Extraction Purification Research Articles

Decolorizing of seaweed extract by electrocoagulation

Electrocoagulation (EC) is a technique commonly used in wastewater treatment to remove biological and chemical contaminants, but the process has the potential to be used in clarifying plant extracts for the isolation and identification of secondary metabolites. Seaweed extracts contain copious amounts of chlorophyll and other pigments that obscure the characterization of secondary metabolites such as phenolic acids and flavonoids. In place of conventional methods that utilize solvents, EC can potentially be applied to clarify and fractionate extracts. In this research, an EC duration of 30 min (22 V, 0.3–0.5A) with aluminum electrodes resulted in a significant decrease, about 76%, of chlorophyll and 70% of carotenoids from seaweed extract measured at 666 nm and 410 nm. The decrease in extract green and yellow color intensity also mirrored a decrease in total phenolic content (TPC) of the extract from 54 ± 1.55 mg GAE/g DW to 3.2 ± 0.01 mg GAE/g DW after 30 min of EC. However, the phenolic acid profile of the extract after electrocoagulation via HPLC-RP indicated the removal of an interference probably caused by polymeric compounds from the extract, thus leaving the simple phenolic acids in solution for detection. The major phenolic acids detected in seaweed crude extract were p-coumaric, o-coumaric, ferulic and syringic acid. Flavonoids detected included catechin, epicatechin, quercetin-3-glucoside and rutin. The results of this study show the potential of replacing conventional plant extract purification methods with a green method that requires no additional solvent.Graphical

Optimized Production of Concanamycins Using a Rational Metabolic Engineering Strategy

Plecomacrolides, such as concanamycin and bafilomycin, are potent and specific inhibitors of vacuolar-type ATPase. Concanamycins are 18-membered macrolides with promising therapeutic potential against multiple diseases, including viral infection, osteoporosis, and cancer. Due to the complexity of their total synthesis, the production of concanamycins is only achieved through microbial fermentation. However, the low titers of concanamycin A and its analogs in the native producing strains are a significant bottleneck for scale-up, robust structure-activity relationship studies, and drug development. To address this challenge, we designed a library of engineered Streptomyces strains for the overproduction of concanamycins by combining the overexpression of target regulatory genes with the optimization of fermentation media. Integration of two endogenous regulators from the concanamycin biosynthetic gene cluster (cms) and one heterologous regulatory gene from the bafilomycin biosynthetic gene cluster into the attB site significantly increased production of concanamycin A and its low abundant analog concanamycin B in Streptomyces eitanensis. The highest titers reported to date were observed in the engineered S. eitanensis DHS10676, which produced over 900 mg/L of concanamycin A and 300 mg/L of concanamycin B. Heterologous overexpression of the identified target regulatory genes across a panel of Streptomyces spp., harboring a putative concanamycin biosynthetic gene cluster confirmed its identity, and significantly improved concanamycin A production in all tested strains. Strain engineering, optimization of fermentation, and extraction purification protocols enabled swift access to these structurally complex plecomacrolides for semi-synthetic medicinal chemistry-based approaches. Together, this work established a platform for robust overproduction of concanamycin analogs across species.

Modulation of Proinflammatory Cytokines by Parmotrema reticulatum Derived Dioxepine Derivative and Benzoic Methyl Ester in Adjuvant-Induced Arthritis.

