Optimized Extraction Procedure Research Articles

Monitoring the Phenolic and Terpenic Profile of Olives, Olive Oils and By-Products throughout the Production Process.

Olive oil is a food of great importance in the Mediterranean diet and culture. However, during its production, the olive oil industry generates a large amount of waste by-products that can be an important source of bioactive compounds, such as phenolic compounds and terpenes, revalorizing them in the context of the circular economy. Therefore, it is of great interest to study the distribution and abundance of these bioactive compounds in the different by-products. This research is a screening focused on phytochemical analysis, with particular emphasis on the identification and quantification of the phenolic and terpenic fractions. Both the main products of the olive industry (olives, olive paste and produced oil) and the by-products generated throughout the oil production process (leaf, "alpeorujo", liquid and solid residues generated during decanting commonly named "borras" and washing water) were analyzed. For this purpose, different optimized extraction procedures were performed for each matrix, followed by high-performance liquid chromatography coupled with electrospray time-of-flight mass spectrometry (HPLC-ESI-TOF/MS) analysis. Although no phenolic alcohols were quantified in the leaf and the presence of secoiridoids was low, this by-product was notable for its flavonoid (720 ± 20 µg/g) and terpene (5000 ± 300 µg/g) contents. "Alpeorujo" presented a complete profile of compounds of interest, being abundant in phenolic alcohols (900 ± 100 µg/g), secoiridoids (4500 ± 500 µg/g) and terpenes (1200 ± 100 µg/g), among others. On the other hand, while the solid residue of the borras was the most abundant in phenolic alcohols (3700 ± 200 µg/g) and secoiridoids (680 ± 20 µg/g), the liquid fraction of this waste was notable for its content of elenolic acid derivatives (1700 ± 100 µg/mL) and phenolic alcohols (3000 ± 300 µg/mL). Furthermore, to our knowledge, this is the first time that the terpene content of this by-product has been monitored, demonstrating that it is an important source of these compounds, especially maslinic acid (120 ± 20 µg/g). Finally, the phytochemical content in wash water was lower than expected, and only elenolic acid derivatives were detected (6 ± 1 µg/mL). The results highlighted the potential of the olive by-products as possible alternative sources of a wide variety of olive bioactive compounds for their revalorization into value-added products.

Metabolomics-based analysis of the diatom Cheatoceros tenuissimus combining NMR and GC–MS techniques

Metabolomics, a recent addition to omics sciences, studies small molecules across plants, animals, humans, and marine organisms. Nuclear magnetic resonance (NMR) and gas chromatography-mass spectrometry (GC–MS) are widely used in those studies, including microalgae metabolomics. NMR is non-destructive and highly reproducible but has limited sensitivity, which could be supplemented by joining GC–MS analysis. Extracting metabolites from macromolecules requires optimization for trustworthy results. Different extraction methods yield distinct profiles, emphasizing the need for optimization. The results indicated that the optimized extraction procedure successfully identified NMR and GC–MS-based metabolites in MeOH, CHCl3, and H2O extraction solvents. The findings represented the spectral information related to carbohydrates, organic molecules, and amino acids from the water-soluble metabolites fraction and a series of fatty acid chains, lipids, and sterols from the lipid fraction. Our study underscores the benefit of combining NMR and GC–MS techniques to comprehensively understand microalgae metabolomes, including high and low metabolite concentrations and abundances.•In this study, we focused on optimizing the extraction procedure and combining NMR and GC–MS techniques to overcome the low NMR sensitivity and the different detected range limits of NMR and GC–MS.•We explored metabolome diversity in a tropical strain of the small cells’ diatom Cheatoceros tenuissimus.

