Allergenic properties of Polish nectar honeys.

BACKGROUND: Small amounts of protein can be found in honey, including well known allergen sources, such as plant pollen and honeybee secretions. Despite this, there are few case reports describing allergic reactions following the consumption of honey. The aim of this study was to examine the allergenic properties of nectar honey collected throughout the entire beekeeping season from different provinces in Poland. METHODS: The immunoreactive properties of 20 Polish nectar honeys were analysed using the sera of IgE pollen allergenic patients (n = 5). The botanical origins and pollen of the anemophilous plants in the studied honeys were identified through palynological analysis. RESULTS: The significant differences in the protein content between the five varieties of honey and the differences in protein pattern and pollen profiles were observed. All of the honey samples contained immunoreactive fractions reacting with IgE present in the sera of patients allergenic to different pollens. CONCLUSIONS: Although honey allergies are reported relatively rarely, all the tested samples of Polish nectar honeys contained many protein fractions which reacted with the IgE antibodies of allergenic patients. In all samples, the immunoreactive protein band with a molecular weight around 60 kDa, probably secreted by bees, was present. The results do not allow the immunoreactive fractions characteristic for particular honey varieties to be identified.

N,N-dimethyl-2-phenylpropan-1-amine quantification in urine: application to excretion study following single oral dietary supplement dose.

N,N-dimethyl-2-phenylpropan-1-amine (NN-DMPPA) is a new designer stimulant prohibited in sport in-competition according to the List of Prohibited Substances and Methods published by the World Anti-Doping Agency (WADA). The first published data on the excretion study of NN-DMPPA to support the knowledge of NN-DMPPA in routine anti-doping control have been presented. The reliable gas chromatography-mass spectrometry quantitative method (GC-MS) has been validated and applied to the excretion study of NN-DMPPA. The validation parameters of the GC-MS method for determination of NN-DMPPA in human urine were the linear calibration range of 100 to 7500 ng/mL, the LOD of 13.9 ng/mL and the LOQ of 42.2 ng/mL. According to the obtained repeatability, intermediate precision, and trueness, the applied GC-MS method was precise and accurate. Urine samples from three volunteers in the excretion study were collected for 5 days after single oral administration of the supplement NOXPUMP containing NN-DMPPA. The obtained results showed the maximum concentration of NN-DMPPA (189-303 ng/mL) in urine samples at a time of 2-3 h post-administration. The NN-DMPPA concentration in urine samples was higher than 50 ng/mL until 22-23 h after the dietary supplement ingestion. This means that according to the WADA rules the use of a supplement containing NN-DMPPA may be related to a positive case when athletes took this supplement in-competition. Moreover, excretion results demonstrate also that NN-DMPPA may be detected in urine samples by the applied GC-MS method till 46 h after supplement administration. Additionally, the excretion study of ß-methylphenethylamine as the second prohibited substance present in the supplement NOXPUMP has been investigated. Graphical Abstract Excretion study of new designer stimulant, N,N-dimethyl-2-phenylpropan-1-amine, and ß-methylphenethylamine following single oral NOXPUMP supplement dose.

Evaluation of Safety of Iron-Fortified Soybean Sprouts, a Potential Component of Functional Food, in Rat.

Ferritin-iron is currently considered as one of the most promising iron forms to prevent iron deficiency anaemia. We found that the cultivation of soybean seeds in a solution of ferrous sulfate results in material with extremely high iron content - 560.6 mg Fe/100 g of dry matter, while ferritin iron content was 420.5 mg/100 g dry matter. To assess the potential adverse effects of a preparation containing such a high concentration of iron, male and female Wistar rats were exposed via diet to 10, 30, 60 g soybean sprouts powder/kg feed for 90 days. There were no differences in final body weight and mean food consumption between controls and rats administered sprouts. No statistically significant differences in haematology and clinical chemistry parameters were found between controls and treated rats. Microscopic examination of 22 tissues did not reveal any pathology due to soybean sprouts intake. Long term administration of the test material did not cause oxidative damage to DNA and protein in the liver as evidenced by the unchanged basal levels of DNA damage as well as carbonyl groups content. Lipid peroxidation was slightly increased only in females. The activity of several antioxidant enzymes: superoxide dismutase, glutathione peroxidase and glutathione S-transferase was increased, which substantially enhanced the antioxidant status in the liver from the rats treated with soybean sprouts. Hence, the material tested can be recommended as a component of food supplements for individuals with iron deficiency anaemia and inflammatory bowel diseases.

Legume seeds and cereal grains' capacity to accumulate iron while sprouting in order to obtain food fortificant.