The present investigation aims to study the therapeutic effect and to identify the lead molecules from lichen Parmotrema reticulatum (Pr) that can solve the complications associated with arthritis. Purification of Pr extract led to isolation of two lead molecules i.e. Compound A (a Dioxepine derivative) as 3-Hydroxy-9-hydroxymethyl-1,8-dimethyl-11-oxo-11H-dibenzo[b,e][1,4]dioxepine-4,7 dicarboxylic acid 7-methyl ester with molecular formula C19H16O9; molecular weight 388.34. Compound B as 3-Hydroxy-5-methoxy-4-methyl-benzoic acid (a Benzoic methyl ester) with molecular formula C9H10O4; molecular weight 182.2. In-silico investigation specific for inflammation revealed good binding affinity of both the compounds with the targeted proteins. Further continuous administration of Pr and novel compound A & B to CFA (Freund's complete adjuvant) induced arthritis rat revealed reduction in arthritis complication possibly by inhibiting the formation of edema. The variations in relative body weight along with paw swelling and arthritic scores were identified. Inhibition of expressions of RA markers like RF, CRP, TNF-𝛼, IL-1𝛽, IL-6, ALP, AST, ALT and LDH were observed there by inhibiting inflammation of synovial cavity and cartilage damage effectively. Hence from our results it is evident that Pr, Compound A & B has down regulated the inflammatory cytokines and can be a potential candidate to treat arthritis.

Screening Anti-Rheumatoid Arthritis Synovitis Effective Ingredients of Total Flavonoid From Artemisia argyi Folium Based on Spectrum-Effect Relationship.

Flavonoids are the main nonvolatile component responsible for the anti-rheumatoid arthritis (RA) synovitis activities of Artemisia argyi Folium. However, the effective ingredient remains unidentified. Spectrum-effect relationships analysis was a reliable and efficient strategy for herbal effective ingredient discovery. This study aimed to screen the effective ingredients within the total flavonoid from Artemisia argyi Folium (TFAA) that exhibit anti-RA synovitis activities based on spectrum-effect relationship. TFAA was obtained through ethanol extraction and subsequent purification with D101 resin from 15 distinct batches of Artemisia argyi Folium. The fingerprint of TFAA was established using HPLC, and its efficacy against RA synovitis was evaluated by determining the inhibition rate of nitric oxide (NO) on MH7A synovioblast induced by TNF-α. Common peaks were identified using HPLC-MS/MS and authentic standards. The spectrum-effect relationships between the fingerprints and efficacy were analyzed by gray relational analysis (GRA), canonical correlation analysis (CCA), and partial least squares regression analysis (PLSR) to pinpoint the peaks responsible for the anti-RA synovitis activity, and the results were further verified by invitro anti-RA synovitis experiments and molecular docking studies. The fingerprint revealed 14 common peaks, and 12 compounds were identified, including four caffeoylquinic acids and eight flavonoids. Among them, five flavonoids-X10 (hispidulin), X11 (jaceosidin), X12 (centaureidin), X13 (eupatilin), and X14 (casticin)-were highly relevant to anti-RA synovitis activity. Verification experiments confirmed their inhibitory effect on NO production and cytokine secretion in MH7A cells, showing anti-RA synovitis potential, which was consistent with the established spectrum effect relationship. The underlying mechanism might be related to the inhibition of iNOS. Hispidulin, jaceosidin, centaureidin, eupatilin, and casticin were the key effective ingredient of TFAA responsible for its anti-RA synovitis effects. These compounds can serve as quality control markers for Artemisia argyi Folium and as lead compounds for anti-RA synovitis treatment.

Preparation of polyacylated anthocyanins of red radish and enhancing its stability and bioaccessibility through encapsulating within double-coated yeast glucan particles

Abstract Although the anthocyanins of red radishes (ARR) rich in polyacylated pelargonidin glucosides are used as commercial food pigment, they are unstable. We found that the anthocyanin purity of the crude ARR extract can be rapidly increased by 5 times using D101 macroporous resin (mass ratio of extract:resin = 1:2.2). Capsule constructed by chitosan (CTS), pectin (PT) and yeast glucan particles (YGP) presented a high efficiency to encapsulate ARR. Liquid chromatography-mass spectrometry analysis verified that polyacylated pelargonidin glucosides in ARR were packed into the capsule. Microstructure observation and Fourier transforms infrared spectroscopy further confirmed the encapsulated structure. Co-encapsulation of CTS, PT and YGP showed effective protection for ARR against heat, oxygen, ascorbic acid, and physiological pH. This encapsulation also significantly improved the gastric and intestinal bioaccessibilities of ARR. These results suggested that the triplex-coated YGPs might be a promising strategy to protect and deliver polyacylated anthocyanin.

