Formulation Of Food Supplements Research Articles

Up-Cycling of Olea europaea L. Ancient Cultivars Side Products: Study of a Combined Cosmetic-Food Supplement Treatment Based on Leaves and Olive Mill Wastewater Extracts.

In recent years, a reversal of the global economic framework has been taking place: from the linear model, there has been a gradual transition to a circular model where by-products from the agri-food industry are taken and transformed into value products (upcycling) rather than being disposed of. Olive tree pruning represents an important biomass currently used for combustion; however, the leaf part of the olive tree is rich in phenolic substances, including hydroxytyrosol. Mill wastewater is also discarded, but it still contains high amounts of hydroxytyrosol. In this study, cosmetic and food supplement formulations were prepared using biophenols extracted from leaves and wastewater and were tested in a placebo-controlled study on healthy volunteers using a combined cosmetic and food supplement treatment. A significant improvement in skin health indicators (collagen density, elasticity, etc.) and a 17% improvement against Photo-induced Irritative Stimulus was observed.

Fomitopsis officinalis: Spatial (Pileus and Hymenophore) Metabolomic Variations Affect Functional Components and Biological Activities.

Fomitopsis officinalis is a holartic polyporous mushroom that forms large fruiting bodies on old standing trees, fallen logs, or stumps. F. officinalis is a medicinal mushroom species that is most commonly used in traditional European medicine. In this study, we explore the spatial metabolic differences in F. officinalis' mushroom parts, i.e., the cap (median and apical parts) and the hymenium. Additionally, chromatographic analysis was conducted in order to unravel the composition of specialized metabolites in the hydroalcoholic mushroom extracts. The potential antifungal and bacterial effects of extracts were tested against pathogen strains of Gram+ and Gram- bacteria, and yeast, dermatophytic, and fungal-pool species. Extracts from the apical part were the richest in terms of phenolic compounds; consistent with this finding, the extracts were also the most effective antiradical and antimicrobial agents with MIC values < 100 µg/mL for most of the tested bacterial and dermatophytic species. According to these findings, F. officinalis extracts are valuable sources of primary and secondary metabolites, thus suggesting potential applications in the formulation of food supplements with biological properties in terms of antioxidant and antimicrobial activities.

Diapin®: A food supplement with diverse therapeutic potentials

Background and Aims: For many centuries herbs and spices have traditionally been used to treat or manage a variety of diseases. The formulation of food supplements containing single or multiple herbs or spices is now popular. These formulations provide biochemical, pharmacological and medicinal benefits due to their diverse phytochemical constituents. Methods: In the present study, Diapin® – a food supplement containing Olea europaea L. leaves extract, Cinnamomum cassia (L.) J. Presl stem extract, Nigella sativa L. seed oil, Cocos nucifera L. oil and vitamin D3 – dissolved in absolute ethanol was evaluated for α-amylase, α-glucosidase, acetylcholinesterase, elastase, neuraminidase, adenosine deaminase and arginase inhibitory activity. Results: The supplement strongly inhibited neuraminidase (IC50 = 0.272 ± 0.007 mg/mL), while adenosine deaminase, acetylcholinesterase, elastase and arginase were moderately inhibited (with an IC50 of 4.562 ± 0.052, 5.396 ± 0.563, 5.783 ± 0.058 and 6.800 ± 0.067 mg/mL respectively). The less inhibition activity was on α-amylase and α-glucosidase (IC50 = 9.593 ± 0.582 and 14.010 ± 2.280 mg/mL respectively). Conclusion: The pharmacological activities of Diapin® can be attributed to its opulent phytochemical composition. The present findings support the folkloric claim of Diapin® supplement having antidiabetic, anticancer, anti-inflammatory, antimicrobial, anti-ageing, and immune bolstering properties, in addition to the mitigation of Alzheimer’s disease and the alleviation of neurological dysfunction.

