Thin-layer Chromatography Research Articles

Genome Analysis of a Polysaccharide-Degrading Bacterium Microbulbifer sp. HZ11 and Degradation of Alginate

Marine bacteria are crucial sources of alginate lyases, which play an essential role in alginate oligosaccharide (AOS) production. This study reports the biochemical characteristics of a new species of the Microbulbifer genus, Microbulbifer sp. HZ11. The strain HZ11 is Gram-negative, aerobic, flagellate-free, and rod-shaped. The genome of strain HZ11 is a 4,248,867 bp circular chromosome with an average GC content of 56.68%. HZ11 can degrade alginate and other polysaccharides. The carbohydrate-active enzyme (CAZyme) genes account for 4.57% of the total protein-coding genes of HZ11. Its alginate metabolism process is consistent with the characteristics of the polysaccharide utilization locus (PUL) system. The alginate lyase produced by strain HZ11 showed the highest activity at 50 °C, pH 8.5, and 0.1 M NaCl. The substrate preference was as follows: sodium alginate > poly mannuronic acid > poly guluronic acid. The thin layer chromatography (TLC) results revealed that the main enzymatic degradation products were monosaccharides or AOSs with a degree of polymerization (DP) of 2–3. These results help clarify the metabolism and utilization mechanism of alginate by marine bacteria and provide a theoretical reference for its application in the degradation of alginate and other polysaccharides.

Exopolygalacturonase Production from the Novel Strain Lichtheimia sp. UV-16 and Enzyme Hydrolysis Properties.

A pectinase-producing strain, Lichtheimia sp. X-8, was isolated from the soil for the first time. Subsequently, Lichtheimia sp. UV-16, with a 1.23-fold increase in pectinase activity, was obtained via UV mutagenesis, and optimization of its liquid fermentation process boosted pectinase activity from 455.6 ± 12.7 to 3202.0 ± 82.1 U/mL. The crude enzyme was purified by salting out and anion exchange resin, with a purification ratio of 2.28-fold and a yield of 36.5%. The optimal reaction temperature for the pure enzyme was 60 °C with an optimal pH of 5.5. Thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) confirmed that the enzyme was an exopolygalacturonase, achieving over 99% efficiency in pectin hydrolysis. Furthermore, incorporating pure enzymes into juice pulps can substantially enhance the juice yield, which makes this polygalacturonase a promising application in the beverage industry.

Sonication Enhancement of Capsaicin Formation in Callus of Chili Pepper, Capsicum annuum L.

The current study investigates the induction of callus from leaf explants of chili pepper Capsicum annuum L. coupled with the isolation of capsaicin from alcoholic extracts. To determine which isolated alkaloid has a positive reaction, the DragenDroff test is used. Alkaloid is identified using conventional diagnostic techniques, such as measuring the absorbance values of the isolated alkaloid with an ultraviolet spectrophotometer, the alkaloid is identified. The results show a complete identity among them, and with control. Thin layer chromatography data showed a 0.8 cm distance between one location from each tested sample with the same rate, which is 0.8 cm from the control’s rate flow value. The chemical structure of studied samples is subsequently determined using nuclear magnetic resonance, which reveals similarities between the isolated alkaloid’s structure and standard capsaicin. A quantitative analysis of the isolated alkaloids revealed variations in the amounts for generated explants relative to other explants. This study shows that fruits are the most effective source of alkaloids. It’s interesting to note that the composition of the explant and the sonicated callus are identical. Since capsaicin discovery, it is used as a homeopathic remedy to treat burning pain using the concept of “treating like with like” or counterirritant, relieve minor pain associated with rheumatoid arthritis or muscle sprains and strains and due to large consumption of this fruit recently, the current study done to find out the structure and quantity.

