Final Extract Research Articles

Risk Assessment of Personal Care Products, Pharmaceuticals, and Stimulants in Mgeni and Msunduzi Rivers, KwaZulu-Natal, South Africa

In this work, environmental occurrence and risk assessment of pharmaceuticals, personal care products (PPCPs), and stimulants are presented. A quantitative technique is described for ultrasonic-assisted solid-phase extraction (SPE) followed by GC-MS after derivatization of PPCPs; propylparaben, triclosan, carbamazepine, chloramphenicol, and stimulant caffeine. Ultrasonic-assisted extraction together with centrifugation were used to extract sediment samples collected from the Mgeni and Msunduzi rivers. An SPE procedure was used for cleanup and to concentrate selected compounds from diluted aqueous extracts. The final extracts were derivatized and analyzed with GC-MS in selected ion monitoring (SIM) mode. The recoveries of the analytes ranged from 66 to 108%. The method detection limits were (0.08–1.82 ng g−1 for solid and 0.08–10 μg L−1 for liquid) and quantification limits (0.42–5.51 ng g−1 for solid and 0.25–25 μg L−1 for liquid). The optimized method was applied in the evaluation of two rivers over 3 months in KwaZulu-Natal, South Africa. All targeted compounds were present in the environment at concentration levels between not detected to 174 ng g−1 and not detected to 30 μg L−1 for solids and aqueous environmental samples, respectively. A comparison of predicted no environmental effect concentration (PNECs) with measured environmental concentration (MECs) showed that these PPCPs present a high ecological risk to the receiving environment (agricultural lands and households). Our work is close to reality because we used MECs as opposed to using predicted environmental concentration (PECs) values, which are normally calculated from consumption, production of compound per year, and various estimated factors.

Paper-based monolith extraction of psychoactive substances from biological fluids

A monolith of poly(methacrylic acid-co-ethylene glycol dimethacrylate) has been immobilised to a nitrocellulose strip by radical photopolymerisation to be used in the extraction of psychoactive substances in biological fluids. Codeine, methylone, amphetamine, methamphetamine, 3,4-methylenedioxymethamphetamine, butylone, norketamine, ketamine, heroin, cocaine, lysergic acid diethylamide and fentanyl were employed as model drugs and final extracts were analysed by ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Polymerisation parameters were adjusted in order to obtain a stable and homogeneous layer of monolith onto the nitrocellulose strip. The resulting sorptive phase was characterized by Fourier-transform infrared spectroscopy and scanning electron microscopy. Extraction conditions were investigated by the evaluation of sample pH, extraction and desorption times and desorption solvent volume, providing enrichment factor values ranging from 5.3 to 39.9. The proposed methodology provided limit of quantification values from 0.013 μg L−1 for methylone to 0.057 μg L−1 for amphetamine, and recoveries from 64 to 120%. Urine and serum certified reference materials were employed in the validation of the proposed methodology, providing results statistically comparable. The developed approach is simple and straightforward for the determination of psychoactive substances in urine and serum samples.

3D Reconstruction of cellular images from microfabricated imagers using fully-adaptive deep neural networks

Millimeter-scale multi-cellular level imagers enable various applications, ranging from intraoperative surgical navigation to implantable sensors. However, the tradeoffs for miniaturization compromise resolution, making extracting 3D cell locations challenging—critical for tumor margin assessment and therapy monitoring. This work presents three machine-learning-based modules that extract spatial information from single image acquisitions using custom-made millimeter-scale imagers. The neural networks were trained on synthetically-generated (using Perlin noise) cell images. The first network is a convolutional neural network estimating the depth of a single layer of cells, the second is a deblurring module correcting for the point spread function (PSF). The final module extracts spatial information from a single image acquisition of a 3D specimen and reconstructs cross-sections, by providing a layered “map” of cell locations. The maximum depth error of the first module is 100 µm, with 87% test accuracy. The second module’s PSF correction achieves a least-square-error of only 4%. The third module generates a binary “cell” or “no cell” per-pixel labeling with an accuracy ranging from 89% to 85%. This work demonstrates the synergy between ultra-small silicon-based imagers that enable in vivo imaging but face a trade-off in spatial resolution, and the processing power of neural networks to achieve enhancements beyond conventional linear optimization techniques.

