Royal Jelly Samples Research Articles

The Effects of Royal Jelly on Autoimmunity in Graves' Disease

Graves' disease is an organ-specific autoimmune disease with unknown etiology. TSHR Ab plays the most important role for the pathogenesis of Graves' disease. Recently, the role of cytokines for the pathogenesis of Graves' disease has been studied extensively. Royal jelly (RJ) is a creamy product secreted by young nurse worker bees (Apis mellifera), and it is synthesized in the hypopharyngeal and mandibular glands. RJ has been reported to have such pharmacological characteristics as antitumor, antibacterial, antihypercholesterolemic, antiallergic, antiinflammatory, and immunomodulatory properties. The major aim of the present study is to evaluate the effect of RJ on autoimmunity in peripheral lymphocyte culture and to establish the therapeutic doses. In the first phase, lymphocyte cell isolation from four voluntary healthy subjects was performed to find the effective concentration of RJ on immunity. Serial dilutions of the RJ were prepared (0-5 mg/mL). All isolated lymphocyte cells were treated with the above diluted samples. MTT test was carried out after incubation of 72 h. In the second phase, six patients with Graves' disease, newly diagnosed by clinical and laboratory methods and admitted to my hospital and untreated were identified. RJ samples of 0 and 4 mg/mL were incubated in a culture medium for 72 h with isolated lymphocytes obtained from the patients. After incubation, MTT test in lymphocyte cell culture, Th1 cytokines IFN-gamma, TNF-alpha, and IL-12, and Th2 cytokines IL-4 and IL-10 levels by the enzyme amplified sensitivity immunoassay (EASIA) method and TSHR Ab by the radioreceptor method were determined. The concentration causing lymphocytes to proliferate was found to be 4 mg/mL by MTT test after incubation of 72 h in cell culture medium. Of the cytokines produced and secreted from lymphocytes, IFN-gamma increased, whereas, other cytokines decreased in RJ concentration of 4 mg/mL. Significant differences were found only for IFN-gamma and TNF-alpha. IL-4 concentrations were kept near the level of significancy. Of Th1/Th2 ratios, IFN-gamma/IL-4 and IFN-gamma/IL-10 ratios also exhibited significant differences between 0 and 4 mg/mL. RJ treatment in lymphocytes from patients with Graves' disease shifted the Th1/Th2 cytokine ratio to the side of Th1 cytokine. Therefore, RJ using the treatment and establishing a remission of Graves' disease may be effective as an antithyroid drug treatment. TSHR Ab levels of lymphocyte cell culture supernatants treated with RJ showed significant decreases. Also, the result may suggest that RJ may exert an effect similar to an antithyroid drug for decreasing TSHR Ab levels. RJ may be effective as an immunomodulatory agent in Graves' disease.

Detection of royal jelly adulteration using carbon and nitrogen stable isotope ratio analysis

Stable isotope ratios ((13)C/(12)C and (15)N/(14)N) were measured in royal jelly (RJ) samples by isotope ratio mass spectrometry (IRMS) to evaluate authenticity and adulteration. Carbon and nitrogen isotope contents (given as delta values relative to a standard, delta(13)C, delta(15)N) of RJ samples from various European origins and samples from commercial sources were analyzed. Uniform delta(13)C values from -26.7 to -24.9 per thousand were observed for authentic RJ from European origins. Values of delta(15)N ranged from -1.1 to 5.8 per thousand depending on the plant sources of nectars and pollen. High delta(13)C values of several commercial RJ samples from -20.8 to -13.3 per thousand indicated adulteration with high fructose corn syrup (HFCS) as a sugar source. Use of biotechnologically produced yeast powder as protein source for the adulterated samples was assumed as delta(15)N values were lower, as described for C(4) or CAM plant sources. RJ samples from authentic and from adulterated production were distinguished. The rapid and reliable method is suitable for urgent actual requirements in food monitoring.