Antiglycation and antioxidant activities of the crude extract and saponin fraction of Tribulus terrestris before and after microcapsule release

ObjectiveThe present study investigated antiglycation and antioxidant activities of crude dry extract and saponin fraction of Tribulus terrestris. It also developed a method of microencapsulation and evaluated antiglycation and antioxidant activities of crude dry extract and saponin fraction before and after microcapsule release. MethodsAntiglycation activity was determined by relative electrophoretic mobility (REM), free amino groups and inhibition of advanced glycation end-product (AGE) formation. Antioxidant activity was determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric ion-reducing antioxidant power (FRAP), nitric oxide (NO) and thiobarbituric acid reactive species (TBARS) tests. Microcapsules were prepared using maltodextrin as wall material and freeze-drying as encapsulation technique. Morphological characterization of microcapsules was evaluated by scanning electron microscopy, and encapsulation efficiency and microcapsule release were determined by total saponins released. Antiglycation and antioxidant assays were performed using crude dry extract and saponin fraction of T. terrestris before and after release. ResultsSaponin fraction showed an increase of 32.8% total saponins. High-performance liquid chromatography-mass spectrometry analysis showed the presence of saponins in the obtained fraction. Antiglycation evaluation by REM demonstrated that samples before and after release presented antiglycation activity similar to bovine serum albumin treated with aminoguanidine. Additionally, samples inhibited AGE formation, highlighting treatment with saponin fraction after release (89.89%). Antioxidant tests demonstrated antioxidant activity of the samples. Crude dry extract before encapsulation presented the highest activities in DPPH (92.00%) and TBARS (32.49%) assays. Saponin fraction before encapsulation in FRAP test (499 μmol Trolox equivalent per gram of dry sample) and NO test (15.13 μmol nitrite formed per gram of extract) presented the highest activities. ConclusionThis study presented antiglycation activity of crude dry extract and saponin fraction of T. terrestris, besides it demonstrated promising antioxidant properties. It also showed that the encapsulation method was efficient and maintained biological activity of bioactive compounds after microcapsule release. These results provide information for further studies on antidiabetic and antiaging potential, and data for new herbal medicine and food supplement formulations containing microcapsules with crude extract and/or saponin fraction of T. terrestris.

Metabolomic Profiling and Biological Activities of Pleurotus columbinus Quél. Cultivated on Different Agri-Food Byproducts.

The genus Pleurotus (Fr.) P. Kumm (Pleurotaceae, Basidiomycota) comprises a cosmopolitan group of mushrooms highly appreciated for their nutritional value and health-promoting benefits. Despite there being many studies about the phytochemical composition of Pleurotus spp., there are very few reports dealing with the phytochemistry, antioxidant and antimicrobial activities of P. columbinus Quél. In this study, a mass spectrometry ultra-performance liquid chromatography mass spectrometry (UHPLC)-QTOF method, coupled with principal component analysis (PCA), was applied to the P. columbinus metabolome in order to investigate the influence of different agri-food residues as growth substrates for P. columbinus cultivation, on the bioactive chemical profile of fruiting bodies and evaluated their potential as antioxidants and antimicrobials. Additionally, a quantitative HPLC-DAD-MS analysis was conducted on phenolic and flavonoid compounds, that could explain, albeit partially, the observed biological effects of P. columbinus extracts. The qualitative metabolic profile identified 97 metabolites, whereas the quantitative HPLC-DAD-MS analysis confirmed the presence of phenolic and flavonoids, in the mushroom extracts, which also showed intrinsic scavenging/reducing and antimicrobial effects. The antibacterial effects were particularly evident against Escherichia coli, whereas Tricophyton and Aspergillus were the dermatophytes more sensitive to the mushroom extracts. The present study supports more in-depth investigations, aimed at evaluating the influence of growth substrate on P. columbinus antimicrobial and antioxidant properties. The extracts from P. columbinus revealed valuable sources of primary and secondary metabolites, thus suggesting potential applications in the formulation of food supplements with biological properties, above all in terms of antioxidant and antimicrobial properties.

Multi-Class UHPLC-MS/MS Method for Plant Toxins and Cyanotoxins in Food Supplements and Application for Belgian Market Samples.

