Fresh Royal Jelly Research Articles

Variation in chemical composition of fresh and commercial Royal Jelly associated with international standards

The chemical composition of fresh (local) and commercial (imported) Royal Jelly (RJ) from Saudi Arabia was investigated. 10-hydroxydec-2-enoic acid (10-HDA), protein content, sugar profile, moisture content, pH, free acidity, electric conductivity (EC), and macro and micro-minerals were analyzed in both RJ groups. Most samples and about 50 % of fresh RJ samples showed 10-HDA and protein content percentages lower or higher than the international standard limits. Differences were significant (p < 0.0001) between fresh and commercial RJ samples in all tested parameters except moisture content. Furthermore, 29 percent of the commercial samples did not pass the 10-HDA standard limits. On the other hand, lactose, a non-RJ sugar, was detected in two commercial samples indicating some contamination or adulteration of these samples. The minerals Zinc (Zn), Iron (Fe), Copper (Cu), Aluminum (Al), Manganese (Mn), Phosphorous (P), Potassium (K), Magnesium (Mg), Sodium (Na), and Calcium (Ca) were detected in all samples. Zinc (Zn), Iron (Fe), Potassium (K), Magnesium (Mg), and Sodium (Na) were significantly higher in fresh RJ samples (P < 0.05). Sodium, potassium, and phosphorus demonstrated significantly higher occurrence in fresh RJ samples from Saudi Arabia and might be good candidates for its characterization. Although 10-HDA and protein ratios are important parameters of RJ authenticity, the current standards of both parameters (ISO,12824:2016) should be updated to cover occurrence limits worldwide. Moreover, mineral analyses might be useful to characterize RJ's originality and explore RJ's adulteration.

Production and Quality Characteristics of Royal Jelly in Relation to Available Natural Food Resources

Royal jelly is a secretion produced from the hypopharyngeal glands of worker bees, which requires significant pollen reserves to stimulate gland secretion. The natural sources of food available to the hive during beekeeping season can greatly affect the quantity and quality of produced royal jelly. In this study, samples of royal jelly were collected throughout the beekeeping season, and their physical and chemical characteristics were analyzed to understand how natural variations in bee diet affect royal jelly production. Before each sample collection, the bees’ food reserves were removed from the experimental colonies so that the royal jelly was produced solely from natural sources. The results showed that the production was significantly lower during the summer months compared with spring and autumn. Additionally, the moisture, protein, fructose, and glucose content of fresh royal jelly also showed significant changes in the summer, and all physical and chemical characteristics decreased when the fresh samples were converted into dry matter. It seems that the quality of pollen entering the hives has a direct impact on the physical and chemical properties of the final product, highlighting the crucial role of available resources in stimulating bees to produce royal jelly.

Development and validation of an analytical methodology based on solvent extraction and gas chromatography for determining pesticides in royal jelly and propolis

We propose a new analytical methodology to determine seven pesticides (atrazine, chlorpyrifos, chlorfenvinphos, α-endosulfan, bromopropylate, coumaphos, and τ-fluvalinate) in royal jelly and propolis products using gas chromatography-mass spectrometry. Sample treatment, with minor modifications for propolis, consisted of a solvent extraction with a hexane and isopropanol mixture, and a further clean-up step. Meanwhile, chromatographic analysis (<25 min) was performed in a DB-5MS column under programmed temperature conditions. In all cases we validated the method in terms of selectivity, limits of detection (0.1–2.8 μg kg−1) and quantification (0.3–9.2 μg kg−1), linearity, matrix effect (<±20 %), trueness (recoveries between 93 % and 118 %), and precision (relative standard deviation < 11 %). All royal jelly liquid dietary supplements were positive for chlorfenvinphos and, in the case of one of them, for α-endosulfan; chlorfenvinphos was determined in some fresh royal jelly samples, and no pesticide residues were detected in the propolis samples analysed.