The Antioxidant and Anti-Inflammatory Activities of the Methanolic Extract, Fractions, and Isolated Compounds from Eriosema montanum Baker f. (Fabaceae)

Background: Inflammation is a natural body’s defense mechanism against harmful stimuli such as pathogens, chemicals, or irradiation. But when the inflammatory response becomes permanent, it can lead to serious health problems. In the present study, the antioxidant and anti-inflammatory potentials of the Eriosema montanum methanolic extract (EMME), as well as its isolated fractions (FA-FJ) and compounds (1–7), were evaluated by using in vitro and cellular models. Methods: The total phenolic and flavonoid contents were determined using, respectively, Folin–Ciocalteu and aluminum chloride colorimetric methods, while 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 2,2′-diphenyl-1-picrylhy-drazyl (DPPH), and ferric ion reducing antioxidant power (FRAP) were used to determine the antioxidant activity. Thin Layer Chromatography (TLC) and column chromatography (CC) were used to isolate and purify the compounds and their elucidation using their NMR spectroscopic data. Results: EMME had moderate antioxidant and anti-inflammatory activities, while fraction FF showed much higher efficacy with IC50 values of 34.64, 30.60, 16.43, and 77.29 μg/mL against DPPH, ABTS, NO, and 15-LOX inhibitory activities, respectively. The EMME fraction was found to be very rich in flavonoids and phenolic compounds, with 82.11 mgQE/g and 86.77 mgGAE/g of dry extract, respectively. Its LC-MS profiling allowed us to identify genistin (5) as the most concentrated constituent in this plant species, which was further isolated together with six other known compounds, namely, n-hexadecane (1), heptacosanoic acid (2), tricosan-1-ol (3), lupinalbin A (4), d-pinitol (6), and stigmasterol glucoside (7). Given these compounds, genistin (5) showed moderate activity against reactive oxygen species (ROS) and NO production in LPS-stimulated RAW264.7 cells compared to EMME, which suggested a synergy of (5) with other compounds. To the best of our knowledge, compounds (1), (2), and (3) were isolated for the first time from this plant species.

Elucidation of Domain Function of a Novel Multifunctional Glycoside Hydrolase and Its Use in Efficient Preparation of Oligosaccharides from Kelp Powder.

Kelp contained laminarin, cellulose, and alginate as major polysaccharides and could be utilized as functional oligosaccharides. A new multifunctional glycoside hydrolase CelA was identified and characterized for the efficient degradation of kelp powder. It displayed cellulase (2308.38 U/mg), alginate lyase (578.68 U/mg), and laminarinase (720.97 U/mg) activities. It exhibited maximal activity on both sodium alginate and laminarin at 50 °C and pH 8.0, while it could degrade sodium carboxymethylcellulose (CMC-Na) by maximal activity at 40 °C and pH 7.0. The action mode analysis by thin layer chromatography and electrospray ionization mass spectrometry indicated that CelA adopted an endolytic manner to degrade CMC-Na, sodium alginate, and laminarin releasing oligosaccharides with degrees of polymerization (Dps) of 2-5. According to domain analysis, CelA contained a GH5 module and a PL6 module, and both of them exhibited glycoside hydrolase and polysaccharide lyase activity. The docking results revealed that Glu163 and Glu250 are essential in cellulose and laminarin degradation. As to the degradation of alginate, Asn376, Lys436, Arg464, Asp496, and Asn551 could bind alginate and Tyr492 and Lys529 acted as catalytic sites. CelA displayed high hydrolysis efficiency for cellulose, β-glucan, laminarin, alginate, and kelp powder. Thus, it has strong potential in food and feed industries as a catalyst for bioconversion of algal biomass into value-added products oligosaccharides.

Undescribed and known phenolic acid derivatives with significant antioxidant activity from the Scorzonera parviflora Jacq. roots.

