Nutraceuticals

Melatonin (MLT), a natural compound found in the animal and vegetable kingdom, participates in several physiological processes. MLT exerts antioxidant and anti-inflammatory activities, among others, but information about its action on lipid metabolism is still scarce. For this reason, mice deficient in apolipoprotein E (ApoE^−/−) fed a Western diet (WD) were intragastrically treated with different concentrations of MLT (2 and 9 mg/kg) for 12 weeks. The lipid parameters were quantified, and, since links between cardiovascular risk and immune function and oxidative stress have been established, we also analyzed the population of leukocytes and the oxidative stress status. Although there was no change in the weight of the mice, a significant reduction in low-density lipoprotein cholesterol (LDL-C) was observed in mice treated with the higher concentration of MLT tested in this study. Additionally, an improvement in cardiovascular risk indexes was observed. A reduction in the hepatic total cholesterol (TC) and LDL-C levels was also observed in the treated mice. Finally, a decrease in leukocytes and lymphocytes in particular, as well as an increase in the antioxidant status, were shown in MLT-treated mice. In conclusion, MLT is a promising candidate that could be considered as a possible functional ingredient capable of preventing cardiovascular risk. Full article

Gamma-aminobutyric acid (GABA), the primary inhibitory neurotransmitter in the central nervous system (CNS), plays a pivotal role in maintaining the delicate balance between inhibitory and excitatory neurotransmission. Dysregulation of the excitatory/inhibitory balance is implicated in various neurological and psychiatric disorders, emphasizing the critical role of GABA in disease-free brain function. The review examines the intricate interplay between the gut–brain axis and CNS function. The potential impact of dietary GABA on the brain, either by traversing the blood–brain barrier (BBB) or indirectly through the gut–brain axis, is explored. While traditional beliefs questioned GABA’s ability to cross the BBB, recent research challenges this notion, proposing specific transporter systems facilitating GABA passage. Animal studies provide some evidence that small amounts of GABA can cross the BBB but there is a lack of human data to support the role of transporter-mediated GABA entry into the brain. This review also explores GABA-containing food supplements, investigating their impact on brain activity and functions. The potential benefits of GABA supplementation on pain management and sleep quality are highlighted, supported by alterations in electroencephalography (EEG) brain responses following oral GABA intake. The comprehensive overview encompasses GABA’s sources in the diet, including brown rice, soy, adzuki beans, and fermented foods. GABA’s presence in various foods and supplements, its association with gut microbiota, and its potential as a therapeutic strategy for neurological disorders are thoroughly examined. The articles were retrieved through a systematic review of the databases: OVID, SCOPUS, and PubMed (keywords “GABA”, “oral GABA“, “sleep”, “cognition”, “neurodegenerative”, “blood-brain barrier”, “gut microbiota”, “supplements” and “therapeutic”, and by searching reference sections from identified studies and review articles). This review presents the relevant literature available on the topic and discusses the mechanisms, effects, and hypotheses that suggest oral GABA benefits range from neuroprotection to blood pressure control. The literature suggests that oral intake of GABA affects the brain illustrated by changes in EEG scans and cognitive performance, with evidence showing that GABA can have beneficial effects for multiple age groups and conditions. The potential clinical and research implications of utilizing GABA supplementation are vast, spanning a spectrum of diseases ranging from neurodegeneration to blood pressure regulation. Importantly, recommendations for the use of oral GABA should consider the dosage, formulation, and duration of treatment as well as potential side effects. Effects of GABA need to be more thoroughly investigated in robust clinical trials to validate efficacy to progress the development of alternative treatments for a variety of disorders. Full article

This study investigated the effect of a commercially available ketone supplement on heart rate (HR), perceived exertion (RPE), blood lactate, glucose, and ketone concentrations, along with time to fatigue (TTF) during a running task to voluntary fatigue. Twelve NCAA Division I cross-country athletes took part in this randomized, double-blind, placebo-controlled cross-over study. Bayesian methodologies were employed for all statistical analyses, and point estimates were determined to be statistically significant if the 95% highest-density intervals (HDI) excluded zero. TTF was not significantly different between conditions with a Mean_diff = 48.7 ± 6.3 s (95% HDI: −335, 424) and a 0.39 probability derived from the posterior distribution, indicating the likelihood that the supplement would increase TTF compared to the placebo control. Lactate concentrations immediately post-exercise were significantly lower in the supplement trial relative to placebo with an estimated Mean_diff = −4.6 ± 1.9 mmol; 95% HDI: −8.3, −0.9. There were no significant interaction effects observed for either blood glucose or ketone concentrations nor HR or RPE. These findings imply that the acute ingestion of ketones before running at lactate threshold pace has a low probability of increasing TTF in highly trained Division I runners. Full article

