Antibacterial Activity against Clinical Strains of a Natural Polyphenolic Extract from Albariño White Grape Marc.

Infections caused by multidrug-resistant bacteria are becoming increasingly frequent and sometimes difficult to treat due to the limited number of antibiotics active against them. In addition, they can spread between countries and/or continents, which is a problem of great relevance worldwide. It is, therefore, urgent to find alternatives to treat infections caused by multidrug-resistant bacteria. This study aimed at exploring a possible therapeutic alternative in the fight against antibiotic resistance. Based on the known antibacterial capacity of polyphenols, we tested the antimicrobial activity of a polyphenolic extract of Albariño white grape marc on clinical strains since research on such bacteria has been very scarce until now. First, the extract was obtained using a medium-scale ambient temperature (MSAT) system, which is an efficient and sustainable extractive method. The determinations of the polyphenolic content of the extract and its antioxidant capacity showed good results. Using chromatographic and mass spectrometric tools, 13 remarkable polyphenols were detected in the extract. The antibacterial activity of our grape marc extract against nineteen clinical strain isolates, some of which are multidrug-resistant, was evaluated by means of the calculation of half of the maximum inhibitory concentration (IC50) and the value of the minimum bactericidal concentrations (MBCs). In conclusion, the extract showed effectiveness against all clinical strains tested, regardless of their level of antibiotic resistance, and shows promise in the fight against antibiotic resistance.

The generally recognized as safe (GRAS) microalgae Haematococcus pluvialis (wet) as a multifunctional additive for coloring and improving the organoleptic and functional properties of foods.

This work proposes the application of astaxanthin-rich H. pluvialis wet paste (HPW) as a partial substitute for wheat flour in the preparation of filloas, a dish that combines the basic ingredients of industrial bakery. The nutritional and color profile of HPW-enriched samples was evaluated by comparative analysis with a mixture of synthetic food dyes. The highest content of carotenoids (798 ± 12 µg g-1) and fatty acids (76 ± 2 mg g-1) was obtained for a filloa fortified with H. pluvialis in contrast to a non-significant dye response. Subsequently, the color stability of the fortified filloa was evaluated over time (3, 6 and 9 days), as well as its physicochemical properties and microbiological profile. As a result, HPW provided filloas with a longer shelf life, brightness (*L), and texture, in comparison with a mixture of synthetic dyes. In addition, an inhibitory effect of HPW towards mesophilic aerobic microorganisms in the food was obtained.

Bioprospecting of Targeted Phenolic Compounds of Dictyota dichotoma, Gongolaria barbata, Ericaria amentacea, Sargassum hornschuchii and Ellisolandia elongata from the Adriatic Sea Extracted by Two Green Methods.

The content of bioactive compounds in four brown and one red algae from the Adriatic Sea (Dictyota dichotoma, Gongolaria barbata, Ericaria amentacea, Sargassum hornschuchii and Ellisolandia elongata) is explored. The efficiency of two different extraction methods viz. ultrasound-assisted extraction (UAE) and matrix solid-phase dispersion (MSPD) to obtain the extracts rich in phenolic compounds was compared. The effect of the extraction solvent to modulate the phenolic profile was assessed. In general, the mixture ethanol/water in an isovolumetric proportion showed the best results. The total phenolic content (TPC) and antioxidant activity (AA), as well as the individual polyphenolic profile, were evaluated for five target algae. TPC values ranged between 0.2 mg GAE/g (for E. elongata) and 38 mg GAE/g (for S. hornschuchii). Regarding the quantification of individual polyphenols by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis, the presence of a high number of hydroxybenzoic acid derivatives (mainly of 3- and 4-hydroxybenzoic acids) in all species was noted. In G. barbata their concentrations reached up to 500 mg/kg. IC50 values (ABTS assay) ranged between 44 mg/L (for S. hornschuchii) and 11,040 mg/L (for E. elongata). This work contributes to the in-depth characterization of these little-explored algae, showing their potential as a natural source of phenolic compounds.

Phytochemical Analysis and Antiproliferative Activity of Ulex gallii Planch. (Fabaceae), a Medicinal Plant from Galicia (Spain).

