TLC Method Research Articles

Development of the method for determining Methandienone in Toxicology and doping-analysis

Anabolic-androgenic steroids are ones of the most frequently detected drugs in amateur and professional sports. Doping control laboratories have developed numerous assays enabling the determination of administered drugs. However, it is relevant today to improve detection methods that are accurate, fast and require small amounts of reagents. The identification and quantification of methandienone from body fluids has been insufficiently developed. The gas chromatography-mass spectrometry (GC-MS) method for the determination of methandienone traces in body fluids was developed and validated in the toxicology laboratory in order to control a non-medical use of an anabolic steroid-methandienone. Extracting procedures were optimized in order to obtain entire amount of the drug and methandienone and its metabolites were determined by sensitive and specific reactions for the compound identification, as well as the system for carrying out the TLC method have been proposed, and conditions for identifying a test sample from the biomaterial by IR spectrophotometry have been selected. The GC-MS method is found to be simple, fast, sensitive, and the strategy proposed can be effortlessly and advantageously applied for the routine analysis of biological fluids for determining methandienone and its metabolites at a level less than 30 ng in toxicology and doping-analysis.

An investigation of aspects of radiochemical purity of 99mTc-labelled human serum albumin nanocolloid

BackgroundNanocolloidal human serum albumin radiolabelled with 99mTc provides a diagnostic radiopharmaceutical for sentinel node lymphoscintigraphy. NanoHSA (Nanotop), a commercially available kit, enables the simple preparation of this radiopharmaceutical via reconstitution with pertechnetate eluted from a generator. Thin-layer chromatography is widely used for determining radiochemical purity in clinical nuclear medicine. Quality control methods recommended by the manufacturer were sometimes reported to yield variable results. Therefore, we proposed and evaluated three alternative thin-layer chromatography methods for the quality control of [99mTc]Tc-NanoHSA from a commercially available kit.ResultsThe radiochemical purity of [99mTc]Tc-NanoHSA determined with all methods was reproducible and met the requirements of the SPC and the European Pharmacopoeia (≥ 95%). Our quality control using iTLC-SG chromatographic paper in methyl ethyl ketone mobile phase identified only free pertechnetate as impurity, resulting in > 99% RCP. The quality control using iTLC-SG in 85% methanol or iTLC-SA in 0.9% NaCl identified an additional small fraction of a hydrophilic impurity, resulting in 95–97% RCP. Glucose was identified as a potential 99mTc-carrying hydrophilic species contributing to hydrophilic impurities.ConclusionOur quality control of [99mTc]Tc-NanoHSA with non-polar mobile phase tended to underestimate the amount of hydrophilic impurities, although without compromising the final quality of the radiopharmaceutical. Alternative TLC methods using aqueous mobile phases enabled a more accurate determination of hydrophilic impurities.

DoE-assisted development and validation of a thin layer chromatography method for optimized separation of major cannabinoids in Cannabis sativa L. samples

As interest in the medical use of cannabis and cannabinoids rises, the need to establish appropriate quality specifications for them also becomes evident. Consequently, various methods applied to the analyses of cannabinoids in C. sativa have been reported. Among the chromatographic techniques used for this purpose, TLC stands out as a suitable alternative for fingerprint assessment. Nevertheless, structural similarity between cannabinoids imposes some analytical challenges. In this context, DoE-assisted optimization may favor achieving an adequate resolution between those analytes. Accordingly, this study aimed at developing and validating a TLC method for cannabinoid profile monitoring in C. sativa inflorescences by employing a DoE-driven strategy. A mixture design was applied to estimate the ideal mobile phase composition. Data analyses focused on maximizing resolutions between eight major cannabinoids and improving band distribution throughout the chromatogram. The overall quality of the chromatographic profiles after method optimization was satisfactory. Method validation covered selectivity, precision, and ruggedness assessment. The Youden design was applied for simultaneous analysis of five experimental factors. The proposed method was shown to be fit-for-purpose, allowing chemotype differentiation, in addition to being stability-indicating. Moreover, the results can support the definition of control strategies to ensure consistent achievement of the intended method's performance.