All over the world, wild edible plants are an essential source of chemical components that justify their use in folk medicine. The aim of this review is to document and summarize the knowledge of ten wild plants analyzed in a previous study for their ethnomedical significance. Achillea millefolium, Borago officinalis, Foeniculum vulgare, Gentiana lutea, Juniperus communis, Laurus nobilis, Malva sylvestris, Satureja montana, Silybum marianum and Urtica dioica were the subjects of our study. They are commonly found in the central Italian Apennines and the Mediterranean basin. Phytochemicals contained in wild plants, such as phenols, polyphenols, flavonoids, condensed tannins, carotenoids, etc., are receiving increasing attention, as they exert a wide range of biological activities with resulting benefits for human health. Based on the 353 studies we reviewed, we focused our study on the following: (a) the ethnobotanical practices and bioactive phytochemicals; (b) the composition of polyphenols and their role as antioxidants; (c) the methodologies commonly used to assess antioxidant activity; (d) the most advanced spectroscopic and spectrometric techniques used to visualize and characterize all components (metabolomic fingerprinting). The potential of pure compounds and extracts to be used as nutraceuticals has also been highlighted through a supposed mechanism of action. Full article

Functional food factors can play a preventive and therapeutic role in several human diseases. The marine alga Sargassum horneri (S. horneri) has restorative effects in several types of metabolic disorders, including osteoporosis, diabetes, inflammatory conditions, and cancer cell growth. Osteoporosis is widely recognized as a major public health problem. Bone loss associated with ageing and diabetic states was prevented through the intake of bioactive compounds from S. horneri water extract in vivo by stimulating osteoblastic bone formation and inhibiting osteoclastic bone resorption in vitro. The intake of S. horneri water extract was found to have preventive effects on diabetic findings with an increase in serum glucose and lipid components. Furthermore, the S. horneri component has been shown to suppress adipogenesis from rat bone marrow cells and inflammatory conditions in vitro. Notably, the growth of bone metastatic human breast cancer MDA-MB-231 cells, which induce bone loss with osteolytic effects, was suppressed through culturing with the S. horneri water extract component in vitro. The S. horneri component, which has a molecular weight of less than 1000, was found to suppress the activation of NF-κB signaling by tumor necrosis factor-α, a cytokine associated with inflammation, in osteoblastic cells and macrophage RAW264.7 cells in vitro, suggesting a molecular mechanism. The bioactive component of S. horneri may play a multifunctional role in the prevention and treatment of metabolic disorders. This review outlines the advanced knowledge of the biological activity of the aqueous extract components of S. horneri and discusses the development of health supplements using this material. Full article

Neuroinflammation plays a crucial role in the development of various neurological diseases, including neurodegenerative disorders, leading to significant neuronal dysfunction. Current treatments involve the use of non-steroidal anti-inflammatory drugs and steroids; however, they are associated with serious adverse effects, limiting their efficacy. Exploring natural products with anti-inflammatory properties appears promising, with resveratrol, a polyphenol found in various plants, standing out for its potential benefits. Studies on resveratrol and its anti-inflammatory properties have been increasing in recent years, and analyzing the profile of this knowledge area can bring benefits to the scientific community. Therefore, this study conducted bibliometric analyses, using “resveratrol AND neuroinflammation” as search terms in the Web of Science Core Collection database. The analysis, performed with VOSviewer software version 1.6.18, encompasses 323 publications. Key terms in the studies include “resveratrol”, “neuroinflammation”, and “oxidative stress”, with China leading in the number of publications. The Federal University of Rio Grande do Sul in Brazil emerges as the institution with the highest contribution, and a phase 2 clinical study on resveratrol was the most cited. These results provide an overview of the global research landscape related to resveratrol and neuroinflammation, aiding decision making for future publications and advancing scientific understanding in this field. Full article