The genus Ulex comprises thirteen accepted species of perennial shrubs in the family Fabaceae. In Galicia (Spain) many of these are considered spontaneous colonizing species, which are easy to establish and maintain. Among them, Ulex gallii Planch. is used in traditional medicine for the same anti-infective, hypotensive and diuretic purposes as Ulex europaeus L., which is the most studied species. Likewise, some studies have described the antitumoral properties of several species. However, there are few scientific studies that justify the use of Ulex gallii Planch. and nothing has been reported about its composition to date. In our study, the entire plant was extracted with methanol and the crude extract was subjected to liquid phase extraction with distinct solvents, yielding three fractions: hexane (H), dichloromethane (D) and methanol (M), which were subsequently fractionated. The dichloromethane (D5, D7 and D8) and methanol (M4) sub-fractions showed antiproliferative activity on A549 (lung cancer) and AGS (stomach cancer) cell lines, and caspase 3/7 activity assessment and DNA quantification were also performed. Targeted analysis via UHPLC-QToF, in combination with untargeted analysis via MS-Dial, MS-Finder and Global Natural Products Social Molecular Networking (GNPS), allowed us to tentatively identify different metabolites in these sub-fractions, mostly flavonoids, that might be involved in their antiproliferative activity.

Identification and Quantification of Polyphenols in Croatian Traditional Apple Varieties.

Apples and apple-based products are particularly interesting due to being a good source of polyphenols in an everyday diet. Recently there has been increased interest in the preservation of traditional apple varieties due to studies that suggest that traditional apple varieties have a higher content of polyphenols and antioxidant activity compared to commercial varieties. This study shows that traditional apple varieties contain higher concentration of polyphenolic compounds than conventional ones, such as chlorogenic acid (1.29-456 mg/kg dw), catechin (0.70-312 mg/kg dw), epicatechin (1.75-244 mg/kg dw), procyanidin B1 + B2 (4.08-358 mg/kg dw) and quercetin-3-glucoside (0.96-231 mg/kg dw). This research underlies the great potential of traditional apple varieties as a source of natural antioxidants and polyphenolics.

The chemistry behind the body art: unveiling the elemental profile and heavy metal content of natural tattoos and dyes by ICP-MS.

Temporary tattoos and dyes are increasing popularity as an alternative to permanent ones. However, knowledge on its elementary composition is scarce and, this scientific gap can potentially make them a source of heavy metal exposure on humans. The present research aimed to explore the possibility of using the inorganic chemical signature to characterize natural pigments-based products and to evaluate the heavy metal content in jagua temporary tattoos and henna temporary hair dyes and tattoos. Thirty-four different samples of commercial products were analyzed for the presence and quantity of eleven trace elements (Al, Cr, Mn, Co, Ni, Cu, Zn, As, Cd, Ba, and Pb) by inductively coupled plasma mass spectrometry (ICP-MS). The overall mean concentrations varied between 0.02 and 973 µg g-1 in solid samples or 0.01-1878 µg g-1 for paste ones, wherein potential fake products were uncovered. None of the 34 samples considered comply with the current European Cosmetics Regulation. Samples were differentiated according to their Cr, Zn, Ba, and Pb content. The overall results revealed highly variable levels in the investigated samples, which leads us to suggest detailed quality controls of these materials, due the fact that their continued use can origin potential threat to human health.

Optimization of bioactives extraction from grape marc via a medium scale ambient temperature system and stability study.

A scalable procedure with minimum energy requirements, MSAT (Medium Scale Ambient Temperature), in combination with solvents generally recognized as safe (GRAS), has been optimized to obtain polyphenolic extracts from white grape (Vitis vinifera) marc. The solvents considered were propylene glycol (Pg), ethanol (Et), and ethyl lactate (Lc), as well as their respective hydro-organic mixtures. In a first approach, the operating parameters were optimized through a response surface matrix: extraction solvent volume (range 10-150 mL), marc mass (range 20-200 g) and marc/dispersant mass ratio (range 0.5-2 g⋅g-1), using the total polyphenol content (TPC) and the antioxidant activity (AA) of the extracts as response parameters. The highest TPC (5,918 mgGAE⋅L-1) and AA (44 mmolTE⋅L-1) values were obtained using 200 g marc and 100 mL solvent. Regarding the type of solvent, a better response was reached with Lc > Et > Pg > H2O obtaining a polyphenol concentration of 252 mg⋅L-1 for the hydro-organic isovolumetric ratio of ethyl lactate. In addition, the stability of the extracts was studied for 62 days. The effect of factors such as temperature, light exposure, and oxidative reactivity was evaluated. The bioactivity indices showed no changes with the storage conditions of the extracts in the first month of analysis, after which 75% of the antioxidant activity as the concentration of the polyphenolic profile (204 mg⋅L-1) remains. The absence of reactive oxygen and the cooling of the extract (4°C) were the most determining factors (p < 0.05) in modulating the stability of the total polyphenolic profile.