The antioxidant activity of Scorzonera parviflora Jacq. roots were assessed by measuring their ability to scavenge ABTS and DPPH radicals. Bioactivity-guided fractionation was utilized to identify the compound(s) responsible for this activity. The most active phase, ethyl acetate, was isolated using column chromatography. The resulting fractions were then purified using preparative TLC on reverse phase and semi-preparative HPLC. The structures of the pure compounds were elucidated by spectral analysis (MS and 1H, 13C, 2D-NMR). Three undescribed phenolic acid derivatives, namely parvifloric acid A (1), B (2), and C (3), and one new sesquiterpene lactone, parviflorin (4) together with seven known compounds were isolated and identified as scopolin (5), scopoletin (6), caffeic acid (7), protocatechuic acid (8), 4,5-O-dicaffeoylquinic acid (9) 3,5-O-dicaffeoylquinic acid (10), and 3,5-O-dicaffeoylquinic acid methyl ester (11). Finally, the pure compounds obtained were tested to evaluate their antioxidant capacities, using ABTS and DPPH radical scavenging potencies. The highest activity was observed with 3,5-O-dicaffeoylquinic acid (10), followed by its methyl ester.

A Review of Principles, Applications, and Recent Developments in HPTLC and HPLC

High-Performance Thin Layer Chromatography (HPTLC) and High-Performance Liquid Chromatography (HPLC) represent fundamental analytical techniques in modern chemical analysis. These sophisticated separation methods have revolutionized the field of analytical chemistry through their precision, reliability, and versatility. HPTLC, an enhancement of traditional thin-layer chromatography, offers advantages such as minimal sample preparation, simultaneous analysis of multiple samples, and cost-effectiveness. It has found extensive applications in pharmaceutical analysis, natural product research, and food quality assessment. HPLC, characterized by its high resolution, sensitivity, and automation capabilities, has become an indispensable tool in various industries, including pharmaceutical, environmental, and biochemical analyses. The technique employs different modes such as reverse phase, normal phase, and ion exchange, enabling the separation of complex mixtures with remarkable accuracy. Recent technological advancements have led to the development of ultra-high-performance liquid chromatography (UHPLC), offering enhanced speed and resolution. Both HPTLC and HPLC complement each other in analytical laboratories, with HPTLC providing rapid screening and HPLC offering detailed quantitative analysis. The continuous evolution of these techniques through technological innovations ensures their pivotal role in modern analytical chemistry.

Chromatographic’s image analysis in phytohomeopathies

Plants signal when establishing interaction with homeopathies in different succussions, however, the type of signaling is not always understandable. Recognizing symptomatic patterns in plants begins by understanding the behavior of homeopathy itself, through varied methods, widely disseminated in scientific literature¹. The present study proposes a new thin-layer chromatography (TLC) analysis method to identify patterns in different phytohomeopathic applications. CCDs were made on Sulfur 9Ch, Nux and Bell 11CH; Calc Ph 29CH; Bry and Cal Carb 30CH; Hyp 50CH; Mag Ph 200CH and Graph Nat 200CH. The analysis of the chromatographic stains was developed with ultraviolet light and the images were analyzed using the Fast Fourier Transform (FFT) method². The statistical design, ANOVA, consisted of 5 completely randomized, double-blind replications for each of the preparations. The results showed significant differences after a few minutes of chromatography, highlighting the order of signals between homeopathies: Bell < Sulph < Mag Ph < Graph Nat < Hyp < Bry < Calc Ph < Cal Carb < Nux. From these results, it is possible to identify phytohomeopathic’ signs and symptoms during the course of the treatment.

СИНТЕЗ N-ГИДРОКСИ-, N-МЕТОКСИ И N-ЭТОКСИБЕНЗОИЛ ПРОИЗВОДНЫХ 5-МЕТИЛТИАЗОЛ-2-АМИНА, КАК ПОТЕНЦИАЛЬНЫХ ЛЕКАРСТВЕННЫХ ПРЕПАРАТОВ