The phytonutraceutical Ormona^® is a product composed of purified oil of Bixa orellana Linné, dry extract of Myrciaria dubia McVaugh, dry extract of Trifolium pratense L., and dry extract of Euterpe oleracea Mart. obtained using Evolve^® technology. This study evaluated the effects of Ormona^® on Wistar rats affected by ovariectomy-induced osteoporosis. Pre-treatment was conducted for 15 days before surgery and continued for a further 45 days after the surgical procedure. The experimental design consisted of five groups (n = 5): OVW: treated with distilled water (1 mL/kg, p.o); ADS: alendronate sodium (4 mg/kg p.o); EST: conjugated estrogen (2 µg/kg, p.o); ORM: Ormona^® (20 mg/kg, p.o); ORM + EST: Ormona^® (20 mg/kg, p.o) + conjugated estrogen (2 µg/kg, p.o). Biochemical and hormonal parameters of bone histopathology and trabecular and femoral diaphysis size were evaluated through scanning electron microscopy (SEM) and bone calcium quantification by atomic absorption spectrophotometry. The results show that ovariectomy caused bone alterations such as loss of femoral microarchitecture, decreased bone homeostasis parameters, and changes in the lipid profile. Estrogen supplementation reduced parameters such as cholesterol, LDL, and Ca²⁺ concentration. However, Ormona^® showed higher serum estradiol levels (p < 0.01), effects on the lipid profile, including parameters that estrogen replacement and alendronate sodium did not affect, with an increase in HDL, and positive modulation of bone metabolism, increasing osteocytes and the presence of osteoblasts. Ormona^®, therefore, produced better results than the groups treated with estrogen and alendronate sodium. Full article

The immune system plays a crucial role in defending the body against foreign invaders, and the balance of various polyunsaturated fatty acids, such as alpha-linolenic acid (ALA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA), can impact immune cell functions and overall immune responses. This study aimed to assess the effectiveness of mussel oil extracts in modulating inflammatory responses by analysing their effects on immune cell lines and cytokine expression. Four different mussel oil extracts were obtained using two extraction methods (organic solvent and supercritical CO₂ extraction) from two tissue sources (fresh and commercial). These extracts were then tested at various concentrations on T lymphocyte (Jurkat) cells, monocytes, and macrophages (THP-1 and U-937). Cytokine levels were quantified using ELISA. The results showed that the solvent-extracted samples had a dose-dependent effect on tumour necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1β) cytokine secretion in THP-1 and U937 cells, with the extract from a commercial mussel powder being more efficient than the extract from fresh powder. However, supercritical CO₂ samples showed elevated cytokine secretion levels despite their high omega-3 content. Furthermore, 100 ug/mL extract from fresh powder successfully reduced interleukin-2 (IL-2) secretion while maintaining cell viability after stimulation. The study demonstrated that solvent-extracted mussel oil can effectively regulate cytokine secretion, modulate immune cell activation, and alleviate inflammation. These findings offer valuable insights into using mussel oil extracts to treat inflammatory disorders and enhance immune responses. Full article

Petroselinum crispum (Mill.) Fuss (Apiaceae), popularly known as parsley, is a ubiquitous aromatic herb used for culinary and medicinal purposes worldwide. In addition to the richness in nutrients, several bioactive secondary metabolites, especially the flavone apigenin and its glycosides, have been described in this species. Parsley has already been shown to have several health-promoting activities, such as antithrombotic, antihypertensive, and hypolipidemic properties. The multiple studies conducted in animal models so far suggest this species is a potential source of cardioprotective agents. This review provides up-to-date information and perspectives on the potential of parsley and its bioactive compounds for the development of nutraceutical products and drugs for promoting cardiovascular health. It includes not only a discussion of parsley’s reported bioactivities but also the knowledge of supplements and food additives developed as innovative parsley-based products. Full article

The objective of this study was to assess the chemical characteristics of cold-pressed beech nut oil. The nuts, gathered from the forest, comprised 25.35% water, 13.19% oil, and 19.40% protein. The predominant fatty acid was linoleic acid (40.5%), followed by oleic acid (35.0%) and gondoic acid (7.7%). All four tocopherols were present in the oil, with γ-tocopherol being the dominant form at 99.38 mg per 100 g of oil. The total sterol content was 2708.73 mg per kg of oil, with β-sitosterol constituting 80.5% of all sterols. The main characteristics of the oil included its relatively high tocopherol and gondoic acid content, a dominant oleic–linoleic fatty acid profile, and elevated levels of carotenoids. Full article