Box–Behnken Design to Optimize Standardized Mangiferin-Rich Mango Peel Extract from Agro-Industrial Waste Product

The light mango or “Ma-Muang Bao” (Mangifere indica L. var.) is a native mango species originating from Malaysia and southern Thailand. The whole Ma-Muang Bao fruit, except peels, is popular as both a raw and ripe fruit for consumption, as well as being used in various processed food products. This study aims to transform the peel of a specific mango variety, which is a byproduct of the agro-industrial sweet, pickled mangoes industry, into a valuable bioactive ingredient for healthcare products. This is achieved through the establishment of a standardized mangiferin-rich mango peel extract (SMPE). Employing the Box–Behnken design (BBD) within the framework of response surface methodology (RSM), an optimal microwave-assisted extraction procedure was developed. A total of 27 experiments, each with four independent variables, including solvent ratio, extraction power, extraction time, and ethanol (EtOH) ratio, were conducted to optimize the extraction method in terms of mangiferin content and extraction yield. The optimized extraction conditions encompassed a solvent ratio of 120 mL EtOH/100 g sample, an extraction power of 450 W, an extraction time of approximately 4.3 min, and an EtOH ratio of 69.44% (EtOH in water). Small-scale extractions were carried out using the following specified parameters: solvent ratio of 120 mL, extraction power of 450 W, extraction time of 4 min, and EtOH ratio of 70% EtOH. These extractions yielded an extract with a mangiferin content of 27.24 ± 2.05 mg/g and an extraction yield of 3.71 ± 0.17% w/w. Notably, these outcomes were better from the mangiferin content of 19.62 mg/g and a yield of fresh peel of 5.61% estimated through BBD analysis. Furthermore, a pilot-scale extraction was performed using 7 kg of fresh mango peel and 70% EtOH (8.4 L) for 4 min, resulting in an extract with a mangiferin content of 51.85 ± 0.35 mg/g and a fresh peel yield of 4.35% w/w. This method emerges as the most suitable for mango peel extraction and forms the basis of the SMPE. The results from biological activities highlight the potential use of SMPE as the active ingredient for cosmeceutical or healthcare products for wound-healing and skin-brightening agents. Additionally, the knowledge from this study presents an alternative approach to various plant sources and sustainable extraction methods for the herbal extract industry.

Linseed mucilage - optimized extraction procedure for genomic research and nutraceutical applications

Linseed mucilage - optimized extraction procedure for genomic research and nutraceutical applications

Multi-component analyses of raspberry: Optimization of extraction procedure and network pharmacology

The contents of ellagic acid and kaempferol-3-O-rutinoside, the chief active components of raspberry, are considered the quality control indices of raspberry. This work employed the ant colony neural network (ACO-BPNN) to optimize their extraction processes, and the combination of network pharmacology and molecular docking technology to unveil the potential pharmacological effects of these components. Based on the single-factor test (ultrasonic time, ethanol concentration, ultrasonic temperature, and solid-liquid ratio), a factorial experiment with 4-factors and 3-levels was conducted in parallel for 3 times. The multi-factor analysis of variance results revealed high-order interactions among the factors. Then, the ACO-BPNN model was established to characterize the complex relationship of experimental data. After further verification, relative errors were all less than 8 %, implying the model's effectiveness and reliability. Moreover, with the network pharmacology, 66 key targets were screened out and mainly concentrated in PI3K-AKT, MAPK, and Ras signal pathways. Molecular docking revealed the binding sites between active components and key targets.

Wheat germ, a byproduct of the wheat milling industry, as a beneficial source of anti-aging polyamines: A quantitative comparison of various forms.

Polyamines have received a lot of attention since the 1990s because of their anti-aging, anti-chronic disease, and proliferative effects. Wheat germ was reported as one of the natural sources of high polyamine, especially spermidine. The current study used three types of wheat germ: group A was industrially separated germ from whole grain, group B was the commercially available germinated wheat germ, and group C was manually separated wheat germ from germinated grain. The polyamine content of putrescine, spermidine, and spermine has been determined using a simplified isocratic LC-MS/MS method. An optimized extraction procedure was performed on all seven samples for obtaining a polyamine-enriched extract. The three dominant carbomylated polyamines were identified by analyzing the extracted samples in order to determine their relative abundance. Wheat germ powders contain the highest amount of polyamines (220-337 μg/g) of which spermidine is one of the most important. Germinated wheat grains, on the other hand, contain the least amount of this polyamine. The commercially available separated wheat germs are suggested as a good nutrition source of these polyamines.