Efficient Simultaneous Isolation of Pinostrobin and Panduratin A from Boesenbergia rotunda Using Centrifugal Partition Chromatography.

The bioactive flavonoids pinostrobin (PN) and panduratin A (PA) from Boesenbergia rotunda are essential for research and therapeutic applications. This study introduces an innovative method utilizing ultrasound-assisted extraction with n-hexane pre-treatment, followed by one-step centrifugal partition chromatography (CPC) purification. Extraction efficiency was evaluated using ultra high-performance liquid chromatography (UHPLC), and the isolated compounds were characterized through 1H-NMR and liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS), adhering to AOAC validation guidelines. Optimal extraction conditions comprised a particle size of 125 μm, a solid-to-liquid ratio of 1:30 g/mL, and a 10 min extraction time, yielding a crude extract of 6.96 ± 0.07%. Using an n-hexane/MeOH/water (5/3.4/1.6, v/v) solvent system in ascending mode, PN (2.16 mg, 98.78% purity) and PA (0.4 mg, 99.69% purity) were isolated from 67 mg of crude extract within 30 min. This streamlined approach enhances purification efficiency, allowing for faster extraction and higher purity, making it a suitable method for commercial applications.

Do commercial dog extracts cross-react with Felis domesticus allergen 1.

Background: Half of U.S. households own a dog despite 10% of individuals being sensitized to dog. Assessment and treatment options for dog allergy include the use of commercial dog extracts which have inconsistent performance, making diagnosing and managing dog allergy challenging. Contamination of dog extracts with other allergens has previously been reported. Objective: We sought to determine whether commercial dog extracts contain other aeroallergens. Methods: An extract purity and quantification study on acetone precipitated dog hair and dander extract (AP dog) was performed, 6 aeroallergens; Alternaria (Alt a 1), Ragweed (Amb a 1), German Cockroach (Bla g 2), Dust Mite (Der p t), Cat (Fel d 1), and Rye Grass (Lol p 1). Following, conventional dog hair and dander extract (CV dog) and the new ultrafiltered dog hair and dander extract (UF dog) were also assessed based on the initial results of AP dog. SDS-PAGE was performed on all three dog extracts to compare allergen content. Lastly, serology results and aeroallergen immunotherapy prescriptions were compared. Results: The ELISA trays with Alt a 1, Amb a 1, Bla g 2, Der p 1, and Lol p 1 antibodies did not capture AP dog, while the ELISA tray with Fel d 1 antibody captured AP dog, CV dog, and UF dog. SDS-PAGE results of the 3 dog extracts did not reveal a band at the molecular weight for Fel d 1. Conculsion: Contamination of commercial dog extracts was not found. However, our findings are supportive of commercial dog extracts containing a Fel d 1-like dog allergen that is cross-reactive to Fel d 1. Cross-reactivity between commercial dog extracts and Fel d 1 could be responsible for double positivity to cat and dog in serology. Additional studies are needed to better illustrate this Fel d 1-like dog allergen.

Optimization of biomimetic fermentation technique for enhanced and eco-friendly extraction of phenolics from pomegranate peel

In this research, we mimicked pomegranate orchard conditions for Aspergillus growth on powdered pomegranate fruit waste (PFW) to extract and purify phenolic compounds. Pomegranate's high ellagitannin content makes it attractive to Aspergillus sp., whose spores are abundant in these environments. Four experiments were conducted with varying PFW:water ratios (1:2.5, 1:5, 1:7, and 1:10), aeration, agitation, and incubation times. The 1:10 ratio achieved the highest antioxidant activity (256.34 mg GAE/g DM) and total phenolic content (300.3 mg GAE/g DM) after 48 h of fermentation, comparable to advanced chemical methods. PCR analysis identified Aspergillus tubingensis, and mycotoxin measurements showed no detectable aflatoxins or ochratoxin A, confirming extract safety and purity. This study introduced a pioneering, natural method for phenolic extraction. Additionally, the very higher protein content of fermented extract (1.18 %) compared to the non-fermented extract (0.03 %), suggests its potential as an attractive topic for future investigations as source of food industrial enzymes.