There has been a rise in popularity of “stimulant-free” or caffeine-free fat loss supplements, but it is not well understood whether those fat loss supplements are effective at enhancing thermogenesis without caffeine’s influence. The purpose of this study was to examine the effects of a caffeinated and non-caffeinated commercially available fat loss supplement on resting energy expenditure (REE), hunger, and hemodynamic variables in healthy adults. Twenty-five healthy male and female participants completed three separate laboratory visits after overnight fasts. Baseline assessments of REE, subjective hunger, heart rate (HR), and blood pressure (BP) were followed by ingestion of a caffeinated (Phoenix, Legion^®; CAF), non-caffeinated (Phoenix Caffeine-Free, Legion^®; NCAF), or placebo (PL) fat loss supplement. REE, hunger, HR, and BP assessments were repeated at 60-, 120-, and 180-min post-ingestion. CAF, but not NCAF, significantly elevated REE greater than PL at all time points (p < 0.05). NCAF significantly reduced hunger compared to CAF and PL at the 120-min time point (p = 0.006). CAF significantly increased diastolic BP 60-min post-ingestion and significantly increased systolic BP 120- and 180-min post-ingestion compared to NCAF and PL. Further research is warranted with respect to investigating non-caffeinated ingredients and their effects on REE. Full article

Plant extracts can be therapeutic alternatives for depression and anxiety. However, some plant-derived preparations can also be toxic. Moringa oleifera leaves are used in human nutrition due to their high nutritional value and antioxidant activity. This study investigated a saline extract from M. oleifera leaves (MoLE) for secondary metabolites, proteins, cytotoxicity, hemolytic activity, in vivo acute oral toxicity, and neurobehavioral effects. MoLE contains flavonoids (rutin and vitexin), lectin, and a trypsin inhibitor. It is neither cytotoxic nor hemolytic for human cells and did not present acute oral toxicity (2000 mg/kg) to mice. The elevated plus maze test showed that MoLE (500, 1000, and 2000 mg/kg, p.o.) significantly increased the number of entries as well as the time spent by mice in open arms, while it decreased the number of entries and the time spent in closed arms when compared to the control. MoLE (500, 1000, and 2000 mg/kg, p.o.) reduced immobility time of mice in the tail suspension and forced swimming tests, compared to the control. The anxiolytic-like effect of MoLE is possibly mediated by a GABA mimetic action once it is prevented by pre-treatment with flumazenil. The present study demonstrated that MoLE has antidepressant and anxiolytic effects in mice and is a promising herbal medicine. Full article

In recent years, the emergence of drug resistance and sensitivity in leading diseases has heightened global interest in natural nutraceuticals as primary health supplements. However, comprehensive scientific scrutiny is essential before marketing these as supplements. In this study, we assessed the nutritional composition, antioxidant activities, and trace metal accumulation in eleven selected Sri Lankan coastal seaweed species. Gracilaria corticata had the highest (p < 0.05) ash and crude fiber content among the species. Protein content ranged from 4.87% to 23.67% (DW), with Ulva rigida displaying the highest (p < 0.05). Crude fat content ranged from 0.09% to 4.13% (DW), with Cladophora herpestica having the highest (p < 0.05) crude fat content. Sargassam cinereum, Turbinaria ornata and Sargassum crassifolium had the highest (p < 0.05) TPC content (51.32 ± 0.61–28.90 ± 2.68 mg/GAE g) and the highest (p < 0.05) radical scavenging antioxidant activity compared to other seaweeds. The study findings indicate that most of the studied metals in seaweeds exceeded the WHO-recommended levels. Aluminum was the highest (p < 0.05) accumulated metal in seaweeds compared to other metals. Toxic heavy metals, such as arsenic, cadmium and chromium, levels in all of the studied seaweeds surpassed the WHO limits. While seaweeds displayed acceptable nutritional and antioxidant properties, heavy metal presence poses a potential health risk to consumers. Products using seaweeds with accumulated heavy metals may have lower nutritional quality. Thus, this study underscores the need for comprehensive scientific investigation before developing high-quality natural food products or supplements from seaweeds. Full article