Understanding the chemical and mineralogical composition of commercial henna and jagua tattoos and dyes-a multi-analytical approach.

Temporary tattoos and dyes constitute a great analytical challenge in relation to the regulatory control of their ingredients. Most of these commercial products are not labeled according to their content and their chemical nature is highly diverse. Therefore, it is necessary to analyze these complex samples to evaluate the potential presence of metallic impurities, to ensure the safety of cosmetic products contributing to health protection. This study proposes a multi-analytical methodology, which includes handheld X-ray fluorescence (h-XRF) and X-ray diffraction (XRD), complemented by variable pressure scanning electron microscopy coupled with energy-dispersive X-ray spectrometry (VP-SEM-EDS) to fully characterize 34 commercial samples of jagua and henna tattoos and dyes. The approach allowed the identification of the main constituents providing complementary compositional data and differences between sample types were established. In addition, information on the degree of natural pigments homogeneity was also obtained. The results' discussion considering the current European cosmetics regulation may be useful to support the drafting of safety requirements and specific regulation.

Occurrence of Patulin and Polyphenol Profile of Croatian Traditional and Conventional Apple Cultivars during Storage.

Apples and apple-based products are among the most consumed fruits around the world. However, they are susceptible to infection with the fungi Penicilium expansum. In addition to the reduction of apple quality, secondary metabolism of this fungus produces a mycotoxin patulin that has a negative effect on human health. Currently, there is no available research in the literature on the resistance of Croatian traditional apple cultivars to contamination with P. expansum, and consequently, on the patulin content in apples and apple juice produced from those apples. Although the mechanism of apple resistance to fungal diseases has not yet been sufficiently investigated, some studies have shown that polyphenolic compounds have some impact on fungi growth. In order to contribute with new knowledge, this research deals with monitoring the growth of P. expansum on apples, patulin detection by LC/MS-MS, determination of polyphenol profile by validated HPLC method, and determining the effect of polyphenolic compounds on fungi growth and patulin production during apple storage. The results of this study have shown that Croatian traditional apple cultivars harvested from family farm Horvatic contain higher concentration of polyphenolic compounds and higher antioxidant activity. At the same time, they showed more resistance to infection by P. expansum than conventional ones. The higher content of dihydrochalcones and flavanols encouraged the biosynthesis of patulin in examined cultivars. However, the higher content of non-flavonoids such as 2-6 dimethoxybenzoic acid, 4-hydroxycinnamic acid and chlorogenic acid leads to decrease in content of patulin. In conclusion, it seems that content of polyphenols and patulin production are correlated.

Multicomponent Polyphenolic Extracts from Vaccinium corymbosum at Lab and Pilot Scale. Characterization and Effectivity against Nosocomial Pathogens.

An extraction method was designed and scaled up to produce multicomponent polyphenolic extracts from blueberries (Vaccinium corymbosum) of three different varieties. The process was specifically drawn up to comply with green chemistry principles. Extracts were obtained for the direct assessment of their antimicrobial and antiadhesive activities, and their direct use in the control of infections caused by concerning multidrug-resistant nosocomial pathogens. Analytical characterization was performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Similar qualitative profiles were obtained in the three studied varieties with some significant quantitative differences. Up to 22 different polyphenols were identified with a clear predominance of anthocyani(di)ns followed by flavanols, non-flavonoids, and far behind by flavan-3-ols and procyanidins. The individual content of the main polyphenols was also discussed. A pilot scale extract has been also produced as a proof-of-concept, showing that scaling-up triples the content of bioactive phytochemicals. The effect of the polyphenolic extracts was analyzed against seven multidrug-resistance bacterial species by performing biofilm formation and growth and killing curves assays. All the studied varieties showed antibacterial and antiadhesive activities, being the extract containing the highest concentration of bioactive polyphenols, the most active with a high bactericidal effect.

Recent Advances in Sample Preparation for Cosmetics and Personal Care Products Analysis.

