Antibacterial Activities of Agaricus bisporus Extracts and Their Synergistic Effects with the Antistaphylococcal Drug AFN-1252.

Agaricus bisporus, commonly known as the button mushroom, has attracted attention for its biological properties, including antimicrobial activities. Here, we evaluated the efficacy of ethanolic and acetonic extracts from white and brown A. bisporus against different bacterial strains, including antibiotic-resistant strains. Bioautography and principal component analysis identified the most active antibacterial compounds for each of the tested bacteria and indicated the main markers responsible for the strain-specific effects. In addition, the mushroom extracts demonstrated a synergistic impact when combined with the antistaphylococcal antibiotic AFN-1252.

A comprehensive study on Geranium robertianum L. antibacterial potential.

AIMS: The research aimed to optimize the ultrasound-assisted extraction of secondary metabolites and the antibacterial activity of the plant species Geranium robertianum. The phytochemical profiles of the optimized extracts, as well as their antibacterial and synergistic activity with an antibiotic and their potential mechanisms of action and cytotoxicity, were examined. METHODS AND RESULTS: Response Surface Methodology was used to optimize extraction conditions. Optimized ethanol and acetone extracts were tested via microdilution, checkerboard, time-kill kinetics, and cell membrane permeability methods. The extracts displayed broad antibacterial activity with minimum inhibitory concentrations ranging from 1.25 to 20 mg ml-1. In addition, the extract synergistically reacted with gentamicin against gentamicin-resistant strains of Escherichia coli and Staphylococcus aureus, enhancing the efficacy of the antibiotic up to 32-fold. The extracts demonstrated strain-dependent bactericidal activity in a 24-h time interval. They increase the permeability of the cell membrane, thus disrupting its normal functioning. The cytotoxic concentration (CC50) on human keratinocytes was 1771.24 ± 5.78 µg ml-1 for ethanol extract, and 958.01 ± 6.14 µg ml-1 for acetone extract. Kaempferol, ellagic acid, quercetin, and rutin were recognized as the main components in both extracts. CONCLUSIONS: The findings of this study indicate that the extracts of G. robertianum can be considered as potential natural antibacterial agents in the control of microorganisms.

Potential application of green extracts rich in phenolics for innovative functional foods: natural deep eutectic solvents as media for isolation of biocompounds from berries.

The health-promoting effects of berries have attracted attention due to the possible application of their extracts as functional ingredients in food products. Natural deep eutectic solvents (NADESs) are a new generation of environmentally friendly solvents for the extraction of natural products, and they are green alternatives to organic solvents, and they can improve the solubility, stability, and bioavailability of isolated biocompounds. In this study, an efficient eco-friendly method was used for the extraction of phenolic compounds from different berries: chokeberries, blueberries, and black goji berries with a range of eutectic solvents consisting of hydrogen bond acceptors (HBAs) such as choline chloride, L-proline, L-glycine, and L-lysine and hydrogen bond donors (HBDs) such as malic, citric, tartaric, lactic and succinic acids, glucose and glycerol. The obtained results indicated the ability of NADESs towards selective extraction of phenolics; the eutectic system choline chloride : malic acid showed selective extraction of anthocyanins, while choline chloride : glycerol and choline chloride : urea showed selectivity towards flavonoids and phenolic acids. The methodology for screening of the NADES extraction performance, which included chromatographic profiling via high-performance thin layer chromatography combined with chemometrics and spectrophotometric essays, allowed effective assessment of optimal eutectic solvents for isolation of different groups of phenolics. Great antioxidant and antimicrobial activities of extracts, along with the green nature of eutectic solvents, enable NADES berry extracts to be used as "green-labelled" functional foods or ingredients.

Bioassay-Guided Assessment of Antioxidative, Anti-Inflammatory and Antimicrobial Activities of Extracts from Medicinal Plants via High-Performance Thin-Layer Chromatography.