Immune cells express Pattern Recognition Receptors (PRRs) to recognize potentially pathogenic microbial forms. Nutraceutical compounds can induce immune cell activation through PRRs. The nutraceutical immune blend (IB), QuickStart™, contains botanical and yeast-derived ligands for PRRs, along with vitamin C and zinc. We evaluated immune-activating effects of the IB and its ingredients in vitro. Human peripheral blood mononuclear cells were treated with either the IB or single ingredients: elderberry extract, the proprietary Saccharomyces cerevisiae fermentate EpiCor™ (Sacc), the plant-based hemicellulose preparation Natramune (PDS-2865)™ (Hemi), vitamin C (VitC), or zinc gluconate (Zinc). The IB triggered sequential waves of immune activation. Initial cytokine induction by the IB at 2 h involved the immune-activating cytokines IL-6, IL-8, MIP-1α, and TNF-α, and the stem cell-mobilizing growth factor G-CSF, as did Sacc and Hemi. The 24 h immune-activation by the IB included increases in IL-1β, IL-17A, IP-10, GM-CSF, Basis FGF, PDGF-BB, and the anti-inflammatory cytokine IL-10. Increased CD69 expression by the IB was also seen for VitC and Sacc. Increased CD25 expression by the IB on monocytes was also seen for Sacc. The IB triggered rapid immune activating events of higher magnitude than the single ingredients, involving immune-activating cytokines and restorative growth factors. Clinical research is warranted to evaluate rapid immune-modulating events upon consumption. Full article

Exercise performance tends to suffer during the morning compared to the evening, which may decrease potential training adaptations. Currently, it is unclear how nutritional interventions may affect this phenomenon and whether supplementation may allow for the attainment of optimal performance regardless of the time of day. The purpose of this study was to investigate the effects of acute yohimbine ingestion on morning-associated decrements in performance and psychophysiological responses to exercise. Physically active females (n = 16) were recruited to participate in three total visits, each with a different treatment: (1) placebo-morning (PL-AM), (2) yohimbine-morning (YHM-AM; oral 2.5 mg), and (3) placebo-afternoon (PM). The morning and afternoon visits occurred between 7:00–8:00 h and 16:00–17:00 h, respectively. The experimental treatments in the morning were ingested 20 min prior to capillary blood collection, which was completed pre- and post-exercise. Following a warm-up, participants completed a 2000 m time trial on a rowing ergometer. Power output, heart rate (HR), and rating of perceived exertion (RPE) were recorded every minute. Time to competition (TTC) and subjective energy, focus, and alertness were documented post-exercise. Pre- and post-exercise blood lactate (La) and plasma hypoxanthine (HX) levels were also assessed. The trials were separated by a 48 h washout period. The results showed that power output (p = 0.010) was lower and TTC (p = 0.003) was significantly slower with PL-AM compared to PM. Furthermore, YHM-AM resulted in higher power output (p = 0.035) and faster TTC (p = 0.007) compared to PL-AM, with no differences compared to PM (p > 0.05). Post-exercise La was significantly lower with YHM-AM compared to PL-AM (p = 0.046) and PM (p = 0.001). Pre-exercise plasma HX, as measured via conversion to xanthine, was significantly higher with PM (p = 0.039), while the levels trended higher with YHM-AM (p = 0.060) compared to PL-AM. Subjective energy was higher with YHM-AM (p = 0.045) and PM (p = 0.009) compared to PL-AM, while alertness was only higher for YHM-AM compared to PL-AM (p = 0.045). No statistical differences between the treatments were found for RPE or HR (p > 0.05). These findings indicate that YHM ingestion attenuates performance decrements in the morning. Improvements in performance may be underpinned by improved feelings of energy and alterations in metabolism. Practically, YHM may represent an effective ergogenic aid to combat a lack of energy and low performance during the morning. Full article

Chondrocyte behavior is critical in osteoarthritis (OA) progression and cartilage deterioration. Recent studies have shown that green-lipped mussel (GLM) oil extracts obtained through solvent extraction can regulate cytokine secretion in cell lines, potentially reducing inflammation and immune cell activation. This study investigates the effects of these extracts on human chondrocyte cell lines to understand their impact on osteoarthritis development. Chondrocyte cultures exposed to osteoarthritis-related inflammatory factors were treated with various concentrations of GLM oil. The results revealed that mussel oils, particularly a commercial mussel powder extract (SOLV.COM), enhance glucose uptake and protect chondrocyte cells. SOLV.COM effectively manages the release of inflammatory markers like interleukin-6 (IL-6) and matrix metalloproteinases-3 (MMP-3), showing increased deoxyglucose uptake. This study highlights the significant influence of extract choice on managing interleukin-6 (IL-6) secretion and cellular activation pathways, demonstrating the potential of SOLV.COM in managing osteoarthritis by controlling reactive oxygen production, regulating glucose metabolism, and inflammatory markers. Full article