The presence of plant toxins and/or cyanotoxins in food supplements implies consumer health risks. Therefore, a targeted ultra-high performance liquid chromatographic-tandem mass spectrometric method to detect/quantify 25 toxins simultaneously in food supplement formulations was developed and validated. Full validation for tablets/powders and secondary validation for a liquid and soft gel capsule indicated that most compounds were efficiently extracted (≥ 75%), while others were only partly extracted (18 - 61%). Trueness was fulfilled (70 - 120%), with some exceptions (mostly at the lowest validation level). Intralaboratory repeatability, intra- and interlaboratory reproducibility values of ≤ 20%, ≤ 25%, and ≤ 25% were obtained for most, respectively. Matrix effects were found to be significant for most compounds. Good sensitivity (µg/kg level) was observed for galegin(e), lycopsamine, lycorine, rubiadin, skimmiamine, and vascin(e), in contrast to helveticoside, lucidin, lucidin-3-primveroside, plumbagin(e), and thujone, which were detected at the mg/kg level. The other compounds were characterized by a sensitivity between 10to 1000 µg/kg. The validated methodology was applied for 52 food supplements (tablets, capsules, liquids/syrup, etc.) purchased from the Belgian market. In more than 25% of the samples, one or more toxins were detected (concentrations determined using standard addition). Lycopsamine, microcystin LR, solamargine, thujone, and vasicin(e) were the most frequently detected toxins. A clear link between the toxins detected and the plant species on the food supplement ingredient list could not always be established. This generic "dilute-and-shoot" procedure can be used for further research on toxins in food supplements and by extension other plant/algae-based food/feed commodities (herbs, edible flowers, etc.).

Evaluation of Dissolution Profiles of a Newly Developed Solid Oral Immediate-Release Formula Containing Alpha-Lipoic Acid

Alpha-lipoic acid (ALA, thioctic acid), a naturally-occurring essential dithiol compound, has become a common ingredient in many pharmaceutical and food supplement products (FSP), used in oxidative stress-dependent pathologies; oral bioavailability of ALA is limited by pharmacokinetic particularities that reduce its therapeutic efficacy-reduced solubility, lack of gastric stability and hepatic degradation, doubled by formulation hinders. The objectives were to develop a solid oral 600 mg ALA FSP to obtain an optimal pharmaceutical profile compared to a reference listed drug (RLD) with a similarity factor f2 50. A comparative dissolution study was performed; an HPLC method was used for ALA quantification. After planning combinatory simulations (formulation stage), two prototype formulas (#1 and #2) were manufactured and further optimized by adjusting ALA physical characteristics and the excipients quantities (#3 and #4) in order to achieve the Quality Target Product Profile. A misshapen of ALA’s in vitro release was observed for #3 Formula (f2 = 31.6); the optimal profile was obtained for Formula #4 (f2 = 58.5). A simple quantitative formula is not enough to assure good ALA bioavailability; the formulation needs multiple compounding modulations under physicochemical compatibility algorithms, with multiple dissolution profiles testing back-ups. It is essential to ensure a formulation with an in vitro dissolution comparable with the RLD, allowing the compound to reach its target level to assure the optimum claimed antioxidant activity of ALA at the cellular level, even for food supplement formulations.

Phytochemistry, nutritional composition, and pharmacological activities of Thaumatococcus daniellii (Benth): a review.

Thaumatococcus daniellii (Benth) is a member of a diverse family of plants known as Marantaceae. Native to the tropical forest zones of West Africa, the plant is globally famous for its low calorie, nondiabetic natural sweetener called thaumatin found in its aril. T. daniellii thrives in deep shade, and it is used locally as a taste modifier and for preparing fish traps, ornamental bags, and mats. Organs of the plant are used in folkloric medicine as a laxative and in treating ailments such as mental disorders, high blood sugar, and lung diseases. The seeds and leaf sap are potent as an antidote against snake venom and bee stings and for preventing dystocia and prolonged child labor. Proximate analysis, phytochemical screening, and gas chromatography-mass spectrophotometry revealed that the plant contains proteins, important macro- and microelements (calcium, magnesium, zinc, sodium, phosphorus, potassium, iron, and manganese), and abundant active principles and compounds such as squalene tannin, alkaloids, saponins, epicatechin, steroids, phlobatannins, anthraquinones, terpenoids, spartein, ribalinidine, rutin, phytic acid, and kaempferol. Biological activities include hypolipidemic, antihyperglycemic, antioxidant, insecticidal, bioremediative, and antimicrobial activities. T. daniellii could be used in the formulation of food supplements and drug development.

Extracts from Myrtle Liqueur Processing Waste Modulate Stem Cells Pluripotency under Stressing Conditions.