Natural products and their analogues have contributed significantly to treatment options, especially for anti-inflammatory and infectious diseases. Thus, the primary objective of this work was to compare the bioactivity profiles of selected medicinal plants that are historically used in folk medicine to treat inflammation and infections in the body. Chemical HPTLC fingerprinting was used to assess antioxidant, phenolic and flavonoid content, while bioassay-guided HPTLC was used to detect compounds with the highest antibacterial and anti-inflammatory activities. The results of this study showed that green tea leaf, walnut leaf, St. John's wort herb, wild thyme herb, European goldenrod herb, chamomile flower, and immortelle flower extracts were strong radical scavengers. Green tea and nettle extracts were the most active extracts against E. coli, while calendula flower extract showed significant potency against S. aureus. Furthermore, green tea, greater celandine, and fumitory extracts exhibited pronounced potential in suppressing COX-1 activity. The bioactive compounds from the green tea extract, as the most bioactive, were isolated by preparative thin-layer chromatography and characterized with their FTIR spectra. Although earlier studies have related green tea's anti-inflammatory properties to the presence of catechins, particularly epigallocatechin-3-gallate, the FTIR spectrum of the compound from the most intense bioactive zone showed the strongest anti-inflammatory activity can be attributed to amino acids and heterocyclic compounds. As expected, antibacterial activity in extracts was related to fatty acids and monoglycerides.

Chemical Composition, Antioxidant Potential, and Nutritional Evaluation of Cultivated Sorghum Grains: A Combined Experimental, Theoretical, and Multivariate Analysis.

Sorghum grain (Sorghum bicolor L. Moench) is a gluten-free cereal with excellent nutritional value and is a good source of antioxidants, including polyphenols, as well as minerals with proven health benefits. Herein, the phenolic composition, elemental profile, and antioxidant activity of sixteen food-grade sorghum grains (S1-S16) grown under agroecological conditions in Serbia were determined. Nine phenolic compounds characteristic of sorghum grains, such as luteolinidin, 5-methoxyluteolinidin, luteolidin derivative, luteolidin glucoside, apigeninidin, 7-methoxyapigeninidin, apigeninidin glucoside, and cyanidin derivative, were quantified. The antioxidant potential of the analyzed sorghum grains was evaluated by UV/Vis (DPPH, ABTS, and FRAP) and Electron Paramagnetic Resonance spectroscopy (hydroxyl and ascorbyl radical scavenging assays). The content of macro- and microelements was determined by Inductively Coupled Plasma Optical Emission spectroscopy. Theoretical daily intakes of selected major and trace elements were assessed and compared with the Recommended Daily Allowance or Adequate Intake. Sample S8 had the highest amount of phenolic compounds, while S4, S6, and S8 exhibited the strongest antioxidative potential. The sorghum studied could completely satisfy the daily needs of macro- (K, Mg, and P) and microelements (Se, Zn, Fe). Pattern recognition techniques confirmed the discrimination of samples based on phenolic profile and elemental analysis and recognized the main markers responsible for differences between the investigated samples. The reaction between hydroxyl radicals and luteolinidin/apigeninidin was investigated by Density Functional Theory and thermodynamically preferred mechanism was determined.

Assessing radical scavenging capacity of Sempervivum tectorum L. leaf extracts: An integrated high-performance thin-layer chromatography/in silico/chemometrics approach.

High-Performance Thin-Layer Chromatography (HPTLC)-radical scavenging capacity (RSC) assays are standard techniques for the separation and identification of antioxidants from complex mixtures. HPTLC coupled with DPPH· visualization of chromatograms allows for the detection of individual antioxidants. However, other HPTLC-RSC assays that recognize compounds exhibiting different mechanisms of radical-scavenging activity are rarely reported. In this study, we developed an integrated approach that combines five HPTLC-RSC assays, principal component analysis (PCA) and quantum chemical calculations to assess the antioxidant capacity of Sempervivum tectorum L. leaf extracts. Two HPTLC assays - potassium hexacyanoferrate(III) total reducing power assay (TRP) and total antioxidant capacity by phosphomolybdenum method (TAC) - were developed for the first time. The method supports a more in-depth study of the RSC of natural products, as it compares the radical scavenging fingerprints of S. tectorum leaf extracts and recognizes differences in their individual bioactive constituents. Kaempferol, kaempferol 3-O-glucoside, quercetin 3-O-glucoside, caffeic acid, and gallic acid were identified as the compounds that discriminate HPTLC-RSC assays according to their mechanism of action and capture the similarities between 20 S. tectorum samples. Additionally, DFT calculations on M06-2X/6-31+G(d,p) level were applied to map thermodynamic feasibility of hydrogen atom transfer (HAT) and single electron transfer (SET) mechanisms of the identified compounds. Based on experimental and theoretical results, a combination of HPTLC-ABTS and HPTLC-TAC assays were proposed as the optimal method for mapping the antioxidants from S. tectorum. This study represents a step forward in identifying and quantifying individual antioxidants from complex food and natural product matrices in a more rational manner.