Study on anti-reflection of deep soft and high gas coal seam floor by deep hole controlled blasting

Addressing the practical challenges of difficult drilling for blasting-induced permeability enhancement in deep, soft, and high-gas coal seams, where fractures remain underdeveloped and prone to re-compaction, this study proposes blasting operations within the floor strata. This approach aims to enhance the permeability of soft coal seams, thereby extending the duration of effective gas extraction. A bidirectional loading gas–solid coupling blasting simulation system was established in the laboratory, enabling multi-faceted analysis of experimental models through macroscopic crack patterns, internal damage mechanisms, and strain data of coal and rock masses. Comparative experiments were conducted, contrasting various control hole spacings with conventional blasting techniques. The findings reveal that as the blasting stress wave traverses the control hole walls, tensile stress waves are reflected, facilitating crack propagation. The guiding effect of the control holes and the spatial compensation they provide significantly increase the extension distance of explosion-induced cracks, resulting in directional failure of the test specimens and heightened damage in the far field of the blast. After the blasting process, the arrangement of control holes can result in an increase of up to 133% in damage to the coal seam and a reduction of up to 167% in damage to the floorboard compared to the model without control holes. Notably, when the control holes are proximal to the coal-rock interface, the near-end coal body experiences the most pronounced effects, with peak damage and tensile strain in the d = 20 mm model being 1.93 and 1.79 times higher, respectively, than those in models without control holes. Conversely, for control holes located further from the interface, the distal coal body experiences the greatest influence, exhibiting 1.53 and 1.55 times higher peak damage and tensile strain, respectively, in the d = 80 mm model compared to uncontrolled counterparts. Field observations at the C13 coal seam of a mine within the Huainan mining area corroborate these findings, where the volume of gas extraction and its concentration experienced a rapid increase following blasting and penetration enhancement. Optimum permeability enhancement occurs when the blasting hole is situated 4 m from the extraction point, resulting in a 131% increase in gas extraction purity from 0.15 × 10–3 m3/min to 1.97 × 10–3 m3/min. Furthermore, gas concentration soars by 373%, from 5.86% to 21.86%. These research outcomes offer valuable insights and hold considerable reference significance for blasting-induced permeability enhancement in deep, soft, and high gas coal seams.

Purification of Glucomannan from Porang (Amorphophallus oncophyllus) Tuber Flour through Hydrolysis of Hydrochloric Acid

Porang (Amorphophallus oncophyllus) is a local perennial plant that commonly grown in Indonesian forest. Its tuber contains a lot of glucomannan that has many benefits as food ingredient or as functional food. Glucomannan has a lot of health benefits, such as anti-diabetic effect, low glycemic index, prebiotic effect, etc. Despite that, the extraction is still a problem because it is difficult to get pure glucomannan extract out of Porang flour. Thus, in this study we are going to use hydrocloric acid to extract the glucomannan from the porang flour through hydrolysis extraction. This research will test the effect of hydrochloric acid concentration on the purity of glucomannan extract obtained from porang flour, to study the effect of nonsolvent, ethanol and isopropyl alcohol on glucomannan concentration, and to obtain the optimal condition to get high concentration of glucomannan using extraction with pretreatment hydrochloric acid hydrolysis.

