiffness in ovariectomized rats (p < 0.01 and p < 0.05, respectively). These findings suggest that the favorable effects of RJ may not be exerted solely by 10H2DA or 10HDAA. However, these effects may be exhibited in combination with other RJ constituents.

Pt(ii) and Pt(iv) complexes with a major component of royal jelly as innovative antitumor drug candidates

The major and unique fatty acid in Royal Jelly, trans-10-hydroxy-2-decenoic acid, which have anti-tumor, immunomodulatory, and other properties, has been combined with the antitumor platinum moiety of clinically used oxaliplatin.

In vitro and in vivo models to study the biological and pharmacological properties of queen bee acid (QBA, 10-hydroxy-2-decenoic acid): A systematic review

• Natural bee products exerts beneficial effects due to their properties. • QBA is the main fatty acid in Royal Jelly and exclusive to this food. • QBA modulates different processes and has a protective effect in vitro and in vivo. • QBA may be a potential treatment due to it immunomodulatory effect among others. Royal jelly (RJ) is one of the most valued natural products and is considered beneficial to human health, mainly due to its many biological and pharmacological properties. 10-Hydroxy-2-decenoic acid (10H 2 DA), also known as queen bee acid (QBA), is exclusive to RJ and represents the main lipid component of this food. Most in vitro studies using QBA have reported its beneficial health properties but only a few in vivo studies have focused on these benefits. Therefore, the focus of the present systematic review (SR) according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement was to analyze the properties of QBA in different diseases such as cardiovascular, age-related or neurodegenerative diseases and cancer, employing in vivo and in vitro models and summarize the beneficial and protective effect of QBA that were observed in most of the studies.

A colorimetric sensing strategy for detecting 10-hydroxy-2-decenoic acid in royal jelly based on Ag(I)-tetramethylbenzidine

Royal jelly as a functional food has great potential to promote human health. As a unique ingredient in royal jelly, 10-hydroxy-2-decenoic acid (10-HDA) is a vital criterion to evaluate the quality of royal jelly. Current approaches for 10-HDA detection mainly rely on high-performance liquid chromatographic (HPLC), but it requires large-scale equipment and skilled technicians that are costly and time-consuming. Herein, we report a colorimetric sensing strategy for 10-HDA detection based on Ag(I) and tetramethylbenzidine (TMB), a widely used chromogenic reagent. Ag(I) can oxidize TMB to generate oxidized TMB (oxTMB) showing a light-blue color. Due to the chelation with Ag (I), the introduction of 10-HDA can prevent the generation of oxTMB that results in a color fading and a decrease in absorbance. This colorimetric sensing strategy enables highly sensitive detection of 10-HDA with a linear range from 1 μM to 10 μM and a limit of detection of 0.13 μM. More importantly, this 10-HDA assay allows for a visual readout either by a smartphone or the naked eye with low cost, convenience, and rapid response that has been successfully applied for evaluating the quality of royal jelly in actual samples. The sensing strategy developed in this work provides a new colorimetric approach that holds great promise as a point-of-care platform for the food industry.

Royal Jelly Protects against Epidermal Stress through Upregulation of the NQO1 Expression.

Royal jelly (RJ) is secreted by honeybees and has been used as an apitherapy to obtain healthy skin since ancient times. However, the mechanism of the protective effects of RJ against skin aging and skin diseases caused by skin stress and its components have not been clarified. In this study, we attempted to understand the effect of RJ on epidermal function and observed that NAD(P)H quinone dehydrogenase 1 (NQO1) is significantly induced by RJ in keratinocytes. The expression of NQO1 was also increased in the 3D epidermal skin model. NQO1 is involved in antioxidation and detoxification metabolism, and we found that RJ protects against the epidermal stress caused by UVB and menadione through the upregulation of NQO1. We identified 10-hydroxy-2-decenoic acid (10H2DA), a major fatty acid in RJ, as an active compound in this reaction as it induced the expression of NQO1 and protected the skin against oxidative stress. We demonstrated that the protective effect of RJ against epidermal stress is mediated through the upregulation of NQO1 by 10H2DA.