Optimization of Phytochemical-Rich Citrus maxima Albedo Extract Using Response Surface Methodology.

In the present study, response surface methodology (RSM) and Box-Behnken design (BBD) were employed to optimize the conditions for the extraction of C. maxima albedo from agricultural waste, to obtain notable phytochemicals. Ethanol concentration, extraction temperature, and extraction time were included as key factors contributing to the extraction. The results showed that the optimum extraction condition for C. maxima albedo was 50% (v/v) aqueous ethanol at 30 °C for 4 h, which provided total phenolic contents and total flavonoid contents at 15.79 mg of gallic equivalent/g dry weight (DW) and 4.50 mg of quercetin equivalent/g DW, respectively. Considerable amounts of hesperidin and naringenin at 161.03 and 3430.41 µg/g DW, respectively, were detected in the optimized extract using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). The extract was later subjected to a test for its enzyme-inhibitory activities against key enzymes relevant to Alzheimer's disease (AD), obesity, and diabetes as well as for its mutagenicity potential. Among enzyme inhibitory activities, the extract showed the highest inhibitory strength against β-secretase (BACE-1), which is a drug target for AD treatment. The extract was also devoid of mutagenicity properties. Overall, this study demonstrated a simple and optimal extraction procedure for C. maxima albedo with a significant quantity of phytochemicals, health benefits, and genome safety.

Optimization of extraction procedure and antioxidant activity of C50 carotenoids from Halorubrum sp. HRM-150

C50 carotenoids are promising bioactive compounds produced by halophilic archaea. Relevant studies have focused on their outstanding antioxidant activity, whereas the extraction process has received less attention. In this study, the effect of factors on carotenoid extraction efficiency was explored through single-factor and orthogonal experiments. The results showed that the C50 carotenoids were effectively extracted by suspending and homogenizing cell pellets in methanol (1:30, w-v) for 10 min, subsequently incubating at 30 °C for 30 min in the dark, finally centrifuging and drying over vacuum rotary evaporation at 30 °C in the dark. The carotenoid content increased 1.84 times by the above procedure than that before optimization. Moreover, the UVVis spectra, TLC and HPLCMS analysis revealed that the main component of carotenoids extracted from Halorubrum sp. HRM-150 was bacterioruberin (84.12 %), followed by monoanhydrobacterioruberin (15.13 %). Both carotenoid extract and bacterioruberin exhibited antioxidant capacity, which were significantly higher than that of astaxanthin (p < 0.05) by scavenging assays of 2,2-azinobis-3-ethyl-benzothiazole-6-sulfonic acid, 2,2-diphenyl-1-picrylhydrazyl and hydroxyl radicals. In addition, the hydroxyl radical scavenging rate of bacterioruberin was significantly higher than that of carotenoid extract (p < 0.01), indicating its potential activity against or delaying diseases caused by reactive oxygen species. C50 carotenoids (especially BR) have great application prospects in human health due to their excellent antioxidant capacity.

New Trend in Toxicological Screening Using Volumetric Absorptive Microsampling (VAMS) and High-Resolution Mass Spectrometry (HR/MS) Combination.