Quality Assessment of Bee Pollen-Honey Mixtures Using Thin-Layer Chromatography in Combination with Chemometrics.

The aim of this study is to develop a rapid, effect-directed screening method for quality assessment of bee pollen-honey mixtures. The comparative antioxidant potential and phenolic content of honey, bee pollen, and the bee pollen-honey mixtures, was performed using spectrophotometry. The total phenolic content and antioxidative activity of bee pollen-honey mixtures with 20 % bee pollen share were in the range 3.03-3.11 mg GAE/g, and 6.02-6.96 mmol TE/kg, respectively, while mixtures with 30 % bee pollen share contained 3.92-4.18 mg GAE/g, and 9.69-10.11 mmol TE/kg. Chromatographic fingerprint of bee pollen-honey mixtures was performed by high-performance thin-layer chromatography with conditions developed by authors and reported for the first time. Fingerprint analysis hyphenated with chemometrics enabled authenticity assessments of honey in mixtures. Results indicate that bee pollen-honey mixtures represent a food with highly, both, nutritious characteristics and health-promoting effect.

Elemental Profile of Propolis from Different Areas of Serbia.

Propolis is a resinous natural substance collected by honeybees from different plant sources. The element content of propolis is influenced by the content of elements in the soil, climatic factors, and the degree of pollution. The aim of the study was to characterize element composition and content of toxic elements in 51 propolis collected from different locations in Serbia and to evaluate the differences among them. Determination of the 21 elements were performed using Inductively coupled plasma - optical emission spectrometry. K was the most abundant element, followed by Ca, Mg, Si, Fe, Zn, and Al. The microelements B, Ba, Mn, Na, Pb, and Ti were also present in propolis, while Co, Cr, Cu, Mo, Ni, Sr, and V were found in trace. Toxic elements such as Pb, As, and Cd were found in the trace. Kruskal-Wallis tests differentiate groups of samples by geographical origin. The presence of B, Fe, Sr, Ti, Zn, and As demonstrated statistical significance between six regions of Serbia.

Authenticity assessment of cultivated berries via phenolic profiles of seeds.

Considering the health-benefits of berry fruits consumption and increased market demands for food authenticity as one of the most important quality assurances, phenolic profiling by high-performance thin layer chromatography and ultra-high-performance liquid chromatography hyphenated with mass spectrometry was combined with multivariate analysis for phytochemical characterization and intercultivar discrimination of cultivated berry seeds. The phenolic profiles of 45 berry seeds from nine genuine Serbian cultivated fruit species (strawberry, raspberry, blackberry, black currant, blueberry, gooseberry, cape gooseberry, chokeberry, and goji berry) revealed a good differentiation according to botanical origin. In order to determine biomarkers responsible for the classification, a total of 103 phenolic compounds were identified, including 53 phenolic acids and their derivatives, 26 flavonoids and 24 glycosides. Biomarkers derived from the phenolic profile of berry seeds proved to be a powerful tool in the authentication of botanical origin, and may be useful in detection of frauds in berry-based seed-containing product.

Effect-Directed Profiling of Strawberry Varieties and Breeding Materials via Planar Chromatography and Chemometrics.

Strawberries are an important fruit in the European diet because of their unique taste and high content of essential nutrients and bioactive compounds. The anthocyanins are known to be colorful phenolics in strawberries. In 17 samples of six strawberry cultivars produced in Serbia, i.e., the common varieties Alba, Asia, and Clery as well as promising breeding materials (11.29.11, 11.34.6, and 11.39.3), the anthocyanin profile as well as antimicrobial and antioxidative activity profiles were determined. All investigated extracts showed antioxidative and antibacterial activities against Gram-negative Aliivibrio fischeri. The responses were quite similar in number and intensity. The HPTLC-DPPH• scavenging assay and HPTLC-Aliivibrio fischeri bioassay coupled with high-resolution mass spectrometry identified pelargonidin-3-O-glucoside (Pg-3-glc) as the main anthocyanin and prominent antioxidative and antimicrobial compound in strawberries. The density functional theory calculations at the M06-2X/6-31+G(d,p) level showed that Pg-3-glc quenches free radicals via sequential proton loss electron transfer mechanism in water and in pentyl ethanoate, where the 5-OH group is the most reactive site for proton and hydrogen atom transfer. The results were confirmed via spectrophotometry. The highest total phenolic content was found in Clery and 11.39.3, while statistically significant differences between the genotypes regarding the antioxidant activity were not confirmed. Although very similar in the anthocyanin, antioxidative, and antimicrobial profile patterns, the strawberry genotypes were successfully classified using principal component analysis.