Investigate phenol and antioxidant properties of plant extract of Dianthus caryophylus and Fumaria vaillanti with TLC and HPLC methods

investigate phenol and antioxidant properties of plant extract of Dianthus caryophylus and Fumaria vaillanti with TLC and HPLC methods

Development of thin-layer chromatographic method for determination of caffeine in black, green, and white tea

Caffeine is naturally present in tea and coffee giving the pleasant and stimulant effect. Several different types of teas, black, green, and white teas bought in market were analysis for caffeine content. The boiled sample tea was filtered through filter paper. Lead(II) acetate was used to separate tannins from caffeine followed by filtration through filter paper with a black ribbon. The liquid-liquid extraction was carried out using dichloromethane (3×5 mL) and sodium sulfate as a drying agent. The TLC method was performed on Merck precoated silica gel plates 5×10 cm (60F254, 200 μm) using either methanol or dichloromethane as solvents and the mobile phases were glacial acetic acid and ethyl acetate (95:5, v/v), while the second one was consisted of ethyl acetate and ethanol (80:20, v/v), respectfully. The Rf values were 0.36 and 0.86 for the first and the second mobile phase, respectively, in comparison to the standard caffeine. The values for pH of boiled sample teas were in the range from 4.85 to 5.80. The most abundant tea sample for caffeine was determined in green tea bought in the grocery store for health nutrition (2.04 %). The yield for tea samples from green market, white tea and two tea black samples were 0.06, 0.71, 0.07, and 0.05%, respectively. The developed TLC method can be used for determination of caffeine content in tea samples.

STUDY OF THE FLAVONOID COMPOSITION AND BIOLOGICAL ACTIVITY OF HAWTHORN`S LEAVES CRATAEGUS PONTICA

The results of studies of the content of polyphenolic compounds and their antioxidant activity in the Pontic hawthorn plant Crataegus pontica K.Koch., growing in the mountainous regions of the Tashkent region of the Republic of Uzbekistan are presented. The collection of raw materials took place in the spring at the beginning of October 2019 and at the end of April 2020. The conditions for the isolation of polyphenolic compounds were selected under various conditions. It was shown that the optimal content of polyphenols is extracted with 70% acetone, followed by fractionation of the aqueous residue with ethyl acetate and precipitation with hexane. It was shown that in the plants harvested in spring, the amount of polyphenols was 4.28%, and the collected volume was 2.6%, of the air-dry mass of the raw material. Chromatographic methods (BC and TLC) revealed that the composition of plant leaves collected in spring contains more polyphenols than those collected in autumn. This plant contains compounds belonging to the class of flavonols, phenolic acids and flavan-3-ols. Polyphenols were identified by rutin, quercetin, quercetin-3-O-β-galactoside, catechin, and gallic acid by BC and TLC methods.
 The antioxidant activity of the sum of polyphenolic compounds was studied in the model of lipid peroxidation (LPO) in rat liver mitochondria. It was found that polyphenols have a protective effect on mitochondria, a reduced damaging effect of Fe2+ / ascorbate and antioxidant activity depends on the concentration of the studied polyphenolic substances. The introduction of rutin into the incubation medium in 5 μM medium inhibits LPO processes by 32.0%, and at 10 μM – by 85.9% and in 20 μM – by 96.8%, compared with the control.

Systemic Optimization and Validation of Normal and Reversed-Phase Eco-Friendly Chromatographic Methods for Simultaneous Determination of Paracetamol and Phenylephrine Hydrochloride in the Presence of Paracetamol Impurities.

Chromatographic behavior of different substances in normal and reversed phases is an interesting area in the scientific community. The work aimed to optimize and validate chromatographic separation methods for simultaneous determination of paracetamol (PAR) and phenylephrine HCl (PHE) in the presence of PAR impurities, namely p-aminophenol, p-nitrophenol, acetanilide, and p-chloroacetanilide with further quantification of these toxic impurities. TLC method based on normal phase separation was carried out on aluminum sheets of silica gel 60 F254 using ethanol:chloroform:ammonia as a developing system, followed by densitometric measurements. While HPLC is based on reversed phase separation using a C18 column against acetonitrile:phosphate buffer pH 5 as a mobile phase. PAR and PHE were determined by the TLC-densitometric method in concentration ranges of 3-25 and 0.1-3 µg/band, respectively, and determined by the HPLC method over concentration ranges of 5-400 and 2-80 µg/mL, respectively. Both methods were optimized and validated. Furthermore, they were successfully applied for pharmaceutical formulations with precision <2%. Moreover, results of a statistical comparison with the official methods confirm the methods' validity claims. Two eco-friendly chromatographic methods were developed to determine PAR and PHE in their binary mixtures, pharmaceutical formulations, and in the presence of PAR-related impurities with further quantification of these toxic impurities. These simple chromatographic methods are the first methods developed for simultaneous determination of PAR and PHE in the presence of PAR-related impurities.

