Development and validation of a versatile analytical method for absolute quantification of seven oligosaccharides in human, bovine, and goat milk.

Oligosaccharides are significant in mammalian milk, where they serve as prebiotics that promote the growth of beneficial gut bacteria in infants. Comprehensive research of milk oligosaccharides requires precise and validated analytical methods for compositional studies. To address this need, the focus of our study was to develop and validate an analytical method using UPLC-MS/MS to quantify seven specific oligosaccharides found in mammalian milk. The developed and optimized method has adequate linearity, accuracy, and precision parameters. The detection (LOD) and quantification (LOQ) limits for the seven compounds ranged from 0.0018 to 0.0030 µg/mL and 0.0054-0.0063 µg/mL, respectively. The sample preparation method yielded recovery rates above 90.5 %. Furthermore, no significant matrix effect was observed. The validated method was successfully applied to human, goat, and bovine milk samples, demonstrating its proficiency in identifying variances in the concentration of oligosaccharides across different mammals. This versatile method will allow future research about factors affecting oligosaccharide composition.

Black Bean Hulls as a Byproduct of an Extraction Process to Enhance Nutraceutical and Glycemic-Related Properties of Nixtamalized Maize Tostadas.

Black bean hulls (BBH) are rich in phenolic compounds, such as anthocyanins, which can be incorporated into common staple foods such as maize tostadas, enhancing the nutraceutical properties of these products. This study incorporates black bean hulls to produce nixtamalized maize tostadas with nutraceutical properties. Nixtamalized corn flour (NCF) and black bean hulls (BBH) were characterized in terms of protein, fat, crude and dietary fiber, anthocyanin concentration, and different starch fractions. NCF and BBH depicted 53.7 and 16.8% of total digestible starch (TDS), respectively, and 1.2 and 7.6% of resistant starch (RS), in the same order. BBH was incorporated into nixtamalized flour at 10, 15, and 20% w/w, and the resulting dough was thermo-mechanically characterized. Tostadas with BBH had higher protein, dietary fiber, and anthocyanin concentrations. Enriched tostadas did not show significant changes in texture or other sensory characteristics. However, a reduction in total digestible starch (61.97 up to 59.07%), an increase in resistant starch (0.46 to 2.3% from control tostadas to 20% BBH tostadas), and a reduction in the predicted glycemic index (52 to 49), among other parameters, indicated that BBH is a suitable alternative for developing nutraceutical food products.

In search of better snacks: ohmic-heating nixtamalized flour and amaranth addition increase the nutraceutical and nutritional potential of vegetable-enriched tortilla chips.

BACKGROUND: Nixtamalized flour snacks such as tortilla chips are widely consumed across the world, but they are nutritionally poor and contribute to obesity and other non-communicable diseases. The production of healthy versions of such snacks, by incorporating vegetables and improving the quality of the flours used in their formulation, could help address these nutritional challenges. This study compared the fortification of baked tortilla chips with vegetable leaf powders (kale and wild amaranth at 0%, 4%, 8%, and 16% w/w) and using two types of nixtamalized flour: traditional (TNF) and with ohmic heating (OHF). RESULTS: Overall, the use of OHF increased 1.88 times the fibre in enriched and non-enriched snacks with respect to TNF, but the latter had 1.85 times more protein. Addition of 16% of vegetable powders increased protein (kale = 1.4-fold; amaranth = 1.3-fold) and dietary fibre (kale = 1.52-fold; amaranth = 1.7-fold). Amaranth enrichment improved total phenolic content (TPC) and total flavonoid content (TFC) of chips at least 1.2 and 1.63 times, respectively. OHF chips also had higher bound TPC than TNF ones, regardless of vegetable addition. Combinations of OHF with 16% amaranth produced chips 1.74-fold higher in antioxidant capacity than non-enriched ones, due to increased content of phenolics such as ferulic acid. CONCLUSION: This work showed that tortilla chips made using nixtamalized flour produced with assisted ohmic heating, alone or in combination with wild amaranth leaf powder, could be used in the production of healthy maize snacks to enhance their prospective antioxidant activity and nutritional value. © 2023 Society of Chemical Industry.

In vitro Evaluation of Anti-Inflammatory Activity of "Habanero" Chili Pepper (Capsicum chinense) Seeds Extracts Pretreated with Cellulase.