Nutraceuticals present in food are molecules able to exert biological activity for the prevention and treatment of various diseases, in form of pharmaceutical preparations, such as capsules, cream, or pills. Myrtus communis L. is a spontaneous Mediterranean evergreen shrub, widely known for the liqueur obtained from its berries rich in phytochemicals such as tannins and flavonoids. In the present study, we aimed to evaluate the properties of myrtle byproducts, residual of the industrial liqueur processing, in Adipose-derived stem cells (ADSCs) induced at oxidative stress by in vitro H2O2 treatment. Cells were exposed for 12-24 and 48h at treatment with extracts and then senescence-induced. ROS production was then determined. The real-time PCR was performed to evaluate the expression of inflammatory cytokines and sirtuin-dependent epigenetic changes, as well the modifications in terms of stem cell pluripotency. The β-galactosidase assay was conducted to analyze stem cell senescence after treatment. Our results show that industrial myrtle byproducts retain a high antioxidant and antisenescence activity, protecting cells from oxidative stress damages. The results obtained suggest that residues from myrtle liqueur production could be used as resource in formulation of food supplements or pharmaceutical preparations with antioxidant, antiaging, and anti-inflammatory activity.

Metabolomic study of bioactive compounds in strawberries preserved under controlled atmosphere based on GC-MS and DI-ESI-QqQ-TOF-MS.

The storage of the vegetables products in a controlled atmosphere (CA) with low O2 and high CO2 concentrations, reduces respiration rates and delays the ripening process, and in some cases, improves their quality and organoleptic properties. To obtain deep insight into strawberry fruit metabolic changes caused by these CA treatments. Freshly harvested strawberries were preserved under different atmospheres enriched with 10%, 20% and 30% of CO2 , for 2days at 0°C, containing in all the cases 5% of O2 and were subjected to a metabolomic analysis based on gas chromatography-mass spectrometry (GC-MS) and direct-infusion with electrospray ionisation source equipped with triple quadrupole coupled to time of flight mass spectrometry (DI-ESI-QqQ-TOF-MS). Partial least square discriminant analysis (PLS-DA) was employed to compare the control and treated samples for the identification of altered metabolites. Several metabolites related to CA treatment could be identified by databases and literature, which are mainly sugars, organic acids and phenolic compounds (bioactive compounds). Good correlation coefficients were obtained between discriminant metabolites and fruit quality parameters. These results suggest that treated strawberries under CA could be considered as bioactive healthy compounds, suggesting that treated strawberries under CA could be used as raw material for the preparation and formulation of food supplements and nutraceutical products.

Identification of l-carnitine and its impurities in food supplement formulations by online column-switching liquid chromatography coupled with linear ion trap mass spectrometry.

The identification of impurities in l-carnitine by mass spectrometry is difficult because derivative reagents or ion pair reagents are usually used to separate and increase the retention of l-carnitine on the reversed-phase column. In this study, four impurities including 3-chloro-2-hydroxy-N,N,N-trimethylpropan-1-aminium, 3-cyano-2-hydroxy-N,N,N-trimethylpropan-1-aminium, 3-carboxy-N,N,N-trimethylprop-2-en-1-aminium, and 4-chloro-2,3,4-trihydroxy-N,N,N-trimethylbutan-1-aminium were identified in l-carnitine and its tablets by using two-dimensional column-switching high-performance liquid chromatography coupled with linear ion trap mass spectrometry. The first column was a C8 column at a flow rate of 0.15 mL/min; the detection wavelength was 220 nm. The second column was an Acclaim Q1 column using a gradient elution program with aqueous 30 mM ammonium acetate (pH 5.0) and acetonitrile as the mobile phase at a flow rate of 0.5 mL/min. The mass fragmentation patterns and structural assignments of impurities were studied, and the quantitative validation of three impurities was further investigated. The linearity (r2 ) was found to be >0.99, with ranges from 0.2 to 50 ng/mL and 0.1 to 10 ng/mL. The method was used successfully for determination of impurities in five samples of l-carnitine and tablets.