The use of cosmetics and personal care products is increasing worldwide. Their high matrix complexity, together with the wide range of products currently marketed under different forms imply a challenge for their analysis, most of them requiring a sample pre-treatment step before analysis. Classical sample preparation methodologies involve large amounts of organic solvents as well as multiple steps resulting in large time consumption. Therefore, in recent years, the trends have been moved towards the development of simple, sustainable, and environmentally friendly methodologies in two ways: (i) the miniaturization of conventional procedures allowing a reduction in the consumption of solvents and reagents; and (ii) the development and application of sorbent- and liquid-based microextraction technologies to obtain a high analyte enrichment, avoiding or significantly reducing the use of organic solvents. This review provides an overview of analytical methodology during the last ten years, placing special emphasis on sample preparation to analyse cosmetics and personal care products. The use of liquid-liquid and solid-liquid extraction (LLE, SLE), ultrasound-assisted extraction (UAE), solid-phase extraction (SPE), pressurized liquid extraction (PLE), matrix solid-phase extraction (MSPD), and liquid- and sorbent-based microextraction techniques will be reviewed. The most recent advances and future trends including the development of new materials and green solvents will be also addressed.

Multi-Target Strategy to Uncover Unexpected Compounds in Rinse-Off and Leave-On Cosmetics.

The wide range and complexity of cosmetic formulations currently available on the market poses a challenge from an analytical point of view. In addition, during cosmetics manufacture, impurities coming from raw materials or formed by reaction of different organic compounds present in the formulation may be present. Their identification is mandatory to assure product quality and consumer health. In this work, micro-matrix solid-phase dispersion (µMSPD) is proposed as a multi-target sample preparation strategy to analyze a wide number of unexpected families of compounds including polycyclic aromatic hydrocarbons (PAHs), pesticides, plasticizers, nitrosamines, alkylphenols (APs), and alkylphenol ethoxylates (APEOs). Analytical determination was performed by gas chromatography-mass spectrometry (GC-MS) for the determination of 51 target compounds in a single run, whereas liquid chromatography tandem mass spectrometry (LC-MS/MS) was employed for the analysis of six APs and APEOs. Both methodologies were successfully validated in terms of linearity, accuracy, and precision in leave-on and rinse-off cosmetics. Limits of detection (LODs) were calculated in the low ng g-1, showing their suitability to determine trace levels of impurities and banned compounds with different chemical natures, providing useful tools to cosmetic control laboratories and companies.

Antimicrobial Activity of Polyphenols and Natural Polyphenolic Extracts on Clinical Isolates.

Antibiotic resistance is a growing global problem that affects people, animals, the environment, and the economy. Many clinically relevant bacteria have become resistant to antibiotics, and this fact is emerging as one of the major threats to public health. The lack of new antibiotics, which is due to their time-consuming and costly development, exacerbates the problem. Therefore, it is necessary to identify new antimicrobial agents to treat bacterial and fungal infections. Plant extracts, which are valuable sources of bioactive compounds, mainly polyphenols, play an important role as a new strategy to combat pathogenic microorganisms. There is an extensive body of supporting evidence for the potent antibacterial and antifungal activities of polyphenols. Furthermore, some polyphenols show a synergistic effect when combined with antibiotics and antifungals, suggesting a promising alternative for therapeutic strategies against antibiotic resistance. However, only a few articles are found when searching the antibacterial or antifungal activities of polyphenols employing clinical isolates. Hence, this review focuses on the antimicrobial activity of polyphenols and extracts rich in polyphenols on clinical isolates, organized according to the World Health Organization priority pathogens classification.

Polyphenolic content and bioactivities of Crataegus oxyacantha L. (Rosaceae).

Solid-Liquid Extraction (SLE) using solvent of different polarities (CHCl3, EtOAc, and n-BuOH) has been applied to leaves and fruits from (Crataegus oxyacantha L.), a deciduous shrub with an expected rich phytochemical profile. The total polyphenols content and the radical scavenging activity of each extract were evaluated. These extracts were analyzed by HPLC-DAD and rutin, quercetin-3-glucoside, caftaric and caffeic acid had been positively identified. The phytochemical study of the ethyl acetate extract of C. oxyacantha, led to the isolation and structural elucidation of quercetin (1); quercetin-3-O-ß-glucoside (2); epicatechin (3); naringenin (4), reported for the first time from this species except caffeic acid and epicatechin. These compounds were identified by 1D and 2D NMR combined analysis as well as by MS and UV.The antimicrobial activity of these extracts has also been tested, showing strong antibacterial activity-solvent dependent-against Gram positive bacteria. Additionally, bactericidal power was demonstrated in fruit extracts.