The synthesis of N-hydroxy-, N-methoxy and N-ethoxybenzoyl derivatives of 5-methylthiazol-2-amine was carried out using the Schotten-Baumann reaction in two stages without isolating the intermediate acyl chloride (one-pot multicomponent synthesis) and conditions were optimized. Thin layer chromatography and nuclear magnetic resonance spectroscopy confirmed the purity and the structure of the compounds obtained. The spectrum of biological activity of compounds was assessed using PASS Online (Predictor of Activity Spectra for Substance). Using the ChemAxon online platform, theoretical calculations of physico-chemical parameters and descriptor values, such as CNS MPO multiparameter, describing the effect of substances on the CNS and cardiotoxicity, as activity against the hERG gene, were carried out. Analysis of the calculated data showed that all the studied substances are safe and will not have a negative effect on cardiac activity. The results of studies using PASS Online indicated that the synthesized new thiazole derivatives combine various types of activities and may turn out to be promising structures for experimental screening of the considered types of biological activities.

A Simple, Cost-Effective, and Sustainable Method for Extraction and Quantification of Chebulagic Acid from Terminalia bellirica (Gaertn.) Roxb. Using RP-HPLC: A Comparative Study

Chebulagic acid (CA), a hydrolysable tannin found in Terminalia bellirica (T. bellirica) (Gaertn.) Roxb., has gained prominence as a significant component in Ayurvedic medicine. This study optimized methods for determining CA in T. bellirica, focusing on fruit samples and prioritizing simplicity, cost-effectiveness, less time duration, and sustainability. We evaluated various extraction techniques, including continuous shaking, microwave assisted extraction, and Soxhlet extraction using green solvents. Quantification of CA was conducted using Reverse Phase High-Performance Liquid Chromatography (RP-HPLC), with confirmation achieved via UV-Visible (UV-Vis) spectroscopy, Thin Layer Chromatography (TLC), and Liquid Chromatography-High Resolution Mass Spectrometry (LC-HRMS). Results indicated that Soxhlet and microwave significant extraction methods, particularly with ethanol as the green solvent. Soxhlet extraction yielded 49607.73±18.82 μg/mL for fresh fruit and 42952.53±19.09 μg/mL for dry fruit, while microwave extraction resulted in 43627.56±113.48 μg/mL for fresh fruit and 40466.33±130.90 μg/mL for dry fruit respectively. These findings conclude that ethanol as a green solvent in Soxhlet or microwave assisted extraction emerge as most efficient, straightforward, and effective approach for detecting and screening CA in T. bellirica samples.

Comparative analysis of biologically active substances in leaves and fruits of European dewberry (Rubus caesius L.)

Introduction. In recent years, the attention of researchers has increasingly been attracted by representatives of the genus Rubus, among which the European dewberry (Rubus caesius L.), as one of the most frequently encountered species. There are a number of studies on the chemical composition and pharmacological activity of raw materials of European dewberry. For example, its fruits, leaves, and shoots are rich in compounds of a phenolic nature, have antitumor, antioxidant, and antidiarrheal activity. Consideration of European dewberry as a producing plant of medicinal plant raw materials is relevant. Objective. The study of the composition and content of some groups of biologically active substances (BAS) in the leaves and fruits of European dewberry. Material and methods. The objects of the study were dried leaves and fruits of European dewberry harvested in the Moscow region and the Republic of Bashkortostan in the summer of 2023. The analysis of BAS was carried out according to the methods set out in the Russian State Pharmacopoeia. BAS was detected by qualitative reactions and thin-layer chromatography (TLC). The quantitative determination of BAS was carried out by titrimetry and spectrophotometry (SPM) methods. Results and discussion. During the work, hydrolyzable tannins (10.01 and 5.6%, respectively), organic acids (1.44 and 3.12%, respectively), ascorbic acid (0.068 and 0.11%, respectively) were found in the leaves and fruits of the European dewberry. The amount of anthocyanins in terms of cyanidin-3-5-diglycoside was determined in the fruits of the European dewberry, the value of which was 0.98%. The content of flavonoids in terms of rutin in the leaves of European dewberry was 0.72%. Conclusion. A comparative analysis of BAS in the leaves and fruits of the European dewberry was carried out. The studied objects are promising sources of substances of primary and secondary metabolism.