Physicochemical characterization of Persian fresh royal jelly samples

Although the health-promoting properties of Royal Jelly (RJ) have been known for many years, only a few countries have set national quality standards or guidance for RJ quality determination. Data about Persian RJ quality are not widely available; therefore, the results presented in this study can be considered pioneering ones for Persian RJ characterization in terms of trans-10-hydroxy-2-decenoic acid (10-HDA), sugars, protein, and moisture content. In this study, high performance liquid chromatographic method (HPLC) was used to determine the amount of main carbohydrates (fructose, glucose, and sucrose), other sugars (maltose, erlose, and turanose) and 10-HDA in fresh RJ samples. Based on our research, the primary sugar compounds were glucose and fructose, with lower concentrations of sucrose. The total amount of these main sugars available in RJ samples ranged from 8.8% to 12.2%, with an average of 10.5%. Sucrose is always present, but often in highly variable concentrations. This study illuminated that fresh Persian samples contain 60–70% water, 13.7% proteins, 7–18% carbohydrates, and up to 1.4% of 10-HDA. The average value for 10-HDA content was 2.3%. Also, it was cleared that the quality of RJ from mountainous areas were higher than the other regions. The results of this study indicated that RJ limits in some countries should be modified and International Honey Commission (IHC) and International Organization of Standardization (ISO) recommendations should be reviewed according to recent data on diversity. We believe that our proposals could be considered to set global standards for RJ.

Lyophilized royal jelly preparation in nanoscale and evaluation of its physicochemical properties and bactericidal activity.

Royal jelly, due to its unique bioactive components, has special biological activities, but a great extent of its nutritional value is lost during processing and storage. Lyophilization, an effective preservation technique, can feasibly preserve the main bioactive compounds present in royal jelly. In this study, fresh royal jelly was subjected to the freeze-drying process at a pressure and temperature of 100 Pa and - 70°C, respectively, for 40 h. The results obtained indicated that the pH, turbidity, total phenol content, and antioxidant activity of the royal jelly powder (RJP), during 3 months of storage at ambient temperature (30°C), were constant with values of 4.30, 1.634 (%A.U.), 0.617 (g/L), and 28.7 (%), respectively. Moisture content of the prepared RJP was less than 1%, while that of the fresh royal jelly was 70%. Furthermore, for the fresh royal jelly, the mentioned parameters were significantly (p < .05) decreased after 2 months of storage at freezer temperature (-20°C). GC-MS analysis indicated that the amount of 10-hydroxy-2-decanoic acid (10H2DA) in RJP was 3.85 times more than that of fresh royal jelly. The obtained results also indicated that prepared RJP had a high bactericidal effect toward Escherichia coli and Staphylococcus aureus, with clear zone diameters of 12 and 15 mm, respectively. The present study provides a foundation for research on the potential application of prepared RJP and the development of dietary supplements and functional foods.

Targeted and Suspect Fatty Acid Profiling of Royal Jelly by Liquid Chromatography-High Resolution Mass Spectrometry.

Royal jelly (RJ) is a bee product produced by the mandibular and hypopharyngeal glands of worker honeybees which has attracted special attention because of its numerous pharmacological activities and its applications to dermatology and cosmetics. In 2020, we demonstrated a liquid chromatography-high resolution mass spectrometry (LC-HRMS) method for the determination of seven medium-chain FFAs in RJ samples. The aim of the present work was to extend our studies on FA profiling of RJ, exploring the presence of common long-chain saturated, mono-unsaturated and poly-unsaturated free FAs in RJ samples using this LC-HRMS method. Among twenty common FAs studied by a targeted approach, palmitic acid, stearic acid and oleic acid were found at concentrations higher than the rest of the FAs (the concentrations of these three acids ranged from 37.4 to 48.0, from 17.7 to 24.0 and from 9.4 to 11.1 mg/100 g of fresh RJ, respectively). The high mass accuracy of LC-HRMS allowed the application of a suspect approach, which enabled the exploration of various C9 and C11 FAs, as well as hydroxylated C12 FAs. Nonenoic acid was indicated as the most abundant among these acids. In addition, for the first time, the presence of a variety of regio-isomers of hydroxymyristic, hydroxypalmitic and hydroxystearic acids was demonstrated in RJ samples.