Trace and mineral elements in royal jelly and homeostatic effects

Royal jelly from Apis mellifera is a highly active natural biological substance and is probably one of the most interesting raw substances in natural product chemistry. Trace elements play a key role in the biomedical activities associated with royal jelly, as these elements have a multitude of known and unknown biological functions. For this reason concentrations of 28 trace (Al, Ba, Sr, Bi, Cd, Hg, Pb, Sn, Te, Tl, W, Sb, Cr, Ni, Ti, V, Co, Mo) and mineral (P, S, Ca, Mg, K, Na, Zn, Fe, Cu, Mn) elements were systematically investigated in botanically and geologically defined royal jelly samples. In addition, concentrations of 14 trace elements were measured in the associated honey samples – honey being the precursor of royal jelly. Concentrations of K, Na, Mg, Ca, P, S, Cu, Fe, Zn, Al, Ba and Sr in royal jelly were determined by inductively coupled plasma optical emission spectroscopy (ICP-OES), while concentrations of Bi, Cd, Hg, Pb, Sn, Te, Tl, W, Sb, Cr, Mn, Ni, Ti, V, Co and Mo in royal jelly were determined by double focusing magnetic sector field inductively coupled plasma mass spectrometry (SF-ICP-MS). In the honey samples, trace and mineral element concentrations strongly depended on botanical and geological origin, and substantial variation was found. In contrast, the concentrations of trace and mineral elements were highly constant in the associated royal jelly samples. The most important results were the homeostatic adjustments of trace and mineral element concentrations in royal jelly. This effect was evidently produced in the endocrine glands of nurse bees, which are adapted for needs of bee larvae. In conclusion, this research yielded a surprising and completely new finding – that royal jelly, as a form of lactation on the insect level, shows the same homeostatic adjustment as mammalian and human breast milk.

Storage Stability Assessment of Freeze-Dried Royal Jelly by Furosine Determination

The effect of freeze-drying and the assessment of the storage stability of freeze-dried royal jelly (RJ) were investigated by the determination of furosine and blocked lysine. The level of furosine in the RJ samples collected from cells at different times (1, 2, and 3 days after grafting) showed that the Maillard reaction had already occurred in the hive as indicated by the increase in furosine: from 9.6 to 20.8 mg/100 g of protein. Freeze-dried RJ was more prone to the early stage of the Maillard reaction than fresh RJ, as confirmed by the significantly higher furosine values found after 12 months, both at 4 degrees C (253.4 versus 54.9 mg/100 g of protein) and at room temperature (884.3 versus 332.5 mg/100 g of protein). After 18 months at room temperature, the lyophilized samples reached a furosine level of 1440.4 mg/100 g of protein, which corresponded to the blocked lysine levels, amounting to 24% of total lysine.

Botanical resources used by Apis mellifera determined by pollen analysis of royal jelly in Minas Gerais, Brazil

SUMMARYPollen analysis of royal jelly samples obtained on the same day from six colonies in an apiary located in the region of Viçosa (Universidade Federal de Viçosa), state of Minas Gerais, Brazil, was carried out using the acetolysis method. Pollen grain types and number varied between the colonies. Three (Arecaceae, Cecropia, Eucalyptus) of the 38 plant taxa identified by the use of pollen morphology were common to all samples. Numerous tree species were represented by a few pollen grains in the royal jelly samples. Shrubs and herbs were also identified. Frequent pollen grains of the forest border plant species confirmed the location of the beehives next to this vegetation. Exotic plant species of Antigonon, Dombeya and Eucalyptus represented important resources for the bees. In accordance with the pollen analysis of propolis and geopropolis samples, pollen spectra of royal jelly samples may constitute a useful tool to indicate the regional origin of the samples and to identify the most important plant taxa for the bees.

Determination of trans-10-hydroxy-2-decenoic acid (10-HDA) in royal jelly from São Paulo State, Brazil

The 10-HDA content in Brazilian samples (São Paulo State) of royal jelly (RJ) was analyzed using an HPLC method based on the work by BLOODWORTH et al. [2]. The chromatographic conditions were: isocratic system, reversed phase C18-H column, auto sampler, diode array UV-VIS detector adjusted to 225nm, mobile phase composed by methanol/water (45:55) at pH= 2.5 adjusted with phosphoric acid; a-naphtol was used as internal standard, and the running time was 30min. By statistical analysis of the results, the 10-HDA contents of the samples analyzed seem to have two ranges: 1.8% and 3% (w/w), that would be useful to qualify the RJ. This is the first data regarding 10-HDA content of Brazilian RJ.

Evaluation of ( E)-10-hydroxydec-2-enoic acid as a freshness parameter for royal jelly

( E)-10-hydroxydec-2-enoic acid (10-HDA) is the main fatty acid present in royal jelly (RJ). Among the criteria for royal jelly quality analysis, 10-HDA content has been proposed as a freshness parameter. In this study, 10-HDA content variations under controlled temperature storage, were evaluated as an aging indicator of RJ samples from two origins (France and Thailand). For a 12 month period, the 10-HDA content loss rates were 0.1 and 0.2% at −18 °C and 4 °C, respectively, whatever the sample origin. At room temperature storage, losses were 0.4 and 0.6% for the French sample and the Thaı̈ sample, respectively. No correlation was found between 10-HDA content and storage duration whatever the storage temperature. Even if 10-HDA is specific to RJ, new criteria should be found to estimate the condition and duration of storage for royal jelly samples.