Renewable sorbent dispersive solid phase extraction automated by Lab-In-Syringe using magnetite-functionalized hydrophilic-lipophilic balanced sorbent coupled online to HPLC for determination of surface water contaminants

An automated methodology for magnetic dispersive solid phase microextraction integrating bead injection approach for renewable sorbent introduction is presented for the first time and was successfully applied to the enrichment of water contaminants. For this purpose, a simple procedure was developed for the functionalization of commercial SupelTM-Select HLB (Hydrophilic modified styrene polymer) sorbent beads that allowed embedding magnetite nanoparticles (Fe3O4). The sorbent was then used in a dispersive solid phase extraction procedure that was carried out entirely inside the void of an automatic syringe pump following the flow-batch concept of Lab-In-Syringe including automated renewal of the sorbent for each analysis. Mixing processes, sorbent dispersion, and sorbent recovery were enabled by using a strong magnetic stirring bar, fabricated from a 3D printed polypropylene casing and neodymium magnets, inside the syringe. The final extract was submitted to online coupled liquid chromatography with spectrometric detection. System and methodology were applied to determine mebendazole, bisphenol A, benzyl 4-hydroxybenzoate, diclofenac, and triclosan selected as models from different groups of environmental contaminants of current concern. Experimental parameters including extraction and elution times, composition and volume of eluent, and bead recollection were optimized. Required system elements were produced by 3D printing. Enlarging the sample volume by repeated extraction to enhance the sensitivity of the method was studied. Using double extraction from 3.5 mL, limits of detection ranged from 1.2 μg L−1 to 6.5 μg L−1 with an RSD (n = 6) value less than 7% for all the analytes at 25 μg L−1 level. The method was linear in the range of 5–200 μg L−1 and was successfully implemented for the analysis of surface waters with analyte recoveries ranging from 78.4% to 105.6%.

A highly sensitive bioanalytical method for the quantification of vinblastine, vincristine, vinorelbine and 4-O-deacetylvinorelbine in human plasma using LC-MS/MS

A highly sensitive method was developed for the quantification of vinblastine, vincristine, vinorelbine, and its active metabolite 4-O-deacetylvinorelbine in human plasma using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Deuterated isotopes were used as internal standard and liquid-liquid extraction with tert-butyl methyl ether (TBME) was used for sample pre-treatment. The final extract was injected on a C18 column (50 × 2.1 mm ID, 5 µm). Gradient elution was used in combination with Reversed Phase chromatography to elute the analytes and internal standards from the column in 5 min and the API4000 triple quadrupole MS detector was operating in the positive ion mode. The calibration model, accuracy and precision, selectivity and specificity, dilution integrity, carryover, matrix factor and recovery, and stability were evaluated over a concentration range from 0.025 to 10 ng/mL for vinblastine, vinorelbine, and 4-O-deacetylvinorelbine and from 0.1 to 40 ng/mL for vincristine. The intra- and inter-assay bias and precisions were within ± 12.4% and ≤ 10.6%, respectively. This method was successfully applied to study the pharmacokinetics of vincristine in paediatrics and vinorelbine and 4-O-deacetylvinorelbine using in vivo mouse models.

A novel method based on saponification coupled to micelle-extraction for recovering valuable bioactive compounds from soybean oil deodorizer distillate

The feasibility of using saponification coupled to extraction with mixed micellar systems to recover bioactive compounds from soybean oil deodorizer distillate, was evaluated for the first time. Under the selected conditions, saponification with KOH 0.6 M and aqueous micellar system prepared with Tergitol 15-S-7 9% w/w, rhamnolipids 0.25 %w/w and sodium citrate 100 mM pH 5.00, at 65 °C, allow the recovery of almost 100% of α- and δ- tocopherols, and 90% of γ-tocopherols. LC-MS measurements demonstrated that the final extract also contained phytosterols and squalene. Additionally, the obtained extract preserved about 100% of the total antioxidant activity. This result was attributable to the fact that 93% of the tocopherols recovered in the micellar phases resulted to be associated with surfactant micelles, environment that is known to improve their antioxidant capacity. These results open perspectives to the use of this methodology to extract these valuable compounds from complex oily sources.