Microencapsulated and Fresh Royal Jelly: Monitoring 10-HDA Content, Antibacterial and Antifungal Activity at Different Storage Periods

Microencapsulated and Fresh Royal Jelly: Monitoring 10-HDA Content, Antibacterial and Antifungal Activity at Different Storage Periods

Physicochemical characteristics of fresh and lyophilized Georgian royal jelly and Formulation bentonite-based cream

Royal Jelly (RJ) is one of the most valued and studied bee products used in medicine and cosmetics due to its diverse pharmacological properties. Chemical composition and biological activity of RJ depends on many factors, including constitution and geographical ​​distribution of plants, the harvest time and bee species. The objective of the present study was physico-chemical characterization and comparison of fresh and lyophilized RJ; and development dermatological formulation by incorporating RJ in Tikha Ascane base. The evaluations of RJ and RJ loaded formulations were performed by oven-dry, titrimetric, potentiometric, FTIR methods; 10-hydroxy-2-decenoic acid (10-HDA) was determined by high-performance liquid chromatography. In addition, the physical stability of the developed formulations was investigated by examining viscosity, spreadability, thermal and colloidal stabilities, pH, homogeneity and appearance. Lyophilized RJ displayed up to similar physico-chemical properties (solubility, pH, acidic number) as fresh RJ. The chromatographic analysis showed alike fingerprints and the existence of 10-HAD in tested samples. No spectral differences (in the range of 3600–2800 cm-1 and 1750–950 cm-1) were observed in FTIR measurements after lyophilization. The study shows that prepared formulations were found to be stable. The tested samples had appealing appearance with smooth texture and no signs of separation. Viscosity, homogeneity, spreadability, pH and release profile of RJ from the formulations were acceptable.

Physicochemical characteristics of local royal jelly produced in Al-Baha region, Saudi Arabia

Royal jelly (RJ) consumption is expanding because of its high nutritional and bioactive content. However, there is a lack of information to standardize the limit of properties of Saudi Royal Jelly for regulation. The aim of this study is to investigate the quality of Saudi royal jelly associated with the international standard. Twelve fresh royal jelly samples were analyzed for moisture, pH value, total acidity, protein, carbohydrate composition, and 10-hydroxydec-2-enoic acid (10-HAD) concentration. The result of physicochemical properties was varied from 61.70 % to76.80 % for moisture; 3.14 to 3.83 for pH; 39.4 to 45.0 mL for free acidity; 0.1 N NaOH/100 g; 571.60 to 745.80 µS/cm for electrical conductivity (EC); 6.73% to 13.27% for crude protein content. Moreover, the 10-HDA, fructose, glucose, sucrose, maltose, and lactose content were ranged from 1.68% to 6.36%, 2.51 to 5.71%, 0.00% to 2.64%, 0.00% to 2.01% and 0.00 to 0.69%, respectively. The obtained results indicated that all samples matched the international standard and Gulf Standardization Organization (GSO ISO 12824:2021) for royal jelly requirements in terms of 10-HDA concentration, one of the most critical quality parameters. The statistical analysis showed a significant positive correlation (r=0.487, P &lt; 0.01) between the EC and the total protein content while negative correlation between EC and sucrose content (r=-0.825, P &lt; 0.01) and 10HDA (r= -0.699, P &lt; 0.01); fructose and glucose content were shown to be statistically significant (r=0.887). The obtained results of present study will assist to the establishment of a standards of the Saudi royal jelly.

The role of microencapsulation in maintaining biological activity of royal jelly: comparison with biological activity and bioaccessibility of microencapsulated, fresh and lyophilized forms during storage.

Royal jelly (RJ) is a unique beehive product and has been recommended for human health since ancient times because of its antioxidant, antimicrobial, antiproliferative, neuroprotective, anti-lipidemic and anti-aging features. However, the biggest obstacle in the use of RJ is the need for cold storage and the instability of bioactive components over time. In the present study, 10-hydroxy-2-decenoic acid (10-HDA) content, as well as antioxidant [using 1,1-diphenyl-2-picrylhydrazy and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) methods] and antimicrobial activity (five Gram-positive, five Gram-negative and three yeasts), were comparatively evaluated for three RJ forms, two of which can be stored at 24 ± 1 °C during storage. Microencapsulated royal jelly (MRJ) stored at room temperature succeeded in preserving its 10-HDA content, a major bioactive compound, during the 6months, with respect to lyophilized royal jelly (LRJ) and fresh RJ stored at 4 °C. The initial 10-HDA contents of RJ, LRJ and MRJ were determined as 1.90%, 5.26% and 2.75%, respectively. Moreover, the total phenolic content, antioxidant capacity and antimicrobial activity mostly remained constant throughout the storage period (P ≥ 0.05). Gram-positive strains were generally more sensitive than Gram-negative strains. In the present study, the in vitro simulated digestion analysis showed that MRJ can tolerate the digestion process. Overall, the encapsulation process was considered as one preservative technique for RJ. The microencapsulation of RJ as shown in the results of the present study are encouraging in terms of enabling the local beekeeping sector to achieve ease of production and increased product diversity. MRJ shows promise as a commercial product with a high export value for producers. © 2022 Society of Chemical Industry.