The Application of Capillary Electrophoresis on the Characterization of Protein in Royal Jelly

The objective of this study was to characterize the protein fractions in royal jelly made from Apis mellifera ligustica from middle and southern areas of Taiwan. The total nitrogen content of fresh royal jelly was 2.46%, and the total amino acid nitrogen was 2.34%, suggesting that the nitrogen compound in royal jelly was mostly derived from protein . The nitrogen content of free amino acid in royal jelly was 0.11%, and the amino type nitrogen was 0.20%, indicating that the protein in royal jelly existed mainly in the form of large moleculars. To characterize the protein, royal jelly was dissolved in 0.1 M phosphate buffer(pH 7.0), followed by centrifugation, ammonia sulfate precipitation and dialysis to separate the protein into water soluble and water insoluble fractions. Water soluble fraction accounts for more than 60 % of the total protein in royal jelly, and was further investigated by DEAE-Sephacel, SDS-PAGE and capillary electrophoresis. By DEAE-Sephacel, two fraction peaks (F1 and F2) were identified and collected. By SDS-PAGE, F1 fraction was further separated into two bands, and the molecular weight was determined to be 50 KDa and 44 KDa, whereas F2 fraction was shown to have only one band with molecular weight of 55 KDa. By capillary zone electrophoresis, four poorly-separated peaks were observed in F1 fraction, and two well-separated peaks in F2 fraction. By capillary gel electrophoresis, two peaks were identified in F1 fraction, of which the molecular weight was estimated to be 59 KDa and 73 KDa. By contrast, only one peak was identified in F2 fraction, of which the molecular weight was estimated to be 118 KDa. However, by capillary isoelectric focusing, 6 peaks were identified in F1 fraction with pI of 6.9, 6.7, 6.3, 5.9, 5.7 and 5.5, respectively. Of that, pI of 4.8 and 4.7 were identified in F2 fraction.

10-Hydroxydecanoic acid inhibits LPS-induced inflammation by targeting p53 in microglial cells

Neuroinflammation, characterized by the activation of microglia and astrocytes, is important in the pathogenesis of many neurological disorders, such as Alzheimer’s disease. Nonsteroidal anti-inflammatory drugs (NSAIDs), a group of chemically heterogenous medications, are used widely in the treatment of inflammation. However, the safety of these drugs is a growing concern due to their side effects on the gastrointestinal tract and liver. Royal jelly (RJ) is a potential functional food produced by the hypopharynx and mandibular salivary glands of nurse bees. In this study, we explored the anti-neuroinflammatory effect of 10-hydroxydecanoic acid (10-HDAA), which is the second most abundant but less studied fatty acid in RJ. We showed that 10-HDAA decreased the lipopolysaccharide (LPS)-induced elevation of inducible nitric oxide synthase (iNOS) and nitric oxide (NO) levels in both microglial BV-2 and N9 cell lines. Compared to the LPS group, the 10-HDAA/LPS treated BV-2 cells had a higher level of the phagocytic receptor TREM2. RNAseq transcriptomic results showed a different transcriptional profile between the LPS group and the 10-HDAA/LPS group in BV-2 cells and the 10-HDAA pre-treatment significantly decreased levels of pro-inflammatory mediators, which were further confirmed by qRT-PCR analysis. Moreover, we found that p53 was a target of 10-HDAA. p53 may mediate the anti-inflammation effect of 10-HDAA in two ways: first by directly deactivating the NLRP3 inflammatory pathway, second by indirectly promoting autophagy. Taken together, our results reveal a novel function of tumor suppressor p53 in the inhibition of neuroinflammation and provide a theoretical basis for broadening the application range of 10-HDAA and RJ.