Development of green production technology and research of harpagophytum procumbens root dry extract

The aim of this work is to study of standardization parameters and pharmacological properties of Harpagophytum procumbens (H. procumbens) root dry extract.&#x0D; Materials and methods – obtaining of H. procumbens root dry extract was performed in accordance with developed scheme that is characterized by simplicity and cost-effectiveness of technology compared to analogous scheme. Study of standardization parameters of obtained extract was performed in accordance with the requirements of the monograph «Harpagophyti extractum siccum» of the State Pharmacopoeia of Ukraine 2.2., harmonized with the monograph «Harpagophyti extractum siccum» of the European Pharmacopoeia 9.5. Pharmacology study of analgesic and anti-inflammatory activities were conducted on the rat paw formalin-induced edema model.&#x0D; Results. Obtained H. procumbens root dry extract is a free flowing, non-hygroscopic, light brown powder with taste and odor that is specific to raw material of H. procumbens. Determined loss on drying was 3.2±0.18 %. Content of heavy metals in dry extract was not more than 0.01 % (100 ppm). Harpagoside was identified by TLC method. Chromatographic zones on the obtained chromatograms of the reference solution and test solutions are the same in intensity of a color, cross-over and sharpness of development. Based on the results of the research performed by HPLC method, it has been established that content of harpagoside in investigated H. procumbens root dry extract is 2.50±0.02 %.&#x0D; According to the results of pharmacological research it was found that investigated dry extract (the content of harpagoside is not less than 37.0 mg/kg) showed a higher activity compared to reference drug («Phong Te Thap»).&#x0D; Conclusions. A new method of obtaining dry extract of H. procumbens (3.5:1) was proposed. The developed green production technology is simple; it does not require special equipment and expensive solvents. A correspondence of quality parameters with the requirements of State Pharmacopoeia of Ukraine and European Pharmacopoeia: appearance, identification of harpagoside and fructose, loss of drying, content of heavy metals, assay of harpagoside (2.50±0.02 %) was set. Analgesic and anti-inflammatory activities of H. procumbens dry extract has been established

Qualitative and Quantitative Analysis of Dexamethasone in Rheumatic Pain Herbal Medicine Using Thin-Layer Chromatography (TLC) – Densitometry

Chemical medicine is chemical substances that is used as the main ingredient of chemical drugs. This compound is usually added to herbal medicine preparation to strengthen the indication of the traditional medicine.Chemical medicine was prohibited from being added to traditional medicinal preparations. But in reality, a lot of medicinal herbs could have contained medicinal chemicals on the market sale. This purpose of this research was to know the chemical contamination of the dexamethasone also the concentration contained in the rheumatic pain herbal medicine. This research was used three kind of rheumatic pain herbal medicine sample to identify dexamethasone, the sample code is S1, S2, and S3. The analysis of qualitative method are organoleptic test, FTIR characteristic test, TLC evaluation. The analysis of quantitative were purposed to know the dexamethasone concentration contained on the rheumatic pain herbal medicine using TLC-Densitometric method. The result show that the three sample of rheumathic pain herbal medicine were contaminated by dexamethasone chemical medicine. Based on organoleptic test, the results show color, smell, and taste. Characterization of the dexamethasone using FTIR were to know functional groups of dexamethasone contained in the rheumatic pain herbal medicine sample, the functional groups of the sample S1, S2, and S3 were compared to dexamethasone standard. To identify of TLC method, obtained Rf value of dexamethasone standard and the sample, visualizing a stain color purple-fluorescence on the UV 254 nm. The analysis of quantitative dexamethasone concentration using TLC-Densitometric showed the presence of dexamethasone in the sample for S1, S2 and S3. The concetration of dexamethasone obtained of sample S1, S2, S3 were 1014.64 µg/g ; 131.15 µg/g ; 135.54 µg/g respectively.

Highbush Blueberry (Vaccinium corymbosum L.) Leaves Extract and Its Modified Arginine Preparation for the Management of Metabolic Syndrome-Chemical Analysis and Bioactivity in Rat Model.

Growing blueberry (Vaccinium corymbosum L., Highbush blueberry) as a berry crop is developing dynamically, especially in warm temperate, subtropical, and tropical regions of the world. When blueberry is cultivated on plantations, the bushes are pruned annually, and tons of leaves become waste. Thus, the aim of the present study was to create a preparation from blueberry leaves, study their chemical composition and determine their potential as a dietary supplement for the prophylactic and correction of the metabolic syndrome. Several schemes for obtaining extracts from blueberry leaves have been developed, including one with addition of arginine. A total of 18 phenolic substances were identified and quantified in the extracts by TLC and HPLC methods. Chlorogenic acid, hyperoside, and rutin were shown to be dominating constituents. Quantitative determination of hydroxycinnamic acid derivatives, flavonoids and other phenolics in the extracts was performed by spectrophotometric method. The extracts administration led to a significant decrease in the level of glucose, insulin and triacylglycerols in blood serum of adult mature inbred rats with insulin resistance induced by the fructose-enriched diet. The most promising one was the extract modified with arginine. The determined hypoglycemic and hypolipidemic activity of chemically standardized extracts from highbush blueberry leaves indicate the potential of this crop residue in utilization as a dietary supplement recommended in prevention of ailments associated with metabolic syndrome.

Differentiation Between Chaenomelis Fructus and its Common Adulterant, Guangpi Mugua.