Static expansion fracturing mechanism for enhancing gas permeability in low permeability coal seams

Extraction of gas from low-permeability thick coal seams poses challenges globally, attributed to low extraction efficiency, limited enrichment content, and extraction complexities. Investigating static fracturing in low-permeability thick coal seams holds substantial engineering significance and practical utility. This research delves into the current conditions in the Western region, characterized by low gas permeability and challenging extraction in thick coal seams. Through the utilization of FLAC3D and COMSOL, simulation schemes are devised to analyze the influence of borehole parameters on fracture efficacy, elucidating the mechanisms by which external loads and internal gas pressure impact coal seam permeability. Field monitoring tests are employed to evaluate and model gas extraction enhancement via borehole positioning. The results suggest that the application of a static fracturing agent inducing 40 MPa expansion stress, along with a 75 mm borehole diameter and 0.5 m spacing, effectively fractures low-permeability thick coal seams. The spacing between extraction and fracturing holes adversely affects gas extraction efficiency due to the limited range of static fracturing. Field experiments demonstrate that static fracturing significantly improves gas extraction from low-permeability thick coal seams, resulting in a twofold increase in average gas extraction purity post-fracturing. This study establishes a robust theoretical foundation for optimizing gas extraction and mining activities in challenging low-permeability thick coal seam environments.

Green extraction and partial purification of roselle (Hibiscus sabdariffa L.) extracts with high amounts of phytochemicals and in vitro antioxidant and antibacterial effects.

The aim of this study is to develop roselle (Hibiscus sabdariffa L.) extracts with high amounts of phytochemicals and in vitro antioxidant and antibacterial effects using green extraction technique. Roselle extract prepared by ultrasonic extraction with 80% ethanol and 1% hydrochloric acid (RSUEH) promoted the highest yield and high total phenolic (5.21±0.05g% gallic acid equivalent of dried extract), total flavonoid (1.29±0.06g% quercetin equivalent of dried extract), and total anthocyanin contents (0.21±0.00 and 0.44±0.00g% cyanidin 3-glucoside equivalent of dried extract determined by pH-differential method with standard calculation and pH-differential method with standard curve of cyanidin 3-glucoside, respectively). From the validated high-performance liquid chromatography analysis, delphinidin-3-sambubioside, delphinidin-3-glucoside, and cyanidin-3-glucoside were found to be the major anthocyanins in roselle extract. This extract exhibited moderate 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging effect (EC50 289.61±0.16µg/mL) and strong inhibitory effects against Escherichia coli, Staphylococcus aureus, Staphylococcus intermedius, Proteus mirabilis, and Pseudomonas aeruginosa with the inhibitory concentration (IC50) values in the range of 0.15±0.05 to 4.64±0.07mg/mL and the minimum inhibitory concentration (MIC) values ranged from 1.66±0.06 to 7.11±1.05mg/mL. From partial purification, fraction 28 (F28) contained higher amounts of all analyzed phytochemicals than roselle extracts and promoted stronger DPPH scavenging effects (EC50 54.88±2.23µg/mL). F28 showed antibacterial activity with IC50 values against all tested bacteria in the range of 0.19±0.07 to 4.69±0.09mg/mL, and the MIC values ranged from 2.46±0.08 to 6.24±0.09mg/mL.

Comparison and Optimization of Three Extraction Methods for Epimedium polysaccharides

Epimedium is used in traditional Chinese medicine. Epimedium polysaccharides have a variety of physiological properties. This study compared three different processes for the extraction of polysaccharides from Epimedium spp., including ultrasonic, aqueous enzymatic, and microwave extraction, to optimize the extraction conditions and determine the optimal extraction method. The optimal parameters for each method were analyzed. The results showed that the optimal process for ultrasonic extraction was an ultrasonic power of 250 W, an extraction time of 60 min, a temperature of 50 °C, and a solid–liquid ratio of 1:35. The optimal conditions for the aqueous enzymatic method were a papain concentration of 70 U/mL, extraction time of 70 min, a temperature of 50 °C, and a material-to-liquid ratio of 1:30, while those for microwave extraction were a microwave power of 650 W, an extraction time of 50 min, a temperature of 40 °C, and a material-to-liquid ratio of 1:25. The polysaccharide yields were 4.85%, 4.72%, and 3.98% for the three methods, respectively, indicating that ultrasonic extraction resulted in the highest yield of polysaccharides from Epimedium brevicornum. After purification with DEAE-cellulose, the polysaccharide yields were 4.13%, 3.67%, and 3.12%, respectively. A comprehensive comparison demonstrated the superiority of the ultrasonic extraction method in terms of both extraction yield and purification efficiency. Characterization of the extracted Epimedium polysaccharides showed the presence of five monosaccharides, i.e., glucose, galactose, mannose, galacturonic acid, and rhamnose, and a number average molecular weight Mn of 1.65 × 105 Da and weight average molecular weight Mw of 6.61 × 105 Da. These results provide a scientific basis for the in-depth study and application of Epimedium polysaccharides.