Determination of 10-Hydroxy-2-Decenoic Acid Content – a Marker in Several Commercial Royal Jelly Products Collected in Vietnam

The optimized capillary electrophoresis (CE) was applied to separate and detect the 10-hydroxy-2-decenoic acid (10-HAD) in royal jelly products. The method only requires that the sample solution need to be centrifuged and filtered before analyzed by the home-made capillary electrophoresis system. Firstly, 10-HDA was separated in a fused silica column with a diameter of 50 um using 20 mM Tris/Acetic buffer (pH 8.5) as a background electrolyte and a separation voltage of -17kV. Then, 10-HDA was detected by capacitively coupled contactless conductivity detection (C4D) with the migration time less than 8 minutes. Nine commercial products of royal jelly with Vietnamese and imported origin including pure royal jelly cream, lyophilized royal jelly (powder and gel) and honey with royal jelly were collected for analysis. The results of this study showed that content of 10-HDA were detected in the range of 0.5 mg/g to 23.1 mg/g. Using paired t test showed that the difference between results obtained from CE-C4D method and from HPLC method as a reference method was not statistically significant.&#x0D; Keywords: 10-HDA, royal jelly products, CE-C4D.&#x0D; References&#x0D; &#x0D; [1] M. Viuda Martos, Y. Ruiz Navajas, J. Fernández López, J.A. Pérez Álvarez, Functional properties of honey, propolis, and royal jelly”, Journal of food science 73 (2007) 117-124. https://doi:10.1111/j. 1750-3841.2008.00966.x.[2] M.F. Ramadan, A. Al-Ghamdi, Bioactive compounds and health-promoting properties of royal jelly: A review, Journal of Functional Foods 4 (2012) 39- 52. https://doi.org/10.1016/j.jff. 2011. 12.007.[3] Dimitrios Kanelis, Chrysoula Tananaki, Vasilis Liolios, Maria Dimou, Georgios Goras, Maria Anna Rodopoulou, Emmanuel Karazafris, and Andreas Thrasyvoulou, A suggestion for royal jelly specifcations, Archives of Industrial Hygiene and Toxicology 66 (2015) 275-284. Https://doi: 10.1515/aiht-2015-66-2651.[4] Brazil Ministério Da Agricultura e Do Abaste Imento. Secretaria De Defesa Agropecuária. Instrução normativa nº3, de 19 de janeiro de 2001. Regulamento técnico para fxação de identidade e qualidade de geléia real, Regulamento técnico para fxação de identidade e qualidade de geléia real liofilizada. Http:// www.engetecno.com.br/port/ legislacao/mel_geleia_real.htm.[5] Mahmut Genc¸ Abdurrahman Aslan, Determination of trans-10-hydroxy-2-decenoic acid content in pure royal jelly and royal jelly products by column liquid chromatography, Journal of Chromatography A. 839 (1999) 265 – 268. Http://doi: 10.1016/s0021-9673(99)00151-x.[6] F. Ferioli, E. Armaforte, M.F. Caboni, Comparison of the Lipid Content, Fatty Acid Profile and Sterol Composition in Local Italian and Commercial Royal Jelly Samples” Journal of the American Oil Chemists' Society, 91(2014) 875-884. Http://doi. org/10.1007/s11746-014-2446-x.[7] J. Kim, J. Lee, Quantitative analysis of trans-10-hydroxy-2-decenoic acid in royal jelly products purchased in USA by high performance liquid chromatography”, Journal of Apicultural Science, 54 (2010) 77-85. Https://pdfs.semanticscholar. org/e85f/dfc1823d778a40c ca2eefa4be8ba4b4d98 6c.pdf.[8] C.I. Pavel, L.A. Mărghitaş, D.S. Dezmirean, L.I. Tomoş, V. Bonta, A. Şapcaliu, A. Buttstedt, Comparison between local and commercial royal jelly-use of antioxidant activity and 10-hydroxy-2-decenoic acid as quality parameter”, Journal of Apicultural Research 53 (2014) 116-123. https:// doi.org/10.3896/IBRA.1.53.1.12.[9] Mahmut Genc¸ Abdurrahman Aslan, Determination of trans-10-hydroxy-2-decenoic acid content in pure royal jelly and royal jelly products by column liquid chromatography, Journal of Chromatography A, 839 (1999) 265 – 268. Https://doi:10.1016/s0021-9673(99)00151-x.[10] Jinhui Zhou, Xiaofeng Xue, Yi Li, Jinzhen Zhang and Jing Zhao, Optimized determination method for trans-10-hydroxy-2-decenoic acid content in royal jelly by high-performance liquid chromatography with an internal standard, Journal of AOAC International, 90(2007) 244-249. https://www.ncbi.nlm.nih.gov/pubmed/17373456.[11] Federico Ferioli, Gian Luigi Marcazzan, Maria Fiorenza Caboni, 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 (2007) 1061-1069. Http://doi:10.1002/jssc.200600416.[12] Orlando Muñoz, Susana Decap, Francisco Ruiz, José Arbildua, Octavio Monasterio, Determination of 10- hydroxy-2-decenoic acid in royal jelly by capillary electrophoresis, J. Chil. Chem. Soc., 56 (2010) 738-740. http://dx.doi.org/10.4067/S0717 97072011000300004.