Enantioseparation of antihistamines using magneto-thin-layer chromatography: The effects of various orientations and intensities of magnetic fields

The effect of magnetic fields produced by magnet and solenoid was investigated for the first time in the enantioseparation of antihistamines using thin-layer chromatography. The racemic mixtures of cetirizine and hydroxyzine were selected as the model drugs for studying the magnetic field effect on enantioseparation. The magnets were fixed to a U-shape blade that rotated around the separation chamber by a motor to apply a magnetic field perpendicular to the direction of mobile phase movement. Also, the separation chamber was positioned within a solenoid, and an electric current was utilized to apply the magnetic field in a parallel direction to the mobile phase movement. L-arginine was added to the mobile phase as a chiral selector to perform the enantioseparation of drugs. When magnets generate the magnetic field, both in attraction and repulsion, the spots become broader and the peak height decreases as the magnetic field intensity and rotation speed of the magnets increase. An increase in peak height was observed in the presence of the magnetic field generated by + 3.25A current in the solenoid. Regardless of the type of applied magnetic field, the retention factor values exhibited slight reductions. It was observed that separated enantiomers were similarly affected by various magnetic fields. The distribution coefficient, mass transfer, equilibrium, and lipophilicity were all potentially influenced by the magnetic field, which could explain the alterations.

Phytochemical characterization and evaluation of anti-amnesic activity of two source plants of Shankhapushpi

BackgroundShankhapushpi is an important Ayurvedic drug used for treating various disease conditions of nervous system. Convolvulus prostratus Forssk. is the genuine source plant for Shankhapushpi as per Ayurvedic Pharmacopoeia of India; however Clitorea ternatea L. is widely used as Shankhapushpi in southern part of India. In this study, comparative phytochemical and pharmacological evaluation has been done in two plants such as Convolvulus prostratus and Clitorea ternatea used as Shankhapushpi. MethodsPhytochemical comparison was done by High Performance Thin Layer Chromatography (HPTLC) and High Performance Liquid Chromatography (HPLC). Detailed metabolite profiling was performed using Q-TOF-LC/MS-MS analysis. Antiamnesic activity of selected plants has been evaluated against scopolamine induced amnesia model in Wistar rats. ResultsPhytochemical studies showed that only a few chemical constituents are common for both species. Most of the phytochemicals are different for selected species. LC/MS analysis showed presence of genipin and 7-hydroxyflavone in both species. Pharmacological study showed that both plants possess antiamnesic activity against Scopolamine induced memory loss; However C. ternatea possesses significant antiamnesic activity than that of C. prostratus. ConclusionThe study provided valuable information about the selected species in terms of its phytochemical composition and pharmacological properties. The study also provided a scientific support for using both species as Shankhapushpi.

Separation of polar and neutral lipids from mammalian cell lines by high-performance thin-layer chromatography

Separation of polar and neutral lipids from mammalian cell lines by high-performance thin-layer chromatography

Identification of Paracetamol in Herbal Medicine for Aches and Pains Using Thin Layer Chromatography (TLC) Method

Jamu adalah salah satu obat tradisional yang banyak digunakan oleh masyarakat untuk mencegah dan atau membantu menyembuhkan penyakit, jamu juga sering kita temukan dalam kehidupan sehari-hari. Peningkatan minat masyarakat serta kebutuhan untuk mengkonsumsi jamu telah disalahgunakan oleh produsen jamu dengan menambahkan Bahan Kimia Obat (BKO), paracetamol merupakan salah satu BKO yang sering ditambahkan dalam jamu yang memiliki khasiat cepat serta keuntungan yang besar. Tujuan penelitian ini untuk mengetahui ada tidaknya kandungan paracetamol dalam jamu pegal linu dianalisis secara kualitatif berupa uji reaksi warna dan uji kromatografi lapis tipis. Tiga pereaksi warna yang digunakan yaitu pereaksi FeCl₃ 10%, K₂Cr₂07, dan NaN0₂. Berdasarkan hasil pengamatan dari sampel yang ditetesi larutan kalium dikromat menghasilkan sampel yang negatif paracetamol, sedangkan larutan FeCl₃ dan natrium nitrit yang dihasilkan sampel jamu positif mengandung paracetamol. Analisis kualitatif menggunakan KLT dan dibaca bercaknya dibawah sinar UV 254 nm lalu di hitung nilai Rf-nya. Hasil analisis kualitatif penelitian ini menunjukkan terdapat dua sampel jamu yang mengandung BKO parasetamol dari 3 sampel yang dikumpulkan. Kesimpulan dari penelitian ini adalah terdapat 2 sampel positif mengandung paracetamol.