Fruitful female fecundity after feeding Gryllodes sigillatus (Orthoptera: Gryllidae) royal jelly

Abstract Dietary honey bee royal jelly increases insect growth rates and adult body size. Royal jelly as a dietary supplement could enhance mass insect production by increasing the body size of mass-reared model species. To determine the effect of royal jelly on a cricket species, Gryllodes sigillatus Walker (Orthoptera: Gryllidae), farmed for human consumption, we ran two experiments. We tested the dose-dependent response of G. sigillatus to royal jelly using a range of diets across 0–30% w/w royal jelly. We also measured the individual-level life history responses to royal jelly over time by individually rearing G. sigillatus nymphs on two separate diets: half were fed a commercial cricket diet, and half were fed the same diet mixed with 15% w/w fresh royal jelly. We found sex-dependent effects: females fed the royal jelly diet were 30% heavier, and this effect was driven by significantly longer abdomens containing 67% more eggs compared to those fed the standard diet. Female mass was optimised at approximately 17% w/w royal jelly. Our results reveal that although a royal jelly dietary supplement can increase the yield of mass-reared insects, the life history responses are species and sex specific.

Formulation of Ocular In Situ Gels with Lithuanian Royal Jelly and Their Biopharmaceutical Evaluation In Vitro.

Royal jelly is a natural substance produced by worker bees that possesses a variety of biological activities, including antioxidant, anti-inflammatory, antibacterial, and protective. Although fresh royal jelly is kept at low temperatures, to increase its stability, it needs to be incorporated into pharmaceutical formulations, such as in situ gels. The aim of this study was to formulate in situ ocular gels containing Lithuanian royal jelly for topical corneal use in order to increase the retention time of the formulation on the ocular surface and bioavailability. Gels were evaluated for physicochemical characteristics (pH, rheological properties, refractive index) and in vitro drug release measuring the amount of 10-hydroxy-2-decenoic acid (10-HDA). An ocular irritation test and cell viability tests were performed using the SIRC (Statens Seruminstitut Rabbit Cornea) cell culture line. Results indicated that all the in situ gels were within an acceptable pH and refractive index range close to corneal properties. Rheology studies have shown that the gelation temperature varies between 25 and 32 °C, depending on the amount of poloxamers. The release studies have shown that the release of 10-HDA from in situ gels is more sustained than royal jelly suspension. All gel formulations were non-irritant according to the short-time exposure test (STE) using the SIRC cell culture line, and long-term cell viability studies indicated that the formulations used in small concentrations did not induce cell death. Prepared in situ gels containing royal jelly have potential for ocular drug delivery, and they may improve the bioavailability, stability of royal jelly, and formation of non-irritant ocular formulations.

Mining the Royal Jelly Proteins: Combinatorial Hexapeptide Ligand Library Significantly Improves the MS-Based Proteomic Identification in Complex Biological Samples.

Royal jelly (RJ) is a complex, creamy secretion produced by the glands of worker bees. Due to its health-promoting properties, it is used by humans as a dietary supplement. However, RJ compounds are not fully characterized yet. Hence, in this research, we aimed to broaden the knowledge of the proteomic composition of fresh RJ. Water extracts of the samples were pre-treated using combinatorial hexapeptide ligand libraries (ProteoMinerTM kit), trypsin-digested, and analyzed by a nanoLC-MALDI-TOF/TOF MS system. To check the ProteoMinerTM performance in the MS-based protein identification, we also examined RJ extracts that were not prepared with the ProteoMinerTM kit. We identified a total of 86 proteins taxonomically classified to Apis spp. (bees). Among them, 74 proteins were detected in RJ extracts pre-treated with ProteoMinerTM kit, and only 50 proteins were found in extracts non-enriched with this technique. Ten of the identified features were hypothetical proteins whose existence has been predicted, but any experimental evidence proves their in vivo expression. Additionally, we detected four uncharacterized proteins of unknown functions. The results of this research indicate that the ProteoMinerTM strategy improves proteomic identification in complex biological samples. Broadening the knowledge of RJ composition may contribute to the development of standards and regulations, enhancing the quality of RJ, and consequently, the safety of its supplementation.