Polyphenolic pattern and in vitro cardioprotective properties of typical red wines from vineyards cultivated in Scafati (Salerno, Italy)

Wines are the subject of increasing numbers of investigations owing to the pharmaceutical usefulness of grape phytochemicals. The aim of the present work was to hypothesize the use of lyophilised red wines for the formulation of food supplements potentially useful against both physiological and induced cardiac oxidative stress. Cardiac derived H9C2 myocytes were incubated with increasing doses (0.01–1μg) of lyophilised Aglianico wine (lioAW). Experiments showed an appreciable direct radical scavenging activity at a maximum lioAW dose of 0.03μg that made the caspase-3 activity decrease by about 41%. Cardiac cells were exposed to 1μM doxorubicin and its combination with different doses of lioAW. Maximum lioAW aliquot of 0.03μg seemed to effectively contrast the induced oxidant injury decreasing the reactive oxygen species (ROSs) levels by about 38%and depressing the caspase-3 activity by about 63%. In both assays, pro-oxidant effects at higher lioAW concentrations were detected.

Antioxidant Profile and in Vitro Cardiac Radical-Scavenging versus Pro-oxidant Effects of Commercial Red Grape Juices (Vitis vinifera L. cv. Aglianico N.)

Several works have reported on the pharmaceutical usefulness of grape phytochemicals. Nevertheless, the scientific literature needs further studies to consider grape extracts as useful dietary supplements. The aim of the present work was to hypothesize for the first time the use of whole commercial red grape juices as food supplements potentially useful against both physiological and induced cardiac oxidative stress. First of all, the results indicated a good antioxidant stability of the juice sample to lyophilization that may be reasonably regarded as a suitable process for the formulation of food supplements. Then, the processed sample (lioRGJ) was tested on cardiac-derived H9C2 myocytes to ascertain its effects on reactive oxygen species (ROS) generation and caspase-3 activity incubating cardiomyocytes with lioRGJ at increasing doses (0.01-1 μg). Experiments showed an appreciable direct radical-scavenging activity at a maximum sample dose of 0.01 μg that made the caspase-3 activity decrease by about 47% (P < 0.001). Cardiac cells were exposed to 1 μM doxorubicin and its combination with different doses of lioRGJ. A maximum sample aliquot of 0.01 μg seemed to effectively contrast the induced oxidant injury, decreasing the ROS levels by about 31% and depressing the caspase-3 activity by about 60% (P < 0.001). In both assays, pro-oxidant effects at higher sample concentrations were detected as indicated by the increase in both ROS generation and apoptotic activity. The data suggested the possible employment of the juice sample as a food supplement with prospective cardioprotective benefits, although further studies are needed to optimize its dosages to avoid harmful pro-oxidant effects.

Quantitative and Qualitative Evaluation of Chondroitin Sulfate in Dietary Supplements

We propose to evaluate the amount and quality of chondroitin sulfate (CS) from several food supplement preparations in the form of tablets, caplets, or capsules containing CS in varying contents and formulations in the presence of various additives and ingredients, with no other pretreatment than centrifugation to remove insoluble material. To quantify CS, two different analytical approaches were applied after their validation: specific and sensitive agarose-gel electrophoresis and strong-anion exchange–high-performance liquid chromatography (SAX-HPLC) determination of the constituent disaccharides after treatment with specific chondroitin lyases. The CS content in finished products evaluated using the two specific validated methods were found to conform to the label specifications. It is worth mentioning that the quantitative determinations have been performed using a very high pure European Pharmacopeia CS reference standard having substantially the same properties as the food supplement CS samples. Furthermore, by means of the specific agarose-gel elctrophoresis approach, we can exclude the presence in the nutraceuticals of other sulfated polysaccharides produced by extraction from tissues, in particular heparin, heparan sulfate, and dermatan sulfate. The SAX-HPLC separation of unsaturated disaccharides for the nutraceutical CS was also used to evaluate its quality and the possible origin. No disulfated disaccharides typical of CS from cartilaginous fishes, making the charge density lower than 1.0, were found, thus confirming the bovine or porcine origin, the most common sources of this “terrestrial” polysaccharide. Finally, high-performance size-exclusion chromatography (HPSEC) analysis was applied to evaluate the CS molecular mass in food supplements also in the presence of additives and other ingredients. This analytical approach confirmed that the nutraceutical CS samples are of high molecular mass and are not degraded during the food supplement preparations. In conclusion, this multianalytical approach can be used in the direct quantitation and evaluation of the quality and the possible origin of CS contained in food supplement formulations.