Valorization of tamarind seeds using high-pressure extraction methods to obtain rich fractions in fatty acid and phenolic compounds

Supercritical fluid extraction (SFE) and pressurized liquid extraction (PLE) were combined, in a biorefinery approach, to recover the nonpolar and polar fractions from tamarind seeds, which represent 25–40% of processed fruit. Total phenolic content (TPC), antioxidant capacity, and chemical composition were evaluated, and compared with values from Soxhlet (SOX) samples. The nonpolar lipid fraction, obtained by SFE with CO2, contains unsaturated and saturated fatty acids, similar to sample by SOX-hexane. The polar phenolic fraction, recovered by PLE-ethanol (60 °C / 10 MPa), presented high TPC (305.1 mg GAE·g-1) and antioxidant capacity, compared to SOX-ethanol sample. Combining SFE and PLE increased the TPC (421.8 mg GAE·g-1) and antioxidant capacity (2208.3–4412.5 µmol TEAC·g-1) of final extract. Chemical analysis confirmed high concentrations of organic acids and phenolic compounds (epicatechin, procyanidin and rutin) from ethanolic extracts. The biorefinery approach provides data for tamarind seeds fractionation, valuing this underestimated by-product by high-pressure methods.

Production, concentration and partial characterization of an enzymatic extract produced by an Aspergillus niger mutant in solid state fermentation

An enzymatic extract from Aspergillus niger 3T5B8 was produced by Solid State Fermentation (SSF) in aerated columns, using wheat bran as substrate. A combination of extracts produced using three different process conditions varying temperature, pH and aeration formed the final extract (Mixture). The Mixture was concentrated by an ultrafiltration process that partially purified and provided an efficient recovery of the enzymatic activities of xylanase (88.89%), polygalacturonase (89.3%), β-glucosidase (93.15%), protease (98.68%) and carboxymethylcellulase (CMCase) (98.93%). SDS-PAGE analysis showed 15 visible protein bands in the crude and concentrated Mixture with molecular weights ranging from 15.1 to 104.6 kDa. Thin layer chromatography confirmed the effective action of β-glucosidase and xylanase hydrolysis activities over cellobiose and xylan, respectively. A central composite design (CCD) with two variables and four replicates at the center points was used to determine the optimal temperature and pH for CMCase and β-glucosidase. The optimal temperature was 78.9 °C and pH 3.8 for CMCase and 52.8 °C and pH 4.8 for β-glucosidase, respectively.

Extract of Trema Orientalis (L.) Stem Bark: A Potential Source of Environmentally Friendly Tanning Agent for Leather Industry

Researchers around the world are in continuous endeavor to develop environment friendly tanning agent due to adverse effect of conventionally used chromium during leather tanning. Recent trend of tanning is thus heading to chrome-free and greener chemical processing options. Vegetable tanning is an exoteric leather processing technique because of their lower pollution load on the environment. Considering the inadequacy and high costing for the commercialized tannins, development of alternative tannins from locally available plants and their characterizations are important. In this research, the stem barks of Trema Orientalis (L.) were extracted at different temperatures employing water solvent with or without additives (sodium hydroxide or sodium sulphite) to attain phenolic-rich extractives. To ascertain appropriateness as a vegetable tanning agent, the obtained extracts were thereafter characterized in respect to yield, total phenolic content, tannin content and molecular structure. The extraction yield for all extracts improves and the quality remains nearly unchanged with temperature rising. The elevated concentration of chemical additives enhances the extraction yield but lessens the quality of extracts. In terms of extraction yield and the quality of extracts the best condition for extraction was discerned at 80°C with water solvent. This water extract has a decent extent of phenolic and tannin content of 266.13 mg Gallic acid equivalent/ gm of dry extract and 30.12 % respectively. The final extract exhibits excellent leather retanning tendency comparable to the commercial quebracho tannins.