Phytochemical analysis and quantification of flavonoid content of ethyl acetate fraction from ‘Alugbati’ (Basella rubra L.) leaves

Basella rubra L. (Alugbati) is a branching, herbaceous vine with heart-shaped leaves widely grown in Asia, with the leaves used as a herb in soups and salads. Several studies have highlighted this plant’s nutritional and biological benefits, which contain various phytochemical substances, including flavonoids. Flavonoids are polyphenolic compounds known to scavenge free radicals through specific mechanisms. This study aimed to extract and quantify flavonoids from the leaves of Basella rubra L. to emphasize its potential benefits as a food or nutraceutical. The dried leaves were pulverized and macerated with 99% methanol before partitioning with ethyl acetate and water. The fractions were tested qualitatively and then analyzed using Fourier Transform Infrared Spectroscopy (FTIR) to detect the presence of flavonoids, followed by thin layer chromatographic (TLC) analysis to confirm the presence of flavonoids. A total flavonoid content (TFC) test was performed to quantify the extracted flavonoids. The positive results from the qualitative tests and the subsequent FTIR spectra reflecting the constituent bonds and functional groups validated the presence of flavonoids in the plant fractions. The aqueous and ethyl acetate fractions yielded a total flavonoid content of 2.71 ± 0.01 and 9.61 ± 0.04 mg QE/g extract, respectively. The plant's physical and environmental background, the extraction process used, and the plant part used are all potential contributors to its flavonoid content. To maximize its nutritional and therapeutic potential, further research into other plant organs, and performing other extraction and purifying procedures, is recommended.

New Antioxidant Caffeate Esters of Fatty Alcohols Identified in Robinia pseudoacacia

The stem bark of black locust (Robinia pseudoacacia L.) was extracted, and nine antioxidant compounds (R1–R9) were detected by high-performance thin-layer chromatography combined with the radical scavenging 2,2-diphenyl-1-picrylhydrazyl (DPPH•) assay, multi-detection, and heated electrospray high-resolution mass spectrometry. For structure elucidation, the methanolic crude extract was fractionated by solid-phase extraction, and the compounds were isolated by reversed-phase high-performance liquid chromatography with diode array detection. The structures of isolated compounds were elucidated by nuclear magnetic resonance and attenuated total reflectance Fourier-transform infrared spectroscopy as well as gas chromatography-mass spectrometry to determine the double bond position. 3-O-Caffeoyl oleanolic acid (R1), oleyl (R2), octadecyl (R3), gadoleyl (R4), eicosanyl (R5), (Z)-9-docosenyl (R6), docosyl (R7), tetracosyl (R8), and hexacosanyl (R9) caffeates were identified. While R1 has been reported in R. pseudoacacia stem bark, the known R3, R5, R7, R8, and R9 are described for the first time in this species, and the R2, R4, and R6 are new natural compounds. All nine caffeates demonstrated antioxidant activity. The antioxidant effects of the isolated compounds R1–R8 were quantified by a microplate DPPH• assay, with values ranging from 0.29 to 1.20 mol of caffeic acid equivalents per mole of isolate.