The aim of this study was to explore the effect of capsaicin and particular phenolic compounds profile from cellulase assisted extracts of Habanero (Capsicum chinense) chili pepper seeds (CPS) on the concentration of cytokines (IL-2, IL-6, TNF-α, IL-1ß) in murine macrophages (RAW 264.7) stimulated with lipopolysaccharides (LPS). Capsaicin was quantified by HPLC-DAD, and the phenolic profile was determined by UPLC-MS-QqQ. Anti-inflammatory activity was evaluated by Mouse Cytokine/Chemokine Magnetic Bead Panel 96-well plate assay. Among the 15 different phenolics found in CPS extracts obtained at 120 or 150 min of maceration with 2,500 UI/L at 30 ºC or 45 ºC in a 1:15 (w:v) proportion, the most abundant was vanillic acid (7.97-12.66 µg/g). The extract obtained at 30 ºC and 120 min, showed similar effects than the observed for synthetic anti-inflammatory drugs indomethacin and dexamethasone, and capsaicin standard. Beyond capsaicin, salicylic, protocatechuic and trans-cinnamic acids as well as vanillin in CPS extracts were correlated with the anti-inflammatory effect. On the other hand, capsaicin and chlorogenic acid contents were potential immunostimulants whose concentration varied depending on the cellulase treatment time.

Enzyme-assisted extraction of anti-inflammatory compounds from habanero chili pepper (Capsicum chinense) seeds.

Capsaicinoids are the main bioactive compounds extracted from chili pepper seeds (CPSs) but other bioactive compounds such as phenolic compounds may be found. Enzyme-assisted extraction (EAE) improves the extraction of bioactive compounds from fruits and seeds. The aim of this study was to establish the cellulase-assisted extraction conditions of capsaicinoids and phenolic compounds from Habanero CPSs (Capsicum chinense) and to evaluate the anti-inflammatory activity of the obtained extracts on murine macrophages. EAE was performed using different temperatures (T1 = 30°C, T2 = 45°C and T3 = 60°C), enzyme concentrations (E1 = 2,500 UI/L and E2 = 250 UI/L), and extraction time periods (0-150 min). Total phenolic compounds were quantified using the Folin-Ciocalteu assay, capsaicin (CAP) and dihydrocapsaicin (DHC) contents were evaluated by HPLC, and anti-inflammatory activity was performed with Griess assay on murine macrophage RAW 264.7 cell culture. The highest phenolic compound content (337.96 mg GAE/L) was achieved at 30°C, 2,500 UI/L, and 150 min of extraction. The highest CAP content (310.23 µg/ml) was obtained at 45°C with 250 UI/L for 150 min, while for DHC (167.72 µg/ml), the conditions were 60°C, 2,500 UI/L, and 120 min. The highest anti-inflammatory response was obtained when 60°C, E2, and 150 min were used for the extraction, and nitric oxide (NO) production was reduced to 22.56%. Based on the results obtained in this research, EAE allowed the recovery of compounds with anti-inflammatory activity from CPS using water as a solvent. There was a correlation between the extraction of CAP and DHC. But although a moderate direct correlation between the concentration of capsaicinoids and total phenolic compounds (TPCs) and an inverse correlation of the presence of the bioactive compounds (TPC, CAP, and DHC) with the NO synthesis, these were not statistically significant. We demonstrated that Habanero seeds are an important raw material to recover anti-inflammatory compounds beyond capsaicinoids using water in EAE.

Engineering and Evaluation of Forcespun Gelatin Nanofibers as an Isorhamnetin Glycosides Delivery System.

Opuntia ficus-indica (L.) Mill (OFI) is considered a natural source of bioactive phytochemicals, mainly isorhamnetin glycosides (IRGs). These compounds have demonstrated antioxidant, anti-inflammatory, and anticancer activities, among others. The development of a suitable delivery system for these compounds is needed to improve their chemical and biological stability. This study aimed to evaluate the feasibility of fabrication and characterization of IRG-loaded gelatin (GL) forcespun fibers and crosslinking with glutaraldehyde (GTA). Two different percentages (25% and 30% w/v) of GL were evaluated with 12% (w/v) OFI flour to obtain nanofibers GL/OFI1 and GL/OFI2, respectively. The morphology and physicochemical properties of the fibers were investigated. The results indicated that the diameters of the fibers were on the nanoscale. The amount of IRGs was determined using high-performance liquid chromatography (HPLC). The IRGs release and the cytocompatibility of the nanofibers were also evaluated. GL concentration significantly affected the IRG release. Among both nanofibers, the GL/OFI2 nanofiber achieved a cumulative IRGs release of 63% after 72 h. Both fibers were shown to be biocompatible with human skin/fibroblast cells. Specifically, GL/OFI1 nanofibers exhibited favorable features for their application as an extract-coupled release system. The IRGs-embedded GL nanofiber mats may become a good alternative for the delivery of phytochemicals for the health sector and biomedical applications.