In toxicology, screenings are routinely performed using chromatographic methods coupled to detection systems such as high-resolution mass spectrometry (HR/MS). The increase in specificity and sensitivity of HRMS is responsible for the development of methods for alternative samples such as Volumetric Adsorptive Micro-Sampling. Whole blood overloaded with 90 drugs was sampled with 20 µL MitraTM to optimize the pre-analytical step as well as to determine the identification limits of drugs. Elution of chemicals was carried out in a solvent mixture through agitation and sonication. After dissolution, 10 μL was injected into the chromatographic system coupled to the OrbitrapTM HR/MS. Compounds were confirmed against the laboratory library. The clinical feasibility was assessed in fifteen poisoned patients using the simultaneous sampling of plasma, whole blood and MitraTM. The optimized extraction procedure allowed us to confirm 87 compounds out of the 90 present in the spiked whole blood. Cannabis derivatives were not detected. For 82.2% of the investigated drugs, the identification limits were below 12.5 ng·mL-1, with the extraction yields ranging from 80.6 to 108.7%. Regarding the patients' analysis, 98% of the compounds in plasma were detected in MitraTM compared to whole blood, with a satisfying concordance (R2 = 0.827). Our novel screening approach opens new insights into different toxicologic fields appropriate for pediatrics, forensics or to perform mass screening.

Discovery, disassembly, depolymerization and derivatization of catechyl lignin in Chinese tallow seed coats

The search for feedstock of catechyl lignin (C-lignin) is great interest and importance, as C-lignin featuring homogeneity and linearity is considered as an “ideal lignin” archetype for valorization and exits in only a few plant seed coats. In this study, naturally occurring C-lignin is first discovered in the seed coats of Chinese tallow, which has the highest content of C-lignin (15.4 wt%) as compared with other known feedstocks. An optimized extraction procedure by ternary deep eutectic solvents (DESs) enables the complete disassembly of C-lignin and G/S-lignin coexisted in Chinese tallow seed coats, and characterizations revealed that the as-separated C-lignin sample is abundant in benzodioxane units with no observation of β-O-4 structures from G/S-lignin. Catalytic depolymerization of C-lignin results in a simplex catechol product in 129 mg per gram seed coats, being higher than other reported feedstocks. Derivatizing the “black” C-lignin via the nucleophilic isocyanation of benzodioxane γ-OH leads to a “whitened C-lignin” with uniform laminar structure and excellent crystallization ability, being conducive to fabricating functional materials. Overall, this contribution showed that Chinses tallow seed coats are suitable feedstock for acquiring C-lignin biopolymer.

Analysis of NSAIDs in Rat Plasma Using 3D-Printed Sorbents by LC-MS/MS: An Approach to Pre-Clinical Pharmacokinetic Studies

Analytical sample preparation techniques are essential for assessing chemicals in various biological matrices. The development of extraction techniques is a modern trend in the bioanalytical sciences. We fabricated customized filaments using hot-melt extrusion techniques followed by fused filament fabrication-mediated 3D printing technology to rapidly prototype sorbents that extract non-steroidal anti-inflammatory drugs from rat plasma for determining pharmacokinetic profiles. The filament was prototyped as a 3D-printed sorbent for extracting small molecules using AffinisolTM, polyvinyl alcohol, and triethyl citrate. The optimized extraction procedure and parameters influencing the sorbent extraction were systematically investigated by the validated LC-MS/MS method. Furthermore, a bioanalytical method was successfully implemented after oral administration to determine the pharmacokinetic profiles of indomethacin and acetaminophen in rat plasma. The Cmax was found to be 0.33 ± 0.04 µg/mL and 27.27 ± 9.9 µg/mL for indomethacin and acetaminophen, respectively, at the maximum time (Tmax) (h) of 0.5-1 h. The mean area under the curve (AUC0-t) for indomethacin was 0.93 ± 0.17 µg h/mL, and for acetaminophen was 32.33± 10.8 µg h/mL. Owing to their newly customizable size and shape, 3D-printed sorbents have opened new opportunities for extracting small molecules from biological matrices in preclinical studies.