Simultaneous liquid chromatographic determination of metals and organic compounds in pharmaceutical and food-supplement formulations using evaporative light scattering detection

A novel method for the non-derivatization liquid chromatographic determination of metals (potassium, aluminium, calcium and magnesium) and organic compounds (ascorbate and aspartate) was developed and validated based on evaporative light scattering detection (ELSD). Separation of calcium, magnesium and aluminium was achieved by the cation exchange column Dionex CS-14 and an aqueous TFA mobile phase according to the following time program: 0–6 min TFA 0.96 mL L −1, 6–7 min linear gradient from TFA 0.96–6.4 mL L −1. Separation of potassium, magnesium and aspartate was achieved by the lipophilic C 18 Waters Spherisorb column and isocratic aqueous 0.2 mL L −1 TFA mobile phase. Separation of sodium, magnesium, ascorbate and citrate was also achieved by the C 18 analytical column, according to the following elution program: 0–2.5 min aqueous nonafluoropentanoic acid (NFPA) 0.5 mL L −1; 2.5–3.5 min linear gradient from 0.5 mL L −1 NFPA to 1.0 mL L −1 TFA. In all cases, evaporation temperature was 70 °C, pressure of the nebulizing gas (nitrogen) 3.5 bar, gain 11 and the flow rate 1.0 mL min −1. Resolution among calcium and magnesium was 1.8, while for all other separations was ≥3.2. Double logarithmic calibration curves were obtained within various ranges from 3–24 to 34–132 μg mL −1, and with good correlation ( r > 0.996). Asymmetry factor ranged from 0.9 to 1.9 and limit of detection from 1.3 (magnesium) to 17 μg mL −1 (ascorbate). The developed method was applied for the assay of potassium, magnesium, calcium, aluminium, aspartate and ascorbate in pharmaceuticals and food-supplements. The accuracy of the method was evaluated using spiked samples (%recovery 95–105%, %R.S.D. < 2) and the absence of constant or proportional errors was confirmed by dilution experiments.

Determination of l-carnitine in food supplement formulations using ion-pair chromatography with indirect conductimetric detection

A novel method for the determination of l-carnitine in food supplement formulations was developed and validated, using ion-pair chromatography with indirect conductimetric detection. The chromatographic method was based on a non-polar (C 18) column and an aqueous octanesulfonate (0.64 mM) eluent, acidified with trifluoroacetic acid (5.2 mM). The retention time was 5.4 min and the asymmetry factor 0.65. A linear calibration curve from 10 to 1000 μg/ml ( r = 0.99998), with a detection limit of 2.7 μg/ml (25 μl injection volume), a repeatability %RSD of 0.8 (40 μg/ml, n = 5) and reproducibility %RSD of 2.6 were achieved. The proposed method was applied for the determination of carnitine in oral solutions and capsules. No interference from excipients was found and the only pretreatment step required was the appropriate dilution with the mobile phase. Recovery from spiked samples was ranged from 97.7 to 99.7% with a precision (%RSD, n = 3) of 0.01–2.1%.

Melatonin comes of age?

Much has been said lately in the lay press about melatonin (Fig. l), the hormone secreted at night from the pineal gland. Science correspondents and even business page reports have highlighted studies indicating the potential benefits of taking melatonin to treat symptoms of jet lag and to promote sleep. Subsequently there has been a large demand for mela- tonin food supplement formulations freely available for sale in health food stores in the USA. This has fuelled the debate about the wisdom of such products remaining unregulated by the Food and Drug Administration, with strong proponents on both sides. Yet, away from this very pub lit debate, a growing body of basic and clinical researchers has con- tinued to examine the actions of this hormone, concentrating in particular on the nature of the cellular targets of melatonin action, its action as an internal zeitgeber (time-giver) for the circadian clock (Fig. 2), and the role that the hormone has to play in the aging process. Recently, significant progress has been made: first, in understanding the receptors at which melatonin acts (for review, seeRef. 1) and second, on the hypothesis that melatonin, or compounds that mimic its action on the human circadian clock in the suprachiasmatic nucleus (SCN) of the hypothalamus, may be used to treat sleep disorders in the elderly. This could satisfy a largely unmet therapeutic need in what is increasingly recognized as a distressing disorder of the aging population.