Development and validation of GC/MS method for determination of residual ethanol in Cannabis extract

Ethanol was found to be the most suitable solvent for optimal extraction of cannabinoids and terpenes from Cannabis herba. Since ethanol use can have a serious impact on human health, purification of pharmaceuticals after ethanol extraction is necessary. Ethanol belongs to Class 3 residual solvents and its concentration is limited to 5000 ppm (0.5%) in the final pharmaceutical dosage forms. Determination (quantification) of ethanol in the final extracts is crucial for the quality and safety of the product. A novel GC/MS method was developed and validated for the determination of residual ethanol in cannabis extracts. The developed method is rapid, specific, accurate and precise. The calibration curve was plotted between the obtained peak areas of ethanol (quantifier ion m/z=31) and its concentration at six different concentration levels. The response was linear over the range of 312.5 ppm – 10000 ppm (R2 =0.9995). The limit of quantification, the limit of detection, system repeatability, method precision and recovery are demonstrated to evaluate the performance of the analytical procedure. The recovery range for added ethanol was 88.44% to 108.57%. The achieved results confirmed that the proposed method is suitable for monitoring the concentration of ethanol in cannabis extracts. Keywords: residual ethanol, cannabis extracts, gas chromatography-mass spectrometry

Organophosphate ester metabolites in human breast milk determined by online solid phase extraction coupled to high pressure liquid chromatography tandem mass spectrometry.

The analysis of metabolites of organophosphate esters (OPEs) in human breast milk is essential to evaluate OPE and OPE metabolite exposure of newborns. In the current study, an analytical method which only needs a small amount of breast milk (100μl) was developed and validated for six diester metabolites and three hydroxylated metabolites applying salt-induced liquid-liquid extraction (SI-LLE) and dispersive solid phase extraction (d-SPE) for sample preparation and online solid phase extraction coupled to high pressure chromatography tandem mass spectrometry (online-SPE-HPLC-MS/MS) for quantitative measurement. The final method consisted of an extraction with formic acid (FA)/acetonitrile (1:200, v/v) and a cleanup with C18 d-SPE. The final extracts were trapped on a C18 cartridge with application of a wash step of 2ml 0.1% FA milli-Q/methanol (98:2, v/v). Method detection limits (MDLs) ranging from 21.7ng/l for BBOEHEP to 500ng/l for BCIPP and average recoveries ranging from 58% for 5-OH-EHDPHP to 120% for BCIPP were achieved. Thirty-three breast milk samples from the LINC (Linking EDCs in maternal Nutrition to Child health) cohort collected in three distinct areas in The Netherlands were analyzed using the validated method. BCEP, BCIPP, BCIPHPP, BDCIPP, and 5-OH-EHDPHP were not detected in any of the samples, while BBOEP was the most frequently detected metabolite with a concentration range of <MDL to l.47ng/ml, followed by DPhP and BBOEHEP, detected in ranges of <MDL to 0.09 and <MDL to 0.027ng/ml. The results indicated that OPEs entering the human body are only to a limited extent excreted via breast milk.

CCAD-Net: A Cascade Cloud Attribute Discrimination Network for Cloud Genera Segmentation in Whole-Sky Images

Cloud detection and recognition are two important tasks usually referring to image binary segmentation and image-level classification individually. Cloud genera segmentation has more practical significance but is much more challenging as a fine-grained pixel-level dense prediction problem. In this letter, a cascade cloud attribute discrimination network (CCAD-Net) is proposed. Based on an improved encoding-decoding model, CCAD-Net adds a binary segmentation branch for cloud detection and a attribute discrimination branch for cloud attribute feature learning in the decoding stage. Especially, in the attribute discrimination branch, several visual attributes are selected to design the attribute discrimination constraint according to prior professional knowledge and corresponding loss function is defined. These two additional branches and the final cloud genera segmentation branch extract their task-specific features successively and form a cascade structure. Due to the fusion of raw feature, binary segmentation feature, attribute discrimination feature, and cloud genera feature, CCAD-Net can achieve significant better performance than the state-of-the-art methods in cloud genera segmentation in whole-sky images.