Germinated chickpea-maize extrudates with high protein content and reduced starch digestibility.

The objective of this study was to produce maize extrudates supplemented with germinated chickpea flour to increase the contents of resistant starch (RS) and protein. Six extrudates were formulated using maize grits (ME), germinated chickpea flour (GCE) and different blends of maize and 10%, 20%, 30%, or 40% of germinated chickpea flour (MGCE-10, MGCE-20, MGCE-30, or MGCE-40). Increase of RS was observed in the defatted samples due to germinated chickpea flour addition. In the nondefatted samples, the highest content of RS was observed in GCE followed by ME and the different MGCE. Interaction between fat, starch, and protein by improved intramolecular association was assessed by Fourier transform- infrared spectroscopy (FTIR). Amylose-lipid complexes in nondefatted samples increased the content of RS in comparison to defatted samples. The highest expansion index was obtained in MGCE-30 and MGCE-40. ME had the highest hardness and crispiness. Germinated chickpea flour increased the water absorption index (WAI), but reduced water solubility index (WSI) when it was combined with maize grits to produce extrudates. The in vitro protein digestibility (IVPD) was higher in the GCE and MGCE with more than 20% of germinated chickpea flour compared to ME. MGCE-20 and MGCE-30 showed the highest acceptability of the supplemented extrudates with 50% more protein than ME, a similar IVPD to that of GCE, and good functional characteristics. PRACTICAL APPLICATION: Combining maize and germinated chickpea flour is a good strategy to have a controlled digestibility of starch and increase the plant based protein content in healthier snacks.

Nanofiber Systems as Herbal Bioactive Compounds Carriers: Current Applications in Healthcare.

Nanofibers have emerged as a potential novel platform due to their physicochemical properties for healthcare applications. Nanofibers' advantages rely on their high specific surface-area-to-volume and highly porous mesh. Their peculiar assembly allows cell accommodation, nutrient infiltration, gas exchange, waste excretion, high drug release rate, and stable structure. This review provided comprehensive information on the design and development of natural-based polymer nanofibers with the incorporation of herbal medicines for the treatment of common diseases and their in vivo studies. Natural and synthetic polymers have been widely used for the fabrication of nanofibers capable of mimicking extracellular matrix structure. Among them, natural polymers are preferred because of their biocompatibility, biodegradability, and similarity with extracellular matrix proteins. Herbal bioactive compounds from natural extracts have raised special interest due to their prominent beneficial properties in healthcare. Nanofiber properties allow these systems to serve as bioactive compound carriers to generate functional matrices with antimicrobial, anti-inflammatory, antioxidant, antiseptic, anti-viral, and other properties which have been studied in vitro and in vivo, mostly to prove their wound healing capacity and anti-inflammation properties.

Modulation of gut microbiota by Mantequilla and Melipona honeys decrease low-grade inflammation caused by high fructose corn syrup or sucrose in rats.

Several studies have shown that consumption of honey is associated with various health benefits. However, there is scarce evidence on whether honeys modify the intestinal microbiota by preventing the inflammatory response in the host. Therefore, the aim of the present work was to study the effect of Melipona (Mel) and Mantequilla (Mtq) honeys, which contain different bioactive compounds and antioxidant capacity on gut microbiota and metabolic consequences in comparison with other sweeteners, in particular sucrose (S) and high fructose corn syrup (HFCS) in rats. The results of the present work showed that both honeys have polyphenols, flavonoids, antioxidant and bactericidal activities. Rats fed with both honeys gained less weight and body fat by increasing energy expenditure compared to S or HFCS and increased gene expression of antioxidant enzymes mediated by the transcription factor Nrf2. Analysis of the gut microbiota showed that consumption of both honeys modified the beta-diversity compared to those fed S or HFCS resulting in increased abundance of a specific cluster of bacteria of the Clostridium genus particularly Coprococcus eutactus, Defluviitalea saccharophila, Ruminicoccus gnavus and Ruminicoccus flavefaciens. As a result of the changes in the gut microbiota, there was a decrease in LPS- and TLR4-mediated low-grade inflammation and an increase in sIgA. Consumption of both honeys prevented glucose intolerance and increased adipocyte size compared to S or HFCS. In conclusion, consumption of MtqH or MelH can reduce metabolic endotoxemia by modifying the gut microbiota to prevent glucose intolerance.