Efficiency of Natural Deep Eutectic Solvents to Extract Phenolic Compounds from Agrimonia eupatoria: Experimental Study and In Silico Modelling.

To replace common organic solvents that present inherent toxicity and have high volatility and to improve the extraction efficiency, a range of natural deep eutectic solvents (NADESs) were evaluated for the extraction of phenolic compounds from Agrimonia eupatoria. Screening of NADES efficiency was carried out based on the total phenolic and flavonoid content and radical-scavenging activity, determined by spectrophotometry, as well as phenolic compounds quantified, obtained using ultra-high-performance liquid chromatography with a diode array detector and a triple-quadrupole mass spectrometer. Increased extraction efficiency when compared with organic solvent was achieved using NADES mixtures choline chloride (ChCl):urea 1:2 and choline chloride:glycerol 1:1. Flavonol glycosides were the most abundant compounds in all extracts. The COSMO-RS model provided insights into the most important intermolecular interactions that drive the extraction process. Moreover, it could explain the extraction efficiency of flavonol glycosides using ChCl:glycerol NADES. The current article offers experimental evidence and mechanistic insights for the selection of optimal NADES to extract bioactive components from Agrimonia eupatoria.

Quality Assessment of Apple and Grape Juices from Serbian and German Markets by Planar Chromatography-Chemometrics.

The high consumption of plant-based foods on a global scale has increased the number of adulterations in the food industry. Along with this, analytical approaches to fraud detection need to be further developed. A nontargeted effect-directed profiling by high-performance thin-layer chromatography hyphenated with five effect-directed assays (free radical scavenging assay, Aliivibrio fischeri bioassay, and acetylcholinesterase, butyrylcholinesterase, and tyrosinase inhibition assays) and multi-imaging provided additional information on the antioxidative, antimicrobial, and enzyme inhibition activities for 18 apple and 18 grape juices from markets in Serbia and Germany. Bioactive zones of interest were eluted using an elution head-based interface and further characterized by electrospray ionization high-resolution mass spectrometry. The different profiles were evaluated chemometrically, and several compounds, which were characteristic of samples from different markets located in Serbia and Germany, were identified in apple juice (such as chlorogenic acid, phloridzin, epicatechin, and caffeic acid) and grape juice (such as chlorogenic acid, epicatechin, and quercetin). The developed rapid and simple method for the quality assessment of fruit juices coming from different (geographic) markets showed clear quality differences. Thus, it could be used to learn more about quality differences, to detect fraud in fruit juice production, and to verify the authenticity of the origin.

Thin-layer chromatography in the authenticity testing of bee-products.

Quality control, nutritional value and the monitoring of hazardous residues in honey bee- products have become major topics for both producers and consumers. Due to its potential role in human health, bee-products rich in bioactive compounds are becoming increasingly popular. This review aims to provide an overview of thin-layer chromatography methods used in quality control,authenticity testing and chemical profiling of bee-products in order to help scientists engaged in the field of bee-products chemistry to utilize the advantages of this technique in the detection and elimination of fraudulent practices in bee-product manufacturing. Recently, hyphenation of thin-layer chromatography, image analysis and chemometrics support bee-products analysisbysimultaneousdeterminationofanalytes with different detection principles, identification of individual bioactive compounds as well as structure elucidation of compounds. Highlighted opportunities of thin-layer chromatography could encourage further investigations that would lead to improvements in the detection and elimination of marketing fraudulent practices.

Fast detection of apricot product frauds by added pumpkin via planar chromatography and chemometrics: Greenness assessment by analytical eco-scale.

The European Commission requires that fruit products distributed on the market meet standards of high quality and authenticity. For the quality assessment of apricot products susceptible to food fraud, an environmentally friendly, simple and cost-effective analytical profiling was developed by high-performance thin-layer chromatography multi-imaging (HPTLC-FLD/Vis). The new phytochemical profiling was applied for analysis of authentic samples (7 apricot and 5pumpkin samples) and simulated adulterated products (11 mixture samples prepared by addition of 2.5-53% pooled pumpkin to pooled apricot). Based on the analytical eco-scale assessment, the HPTLC-FLD/Vis method was proven as an excellent green analytical method with low energy and solvent consumption. Chemometric data analysis confirmed the difference between apricot and apricot-pumpkin mixtures based on the phytochemical profile. Chemical markers responsible for their differentiation were identified. The results indicated that frauds by adding pumpkin to apricot products can be detected at added contents as low as 2.5%.