Quantify biomarkers in Asthimajja Pachak Kwath by High Performance Thin Layer Chromatography

Quantify biomarkers in Asthimajja Pachak Kwath by High Performance Thin Layer Chromatography

Red, green, and blue model assessment and AQbD approach to HPTLC method for concomitant analysis of metformin, pioglitazone, and teneligliptin

BackgroundThe CDSCO of India has authorized a combination of metformin hydrochloride, teneligliptin hydrochloride, and pioglitazone hydrochloride for the treatment of insulin-independent diabetes. For the purpose of estimating metformin, teneligliptin, and pioglitazone combinations as well as individual commercial formulations, there are a plethora of publicly accessible chromatographic techniques. More importantly, the development of these chromatographic procedures has included the use of chemical solvents that are dangerous to both animals and the environment.ObjectivesHowever, to date, there has been no documented chromatographic technique that can concomitantly estimate various commercial formulations of drugs under study employing a uniform chromatographic condition and environmentally friendly solvents. In order to concomitantly estimate drugs under study utilizing unified chromatographic conditions, a green HPTLC method was developed.MethodThe AQbD approach was used to carry out the method development. To determine the most important method parameters and response variables, the analytical risk assessment was conducted using the risk priority number ranking and screening approach. Critical method parameters and response variables were modeled using the response surface modeling approach, which relies on the central composite design. Optimal ranges for the intended method operable design region were determined, and control strategy was framed. The chromatographic separation was carried out on preparative TLC plate precoated with silica gel G-60 F254 using 1.0%W/V ammonium acetate in ethanol: water: triethylamine (6.5:0.4:0.6, V/V) as mobile phase. The detection of the anti-diabetic drugs under study was carried out at 267 nm wavelength.ResultsThe linearity of metformin, teneligliptin, and pioglitazone was found to be 5000–25000 ng/band, 200–1000 ng/band, and 150–750 ng/band, respectively. The %RSD for robustness and precision study was found to be less than 2.0%. The %recovery of method was found to be 98–102%. The assay results were shown to be in compliance with respective labeled claims of anti-diabetic medications when the suggested method was used for concurrent analysis of several formulations and combinations of drugs under study.ConclusionThe suggested technique was evaluated utilizing red–green–blue model scoring tools. The suggested technique was determined to be precise, accurate, rapid, cost-effective, and easy to apply for the estimation of drugs under study.

Extracting Quercetin from Different Plant Sources, Purifying It Using Different Extraction Methods (Chemical, Physical, and Enzymatic), and Measuring Its Antioxidant Activity

Background: Flavonoids are among the most important compounds found in plants, since laboratory studies have shown them to be a daily requirement in human diets due to their various health benefits. Therefore, this study focused on extracting, purifying, and measuring the antioxidant activity of the flavonoid quercetin, which is widely found in plants and possesses a variety of biological activities, from different plant sources. Methods: The extraction of quercetin was performed using several methods (chemical, physical, and enzymatic) and several extraction solutions (water, ethanol, and chloroform) from several plants (spinach, dill, Onion Skin, Pistacia eurycarpa, sumac, digalkhasab chemri, and leelwi chemri). The alcoholic extract extracted by chemical method was purified and the content of total flavonoids based on quercetin in all plant extracts was determined using adsorption chromatography on a silica gel column (100–200 mesh), followed by thin layer chromatography (TLC). TLC and high performance liquid chromatography (HPLC) were used to assess the purity of quercetin. The ability of quercetin to capture free radicals using 2,2-diphenyl-1-picrylhydrazyl (DPPH) was compared to that of butylated hydroxytoluene (BHT). Statistical analyses were performed using completely randomized designs (CRD) for factorial experiments, and the least significant difference (LSD) test was used to calculate the significant differences between the averages of the coefficients at the 0.05 probability level. Results: The alcoholic Pistacia extract extracted by chemical method yielded the highest concentration of quercetin (84.037 mg/g). Furthermore, it was found that quercetin purified from Pistacia possessed strong antioxidant activity, and its antioxidant activity increased with increased concentration. Conclusions: Pistacia eurycarpa showed the highest quercetin content among the assessed plants. Moreover, solvents played a major role in extracting plant components due to the high polarity of flavonoids. Quercetin purified using a silica gel column demonstrated antioxidant activity.