BACKGROUND: Prepared sprouts, after culturing in a medium with an increased iron concentration, could become a beneficial food iron fortificant. However, the efficient iron accumulation depends on the plants genus, species and/or varieties. The aim of the study was to indicate the seeds or grains which accumulate iron most efficiently during the sprouting process. METHODS: Alfalfa, lentil, lupine and soybean seeds as well as wheat grains were sprouted in abiotic stress conditions induced by the excess of iron(II) in culture media. The tolerance of these plants to iron concentration and its accumulation in the material obtained (with FAAS method) were analyzed. RESULTS: The smallest tolerance was noted for lentil seeds and wheat grains. Other plants developed in 25 mM solution of FeSO4. The highest accumulation of iron was observed in alfalfa sprouts. However, lupine and soybean seeds are the most recommended raw material for the production of the sprouts on an industrial scale.

Determination of designer doping agent--2-ethylamino-1-phenylbutane--in dietary supplements and excretion study following single oral supplement dose.

The quantitative analysis of a new designer doping agent, 2-ethylamino-1-phenylbutane (EAPB) and its metabolite, 2-amino-1-phenylbutane (APB) in urine samples, and the determination of EAPB in dietary supplement samples, have been presented. The main purpose of the present study was to develop simple and reliable gas chromatography-mass spectrometry method (GC-MS) for excretion study following a single oral administration of dietary supplements containing EAPB. Three analytical methods for the determination of EAPB in urine and supplement samples, and APB in urine samples using the GC-MS system, have been validated. The method of the determination of EAPB in supplement samples was applied to analyze seventeen dietary supplements, CRAZE and DETONATE. Two other methods were used to determine the urinary excretion profile of EAPB and APB in the case of three healthy volunteers and, on further investigation, it was applied to the anti-doping control in sport. Quantification was obtained on the basis of the ions at m/z 86, 58 and 169, monitored for EAPB, APB and diphenylamine (used as an internal standard), respectively. The limits of detection and quantification were 2.4 and 7.3µg/g for EAPB in the case of supplement analysis, 2.9 and 8.8ng/mL for EAPB in the case of urine analysis, and 3.2 and 9.7ng/mL for APB. The other validation parameters as linearity, precision and trueness have been also investigated with the acceptable results. The extraction yield of all presented methods was above 69%. EAPB was detected in fourteen analyzed supplements (not included EAPB in their labels) and its content varied between 1.8 and 16.1mg/g. Following oral administration of three supplements with EAPB to one male and two female volunteers, the parent compound of EAPB and its metabolite were monitored and the excretion parameters as the maximum concentration of the analyte in urine (2.2-4.2µg/mL for EAPB; 1.1-5.1µg/mL for APB) and the time for the maximum height of the excretion peak (2-8h and 22h in one case for EAPB; 20-22h and 4h in one case for APB) have been indicated. EAPB and APB were detected at the level above 50ng/mL (50% of the minimum required performance level for stimulants in the anti-doping control in-competition in sport) in the urine up to 46-106h and 58-120h, respectively. Additionally, the result of the anti-doping control during swimming competition of one athlete, whose urine sample was analyzed for stimulants and narcotics, has been presented. The qualitative and quantitative analyses of new designer agents in urine samples and the excretion studies of these substances are of a great importance in the anti-doping control in sport. Moreover, the presentation of detection examples of these agents in supplements that haven't got included an information about them in the labeling, make athletes (and other supplement customers) more and more aware of the risk of the supplement use and possible health and doping consequences.

N,N-dimethyl-2-phenylpropan-1-amine - new designer agent found in athlete urine and nutritional supplement.

Reports of new designer agents banned in sport being detected in supplements widely available for athletes are constantly emerging. The task of anti-doping laboratories is to control athletes for the presence of substances listed by the World Anti-Doping Agency (WADA) and those that are structurally/biologically similar to them. Recently, a new designer stimulant, N,N-dimethyl-2-phenylpropan-1-amine (NN-DMPPA), was detected by the WADA accredited anti-doping laboratory in Warsaw during routine anti-doping control. The urine samples from four athletes were analyzed in the screening method for stimulants and narcotics and the presence of NN-DMPPA was detected. The identity of NN-DMPPA was confirmed by gas chromatography-mass spectrometry using a synthesized reference standard. The measured concentrations of NN-DMPPA were between 0.51 and 6.51 µg/mL. The presence of the NN-DMPPA compound has been detected in the 'nutritional supplement' NOXPUMP that had been purchased in a store in Poland. NN-DMPPA at 121.7 µg/g was indicated in the investigated supplement together with another banned stimulant ß-methylphenethylamine. The presence of this new stimulant was not indicated on the labelling of the supplement, a situation which is not unusual within this market. Thus, it is important to make athletes aware of the risk related to the use of supplements. Moreover, specific legistation dealing with the commercialization of drugs banned for sport should be undertaken.

Study on iron availability from prepared soybean sprouts using an iron-deficient rat model.