Chemical Content of Five Molluscan Bivalve Species Collected from South Korea: Multivariate Study and Safety Evaluation.

Bivalves are a good source of nutrients but also a potential source of environmental contaminants, which could pose a risk for consumers. The aims of this study were: the determination of 16 elements by ICP-MS in 48 samples of five bivalve species purchased from market in Korea; the identification of elements useful for species classification using multivariate analyses; and the benefit-risk evaluation associated to the consumption of these bivalves. The highest difference among content of elements between species was found for Cd, Mn, Ni, Zn, and Fe. Partial last squares discriminant analysis revealed elements with a VIP score >1 which were considered as the most relevant for explaining certain species. As, Cd, Co, and Ni were found as taxonomical markers of V. philippinarum; Mn, Zn, Mg, and Na of A. irradians; and Cd, Ni, and Fe of M. yessoensis. These species could serve as good dietary sources of essential elements. Cd exposure by consumption of Manila clams is not representing a health risk for the Korean population; however, through consumption of Yesso scallops, 5.3% of the Korean population has a potential health risk. Removal of the digestive gland before eating will drastically reduce the amount of Cd ingested.

Rapid analytical approach for bioprofiling compounds with radical scavenging and antimicrobial activities from seaweeds.

Brown seaweeds are traditionally used as food in Asian countries, and they are a valuable source of bioactive compounds. Herein, a novel high-throughput methodological approach was developed for the tracing of compounds with radical scavenging and antimicrobial activities in Saccharina japonica and Undaria pinnatifida methanol extracts. The seaweed metabolites were separated by a novel high-performance thin-layer chromatography method, the bioactive bands were identified by bioautography assays. The bioactive compounds were characterized with ultra-high-performance liquid chromatography coupled with linear trap quadrupole tandem mass spectrometry. Stearidonic, eicosapentaenoic, and arachidonic acids were identified as major components having radical scavenging and antimicrobial activities. The suggested method provides a fast identification and quantification of bioactive compounds in multicomponent biological samples.

New perspectives of purple starthistle (Centaurea calcitrapa) leaf extracts: phytochemical analysis, cytotoxicity and antimicrobial activity.

Ethnobotanical and ethnopharmacological studies of many Centaurea species indicated their potential in folk medicine so far. However, investigations of different Centaurea calcitrapa L. extracts in terms of cytotoxicity and antimicrobial activity against phytopathogens are generally scarce. The phenolic profile and broad antimicrobial activity (especially towards bacterial phytopathogens) of methanol (MeOH), 70% ethanol (EtOH), ethyl-acetate (EtOAc), 50% acetone (Me2CO) and dichloromethane: methanol (DCM: MeOH, 1: 1) extracts of C. calcitrapa leaves and their potential toxicity on MRC-5 cell line were investigated for the first time. A total of 55 phenolic compounds were identified: 30 phenolic acids and their derivatives, 25 flavonoid glycosides and aglycones. This is also the first report of the presence of centaureidin, jaceidin, kaempferide, nepetin, flavonoid glycosides, phenolic acids and their esters in C. calcitrapa extracts. The best results were obtained with EtOAc extract with lowest MIC values expressed in µg/mL ranging from 13 to 25, while methicillin resistant Staphylococcus aureus was the most susceptible strain. The most susceptible phytopathogens were Pseudomonas syringae pv. syringae, Xanthomonas campestris pv. campestris and Agrobacterium tumefaciens. The highest cytotoxicity was recorded for EtOAc and Me2CO extracts with the lowest relative and absolute IC50 values between 88 and 102 µg/mL, while EtOH extract was the least toxic with predicted relative IC50 value of 1578 µg/mL. Our results indicate that all tested extracts at concentration considered as non-toxic can be one of great importance in combat towards phytopathogenic and human pathogenic strains, as well as natural sources of antimicrobials.

Radical Scavenging and Antimicrobial Properties of Polyphenol Rich Waste Wood Extracts.