Exploring the powerful phytoarsenal of white grape marc against bacteria and parasites causing significant diseases.

Natural extracts containing high polyphenolic concentration possess antibacterial, anti-parasitic and fungicidal activities. The present research characterises two extracts based on white grape marc, a winemaking by-product, describing their physicochemical features and antimicrobial capacities. The main components of these extracts are phenolic acids, flavan-3-ols and their gallates and flavonols and their glycosides. As a result of this complex composition, the extracts showed pronounced bioactivities with potential uses in agricultural, pharmaceutical and cosmetic industries. Polyphenol compounds were extracted by using hydro-organic solvent mixtures from the by-product of Albariño white wines (Galicia, NW Spain) production. The in vitro antimicrobial activity of these extracts was evaluated on Gram-positive and Gram-negative bacteria and Apicomplexan and Oomycota parasites. Microbial species investigated are causing agents of several human and animal diseases, such as foodborne illnesses (Bacillus cereus, Escherichia coli, Salmonella enterica, and Toxoplasma gondii), skin infections and/or mastitis (Staphylococcus aureus and Streptococcus uberis), malaria (Plasmodium falciparum) and plant infections as "chestnut ink" or "root rot" (Phytophthora cinnamomi). Both extracts showed activity against all the tested species, being nontoxic for the host. So, they could be used for the development of biocides to control a wide range of pathogenic agents and contribute to the enhancement of winemaking industry by-products.

Anti-adhesive activity of a Vaccinium corymbosum polyphenolic extract targeting intestinal colonization by Klebsiella pneumoniae.

The therapeutic effect of Vaccinium polyphenols against uropathogens has been widely studied. Most attention has focused on the antimicrobial activity against P-fimbriated Escherichia coli strains. The present study investigated the anti-adhesive and anti-biofilm activity of a saline extract of blueberry (Vaccinium corymbosum) targeting intestinal colonization by a highly adherent Klebsiella pneumoniae strain. This strain, responsible for a large outbreak of infection in Spain, was selected on the basis of its remarkable capacity to colonize the gastrointestinal tract of patients. The blueberry extract was obtained using a medium scale ambient temperature system (MSAT) in a novel approach based on the use of an aqueous solvent and addition of mineral salts. The polyphenolic content was determined by liquid chromatography coupled to tandem mass-spectrometry (LC-MS/MS). The findings confirmed that the blueberry extract is a rich source of phenolic compounds, including the most polar polyphenols (mostly non-flavonoids), intermediate polarity compounds (flavan-3-ols and most procyanidins) and low polarity compounds (flavonols and anthocyanins). The extract significantly inhibited biofilm formation and bacterial adhesion to HT-29 colorectal cells by a highly adherent multidrug-resistant K. pneumoniae. Although some individual anthocyanidins (malvidin, delphinidin and cyanidin) and one hydroxycinnamic acid (caffeic acid) proved capable of reducing bacterial adhesion, the unfractionated extract was more active than any of the individual polyphenolic compounds. In addition, the extract displayed considerable potential as an intestinal decolonization treatment in a murine model. The study findings demonstrate the potential value of the V. corymbosum extract as an alternative treatment for K. pneumoniae infections.

Green Extraction Methods for Extraction of Polyphenolic Compounds from Blueberry Pomace.

In this study, green extraction methods-high voltage electrical discharges (HVED), pulsed electric field (PEF), and ultrasound-assisted extraction (UAE)-were compared in terms of extraction yield of total and individual polyphenolic compounds, as well as the antioxidant capacity of blueberry pomace extracts. All extractions were performed with methanol- and ethanol-based solvents. The highest total polyphenols content (TPC) (10.52 mg of gallic acid equivalent (GAE) per g of dry weight (dw)) and antioxidant activity (AA) (0.83 mmol TE/g dw) were obtained by PEF-assisted extraction in the ethanol-based solvent after 100 pulses and 20 kV/cm, which corresponds to an energy input of 41.03 kJ/kg. A total of eighteen individual polyphenols were identified in all investigated blueberry pomace extracts by high-performance liquid chromatography with the diode-array detector (HPLC-DAD) and liquid chromatography electrospray ionization tandem mass spectrometric (LC-(HESI)-MS/MS). The highest anthocyanin (1757.32 µg/g of dw) and flavanol (297.86 µg/g of dw) yields were obtained in the methanol-based solvent, while the highest phenolic acid (625.47 µg/g of dw) and flavonol (157.54 µg/g of dw) yields were obtained in the ethanol-based solvent by PEF-assisted extraction at the energy input of 41.03 kJ/kg. These results indicated that PEF is a promising green extraction method which can improve the blueberry pomace's polyphenol extraction yield.