Speciation analysis of iodine in seaweed: optimisation of extraction procedure and chromatographic separation

Environmental context Seaweed is a good natural dietary source of iodine and some types of seaweed are rich in iodine. Iodine has a diverse chemistry in seaweeds and may exist as different chemical species; however, the occurrence and identity of the individual species are still not fully elucidated. Hence, development of sensitive and selective iodine speciation methods for studies of iodine chemistry and biotransformation in seaweeds are needed.Rationale Iodine is an essential element required for human health and metabolism. Seafood and especially seaweed can accumulate iodine to high amounts. Iodine may exist in different chemical forms (species) in seaweeds.Methodology The present study describes the development and optimisation of a method for iodine speciation analysis in seaweed based on high performance liquid chromatography–inductively coupled plasma–mass spectrometry (HPLC-ICP-MS). The extraction procedure was conducted in two steps, pancreatic enzymatic extraction followed by alkaline extraction with tetramethylammonium hydroxide for optimum extraction efficiency without compromising species integrity.Results and discussion Total iodine and iodine species were determined in a range of brown (6 samples), red (6 samples) and green (3 samples) seaweeds. A large variation in the total iodine content of the different seaweeds was observed (33–5611 µg g–1 dry weight) with the highest levels encountered in brown seaweed. Iodine speciation analysis revealed differences in the speciation profile of the different types of seaweed. In all seaweeds iodide was the predominant species, and minor contents of MIT (monoiodotyrosine) and DIT (diiodotyrosine) were found in most seaweeds. Furthermore, peaks originating from six unknown iodine-containing species were observed in the chromatograms, especially in red and green seaweeds, while less abundant in brown seaweeds. The speciation method presented here will be valuable in future studies on iodine speciation in seaweed and an important tool for the investigation of iodine speciation and biotransformation in marine algae.

Metabolic fate of black raspberry polyphenols in association with gut microbiota of different origins in vitro

Polyphenols, ubiquitous in plant-originated foods, exhibit multifarious health benefits but are notorious for their low bioavailability. As the majority of polyphenols enter the colon where they are metabolized into the more bioavailable metabolites with multifarious bioactivities, gut microbiota composition is critical for realizing the health benefits of polyphenols. Black raspberries (BRB) are highly abundant in health-promoting but poorly absorbable polyphenols, primarily ellagitannins and anthocyanins. This study aimed to dissect the bilateral interactions between polyphenols and gut microbiotas of diverse origins. Using an optimized extraction procedure, BRB polyphenols were first comprehensively characterized by UHPLC-QTOF-MS/MS. Next, in vitro anaerobic fermentations with microbiota from human adults, infants, rats, and mice were conducted for comparison of phenolic profile (by metabolomics) and microbiota composition (by 16S rDNA sequencing) before and after fermentation. Bioinformatics analysis further revealed unique polyphenol-microbiota interactions and identified several bacterial species that could potentially produce bioactive and bioavailable phenolic metabolites following BRB consumption.

A New HPLC-MS/MS Method for the Simultaneous Determination of Quercetin and Its Derivatives in Green Coffee Beans.

Green coffee (Coffee arabica and Coffee robusta) is one of the most commonly traded goods globally. Their beans are enriched with polyphenols and numerous health benefits are associated with their consumption. The main aim of this work was to develop a new and fast analytical HPLC-MS/MS method to simultaneously determine six flavonoid polyphenolic compounds (quercetin, rutin, isorhamnetin, quercetin-3-glucouronide, hyperoside, and quercitrin) in 22 green coffee samples from six different geographical origins (Ethiopia, Brazil, Guatemala, Nicaragua, India and Colombia). In addition, by adjusting pH, temperature, solvent type, and extraction duration, several extraction methods such as acidic and alkaline hydrolysis, and extraction without hydrolysis were evaluated. The optimal extraction procedure in terms of recovery percentages (78.67–94.09%)was acidic hydrolysis at pH 2, extraction temperature of 60 °C, extraction solvent of 70% ethanol, and extraction duration of 1.5 h. Hyperoside (878–75 μg/kg) was the most abundant compound followed by quercitrin (408–38 μg/kg), quercetin (300–36 μg/kg), rutin (238–21 μg/kg), and quercetin-3-glucouronide (225–7 μg/kg), while isorhamnetin (34–3 μg/kg) showed the lowest amount. Overall, green coffee beans are rich in flavonoid polyphenolic compounds and could be used as part of a healthy diet.