Geranylgeraniol (GG), an ingredient extracted from the South American annatto plant, has been shown to benefit bone and muscle health, is crucial in the biosynthesis of menaquinone-4 and coenzyme Q10, and has pain and inflammation reduction activities. However, no known studies to date have demonstrated the safety and impact of GG supplementation in humans. This 8-week, randomized, placebo-controlled, dose-escalated trial was conducted to determine the effect of GG on blood safety and hormone markers in healthy adults. Sixty-six males and females between 30 and 49 years of age were supplemented with either GG or a placebo (PLA) for 8 weeks, with dose escalation from 150 mg to 300 mg occurring after 4 weeks in the treatment group. Changes in complete blood count and comprehensive metabolic panels were analyzed for whole study groups (males and females) while changes in sex hormone panels were analyzed for males and females independently. There were no significant changes in complete blood count, comprehensive metabolic panel, progesterone, estradiol, sex–hormone binding globulin, or dihydrotestosterone (p > 0.05). An exploratory analysis of testosterone levels in a subgroup of males with baseline (Pre) total testosterone < 700 ng/dL (GG = 15; PLA = 13) demonstrated a significant increase (p < 0.05) from Pre to Week 8 in total-, free-, and bioavailable testosterone (+7.5%, +15.0%, and +14.8%, respectively). This study demonstrates that GG does not significantly change the composition of blood chemistry, hematology, or sex hormone profiles in adult males or females. Given the effects observed in the exploratory analysis in a subgroup of males, GG supplementation may be beneficial for testosterone enhancement in male populations subject to low testosterone (i.e., aging males and those with late-onset hypogonadism), but further research is needed. Full article

As the spread of bacterial resistance to clinically available antibiotics has become a global public health problem, the scientific community has intensified its studies in the search for natural compounds and their derivatives to combat bacterial resistance. In this work, a circadian study of the essential oil extracted from the leaves of Croton piauhiensis (EOCP) was carried out. We also sought to evaluate its antibacterial activity, modulatory potential and if it acts as a possible inhibitor of the efflux pump by determining the minimum inhibitory concentration (MIC) and the association of the oil in subinhibitory concentrations with the antibiotic ciprofloxacin and with ethidium bromide (EtBr) against the strain of Staphylococcus aureus K2068 strain. The assays used to obtain the MIC of the EOCP were performed by broth microdilution, while the efflux pump inhibitory test was performed by the MIC modification method. According to the results, the circadian study showed differences in the chemical composition and percentage of oils collected at different times of the day, which can be attributed to environmental conditions. The main components of the EOCP were β-caryophyllene (6 h—21.23%; 12 h—22.86% and 18 h—16.95%), followed by D-Limonene (6 h—13.27% and 18 h—15.95%) and γ-Elemene (12 h)—12.61%). The EOCP collected at 12 h had a better profile in reducing MIC, presenting antibacterial activity for Staphylococcus aureus and Escherichia coli. In the efflux pump test, it was observed that the oil was able to potentiate the action of ethidium bromide against the S. aureus K2068 strain, which can contribute to the prevention or treatment of infectious diseases caused by multidrug-resistant (MDR) strains. Full article

Gemmotherapy, a natural therapy based on bud macerates, has recently gained importance in the field of food supplements. However, two coexisting extraction methods employ a glycerin-based solvent, either in a binary or ternary solvent mixture. The absence of an official method for bud preparation leads to non-standardized bud macerates. Given this context, this study aimed to (i) assess the influence of solvent composition on the chemical profile of olive young shoot macerates obtained using glycerin-based solvents or using different solvent extractions and (ii) to compare the two coexisting traditional bud extraction methods described by Dr Pol Henry and by the European Pharmacopoeia. A comprehensive phytochemical analysis of all macerates was conducted using HPLC-DAD-ELSD-MS², identifying 50 metabolites divided into 7 classes through dereplication. The extracts obtained with the solvent described by the European Pharmacopoeia (ethanol/glycerin) and by Dr Pol Henry (water/ethanol/glycerin) appeared to be the most diversified in terms of metabolite distribution and possessed higher rates of secondary metabolites. These observations reinforce the interest in a glycerin-based solvent mixture for bud extraction in gemmotherapy. In addition, the difference in composition between the two traditional solvents was highlighted. Indeed, iridoids were predominant in both macerates, representing about 50% of the chemical composition, but differences were observed from one macerate to another regarding the proportions of the other chemical classes. This emphasizes the necessity for standardized gemmotherapy macerates. Full article