Comparação de metodologias para a determinação de umidade em geléia real

The aim of this paper was to compare methods of moisture determination to choose the best one for the determination of this parameter in royal jelly samples. Royal jelly is sensitive to high temperatures becoming dark and loosing volatiles in high temperatures. The methods were: vacuum oven at 60 oC, 70 oC, conventional oven at 105 oC, Karl Fisher, dissecator with sulfuric acid and dryness with infrared light at 105 oC. Based on the results, the best method was the dissecator with sulfuric acid for moisture determination in royal jelly.

Furosine: a Suitable Marker for Assessing the Freshness of Royal Jelly

Fifteen commercial samples of royal jelly, consisting of 10 imported samples, and 5 samples of known origin obtained freshly harvested from beekeepers, were analyzed for protein, lysine, and furosine content. In addition, a commercial sample of royal jelly, at the beginning of its commercial shelf life, was stored for 10 months both at 4 degrees C and at room temperature in order to assess the development of the Maillard reaction (furosine) and relative nutritional damage (blocked lysine). The commercial royal jelly products contained different amounts of furosine, ranging from 37.1 to 113.3 mg/100 g protein, evidence of different storage times and conditions. The average furosine content of the royal jelly samples of known origin and harvesting was significantly lower than that of the imported samples (41.7 versus 73.6 mg/100 g protein, respectively). With regard to shelf life, furosine content increased significantly from 72.0 mg/100 g protein to 500.8 mg/100 g protein after 10 months of storage at room temperature, while it increased to a much lower level (100.5 mg/100 g protein) when the royal jelly was stored at 4 degrees C. However, nutritional damage, expressed as blocked lysine (calculated indirectly from the furosine content), was minor or negligible, 11.9 and 2.3% of total lysine, in samples stored at room temperature and at 4 degrees C, respectively. Lysine was determined by an innovative procedure based on high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). The results showed that furosine is a suitable index for assessing the quality and freshness of royal jelly.

Colitis in a human consuming royal jelly

Using headspace solid-phase microextraction (HS-SPME) followed by diethyl ether and methanol extraction, it was possible to isolate as many as 185 organic compounds out of 17 samples of crude royal jelly (RJ). Of the above compound number, 169 compounds were positively identified by means of gas chromatography–mass spectrometry. The volatile fraction of RJ consists of 25 different compounds where approximately 47% of the total ion current (TIC) of volatile compound chromatograms were composed of substances characterized by bactericidal (phenols) and repelling (octanoic acid and 2-heptanone) activities. Preliminary investigations have shown that RJ stored for 10 months at −18 °C and 4 °C keeps its composition of volatile compounds unchanged, however, at the same time at room temperature RJ phenol contents is decreased twice, whereas the fraction of aliphatic acids is increased 2.8 times due to the presence of both acetic and butyric acids. The chromatogram of RJ ether extracts showed 85 different compounds, however about 88% of TIC consisted exclusively of 8 compounds, i.e. 10-hydroxy-2-decenoic, 10-hydroxydecanoic, 3,10-dihydroxydecanoic, 8-hydroxyoctanoic, 2-decene-1,10-dioc and (Z)-9-hydroxy-2-decenoic acids. Nine aliphatic acids, which were detected for the first time, are the homologues of hydroxy- and oxo-acids identified earlier in RJ. In the RJ methanol extracts 82 compounds were identified, mainly carbohydrates and their derivatives. Approximately 87% of TIC consisted of fructose, glucose and sucrose. Special attention was paid to discrepancies between obtained and literature data concerning the presence of free amino acids in RJ. It was suggested that these inconsistencies can be explained by the differences in the methods of RJ collection and/or sample preparation.

Determination of trans-10-Hydroxydecenoic Acid in Royal Jelly by Capillary Gas Chromatography.

The determination and confirmation of trans-10-hydroxydecenoic acid (10-HDA) in royal jelly (RJ) were developed by capillary gas chromatography (GC) and GC-chemical ionization mass spectrometry (CI-MS) equipped with an ion trap detector (ITD) as a mass spectrometer. It was found that 10-HDA extracted with dichloromethane at pH 2.5 was converted into trimethylsilyl (TMS) derivative in a constant yield by the reaction with N, O-bis (trimethylsilyl) acetamide at room temperature for 15 min. The recovery of 10-HDA (50 μg) was 99.5%. The TMS derivatives of raw RJ tested showed a satisfactory result of separation and sensitivity by GC using a DB-1 capillary column with a flame ionization detector. The linear dynamic detection range was approximately from 50 ng to 500 μg (16.7-167000 ppm) determined using n-eicosane as an internal standard. The minimum detectable amount of 10-HDA was found to be 50 ng (16.7 ppm) at the split ratio of 40 : 1 of the split injection. The amount of 10-HDA in the raw RJ sample was 2.28±0.04% determined by the GC method and 2.25±0.01% by the HPLC method as described previously, respectively. The TMS derivatives of 10-HDA and its related compounds in the raw RJ were confirmed by ITD-GC. The diagnostic ions of TMS-HDA were presented at m/z 331 (M+1), 315 (M-Me), and 241 (M-OTMS) in isobutane CI mode. The present GC method is an easier and more convenient method for the analysis of 10-HDA in RJ and is applicable for the determination and confirmation of 10-HDA in forensic and food samples.