Carotenoid-Producing Yeasts: Selection of the Best-Performing Strain and the Total Carotenoid Extraction Procedure

Yeasts are considered an extraordinary alternative source of natural carotenoids and pigmented terpenoids with multiple applications. Production of carotenoids by yeast fermentation technology has many benefits; it is cost-effective, easily scalable, and safe. The aim of this research is the isolation of yeasts from natural resources and selection of the most potent bioagent for carotenoid production. Additionally, an upgraded carotenoid extraction protocol we established, which implies the testing of four methods for cell lysis (hydrochloric acid treatment, ultrasound treatment, milling treatment, and osmotic pressure treatment), three extraction methods (conventional extraction, ultrasound extraction, and conventional + ultrasound extraction), and three extraction solvents (acetone, isopropanol/methanol (50:50), and ethanol). For the first time, the obtained results were further modeled by an artificial neural network (ANN). Based on the obtained maximal carotenoid yield (253.74 ± 9.74 mg/100 g d.w) for the best-performing Rhodotorula mucilaginosa, the optimized extraction procedure involving milling treatment (for cell lysis) and conventional extraction with acetone (for carotenoid extraction) convincingly stood out compared to the other 35 tested protocols. Therefore, the selected carotenoid extraction protocol was verified with respect to its universality for all other yeast isolates, demonstrating its simplicity and effectiveness.

Multi-panel detection of drugs and drug metabolites in hair samples using a comprehensive extraction method

The aim of this presentation is to demonstrate a robust and sensitive LC-MS/MS-based method for the detection of a wide range of drugs and drug metabolites in hair. The presentation will demonstrate that the use of an optimize extraction method enabled accurate detection and sensitive quantification of a panel of 77 compounds including novel psychoactive substance (NPS) and metabolites with a wide range of physical and chemical properties. A total of 77 compounds were included in the novel psychoactive substance (NPS) panel. The drugs of abuse (DOA) panel consisted of 23 compounds and the third panel consisted of Ethyl glucuronide (EtG) only. Hair samples were washed thoroughly with methanol for 5 minutes then diethyl ether for another 5 minutes, dried and spiked with the various drug calibrator mixtures. 500 μL of a proprietary aqueous extracting solution was added to the spiked hair samples, gently vortexed, centrifuged at 5000 rpm for 1 minute and incubated for 60 minutes at 100 °C. Following the incubation step, the liquid extract was transferred to a new tube and centrifuged at 5000 rpm for 5 minutes. The organic phase was transferred to a sample vial for analysis. Analytes were chromatographically separated at 50 °C using a Phenomenex Synergi 2.5 μm Hydro-RP column (50 × 3 mm, 2.5 μm, P/N 00D-4387-Y0). Mobile phases used consisted of HPLC grade water with 0.1% formic acid (mobile phase A) and acetonitrile with 0.1% formic acid (mobile phase B). Three separate gradients, flow rates and injection volumes were were used for each of the LC methods. MS data were collected in positive mode on the SCIEX QTRAP 6500+ System. Three separate acquisition methods were created using the Scheduled multiple reaction monitoring (MRM) Algorithm. The acquisition methods consisted of 156 MRM transitions for the NPS panel, 71 MRM transitions for the DOA panel and 3 MRM transitions for the EtG experiment, respectively. The calibration curves resulting from the NPS panel demonstrated excellent linearity for all the analytes included in the panel with R2 values > 0.98. The robustness of the NPS screening method was further investigated by analyzing 10 authentic hair samples in which two synthetic cannabinoids JWH-122 and AM-2201 were detected in an authentic case sample at concentrations of 111.8 and 11.72 pg/mg, respectively. Ketamine, oxycodone, tramadol and zolpidem were detected in another authentic case sample at concentrations of 0.39, 1602.5, 248.4 and 0.04 ng/mg, respectively. The results demonstrate that this robust method enable sensitive quantification of a variety of drugs and metabolites from authentic case samples. A comprehensive workflow combining the sensitivity of the SCIEX QTRAP 6500+ LC-MS/MS System with a simple and optimized extraction procedure enabled detection of a wide variety of drugs and drug metabolites in hair samples. The developed workflow was shown to enable accurate identification and sensitive quantification of a wide range of chemically diverse analytes included in three different panels. A number of analytes were accurately detected in authentic hair samples, demonstrating the broad applicability of the sample preparation procedure for the detection of the analytes included in the panel.