Quantification of Branched-Chain Amino Acids in Plasma by High-Performance Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS).

Branched-chain amino acids (BCAA), including valine, alloisoleucine, isoleucine, and leucine, play significant roles in a number of metabolic pathways in the body. Deficiency in branched-chain ketoacid dehydrogenase complex, an enzyme required for metabolism of those amino acids, will lead to elevation and accumulation of BCAA and ketoacids in bodily fluids. This results in maple syrup urine disease (MSUD), a condition estimated to affect 1 in 100,000-300,000 births. If MSUD is not diagnosed in the first few days of life, progression of this disease can lead to intellectual disability, coma, irreversible brain damage, seizures, or even death. If diagnosed early, MSUD can be managed by monitoring the blood concentrations of BCAA and adjusting the patient's dietary intake accordingly. Therefore, it is critical to have a rapid, accurate, and reliable BCAA assay for confirmation of MSUD in newborns as well as routine monitoring of MSUD patients. Here, we describe a high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) method for BCAA measurement which requires only 20μL of plasma. The sample preparation does not require derivatization and only involves protein precipitation with LC/MS-grade methanol, which contains leucine(13C6;15N), isoleucine(13C6;15N), and valine(13C5;15N) as the internal standards. The final sample extracts do not require dry-down and reconstitution and are readily compatible with the liquid chromatography (LC) method. BCAA are separated using the isocratic gradient method on a mixed-mode Intrada column. Multiple-reaction monitoring (MRM) mode is used for MS/MS detection to monitor the parent-to-daughter transitions m/z 132.2 to 86.4 for leucine, isoleucine, and alloisoleucine; m/z 118.2 to 72.4 for valine; m/z 139.2 to 92.4 for leucine(13C6;15N) and isoleucine(13C6;15N); and m/z 124.2 to 77.4 for valine(13C5;15N).

Use of ultrasound and acerola (Malpighia emarginata) residue extract tenderness and lipid oxidation of pork meat

This work aims to evaluate the application of ultrasound and marinate with extract of acerola (Malpighia emarginata) residue on the tenderness and lipid oxidation of the meat pork. Samples of loin (Longissimus dorsi) containing approximately 90 g, 10 cm long, 5 cm wide, and 2 cm high were extracted from the pig carcass. The samples were coded and randomly allocated under a factorial scheme 2 (with and without antioxidant extract) x 3 (ultrasonic bath times 0, 5, and 10 minutes); each treatment had four replicates. The final extract volume was adjusted with distilled water, ensuring 200 mg of gallic acid equivalents per kilo of meat. Then, the samples were placed in the ultrasonic bath at a fixed frequency of 170 W, 35 kHz. During the ultrasound application, the bath temperature was maintained at 10 °C with ice/ice water addition when necessary. The acerola´s extract showed a significant effect (P < 0.05) on the meat Cohesiveness and to a lesser extent before rupture. With the modification of the microstructure in the meat, there was a decrease in elasticity. The application of ultrasound to meat (170 W, 35 kHz) in times (5 and 10 minutes) combined with the marinating in a natural antioxidant acerolas residue extract improves the meat's quality characteristics, decreasing the hardness and chewiness.

Recycled Pequi (Caryocar brasiliense, Camb.) Shell Ethanolic Extract Induces Apoptosis in Canine Osteosarcoma Cells