The main focus of this study is to assess radical scavenging and antimicrobial activities of the 11 wood extracts: oak (Quercus petraea (Matt.) Liebl., Q. robur L., and Q. cerris L.), mulberry (Morus alba L.), myrobalan plum (Prunus cerasifera Ehrh.), black locust (Robinia pseudoacacia L.), and wild cherry (Prunus avium L.). High-performance thin-layer chromatography (HPTLC) provided initial phenolic screening and revealed different chemical patterns among investigated wood extracts. To identify individual compounds with radical scavenging activity DPPH-HPTLC, assay was applied. Gallic acid, ferulic and/or caffeic acids were identified as the compounds with the highest contribution of total radical scavenging activity. Principal component analysis was applied on the data set obtained from HPTLC chromatogram to classify samples based on chemical fingerprints: Quercus spp. formed separate clusters from the other wood samples. The wood extracts were evaluated for their antimicrobial activity against eight representative human and opportunistic pathogens. The lowest minimum inhibitory concentration (MIC) was recorded against Staphylococcus aureus for black locust, cherry and mulberry wood extracts. This work provided simple, low-cost and high-throughput screening of phenolic compounds and assessments of the radical scavenging properties of selected individual metabolites from natural matrix that contributed to scavenge free radicals.

Extraction as a Critical Step in Phytochemical Analysis.

Phytochemical analysis, the chemical investigation of plants, is a rapidly developing chemical discipline with various goals such as the determination of the substance classes and the isolation and qualitative and/or quantitative analyses of bioactive compounds or their structural analysis. The analysis of plants begins with optimizing extraction procedures because they play a key role in extraction outcomes, i.e., the isolation of bioactive compounds from natural products and the selection of the subsequent tests used in the analyses. The extraction is a critical step because it is necessary to extract the desired components of the complex natural matrix without destroying them. A wide range of techniques for the extraction of bioactive compounds from natural matrixes are currently available. This review aims to describe and compare the most commonly used methods, traditional/conventional and innovative/green. It will focus on the principles behind each method and on their strengths and limitations in order to help evaluate their suitability and economic feasibility, highlighting the advantages of new innovative techniques over conventional extraction procedures. Finally, the recent application of deep eutectic solvents as a new, eco-friendly method for the extraction of bioactive compounds from natural matrixes is also discussed.

Effect-directed screening of Bacillus lipopeptide extracts via hyphenated high-performance thin-layer chromatography.

Bacillus species produce a wide array of biologically active metabolites, including nonribosomaly synthesized lipopeptides (LPs). The high-performance thin-layer chromatography (HPTLC) technique hyphenated with different bioassays and mass spectrometry was demonstrated as a valuable tool for effect-directed analysis of iturins, surfactins, fengycins and kurstakins homologues from complex mixtures of LPs. As proof of this straightforward strategy, the found surfactin and iturin A homologues were characterized and compared with reference substances. This study considered two different extraction methods for LPs produced by five Bacillus strains. The ethyl acetate extraction (Ex-1), and the acidic precipitation followed by methanol extraction (Ex-2) were investigated. Diverse enzyme inhibitions and antimicrobial potentials of LPs were analyzed, and in parallel, high-resolution mass spectra (HRMS) were online recorded from the HPTLC zones of interest. No antimicrobial effect against Gram-positive B. subtilis was evident for iturin, whereas a response was detected for surfactin. The nonpolar kurstakin compounds showed a pronounced B. subtilis antimicrobial activity in Ex-1 of almost all strains, whereas the fengycin homologues were detected in Ex-2 of SS-10.7 and SS-27.2. Iturin had also no activity against Gram-negative Aliivibrio fischeri, while again surfactin showed an enhancing luminescent activity. Contrary, kurstakin compounds caused a decrease in the luminescence in Ex-1 of all strains, particularly for SS-13.1. Both, iturin and surfactin showed a strong acetylcholinesterase (AChE) and α-glucosidase inhibition, but surfactin caused a much stronger inhibition. This was evident in all bacterial strains, except for SS-13.1 in Ex-1 and for SS-38.4 in Ex-2. Although, iturin and surfactin exhibited no DPPH˙ scavenging activity, Ex-1 of all strains contained more intense DPPH˙ scavenging compounds compared to Ex-2, and surfactin methyl esters showed a pronounced DPPH˙ activity, particularly in SS-12.6 in Ex-1. This study pointed to active metabolites of strains that can be used as therapeutics and biocontrol agents with beneficial effects on human health. The straightforward HPTLC profiling served as an excellent bioanalytical tool to control the formed bioactive metabolites. As the fermentation process is very sensitive to external influences, it could be a helpful control tool for standardization of the biotechnological processing.