Immunomodulatory effect of metabolites from digested and fermented fractions from irradiated pineapple (Annanas comosus L.) snack-bars.

Phenolic compounds (PC), can modulate the immune response. UV-C irradiation, commonly used as a minimal processing method in fresh-foods to reduce the microbial load, increase shelf-life, provide a minimal processing and facilitate the release of PC. This study aimed to evaluate the effect of intestinal (IF) and fermented (FF) fractions of non-irradiated (NIPB) and irradiated (IPB) pineapple snack-bars on the production of nitric oxide (NO), interleukin 6 (IL-6), cyclooxygenase 2 (COX-2), and tumor necrosis factor-alpha (TNF-α) in mice macrophages. IF of NIPB and IPB exerted an immunomodulatory effect by promoting the production of NO (26 pg/mL) in both treatments, COX-2 (438 and 399 pg/mL), and TNF-α (778 and 802 pg/mL) for NIPB and IPB respectively. The TNF-α increased in IF of NIPB and IPB approximately 371 %, and in FF, only increased 132 %. The NO production was not different between IF and FF. COX-2 production was higher in FF.

Microencapsulation of steroidal saponins from agave sap concentrate using different carriers in spray drying.

Concentrated agave sap is a product with in vivo proven hypocholesterolemic and hypoglycemic activities, as well as in vitro anticancer potential. In the present work, a factorial design was used to determine the suitable drying conditions of concentrated agave by studying the effect of inlet temperature (150 °C, 180 °C and 210 °C) and the type of carrier agent (maltodextrin, hydroxypropyl methylcellulose, guar gum and xanthan gum). The response variables for each treatment were the product recovery and microencapsulated saponins. Further characterization of concentrated agave powders was performed: solubility in water, hygroscopicity, moisture content, tap density, bulk density, Carr's index followability and morphology by scanning electron microscopy analysis. The hydroxypropyl methylcellulose proved to improve physicochemical properties and enhance product yield, using 210 °C inlet temperature and a mix of carrier agents of maltodextrin/hydroxypropyl methylcellulose/xanthan gum at 50/48.5/1.5 (w/w/w) proportion exhibited the highest saponin recovery of 53.81%. Moreover, different carrier agents in powders revealed two shapes, regular spherical shape with smooth surface and collapsed shapes. The use of polymers excipients helped to decrease the stickiness of the desired product and enhanced the powder stability and microencapsulation of the steroidal saponins.

Is Apo-CIII the new cardiovascular target? An analysis of its current clinical and dietetic therapies.

AIMS: Recently, Apolipoprotein CIII (Apo-CIII) has gained remarkable attention since its overexpression has been strongly correlated to cardiovascular disease (CVD) occurrence. The aim of this review was to summarize the latest findings of Apo-CIII as a CVDs and diabetes risk factor, as well as the plausible mechanisms involved in the development of these pathologies, with particular emphasis on current clinical and dietetic therapies. DATA SYNTHESIS: Apo-CIII is a small protein (∼8.8 kDa) that, among other functions, inhibits lipoprotein lipase, a key enzyme in lipid metabolism. Apo-CIII plays a fundamental role in the physiopathology of atherosclerosis, type-1, and type-2 diabetes. Apo-CIII has become a potential clinical target to tackle these multifactorial diseases. Dietetic (omega-3 fatty acids, stanols, polyphenols, lycopene) and non-dietetic (fibrates, statins, and antisense oligonucleotides) therapies have shown promising results to regulate Apo-CIII and triglyceride levels. However, more information from clinical trials is required to validate it as a new target for atherosclerosis and diabetes types 1 and 2. CONCLUSIONS: There are still several pathways involving Apo-CIII regulation that might be affected by bioactive compounds that need further research. The mechanisms that trigger metabolic responses following bioactive compounds consumption are mainly related to higher LPL expression and PPARα activation, although the complete pathways are yet to be elucidated.

Influence of extrusion process on the release of phenolic compounds from mango (Mangifera indica L.) bagasse-added confections and evaluation of their bioaccessibility, intestinal permeability, and antioxidant capacity.