Royal jelly alleviates the detrimental effects of aging on immune functions by enhancing the in vitro cellular proliferation, cytokines, and nitric oxide release in aged human PBMCS

Aging strongly delays the immunity. Our research aims to assess the in vitro effects of royal jelly (RJ) on the immune function of aged PBMCs. PBMCs were obtained from 10 healthy aged and young donors by the gradient density centrifugation method and further cultured in RPMI-1640 medium supplemented with or without RJ in the presence of Con A. Cell proliferation was assessed by MTT assay along with the measurement of interleukins, Nitric oxide (NO), Glutathione (GSH), and Malondialdehydes (MDA). Our results showed that RJ improved PBMCs proliferation significantly in the elderly subjects, accompanied by the increase in NO (p=.001) and the release of IL-2, IL-4, and IL-6 cytokines. RJ also increased the intracellular GSH (p= .001) and MDA (p=.001) levels in aged PBMCs. In young subjects, RJ enhanced PBMCs proliferation potency, IL-4, IL-6, GSH, and intracellular MDA levels but with a concomitant decrease in NO and IL-2 cytokine secretion as compared with non RJ-treated cells. In conclusion, RJ restored functions of the aged PBMCs as well as the young control subjects, indicating a beneficial effect on immune status during the aging process. PRACTICAL APPLICATIONS: Royal jelly is a well-known edible dietary compound, used traditionally to treat many diseases throughout the world. Since antiquity, it was shown to have medicinal importance. The immuno-enhancing potential of this food was largely and scientifically established by the lipid and protein fractions. The present study illustrates the anti-aging and stimulatory effects of the fresh RJ whole extract, from local Algerian honey bee: Apis mellifera intermissa, on the immunity of aged men. This study provides the experimental evidence supporting anti-immunosenesence effects of royal jelly. RJ supplementation can be used in the old age management and human age-related complications, especially, associated with the weaknesses of the immune response.

Abstract 14476: A Randomized, Double-blind Comparison Study of Royal Jelly to Improve Endothelial Function in Healthy Volunteers

Backgrounds: Royal jelly is a creamy substance secreted by honeybees. It has been reported that royal jelly has benefits on dyslipidemia and metabolic syndrome. However, the effects of royal jelly on atherogenesis remains unknown. We prospectively evaluated the effects of royal jelly on endothelial function, which can reflect early atherogenesis, in healthy volunteers. Methods: This was a single center, double-blind, 1:1 randomized placebo-controlled study. One-hundred healthy volunteers were randomly assigned to receive royal jelly 690 mg daily (equivalent 2040 mg of fresh royal jelly) or placebo from October 2018 and December 2019. Royal jelly or placebo were administered for 28 days. Endothelial function assessed by reactive hyperemia peripheral arterial tonometry (RH-PAT) and blood chemistries were compared between baseline and 28 days after enrollment. Primary endpoint was the change of RH-PAT index. Secondary endpoint was the change of liver function and lipid profiles. Results: The mean (SD) age of the participants was 35.0 (9.3) and 36.1 (9.1) years, and male was 45% and 50% in the placebo and the royal jelly group, respectively. The relative change of RH-PAT index was significantly higher in the royal jelly group compared with placebo control (21.4 ± 53.1% vs. 0.05 ± 40.9%, P = 0.037). The relative change of alanine aminotransferase and γ-glutamyl transpeptidase were significantly lower in the royal jelly group than the placebo group (alanine aminotransferase: –6.06 ± 22.2% vs. 11.6 ± 46.5%, P = 0.02; γ-glutamyl transpeptidase: -3.45 ± 17.8% vs. 4.62 ± 19.4%, P = 0.045). Lipid profiles were not significantly different between two study groups. Conclusions: Royal jelly improved endothelial function assessed by RH-PAT and liver function in healthy volunteer. It can be expected that royal jelly has anti-atherogenic property through improving endothelial function.