Abstract Osteosarcoma is a mesenchymal cancer associated with high mortality in dogs and in humans. The biodiversity-rich Cerrado, the predominant biome in the Midwest region of Brazil, is native to locally appreciated fruits such as pequi (Caryocar brasiliense, Camb). Although this plant has been frequently used in folk medicine, the pharmacological properties of pequi fruit shells have not been fully evaluated. Thus, this study aimed to determine the cytotoxic activity of ethanol extract of recycled pequi shells on canine osteosarcoma cells in vitro. Cells were cultured and treated with final extract concentrations of 0, 0.029 µg/µL, 0.29 µg/µL, and 2.91 µg/µL for 24, 48, or 72 hours. Cell viability assay using trypan blue exclusion method and tetrazolium reduction method, cell survival assay, and double labeling with annexin V and propidium iodide were performed in the treated osteosarcoma cells. These allowed the determination of IC50, survival fraction, and type of cell death, respectively. Pequi shell ethanol extract at a concentration of 2.91 µg/µL showed the greatest inhibition of osteosarcoma cell growth in vitro, resulting in a 71.80% decrease in growth compared to the control. The mean IC50 was 155.2 μg/mL at 72 hours. The calculated survival fractions showed that cell growth at 72 hours was 3.33% lower in cells treated with 2.91 µg/µL extract. Results from the double labeling experiment suggest that apoptosis was the predominant type of cell death in cells treated with 2.91 µg/µL extract. These results demonstrate that ethanol extract of recycled pequi shells promotes apoptosis in canine osteosarcoma cells.

Herbal formuLation of Azadirachta indica and Stevia rebaudiana and Its Anti-inflammatory and Anti-Diabetic Activity

Introduction: Azadirachta indica, a natural herb also called Neem, and its seed extract has various medicinal uses. Neem also possesses notable insecticidal and pesticide activities. On the other hand, Stevia rebaudiana extract has alpha amylase glycosidase inhibitory activity, which contrasts with traditional wheat bread. This last plant can also regulate blood glucose levels, and prevent hypertension and caries.&#x0D; Aim: This study aimed to analyse the anti-diabetic and anti-inflammatory activities of Azadirachta indica and stevia rebaudiana.&#x0D; Materials and Methods: 2.5 g from Azadirachta indica and Stevia rebaudiana plants mixed with 100 mL of distilled water were boiled. The final extract was then filtered using Whatman filter paper. Spearman correlation analysis was done by SPSS. Positive correlation analysis level of significance was set as r=1.&#x0D; Results: The anti-diabetic activity and anti-inflammatory activities of Azadirachta indica and Stevia rebaudiana extracts showed positive spearman correlations (r = 1). Both activities happen in a dose-dependent manner, being higher with the increasing of the concentration and of the absorbance values.&#x0D; Conclusion: Through this study, it was possible to conclude that Azadirachta indica and Stevia rebaudiana have great anti-inflammatory and anti-diabetic activities and can be used as a possible alternative drug for attenuating inflammation and preventing diabetic complications.

Multi-residue method validation and safety evaluation of pesticide residues in seed spices cumin (Cuminum cyminum) and coriander (Coriandrum sativum) by gas chromatography tandem mass spectrometry (GC–MS/MS)

The manuscript reports comprehensive multi-residue determination of 215 pesticides in two commercially important Indian spices, Cumin (Cuminum cyminum) and Coriander (Coriandrum sativum) by GC–MS/MS analysis. The proposed method involved liquid–liquid extraction with acetonitrile, d-SPE clean-up and final reconstitution of extract in ethyl acetate. d-SPE clean-up with PSA and C18 minimized the matrix effects by 40 and 16%, respectively. Reconstitution of final extract reduced the non-volatile matrix co-extractives by 36–40%. The method was validated as per SANTE/12682/2019 and recoveries at 10, 25 and 50 μg kg−1 were within 70–120% with RSD ≤ 20%. A fit for purpose method LOQ of 10 μgkg−1 was achieved for 85% of analytes. The method was successfully applied for comprehensive screening of cumin and coriander market samples. The calculated TMDI for acute and chronic exposure assessment were less than calculated MPI in respective matrices and therefore did not cause any adverse effect to consumers.

In vitro acaricidal activity of extracts of red cultivar Allium cepa L. on engorged female Rhipicephalus (Boophilus) annulatus from naturally infested cattle