Trans-10-hydroxy-2-decenoic acid protects against LPS-induced neuroinflammation through FOXO1-mediated activation of autophagy.

Neuroinflammation is thought to be associated with the pathogenesis of a series of neurodegenerative diseases. We have previously reported that royal jelly (RJ) has an anti-inflammatory effect on microglial BV-2 cells. However, components contributing to the effect of RJ were largely unexplored. The aim of this study was to assess whether trans-10-hydroxy-2-decenoic acid (10-HDA), the exclusive fatty acid in RJ, can alleviate neuroinflammation and to further explore the underlying mechanisms. Immunohistochemistry staining, ELISA, qRT-PCR and Western blot were used to assess the effect of 10-HDA on LPS-induced neuroinflammation both in vivo and in vitro. To determine the extent of inflammatory changes after 10-HDA treatment, RNAseq transcriptomic analysis was conducted. 10-HDA pretreatment significantly reduced the production of pro-inflammatory mediators in LPS-treated C57BL/6J mice and microglial BV-2 cells. 10-HDA inhibited the activation of the TNF-α/NF-κB axis and NLRP3 inflammasome-IL-1β pathway, which may be the anti-neuroinflammatory mechanism of 10-HDA. We also demonstrated that 10-HDA triggered cell autophagy, as evidenced by elevated levels of microtubule-associated protein 1 light chain 3-II (LC3-II) and decreased expression of SQSTM1. More importantly, 10-HDA increased the transcriptional activity of FOXO1 by increasing FOXO1 nuclear localization. Inhibition of FOXO1 and autophagy using chemical inhibitors markedly blunted the effect of 10-HDA on the TNF-α pathway and NLRP3 inflammasome-IL-1β pathway, indicating that 10-HDA alleviates neuroinflammation in BV-2 cells by modulating FOXO1-mediated autophagy. 10-HDA may be a promising agent for various neuroinflammation-associated diseases.

Down-regulation of aquaporin 9 gene transcription by 10-hydroxy-2-decenoic acid: A major fatty acid in royal jelly.

10‐Hydroxy‐trans‐2‐decenoic acid (10H2DA) is a unique lipid component of royal jelly produced by worker honeybees that exerts insulin‐like effects. We herein investigated the effects of 10H2DA on the gene expression of aquaporin 9 (AQP9), which functions as a glycerol transporter in the liver, to clarify whether 10H2DA modulates energy metabolism. 10H2DA suppressed AQP9 gene expression in HepG2 cells by promoting the phosphorylation of Akt and AMP‐activated protein kinase (AMPK). This suppression was partially recovered by the treatment of cells with inhibitors for Akt and AMPK. Based on the result showing that leptomycin B partially recovered the suppression of AQP9 gene expression, 10H2DA inhibited the expression of Foxa2, a transcription factor for the AQP9 gene, and also induced its nuclear exclusion. Although 10H2DA up‐regulated phosphoenolpyruvate carboxykinase and glucose‐6‐phosphatase gene expression, this was suppressed through the modulation of Foxa2 by insulin. These results suggest that 10H2DA suppresses AQP9 gene expression through the phosphorylation of Akt and AMPK and down‐regulation of Foxa2 expression.