Comparative transcriptome analysis of hypopharyngeal glands from nurse and forager bees of Apis mellifera with the same age

The hypoharyngeal glands (HGs) are the major organ for synthesizing and secreting royal jelly (RJ) in nurse honeybees. To eliminate the influence of age, we explored the molecular basis of RJ production by determining the morphology and protein content, and comprehensively comparing the transcriptome of HGs of age-matched nurses and foragers obtained from an established colony. Acinus area and total protein content of HGs in nurses were significantly increased compared with aged-matched foragers. RNA-seq analysis showed that 510 up-regulated genes in HGs of nurses, mainly enriched on translation, transcription, DNA replication, and energy metabolism, were associated with synthesis and secretion of RJ, suggesting that HGs of nurses might employ diverse strategies to drive RJ synthesis. Particularly, MRJP1, MRJP4, and MRJP5, the primary components of fresh RJ, were extremely higher in HGs of nurses than those in the corresponding age foragers. This study will provide considerable insight into the strategies employed by nurses for RJ secretion.

Effects of royal jelly on bone metabolism in postmenopausal women: a randomized, controlled study

Objectives This study aimed to evaluate the effects of royal jelly (RJ) supplementation on bone metabolism in postmenopausal women. Methods This was a randomized, double-blind, placebo-controlled clinical trial. Seventy-two healthy postmenopausal women aged 45–60 years within 5 years after menopause were randomized into two groups: women in the RJ group (n = 36) received capsules containing dried RJ (equivalent to 3000 mg of fresh RJ); and women in the placebo group (n = 36) received placebo daily for 6 months. Bone mineral density (BMD) of the lumbar spine (L2–L4) and left proximal femur, hip structural analysis (HSA) of the left hip, and bone turnover markers were measured. Results Although women in the placebo group experienced a significant loss of BMD and deterioration in HSA parameters of the femur, no significant differences were found in these parameters in women in the RJ group. The levels of total procollagen type 1 N-terminal propeptide (P1NP) and tartrate-resistant acid phosphatase decreased significantly in the placebo group; however, the total P1NP level, a marker of bone formation, was not significantly different in the RJ group at postintervention compared with baseline. Conclusion RJ consumption may ameliorate decreases in femoral BMD and strength in postmenopausal women.

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.

Study of the Royal Jelly Free Fatty Acids by Liquid Chromatography-High Resolution Mass Spectrometry (LC-HRMS).

The lipidome of royal jelly (RJ) consists of medium-chained (8–12 carbon atoms) free fatty acids. We present herein a liquid chromatography-high resolution mass spectrometry (HRMS) method that permits the determination of RJ fatty acids and at the same time the detection of suspect fatty acids. The method allows for the direct quantification of seven free fatty acids of RJ, avoiding any derivatization step. It was validated and applied in seven RJ samples, where the major RJ fatty acid trans-10-hydroxy-2-decenoic acid (10-HDA) was found to vary from 0.771 ± 0.08 to 0.928 ± 0.04 g/100 g fresh RJ. Four additional suspect fatty acids were simultaneously detected taking advantage of the HRMS detection.

Quantification of Major Royal Jelly Protein 1 in Fresh Royal Jelly by Ultraperformance Liquid Chromatography-Tandem Mass Spectrometry.

Major royal jelly protein 1 (MRJP1) is the most abundant protein in royal jelly (RJ), and the level of MRJP1 has been suggested as a promising parameter for standardization and evaluation of RJ authenticity in quality. Here, a quantitative method was developed for the quantification of MRJP1 in RJ based on a signature peptide and a stable isotope-labeled internal standard peptide FFDYDFGSDER*(R*, 13C6, 15N4) by ultraperformance liquid chromatography-tandem mass spectrometry. Recoveries of the established method ranged from 85.33 to 95.80%, and both the intra- and interday precision were RSD < 4.97%. Quantification results showed that content of MRJP1 in fresh RJ was 41.96-55.01 mg/g. Abnormal levels of MRJP1 were found in three commercial RJs and implied that these samples were of low quality and might be adulterated. Results of the present work suggested that the developed method could be successfully applied to quantify MRJP1 in RJ and also could evaluate the quality of RJ.