Unraveling the environmental impacts of bioactive compounds and organic amendment from grape marc.

In a society that produces large amounts of solid waste, the search for new methods of valorisation has led to the development of techniques that make it possible to obtain new products from waste. In the case of bio-waste, biological treatment such as anaerobic digestion or composting appear to be suitable options for producing bio-energy or bio-fertilizers respectively. Vermicomposting is a method of converting solid organic waste into resources through bio-oxidation and stabilization of the organic waste by earthworms. The purpose of this study is to establish the environmental impacts of a complete route for the valorisation of grape pomace in order to identify environmental hotspots. In this valorisation route, different value-added products are produced with potential application in the cosmetic, food and pharmaceutical sectors. Priority was given to the use of primary data in the elaboration of the data inventories needed to perform the life cycle assessment (LCA). The main findings from this study reported that the energy requirement of the distillation process is an important hot spot of the process. Although the valorisation route has some poor results in terms of the two environmental indicators (carbon footprint and normalised impact index), when economic revenues were included in this analysis, its environmental performance was better than that of other alternatives for bio-waste recovery.

Peel of Traditional Apple Varieties as a Great Source of Bioactive Compounds: Extraction by Micro-Matrix Solid-Phase Dispersion.

Micro matrix solid phase dispersion (micro-MSPD) was optimized by response surface methodology for the extraction of polyphenols from the peel of twelve traditional and eight commercial apple varieties grown in Croatia. The optimized micro-MSPD procedure includes the use of 0.2 g of sample, 0.8 g of dispersant, a 57% solution of methanol in water as the solvent and 5 mL of extract volume. The total polyphenolic index (TPI) and antioxidant activity (AA) were measured by spectrophotometric assays. Eighteen polyphenolic compounds were identified in all investigated apples by HPLC-DAD and LC-(ESI)-MS. The peel of traditional apple varieties had higher contents of all investigated polyphenols. Calculated relative contribution of polyphenol groups indicated non-flavonoids (28.6%) and flavanols (46.2%) as the major contributors to the total polyphenolic content in traditional and commercial apple varieties, respectively. The most abundant polyphenol in traditional apple peel was chlorogenic acid, procyanidin B2 and epicatechin (1143 ± 755 µg/g dw, 954 ± 343 µg/g dw and 560 ± 362 µg/g dw, respectively). The peel of varieties 'Apistar', 'Bobovac' and 'Bozicnica' could be highlighted as an important source of polyphenols.

Matrix solid-phase dispersion as a greener alternative to obtain bioactive extracts from Haematococcus pluvialis. Characterization by UHPLC-QToF.

So far, research on the microalga Haematococcus pluvialis has been focused mainly on the exploitation of its high astaxanthin content, leaving aside the use of other bioactive compounds present. This study is focused on obtaining and characterizing extracts enriched in bioactive compounds from this microalga red aplanospores. This is performed by means of Matrix Solid-Phase Dispersion (MSPD) extraction process, in an environmentally friendly way with low energy consumption and GRAS solvents. The effects of extraction parameters, particularly the extraction solvents (ethanol, ethyl lactate and water) are studied, in order to obtain maximum recovery of the main antioxidant compounds of interest (carotenoids, fatty acids and derivatives). Characterization of extracts is carried out by HPLC-DAD (High Performance Liquid Chromatography Diode Array Detector) and UHPLC-QToF (Ultra High-Performance Liquid Chromatography Quadrupole Time-of-Flight). The results show that MSPD produced extracts with higher bioactive compound recoveries than conventional cell disruption extractions. At the same time, a novel untargeted characterization for this species is performed, identifying compounds not previously dated in H. pluvialis, which include 10-phenyldecanoic acid and the -oxo and -hydroxy derivatives of palmitic acid. This approach, first applied to a freshwater microalgae, characterized by rigid and resistant aplanospores, provided a synergistic and sustainable extract, giving a broader focus on the use of this microalga.