10-Hydroxy-trans-2-decenoic acid attenuates angiotensin II-induced inflammatory responses in rat vascular smooth muscle cells

10-Hydroxy-trans-2-decenoic acid (10-HDA) is a major medium chain fatty acid in royal jelly. In this study, the rat vascular smooth muscle cells (VSMCs) were pretreated with 10-HDA (250 μM, 500 μM, 1000 μM), followed by incubation with Angiotensin II (Ang II, 100 nM). 10-HDA significantly reduced the productions of Ang II induced inflammatory cytokines interleukin 6 and C-reactive protein, through downregulating the nuclear factor kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways. 10-HDA also attenuated the Ang II-stimulated reactive oxygen species production and the NADPH oxidase 4 (NOX4) expression. Further mechanistic studies showed that 10-HDA reduced the elevated abundances of the angiotensin II type 1 receptor, and inhibited cAMP-response element-binding protein binding to the cAMP-response element site in the IL-6 promoter region. The results suggested that 10-HDA effectively attenuated Ang II-induced inflammatory responses and ROS production via regulating multiple steps in NF-κB and MAPK signaling pathways in VSMCs.

Long-Term Administration of Queen Bee Acid (QBA) to Rodents Reduces Anxiety-Like Behavior, Promotes Neuronal Health and Improves Body Composition.

Background: Queen bee acid (QBA; 10-hydroxy-2-decenoic acid) is the predominant fatty acid in royal jelly (RJ) and has activity at estrogen receptors, which affect brain function and body composition. However, few, long-term studies have assessed QBA effects in brain health and body composition. Methods: Primary hippocampal neurons were treated with QBA (0–30 µM) and challenged with glutamate or hypoxia. QBA was fed to aged, male Sprague-Dawley rats (12–24 mg/kg/day) and to adult male and female Balb/C mice (30–60 mg/kg/day) for ≥3.5 months. Rats were evaluated in a behavioral test battery of brain function. Mice were measured for fat and muscle composition, as well as bone density. Results: QBA increased neuron growth and protected against glutamate challenge and hypoxia challenge. Rats receiving QBA had reduced anxiety-like behavior, increased body weight, and better maintenance of body weight with age. Mice receiving QBA exhibited increased body weight, muscle mass, and adiposity in males, and increased bone density, but decreased adiposity, in females. Conclusions: QBA is an active component of RJ that promotes the growth and protection of neurons, reduces anxiety-like phenotypes, and benefits bone, muscle and adipose tissues in a sex-dependent manner, which further implicates estrogen receptors in the effects of QBA.

Comparison of salting-out and sugaring-out liquid–liquid extraction methods for the partition of 10-hydroxy-2-decenoic acid in royal jelly and their co-extracted protein content

Homogeneous liquid–liquid extraction (h-LLE) has been receiving considerable attention as a sample preparation method due to its simple and fast partition of compounds with a wide range of polarities. To better understand the differences between the two h-LLE extraction approaches, salting-out assisted liquid–liquid extraction (SALLE) and sugaring-out assisted liquid–liquid extraction (SULLE), have been compared for the partition of 10-hydroxy-2-decenoic acid (10-HDA) from royal jelly, and for the co-extraction of proteins. Effects of the amount of phase partition agents and the concentration of acetonitrile (ACN) on the h-LLE were discussed. Results showed that partition efficiency of 10-HDA depends on the phase ratio in both SALLE and SULLE. Though the partition triggered by NaCl and glucose is less efficient than MgSO4 in the 50% (v/v) ACN-water mixture, their extraction yields can be improved to be similar with that in MgSO4 SALLE by increasing the initial concentration of ACN in the ACN-water mixture. The content of co-extracted protein was correlated with water concentration in the obtained upper phase. MgSO4 showed the largest protein co-extraction at the low concentration of salt. Glucose exhibited a large protein co-extraction in the high phase ratio condition. Furthermore, NaCl with high initial ACN concentration is recommended because it produced high extraction yield for 10-HDA and the lowest amount of co-extracted protein. These observations would be valuable for the sample preparation of royal jelly.