Extruded polyphenol-rich by-products like mango bagasse (MB) could be used to manufacture functional confections. However, few reports have assessed the extrusion impact on MB polyphenols within a food matrix. This research aimed to evaluate the impact of extrusion on the bioaccessibility, intestinal permeability, and antioxidant capacity of phenolic compounds (PC) from non-extruded and extruded MB-added confections (EMBC and MBC, respectively). The inhibition of 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl radicals and in silico approaches were used to evaluate the antioxidant capacity. MBC displayed the highest gastric bioaccessibility (%) of xanthones and flavonoids, whereas selective release of gallic acid, mangiferin, and quercetin glucoside was shown for EMBC. Lower PC' apparent permeability coefficients were found in EMBC compared to MB (0.11 to 0.44-fold change, p < 0.05). EMBC displayed the highest antioxidant capacity by the DPPH method for the non-digestible fraction, being mangiferin the highest in silico contributor (-4 kcal/mol). Our results showed that the extrusion process helps release selective phenolics from MBC, which increases their bioaccessibility and intestinal permeability.

Assessment of the bacterial diversity of agave sap concentrate, resistance to in vitro gastrointestinal conditions and short-chain fatty acids production.

Low bacterial diversity in the gut has been associated with the development of several diseases. Agave sap concentrate (ASC) is obtained from the thermal treatment of the fresh sap called "aguamiel", an artisanal Mexican food. In this study, we assessed the microbial diversity from three different ASC producing regions in Mexico using high-throughput sequencing of the 16S rRNA gene and evaluated their resistance to an in vitro gastrointestinal process as well as their ability to produce short-chain fatty acids (SCFA). Seven phyla and 120 genera were detected in ASC samples; Firmicutes had the highest relative read abundance at the phylum level, whereas Bacillus was the most abundant genus. Bacterial diversity at phylum and genus levels was highly dependent on the region where ASC was produced. The microbiota from a selected sample was resistant to low pH conditions, bile salts and intestinal enzymes. Moreover, bacteria were able to survive and grow in the colonic environment. SCFA production was comparable with that observed for a well-known probiotic, Lactobacillus plantarum 299v, that was used as control. These findings demonstrate that ASC contains a bacterial ecosystem with potential probiotic benefits.

Inhibition of miR31 and miR92a as Oncological Biomarkers in RKO Colon Cancer Cells Treated with Kaempferol-3-O-Glycoside Isolated from Black Bean.

MicroRNAs (miRNAs) are small molecules of 19-23 nucleotides of RNA that act as regulators of the expression of proteins in eukaryotic cells. Currently, the participation of miRNAs in the development of different types of cancer has been observed. To evaluate the inhibitory effect of kaempferol-3-O-glycoside on the expression of oncological biomarkers, miR31 and miR92a in a colon cancer cell line (RKO) were analyzed. Cells were cultured and treated with 1 mM kaempferol-3-O-glycoside isolated from black bean. Expression levels of miR31 and miR92a were evaluated by real-time PCR using TaqMan probes; in addition, two oncogenes (KRAS and c-MYC) and two tumor suppressors (AMP-activated protein kinase [AMPK] and adenomatous tumors of polyposis coli [APC]) were quantified to validate the biological effects; normalization of expression levels were carried out by 2-ΔΔCt. Results were analyzed by one-way ANOVA. The expression levels of miR31, miR92a, KRAS oncogene, and the c-MYC transcription factor were subexpressed upon 72 h post-treatment with kaempferol-3-O-glycoside compared with the control without treatment (P < .05); in contrast, the tumor suppressor genes AMPK (∼4.85, P = .005) and APC (∼2.71, P = .066) tumor suppressors genes were overexpressed. Our results showed the inhibitory effect of isolated black bean flavonoid kaempferol-3-O-glycoside on cancer biomarkers: miR31 and miR92a; based on our results, this flavonoid may have interesting nutritional, therapeutic, and/or prophylactic applications to combat colon cancer.

Isorhamnetin glycoside isolated from Opuntia ficus-indica (L.) MilI induces apoptosis in human colon cancer cells through mitochondrial damage.

This work aimed to evaluate the mechanisms involved in the apoptosis induction of isorhamnetin-3-O-glucosyl-pentoside (IGP) in metastatic human colon cancer cells (HT-29). To achieve this, we assessed phosphatidylserine (PS) exposure, cell membrane disruption, chromatin condensation, cell cycle alterations, mitochondrial damage, ROS production, and caspase-dependence on cell death. Our results showed that IGP induced cell death on HT-29 cells through PS exposure (48%) and membrane permeabilization (30%) as well as nuclear condensation (54%) compared with control cells. Moreover, IGP treatment induced cell cycle arrest in G2/M phase. Bax/Bcl-2 ratio increased and the loss of mitochondrial membrane potential (63%) was observed in IGP-treated cells. Finally, as apoptosis is a caspase-dependent cell death mechanism, we used a pancaspase-inhibitor (Q-VD-OPh) to demonstrate that the cell death induced by IGP was caspase-dependent. Overall these results indicated that IGP induced apoptosis through caspase-dependent mitochondrial damage in HT-29 colon cancer cells.