Introduction: Tick infestation is a major challenge facing livestock production in the Sub-Saharan zone. Most species of ticks have developed resistance to most of the available synthetic acaricides. Hence, the need to search for and develop ethno-acaricidal products. This study investigated the in-vitro acaricidal activity of aqueous and methanol extracts of red cultivar Allium cepa L. on engorged female Rhipicephalus (Bophillus) annulatus ticks. Methods: The extracts were prepared using standard method and the final brownish black extracts were kept in the refrigerator at 4oC. In-vitro acaricidal bioassay was done using the adult immersion test. Engorged ticks were randomly assigned into six groups (I-VI) (50mg/ml, 100mg/ml, 200mg/ml, and 400mg/ml (of the extracts), Distilled water (10 ml/kg) and Cypermethrin (10% EC) having three (3) replicates per group and containing 10 ticks per replicate administered. Results: Tick mortality of 66.66 % was effected by immersion in 400 mg/ml aqueous extract of red cultivar A. cepa as against 60 % obtained with cypermethrin within the same period. Aqueous extract caused 80 % inhibition of oviposition 72 hours post immersion (PI) as against 40 % caused by appli-cation of cypermethrin. Significance: The tickicidal and anti-oviposition activities of crude aqueous and methanol extract of red cultivar A. cepa L. observed in this present study revealed that the extracts possess some acaricidal properties/principles. Therefore, the plant could be an alternative source of tick control method with similar benefits compare to chemical acaricides such as cypermethrin, but without exposing our environment, animal and human to hazard.

Activity of antifungal drugs and Brazilian red and green propolis extracted with different methodologies against oral isolates of Candida spp.

BackgroundOral candidiasis is an opportunistic disease caused by fungi of the Candida genus. The occurrence of Candida spp. resistance to the commercial antifungal drugs points to the search for alternative treatments. Propolis has been successfully used in the treatment of infectious diseases for centuries. It has been proposed that an ultrasound pretreatment in the propolis extraction protocol can enhance the concentrations of molecules with antimicrobial activities in the final extract. Thus, this study aimed to compare the antifungal activity against oral Candida spp. isolates of green and red propolis extracts submitted or not to an ultrasound pretreatment before the extraction procedure.MethodsCandida spp. were isolated from denture stomatitis lesions and identified by sequencing. Oral Candida spp. isolates and reference strains were submitted to broth microdilution assays using commercial antifungals and Brazilian green and red propolis extracts submitted or not to an ultrasound pretreatment. Minimal Inhibitory Concentrations (MIC) and Minimal Fungicide Concentrations (MFC) were determined and biofilm formation interference was evaluated for resistant isolates.ResultsC. albicans, Candida tropicalis and Candida dubliniensis were isolated from denture stomatitis lesions. Growth inhibition was observed in all Candida isolates incubated with all green and red propolis extracts. At lower doses, red propolis extracts presented significant antifungal activity. The ultrasound pretreatment did not promote an increase in the antifungal activity of green or red propolis. Three isolates, which were highly resistant to fluconazole and itraconazole, were susceptible to low doses of red propolis extracts. These same three specimens had their biofilm formation inhibted by red propolis ethanolic extract.ConclusionsThus, red propolis can be faced as a promising natural product to be used in the auxiliary antifungal therapy of denture stomatitis.

Glucosinolates and Isothiocyantes in Processed Rapeseed Determined by HPLC-DAD-qTOF.

Glucosinolates are well known as natural antimicrobials and anticarcinogenic agents. However, these compounds can lose their properties and transform into antinutrients, depending on processing conditions. In addition, the bitterness of some glucosinolate in rapeseed meal can affect the likability of the final product. Therefore, it is important to identify and determine each glucosinolate and its derived form, not just the total glucosinolate content, in order to evaluate the potential of the final rapeseed protein product. This study provides a comprehensive report of the types and quantities of glucosinolates and their derived forms (isothiocyanates) associated with different rapeseed processing conditions. Glucosinolates and isothiocyanates were determined by HPLC-DAD-qTOF. In our study, the enzymatic degradation of glucosinolates by myrosinase was the main factor affecting either glucosinolate or isothiocyanate content. Other factors such as pH seemed to influence the concentration and the presence of glucosinolates. In addition, process parameters, such as extraction time and separation technology, seemed to affect the amount and type of isothiocyanates in the final protein extracts. Overall, both determined intact glucosinolates and their derived forms of isothiocyanates can give different types of biological effects. More studies should be performed to evaluate the impact of glucosinolates and isothiocyanates on human health.