Quality Parameters for Commercial Royal Jelly

Royal jelly has become a high-value commercial product and the standardization of this product is required to guarantee its quality on the market. The objective of the research activity was to pursue the chemical composition of commercial samples of Royal Jelly in Romania in order to propose standardization for this product. The physico-chemical composition of commercial Royal Jelly samples was analysed by determining quality parameters like: carbohydrates, lipids, proteins, 10-hydroxy-2-decenoic acid (10-HDA) and mineral elements. Carbohydrates analysis showed values between 3.4 % and 5.87 % for fructose, 4.12 % and 7.05 % for glucose, while for sucrose the values ranged between 0.95 % and 2.56 % (determined by HPLC-RI). The lipids content ranged between 1.85 % and 6.32 % (determined by the Soxhlet method). The protein values extended from 13.10 % (RJ2) to 17.04 % (RJ10) (the total protein content was determined by the Kjeldahl method). The values for the major fatty acid in Royal Jelly, 10-HDA, ranged between 1.35 % (RJ8) and 2.03 % (RJ10) (determined by high-performance liquid chromatography). The concentration of minerals varied between 3188.70 mg/kg and 4023.39 mg/kg (the concentration of minerals was measured using flame atomic absorption spectrometry). Potassium, followed by magnesium, sodium and calcium, occurs in the highest concentrations. The commercial Royal Jelly samples analysed presented variable physico-chemical characteristics that correspond with the values given by international quality standard proposals for Royal Jelly.

Isolation and characterization of proteins and lipids from honeybee (Apis mellifera L.) queen larvae and royal jelly

Honeybee queen larvae (QL), the byproduct of royal jelly (RJ) production, have not been fully utilized as a food resource. Proteins in QL were characterized by gel electrophoresis and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry. Qualitative and quantitative analysis of fatty acid and sterol composition in QL and RJ were performed. The molecular weight of proteins in QL was generally lower than those in RJ. QL were dominated by major royal jelly protein 1 (MRJP1) and MRJP2 as well as their degradation proteins. High amounts of triacylglycerols (797.5mg/g lipid) and phospholipids (89.8mg/g lipid) were presented in QL, whereas the main fatty acids in RJ were composed of free middle carbon chain fatty acids (>91%). Twelve sterols were identified and clerosterol, ∆7-sitosterol and cycloartenol were reported for the first time in QL. Noteworthy, little difference was observed in the sterol profiles of QL and RJ. Sterols were found to be integrated with MRJP1 oligomer in QL and RJ. This study provides new insight into the nutritional values of QL. The results suggest overall that proteins and lipids of QL compare favorably with those of RJ and could be potential used in food industry.

Contribution of Lipids in Honeybee (Apis mellifera) Royal Jelly to Health

Honeybee (Apis mellifera) royal jelly (RJ) has a long history in human medicine because of its health-protecting properties. To develop a fundamental and comprehensive understanding of lipids in RJ, this article reviews the available literature on lipid compounds identified from RJ extracts and in vitro pharmacological effects of 10-hydroxy-2-decenoic acid in RJ and other closely related compounds, some of which are also identified as lipid compounds in RJ. Overall, the lipids in RJ are composed of mostly (aliphatic) fatty acids, almost all of which are present as free fatty acids and scarcely any as esters. Most fatty acids in RJ are medium-chain fatty acids, whether hydroxylated in terminal and/or internal positions, terminated with mono- or dicarboxylic acid groups, and saturated or monounsaturated at the 2-position. Besides these fatty acids, lipids in RJ contain sterols in minor amounts. Lipids in RJ are useful as preventive and supportive medicines with functionalities that include potential inhibitors of cancer growth, immune system modulators, alternative therapies for menopause, skin-aging protectors, neurogenesis inducers, and more. Taken together, the evidence suggests that health-protecting properties of RJ can be, in part, ascribed to actions of lipids in RJ.