Paralytic Shellfish Toxin Extraction from Bivalve Meat for Analysis Using Potentiometric Chemical Sensors.

A simple and reliable methodology for the detection of paralytic shellfish toxins (PSTs) in bivalve tissues using potentiometric chemical sensors was developed. Five methods of PST extraction from mussel and oyster tissues were evaluated, including the AOAC-recommended method, which served as the reference. The main objective was to minimize the matrix effect of the extracts on the sensors' responses and ensure efficient toxin recovery. Extraction procedures using acetic acid with heating and water yielded the highest responses from the potentiometric chemical sensors to PSTs. The highest recovery of PSTs from bivalve tissues was achieved with extraction using acetic acid and heating. Further extract purification, which is indispensable for liquid chromatography with fluorometric detection (LC-FLD) analysis, was found to be unnecessary for analysis with chemical sensors. While water extraction can also be used as a rapid and simple PST extraction method, the lower recoveries should be considered when interpreting the results. Further research is needed to identify the compounds remaining in the extracts that cause a decrease in sensor responses and to develop procedures for their elimination.

Innovative Techniques for Pharmaceutical Waste Management: Enhancing Drug Recovery and Environmental Sustainability

The pharmaceutical sector is a major component of current healthcare, manufacturing and distributing drugs, biological substances, and medical equipment. Despite its advantages, the sector creates enormous waste, including materials for packaging, production by-products, expired or unused drugs, and other residues, creating health and environmental issues. Appropriate pharmaceutical waste handling and medication recovery strategies are vital for limiting these problems. This article aims to investigate and evaluate multiple techniques for recovering pharmaceuticals from pharmaceutical waste, highlighting the significance of sustainable waste management in the pharmaceutical sector. The paper emphasizes the need to use modern methods such as liquid-liquid extraction, membrane crystallization, solid-liquid extraction, and adsorption to recover drugs from pharmaceutical waste. Liquid-liquid extraction exhibits excellent adaptability and efficiency for varied Active Pharmaceutical Ingredients (APIs), whereas membrane crystallization provides low-energy solutions for thermally sensitive compounds. Solid-liquid extraction is useful for recovering APIs from solid dosage forms, while adsorption approaches exploit substances like activated carbon for organic component recovery. Each process has particular benefits and disadvantages, with the selection of methodology based on waste properties and recovery objectives. It emphasizes the promise of these technologies for high extraction yields, purity, and environmental sustainability, supporting effective pharmaceutical waste management procedures. Additionally, difficulties such as cost-effectiveness, scalability, and regulatory compliance are addressed, pointing to opportunities for future research and development to improve the efficacy of drug recovery procedures. In conclusion, using advanced techniques to recover pharmaceuticals from pharmaceutical waste offers a viable way to implement sustainable waste recovery procedures and lessen the pharmaceutical industry's negative environmental effects.