Immunoassay for Natamycin Trace Screening: Bread, Wine and Other Edibles Analysis.

The antifungal drug natamycin (NAT) is widely used in medicine and in the food industry as preservative E235 for a wide variety of foods. The risk of the development of resistance to NAT and its spread in relation to other polyene antibiotics is fraught with the emergence of incurable infections. This work is devoted to the development of an immunoassay to investigate the prevalence of NAT use for food preservation. Two immunogen designs based on tetanus toxoid, conjugated to NAT through different sites of hapten molecules, were compared in antibody generation. Assay formats using heterologous coating antigens were superior for both antibodies. The ELISA variant demonstrated the highest sensitivity (IC50 = 0.12 ng/mL), and a limit of detection of 0.02 ng/mL was selected for NAT determination. The optimized extraction procedure provided a recovery rate of 72–106% for various food matrixes with variations below 12%. Cyclodextrins, as well as NAT–cyclodextrin complex formulations, showed no interference with the quantification of NAT. One hundred and six food product brands, including baked goods, wines, beers, drinks, sauces, and yogurts, were tested to assess the prevalence of the undeclared use of NAT as a preservative. The screening examination revealed three positive yogurts with an undeclared NAT incorporation of 1.1–9.3 mg/kg.

Carbon dioxide expanded liquid: an effective solvent for the extraction of quercetin from South African medicinal plants

BackgroundQuercetin is one of the most important bioflavonoids having positive effects on the biological processes and human health. Typically, it is extracted from plant matrices using conventional methods such as maceration, sonication, infusion, and Soxhlet extraction with high solvent consumption. Our study aimed to optimize the environmentally friendly carbon dioxide-based method for the extraction of quercetin from quince fruit with an emphasis on extraction yield, repeatability, and short extraction time.ResultsA two-step design of experiments was used for the optimization of the key parameters affecting physicochemical properties, including CO2/co-solvent ratio, co-solvent type, temperature, and pressure. Finally, gas expanded liquid combining CO2/ethanol/H2O in a ratio of 10/81/9 (v/v/v) provided the best extraction yield. Extraction temperature 66 °C and pressure 22.3 MPa were the most suitable conditions after careful optimization, although both parameters did not significantly affect the process. It was confirmed by experiments in various pressure and temperature conditions and statistical comparison of obtained data. The optimized extraction procedure at a flow rate of 3 mL/min took 30 min. The repeatability of the extraction method exhibited an RSD of 20.8%.ConclusionsThe optimized procedure enabled very fast extraction in 30 min using environmentally friendly solvents and it was successfully applied to 16 different plant samples, including 14 bulbs and 2 fruits from South Africa. The quercetin content in extracts was quantified using ultra-high performance liquid chromatography (UHPLC) with tandem mass spectrometry. UHPLC hyphenated with high-resolution mass spectrometry was used to confirm chemical identity of quercetin in the analyzed samples. We quantified quercetin in 11 samples of all 16 tested plants. The quercetin was found in Agapanthus praecox from the Amaryllidaceae family and its presence in this specie was reported for the first time.Graphical

Approaches for the Elimination of Microbial Contaminants from Lippia multiflora Mold. Leaves Intended for Tea Bagging and Evaluation of Formulation.