HPLC determination of pantothenic acid in royal jelly

Although royal jelly constitutes one of the richest natural sources of pantothenic acid (vitamin B5), a reliable and validated chromatographic method to determine this analyte in this matrix has yet to be described in the literature. In this work we present an original RP-HPLC procedure to measure the concentration of pantothenic acid in royal jelly. A sample pre-treatment is needed to prevent the interference of high protein concentration in the matrix. The method has been validated in terms of LoD, LoQ, linearity, precision (repeatability and reproducibility) and bias. Finally, the whole procedure was tested on a number of samples of royal jelly from different origins, providing concentration values ranging from 120 ± 30 to 565 ± 40 mg kg−1 of pantothenic acid.

SIMULTANEOUS DETERMINATION OF 14 QUINOLONES IN ROYAL JELLY BY LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY USING ANION-EXCHANGE SOLID-PHASE EXTRACTION

A new method based on liquid chromatography-tandem mass spectrometry has been developed for simultaneous determination of 14 quinolones (QNs) residues, including ciprofloxacin, danofloxacin, difloxacin, enoxacin, enrofloxacin, flumequine, lomefloxacin, marbofloxacin, norfloxacin, orbifloxacin, ofloxacin, pipemidic acid, pefloxacin, and sarafloxacin in royal jelly. The proposed analytical procedure involves extraction of the QNs from samples by 0.1 M sodium hydroxide aqueous solution, a step for clean-up and preconcentration of the analytes by anion-exchange solid-phase extraction (SPE) and liquid chromatographic separation with mass spectrometric detection. Internal standard calibration was applied with norfloxacin-D5 as internal standard (I.S.). Satisfactory recovery (from 64% to 113%), excellent repeatability precision (relative standard deviations, RSDs below 6%), reproducibility precision (RSDs below 10%), and low limits of quantifications (LOQs from 0.3 to 2.5 µg kg−1) were evaluated from spiked royal jelly samples at 2.5, 5.0, and 10.0 µg kg−1 three concentration levels. This protocol has been validated by five authoritative laboratories and successfully used for analysis of a large number of import–export samples (n > 2000). Norfloxacin and ciprofloxacin were found to be the most common contamination in royal jelly in the ranges of 3.5–30 µg kg−1 and 4.5–20 µg kg−1, respectively.

Determination of 10-hydoxy -2-decenoic acid in pure Iranian royal jelly by HPLC method

Royal jelly (RJ) is generally regarded as the major reason of the significant morphological and functional differences between queen and worker bees. The unique and chemically most interesting feature of RJ is its fatty acids. RJ fatty acids are short-chained, 8-10 carbon free acids that are usually either hydroxy fatty acids or dicarboxylic acids. The major fatty acid in RJ is 10-hydroxy-2-deconic acid (10-HDA) and the amount of this fatty acid in Pure RJ varies depending on the origin of the jelly and characteristics of the bee. Even other bee products do not contain 10-HDA. Thus, the presence of 10-HDA can be used as a marker to differentiate RJ from other bee products. The pure RJ samples from different region of Iran and three RJ capsules from China, Australia and Canada were purchased. The obtained crystals, 10-HDA standard from pure RJ were analyzed by RP-HPLC. The eluent was CH3CN-THF-H2O (50.4:21.6:26 v/v/v with pH=2.5). The flow-rate and wavelength were adjusted at 1 ml/min and 215 nm respectively. A LC run 30 min was conducted so that no other peaks were present. The external calibration method was used for determination of 10-HDA in samples. The R2 value for calibration cure was 0.998. The amounts of 10-HDA found in RJ samples was between 0.75 and 2.54%. The minimum recovery determined found to be 98%. The results show that 10-HDA crystals are pure and the method can be used for RJ evaluation.

Quantitative determination of trans-10- Hydroxy-2-Decenoic Acid (10-HDA) in Brazilian royal jelly and commercial products containing royal jelly

Quantitative determination of trans-10- Hydroxy-2-Decenoic Acid (10-HDA) in Brazilian royal jelly and commercial products containing royal jelly