Extraction and partial purification for adhesive protein FimH from type-1 pili which isolated from uorpathogenic E.coli

استخلص خمل النمط الاول من عزلة لبكتريا الـ E.coli معزولة من دراسة سابقة حسب طريقة Brinton المحورة والتي امكن من خلالها الحصول على 2.8 ملغرام خمل / 10 غم بكتريا ، اتبعت طريقة الاذابة واعادة البلورة ، واستعمال سلسلة من المحاليل التي ساعدت على عملية اذابة الشوائب المرافقة لبروتينات الخمل لغرض التنقية وكان ناتج عمليات التنقية الجزئية هذه الحصول على 2.1 ملغم/10غم بكتيريا . استخلص بروتين الالتصاق FimH وتنقيته جزئيا باستعمال كروموتوغرافيا الترشيح الهلامي .فوجد ان تركيز بروتين الالتصاق كان 250 مايكروغرام / مل ،في حين بلغ تركيز بروتين الخمل 50.6 مايكرومايكوغرام / مل . ومن خلال تقنية الترحيل الكهربائي باستعمال عديد الاكريل امايد بوجود المادة الماسخة SDS ، تم تحديد الوزن الجزيئي لبروتين الالتصاق وبروتين الخمل حيث بلغ 28300 ، 17500 دالتن على التوالي عند المقارنة بمنحنى البروتينات القياسية .

Accurate determination of three new antihypertensive drugs illegally added to food using ultra-high performance liquid chromatography-tandem mass spectrometry

Effective strategies are required to address food safety issues related to the illegal addition of antihypertensive drugs to food and claims of antihypertensive function. In this study, a novel ultra-high performance liquid chromatography-triple-quadrupole mass spectrometry (UHPLC-MS/MS) method was developed for the simultaneous determination of three antihypertensive drugs (azilsartan, candesartan cilexetil, and lacidipine) in 12 food matrices (pressed candies, solid beverages, alternative teas, tea drinks, biscuits, jellies, mixed liquors, oral liquids, medicinal teas, tablets, hard capsules, and soft capsules). Initially, mass spectrometry parameters, such as the collision energies of the three antihypertensive drugs, were optimized. Subsequently, the response intensities and chromatographic separation conditions of the three drugs in different mobile phases were compared. In addition, to enhance the recoveries, various extraction solvents and purification methods, including solid-phase extraction (SPE) columns and the QuEChERS technique, were investigated. In the developed method, sample determination involved three steps. First, the sample was extracted using 0.2% (v/v) formic acid in acetonitrile and then filtered using high-speed centrifugation, in addition, the extracted solution of alternative tea and medicinal tea was purified using the QuEChERS technique. Second, the supernatant was diluted with water, and filtered through a 0.22 μm polytetrafluoroethylene (PTFE) membrane. Finally, the analytes were separated on an Agilent Eclipse Plus RRHD C18 column (50 mm×2.1 mm, 1.8 μm) using a 5 mmol/L ammonium formate aqueous solution and acetonitrile as the mobile phases under gradient elution conditions and then detected using UHPLC-MS/MS with positive electrospray ionization (ESI) in the multiple reaction monitoring (MRM) mode. Quantitative analysis was performed using a matrix-matched external standard method. Methodological validation showed good linear relationships for all three antihypertensive drugs in the investigated concentration ranges, with correlation coefficients (r2) greater than 0.996. The limit of detection (LOD) and limit of quantification (LOQ) of lacidipine were 0.02 mg/kg and 0.04 mg/kg, respectively, whereas those of the other two drugs were 0.01 mg/kg and 0.02 mg/kg, respectively. In the 12 food matrices, the average recoveries of the drugs ranged from 86.6% to 107.5% with relative standard deviations (RSDs) of 1.1%-10.9% (n=6) at low, medium, and high spiked levels. Furthermore, this method was successfully applied to the analysis of real food samples. Overall, the newly developed method is simple, rapid, sensitive, accurate, and suitable for the qualitative and quantitative determination of antihypertensive drugs in different food matrices. This work could provide technical support for food safety agencies in implementing measures against the illegal addition of antihypertensive drugs to food.

Application of Response Surface Methodology to Design and Optimize Purification of Acetone or Aqueous Acetone Extracts of Hop Cones (Humulus lupulus L.) Using Superparamagnetic Iron Oxide Nanoparticles for Xanthohumol Isolation.