Elimination of microorganisms from herbal products has been a major concern due to its implicated health risk to consumers. Drying of herbal materials has been employed for centuries to reduce the risk of contamination and spoilage. The present study adopted three drying approaches in an attempt to eliminate microorganisms from Lippia multiflora tea bag formulation. This study also evaluated the tea bags and optimized the extraction procedure. The L. multiflora leaves for tea bagging were air-dried and milled (A), oven-dried and milled (B), and microwaved (the milled air-dried leaves) (C). The moisture contents were determined at 105°C ± 2°C for 2 hours to constant weight. Phytochemical parameters such as phytochemical constituents, total water extractive, and pH were assessed. The microbial safety and quality of the L. multiflora tea bags were evaluated using the British Pharmacopoeia 2019 specifications. The uniformity of the mass of the formulated tea bags was also determined. Extraction from the Lippia tea bags was optimized. The results showed that using the approaches (A, B, and C) adopted for drying and processing, the moisture contents of the formulated tea bags were in the range of 9.75–10.71% w/w. All the formulated tea bags contained reducing sugars, phenolic compounds, polyuronides, flavonoids, anthracenosides, alkaloids, saponins, and phytosterols. The pH range of the formulations was 7.11–7.54, whereas the total water extractive values were in the range of 19.12–20.41% w/w. The one-way analysis of variance demonstrated no significant difference in the data obtained from the results from A, B, and C. The formulation from A was found to be unsafe for consumption due to unacceptable microbial contamination limits. Microbial load of the formulations from B and C were within the BP specifications. All the batches of the formulations passed the uniformity of mass test. An optimized extraction procedure was obtained when one tea bag was extracted in 250 mL of hot water within the specified time. L. multiflora leaves meant for tea bagging should be oven-dried or microwaved before tea bagging for safe consumption.

Optimization for ultrasonic-microwave synergetic extraction of total iridoid glycosides and screening of analgesic and anti-inflammatory active fractions from patrinia scabra Bunge (Valerianaceae)

BackgroundPatrinia scabra Bunge is a well-known herbal medicine for its favorable treatment on inflammatory diseases owing to its effective ingredients, in which iridoid glycoside plays an extremely significant role. This article aimed to improve the content of total iridoid glycosides in crude extract through a series optimization of extraction procedure. Moreover, considering that both pain and inflammation are two correlated responses triggered in response to injury, irritants or pathogen, the article investigated the anti-inflammatory and analgesic activities of P. scabra to screen out the active fraction.MethodP. scabra was extracted by ultrasonic-microwave synergistic extraction (UMSE) to obtain total iridoid glycosides (PSI), during which a series of conditions were investigated based on single-factor experiments. The extraction process was further optimized by a reliable statistical method of response surface methodology (RSM). The elution fractions of P. scabra extract were prepared by macroporous resin column chromatography. Through the various animal experiment including acetic acid-induced writhing test, formalin induced licking and flinching, carrageenan-induced mice paw oedema test and xylene-induced ear edema in mice, the active fractions with favorable analgesic and anti-inflammatory effect were reasonably screen out.ResultsThe content of PSI could reach up to 81.42 ± 0.31 mg/g under the optimum conditions as follows: ethanol concentration of 52%, material-to-liquid ratio of 1:18 g/mL, microwave power at 610 W and extraction time of 45 min. After gradient elution by the macroporous resin, the content of PSI increased significantly. Compared with other concentrations of elution liquid, the content of PSI in 30 and 50% ethanol eluate was increased to reach 497.65 and 506.90 mg/g, respectively. Owing to the pharmacology experiment, it was reasonably revealed that 30 and 50% ethanol elution fractions of P. scabra could relieve pain centrally and peripherally, exhibiting good analgesic and anti-inflammatory activities.ConclusionPatrinia scabra possessed rich iridoids and exhibited significant analgesic and anti-inflammatory activities.