Dissipation of Oxytetracycline Residue in Royal Jelly

Oxytetracycline (OTC) in royal jelly can be extracted by organic solvents and Amberlite XAD-2 resine column chromotography, and the average of OTC recovery was 64.86±6.14%. The cylinder-plate method is generally used for the assay of OTC residue with Bacillus cereus var. mycoides (ATCC 11778). Different dosages of OTC were applied to honeybee colonies to control foulbrood. The dissipation curves of OTC residue were achieved by periodically detecting samples of royal jelly. The curve showed that the OTC residue reduced to about 10% of initial residual level within 13 days (4 samplings). After then, the OTC residue dissipated slowly to undetectable level. For hives received 200ppm (0.125g) OTC twice, 100ppm (0.063g) twice and 50ppm (0.031g) once, the periods needed for residual dissiption to lower than 0.05ppm were 45, 35 and 24 days, respectively.

Studies on Composition, Freeze-Drying and Storage of Turkish Royal Jelly

SummaryThis work characterizes the composition of Turkish royal jelly as determined from a sample provided by a local commercial producer. The constituents analysed were moisture, total sugars, lipids, proteins, amino acids, ash, minerals (iron and calcium) and vitamins (thiamin riboflavin, niacin). The same royal jelly sample was freeze-dried, both at a laboratory and an industrial scale. The compositions of freeze-dried products were investigated comparatively through determination of total acidity over a 6-month period. Various formulations were developed for tablet-making and the most favourable formulation was recommended for industrial production.

Decreasing Effect of Honey on Hydroxy Acids in Royal Jelly Products

Royal jelly has been used as a material for “health food” and crude drugs. In order to estimate the royal jelly content in various royal jelly products, 10-hydroxy-2-decenoic acid (10-HDA), one of the specific fatty acids of royal jelly, has been analyzed by gas chromatography. A survey on commercial royal jelly products containing honey revealed 10-HDA concentrations in the range from 1.03-1.60%, though 10-HDA in fresh royal jelly samples was in the range from 1.84 to 2.50%. 10-Hydroxy decanoic acid contents were 0.60 to 0.78%.The 10-HDA concentration in honey-containing royal jelly products was less than in fresh royal jelly samples. Therefore, experiments were conducted to investigate the relationship between the decrease of 10-HDA concentration and time in samples of 0.20% 10-HDA with 10.0% honey in neutral aqueous solution and 10.0% fresh royal jelly with 10.0% honey in neutral aqueous solution. The concentration of 10-HDA in the former sample decreased to 0.04% (initially 0.20%) after 90 days, while that in the latter sample decreased to 0.14% (initially 0.20%).

Stable Carbon-Isotope Analysis of Harvested and Commercial Royal Jelly

SummaryDel 13C-isotope ratios were determined in royal jelly samples harvested from 4 Apis meillfera colonies at Pennsylvania State University. One sample, collected in July 1981, had a 13C-isotope ratio of −13·3%0. This ratio indicates that the carbon of the dietary organic material utilized by the bees was predominantly from C4 plants. The other samples, collected during August of the same year, had 13C-isotope ratios ranging from—25·2 to −25·6%0; thus the carbon of the dietary sources of these bees was primarily from C3 plants. The isotope ratios for 6 samples of commercial royal jelly products (pure royal jelly or royal jelly + honey) ranged from −6·9%0 to −28·9%0. The possibility that some of these products were adulterated is discussed.

Components of Royal Jelly II. The Lipid Fraction, Hydrocarbons and Sterols

SummaryQuantitative differences were found in the chemical composition of royal jelly samples collected in spring and summer by the same producer. The main difference was in the free fatty acids, which showed a marked increase in 10-hydroxydecanoic acid in summer. The sterol and hydrocarbon fractions were also investigated. The most significant sterols were identified by gas chromatography—mass spectrometry. The hydrocarbon components, identified by their retention times, were a homologous series of straight-chain compounds, from C16 to C33, with even and odd numbers of carbon atoms. Considerable amounts of some hydrocarbons were found. Of the sterols identified, the most abundant was 24-methylene cholesterol; stigmasterol, β-sitosterol, Δ5-avenasterol, Δ7-avenasterol and cholesterol were also present.