Iron oxide nanoparticles (IONPs) are an ideal sorbent for magnetic dispersion extraction due to their superparamagnetic properties and developed and active surface. This work aims to use IONPs, obtained by chemical co-precipitation, to purify 100% acetone and 50% acetone extracts from hop cones (Humulus lupulus L.) obtained by ultrasonic-assisted solvent extraction. The extracts were purified from bitter acids (i.e., humulones, lupulones) to isolate xanthohumol. The sorption conditions were optimized depending on the composition of the extraction mixture, specifically the mass of IONPs and the time needed to achieve effective sorption using response surface methodology (RSM). An analysis of variance (ANOVA) was performed to assess the adequacy of the developed model, and a good agreement was found between the experimental data and the proposed model. The polynomial equation describing the model is highly significant (p < 0.05), with a precision of Adeq (above 4). This indicates the usefulness of the polynomial regression model for prediction in experimental design. The final products of the purification for 100% acetone extracts and 50% acetone contain 40.58 ± 2.84 µg mL-1 and 57.64 ± 0.83 µg mL-1 of xanthohumol, respectively. The use of 50% acetone extract provides more favorable conditions due to the smaller amount of nanoparticles required for extract purification and a higher recovery of xanthohumol. The development of a reliable multivariate model allowed for the optimization of the extract purification process, resulting in high-purity xanthohumol from natural sources.

The assessment of several extraction methods, purification, and characterization of Pseudomonas aeruginosa lipopolysaccharide (LPS) and studying its biological activity

Many Gram-negative bacteria have LPS, a virulent component, on their outer membranes. LPS, a surface structural component protects the outside layer while intoxicating host cells. Research linked LPS to antibiotic tolerance and biofilm formation. LPS induces pathophysiological effects, LPS is pyrogenic, it causes endotoxin shock and induces the clinical manifestations of infections. Therefore, several methods for isolating and purifying LPS have been developed. Extraction, purification, characterization and yields of Lipopolysaccharide (LPS) were obtained from P. aeruginosa through various methods to determine the pyrogenic effect of extracted LPS (ELPS) in rabbits. The study included the extraction of LPS from P. aeruginosa using manual methods including the modified hot (phenol-water) method, the methanol/ chloroform extraction method and an LPS extraction kit. The extracted LPS was detected and characterized by SDS-PAGE with silver staining and purified with HPLC. The protein and nucleic acid content of Extracted LPS were detected using SDS-PAGE with Coomassie blue staining and gel electrophoresis respectively. The study also determined the pyrogenic effect of extracted LPS (ELPS) in rabbits. Extracted LPS, demonstrated a typical ladder pattern upon silver staining and no contaminating protein was observed by coomassie blue staining and contamination of nucleic acid. In contrast, the extracted LPS without the enzymatic treatments revealed lighter protein and nucleic acid contamination bands. The elevated body temperature of rabbits following intraperitoneal administration of purified LPSs confirmed the functional activity of extracted LPSs. It was concluded from the study that the LPS extracted with manual methods and with an extraction kit showed high purity and yield compared to standard LPS. The modified hot (phenol-water) method showed the highest yield among all three methods. Rabbit pyrogencity testing of extracted LPS demonstrated the endotoxin's biological activity and pyrogenic nature.

Red beet betalains extraction process: A comprehensive review of methods, applications, and physicochemical properties.

Red beet extract is rich in bioactive compounds and possesses health-promoting properties. Moreover, the stability of red beet extract over a broad acidic pH range has given them great potential in developing new functional foods and drinks. The choice of extraction solvent and methodology significantly influences the efficiency of betalain extraction from plant vacuoles. Although the conventional solvent extraction method has been widely employed for betalain extraction, recent innovations have introduced alternative methods that offer advantages, such as reduced solvent consumption, energy efficiency, and minimized exposure to high temperatures. This paper aims to summarize the current knowledge about conventional and novel extraction methods, applications, biological activities, and purification of red beet betalains. Furthermore, the physicochemical properties of betalain-rich extract of red beet and associated safety considerations have been investigated.