During soya seeds germination in FeSO(4) solutions their phytoferritin content is multiplied. Prepared soybean sprouts have been proposed as a safe and easily available source of iron supplementation. The preparation was compared with FeSO(4) and ferritin isolates, using rats with induced iron deficiency anaemia. After the end of the 2-week supplementation experiment, it was observed that no statistically significant differences in haemoglobin concentration, mean corpuscular volume, mean corpuscular haemoglobin, and mean corpuscular haemoglobin concentration existed between those animals supplemented with sprouts enriched in ferritin, ferritin isolate and FeSO(4) and healthy animals forming the control group. Moreover, the examined preparation had a beneficial influence on the recreation of ferritin reserves in both the liver and the blood serum, and also did not induce negative alterations in general growth parameters of animals. Use of an easily obtainable ferritin iron source may be a profitable alternative in supplementation due to its wide availability and food preservative properties.

Digestive enzyme inhibitors from grains as potential components of nutraceuticals.

Inhibitor activity against digestive enzymes, such as α-amylase from human saliva and porcine pancreas and trypsin from bovine pancreas, of three cereal grains species were studied as potential components of nutraceuticals strengthening diabetes and obesity treatment. Significant differences were demonstrated: the highest antitryptic activity was found in the grain of the rye varieties studied, whereas the grain of the wheat varieties had significantly higher ability to inhibit α-amylases from human saliva and porcine pancreas. Additionally, seeds of Puma, one of the studied wheat varieties, demonstrated especially low antitryptic activity. Such a beneficial arrangement of inhibitors, i.e. high level of inhibitors of α-amylase from human saliva and porcine pancreas and simultaneously low level of trypsin inhibitors from bovine pancreas, indicate the possibility of the application of seeds with such properties to the preparation of nutraceuticals for people with obesity or suffering from diabetes.

Seized designer supplement named "1-Androsterone": identification as 3ß-hydroxy-5α-androst-1-en-17-one and its urinary elimination.

New analogues of androgens that had never been available as approved drugs are marketed as "dietary supplement" recently. They are mainly advertised to promote muscle mass and are considered by the governmental authorities in various countries, as well as by the World Anti-doping Agency for sport, as being pharmacologically and/or chemically related to anabolic steroids. In the present study, we report the detection of a steroid in a product seized by the State Bureau of Criminal Investigation Schleswig-Holstein, Germany. The product "1-Androsterone" of the brand name "Advanced Muscle Science" was labeled to contain 100mg of "1-Androstene-3b-ol,17-one" per capsule. The product was analyzed underivatized and as bis-TMS derivative by GC-MS. The steroid was identified by comparison with chemically synthesized 3ß-hydroxy-5α-androst-1-en-17-one, prepared by reduction of 5α-androst-1-ene-3,17-dione with LS-Selectride (Lithium tris-isoamylborohydride), and by nuclear magnetic resonance. Semi-quantitation revealed an amount of 3ß-hydroxy-5α-androst-1-en-17-one in the capsules as labeled. Following oral administration to a male volunteer, the main urinary metabolites were monitored. 1-Testosterone (17ß-hydroxy-5α-androst-1-en-3-one), 1-androstenedione (5α-androst-1-ene-3,17-dione), 3α-hydroxy-5α-androst-1-en-17-one, 5α-androst-1-ene-3α,17ß-diol, and 5α-androst-1-ene-3ß,17ß-diol were detected besides the parent compound and two more metabolites (up to now not finally identified but most likely C-18 and C-19 hydroxylated 5α-androst-1-ene-3,17-diones). Additionally, common steroids of the urinary steroid profile were altered after the administration of "1-Androsterone". Especially the ratios of androsterone/etiocholanolone and 5α-/5ß-androstane-3α,17ß-diol and the concentration of 5α-dihydrotestosterone were influenced. 3α-Hydroxy-5α-androst-1-en-17-one appears to be suitable for the long-term detection of the steroid (ab-)use, as this characteristic metabolite was detectable in screening up to nine days after a single administration of one capsule.

Characteristics of chelation ability of chosen metal ions by protective ointments containing Na2H2EDTA.

Allergy contact dermatitis is a common occupational disease and the protective ointments are often used by the sensitized subjects. The efficacy of the chelation ability of the barrier creams containing Na2H2EDTA was evaluated. The in vitro test with the diffusion chamber and artificial membrane was performed. The effect of the Na2H2EDTA concentration (3, 5 or 10%), pH of the buffer for Na2H2EDTA dissolving and the vehicle of the ointment on the chelation of Ni2+ and Co2+ were assessed. The ointment with 10% Na2H2EDTA dissolved in the buffer of pH 7.0 or 7.4 buffer was found as optimal for the protection ability. There was no influence of the formula of the ointements on the efficiency of chelation.