Selenium in Germinated Chickpea (Cicer arietinum L.) Increases the Stability of Its Oil Fraction.

Selenium is an essential mineral in human nutrition. In order to assess its effect on the stability of chickpea oil, seeds were germinated and tested with different amounts of sodium selenite (0.0, 0.5, 1.0 and 2.0 mg/100g seeds) for four days. Oil was extracted from sprouted chickpea and its physical properties, fatty acid profile (FAME), oxidative stability index (OSI), lipase and lipoxygenase (LOX) activities, cellular antioxidant activity (CAA), and phenolics and carotenoids were assessed and compared to chickpea seed oil. The amount of chickpea oil and its acid value (AV) increased during germination. The OSI increased by 28%, 46% and 14% for 0.5, 1.0 and 2.0 mg/100g compared with non-selenium treated sprouts. Phenolics increased up to 36% and carotenoids reduced by half in germinated sprouts with and without selenium compared to seeds. Carotenoids increased by 16% in sprouts treated with 1.0 mg/100 g selenium compared to their counterparts without selenium. FAME was not affected by treatments but samples with the highest selenium concentration increased lipase activity by 19% and decreased lipoxygenase activity by 55% compared with untreated sprouts. The CAA of oils increased by 43% to 66% in all germinated treatments compared with seeds. Results suggest that Se-enriched chickpea sprouts could represent an excellent source of oil with a high OSI and CAA, associated with a reduction in LOX activity and an increase in phenolics, respectively.

Changes in digestibility of proteins from chickpeas (Cicer arietinum L.) germinated in presence of selenium and antioxidant capacity of hydrolysates.

Germination in the presence of selenium (Se) is an alternative to increase the healthy properties of seeds. This study aimed to compare the Se accumulation in different protein fractions from germinated chickpea (Cicer arietinum L.) and the effect on digestibility and cellular antioxidant activity (CAA) of protein hydrolysates. Chickpeas were germinated during four days after soaking with sodium selenite (0, 1, or 2 mg/100 g seeds). Total protein (TP) and glutelin (Glu), albumin (Alb) and globulin (Glo) fractions were digested and ultrafiltrated through a 10 kDa membrane. Se accumulated in the order of Glu > Alb > Glo. Ultrafiltrated Glu hydrolysate of four days germinated chickpeas treated with 2 mg Na2SeO3/100 g increased CAA (51.47%), demonstrating the potential health benefits of selenization. The intensity of vicilin bands (34-37 kDa) increased from the second to the fourth day compared with the control samples. Glo digestibility was higher in selenized chickpea sprouts.

Enhanced production and identification of antioxidants in in vitro cultures of the cacti Mammillaria candida and Turbinicarpus laui.

Cacti are an important source of metabolites but present limitations for their commercial exploitation, like slow growth and a decrease of wild populations. An alternative to obtain their biocompounds without affecting the natural environment are the in vitro culture techniques. We established in vitro cultures from Mammillaria candida Scheidweiler and Turbinicarpus laui Glass and Foster and used different stresses to increase metabolites and antioxidant activity. The cultures were exposed to 1.25% polyethylene glycol to induce a moderate drought stress, 50 g L-1 sucrose to generate an osmotic stress, chitosan (1.25 to 5 mg mL-1) to simulate a biotic attack, or to UV light. Chitosan was the best elicitor improving 1.5 times the concentration of phenolics, 9 to 10 times the content of flavonoids and betalains, and 16% the antioxidant activity in M. candida suspensions. In T. laui suspensions, this elicitor duplicates the flavonoids content and antioxidant activity. The antioxidant levels in elicited suspensions increased 5 to 10 times in relation to plant tubercles. Eleven compounds were identified in M. candida suspensions being digalloyl rhamnoside and epicatequin gallate the most abundant; in the T. laui suspensions, 16 compounds were detected and the most abundant were 17-decarboxi neobetanin and derivatives of luteolin. Thus, cacti in vitro culture is an efficient system to obtain high level of metabolites of biological interest.