The dried fruit of Chaenomeles speciosa, known as Chaenomelis Fructus or Zhoupi Mugua, is a type of traditional Chinese medicine (TCM) that is widely used to treat many diseases. Guangpi Mugua, the dried fruit of the Chaenomeles sinensis, is its most commonly known adulterant. To establish a robust approach for the quality control and identification of Chaenomelis Fructus. Thin-layer chromatography (TLC) was optimized and used to discriminate Chaenomelis Fructus from Guangpi Mugua. High-performance liquid chromatography (HPLC) combined with fingerprint analysis and partial least-squares (PLS) discrimination analysis (DA) was employed to study the chemical differences between Chaenomelis Fructus and Guangpi Mugua. The single standard to determine multi-components (SSDMC) method, with credible precision, repeatability, stability, and durability, was developed for quantitative analysis of the abundant markers. The developed TLC and HPLC methods were effective in the authentication of Chaenomelis Fructus. Moreover, oleanolic acid, ursolic acid, pomolic acid, corosolic acid, 3-O-acetylpomolic acid, and one unknown compound were identified to be critical markers for the discrimination of Chaenomelis Fructus from Guangpi Mugua. Adulteration has always been a challenge in the development of TCM. This study presents useful insights that may help solve the problem of adulteration during the preparation of Chaenomelis Fructus. The present study provides a systematic method for the quality control of Chaenomelis Fructus. This is therefore the first step towards solving the problem of adulteration to improve the clinical safety and effectiveness of Chaenomelis Fructus.

Effect of Long-Term Storage on Mycobiota of Barley Grain and Malt

Contamination of malting barley grain and malt with micromycetes sampled at various periods post-harvest (3rd, 6th, and 9th month of storage) and types of storage (storage silo and floor warehouse) was investigated. Each of these barley grain samples was malted. This article reports on the changes in the fungal microbiome composition and their overall count in barley grain and malt. From the surface-disinfected barley grain samples collected immediately after harvest, there were eight genera isolated, with a predominance of Alternaria. A small increase of isolated microfungi was detected in barley stored in silo for 3 and 6 months (from 142 isolates to 149) and decreased below the number of isolates in barley before storage (133 isolates). Fungal count during storage gradually decreased up to 9 month in barley stored in floor warehouse (from 142 isolates to 84). The initial total count of microscopic fungi in malt before storage was the highest (112 isolates) with 7 genera detected, compared to malts prepared from barley stored for longer time (54 isolates, 7 genera, 9th month of storage). Alternaria was the most abundant and frequent genus. Quantitative representation of the filamentous microscopic fungi was lower compared to yeasts especially in barley and malt prepared from barley stored at third month of storage in both type of storage. Yeasts were identified from all grain samples and malt samples with mass spectrometry. Most attention was given to the widely distributed fungus Penicillium, 79% of strains produced at least one mycotoxin detected under in vitro assays using the TLC method (97% of them produced griseofulvin, 94% CPA, 79% patulin, 14% roquefortin C, and penitrem A was produced by two screening strains under laboratory conditions). It is therefore important to monitor the microflora throughout the production cycle of “barley to beer”.

In vitro antiviral activity of peptide-rich extracts from seven Nigerian plants against three non-polio enterovirus species C serotypes

BackgroundAs frequent viral outbreaks continue to pose threat to public health, the unavailability of antiviral drugs and challenges associated with vaccine development underscore the need for antiviral drugs discovery in emergent moments (endemic or pandemic). Plants in response to microbial and pest attacks are able to produce defence molecules such as antimicrobial peptides as components of their innate immunity, which can be explored for viral therapeutics.MethodsIn this study, partially purified peptide-rich fraction (P-PPf) were obtained from aqueous extracts of seven plants by reverse-phase solid-phase extraction and cysteine-rich peptides detected by a modified TLC method. The peptide-enriched fractions and the aqueous (crude polar) were screened for antiviral effect against three non-polio enterovirus species C members using cytopathic effect reduction assay.ResultsIn this study, peptide fraction obtained from Euphorbia hirta leaf showed most potent antiviral effect against Coxsackievirus A13, Coxsackievirus A20, and Enterovirus C99 (EV-C99) with IC50 < 2.0 µg/mL and selective index ≥ 81. EV-C99 was susceptible to all partially purified peptide fractions except Allamanda blanchetii leaf.ConclusionThese findings establish the antiviral potentials of plants antimicrobial peptides and provides evidence for the anti-infective use of E. hirta in ethnomedicine. This study provides basis for further scientific investigation geared towards the isolation, characterization and mechanistic pharmacological study of the detected cysteine-rich peptides.

Optimization of the culture medium and characterization of antioxidant compounds of a marine isolated microalga as a promising source in aquaculture feed

Microalgae as photosynthetic microorganisms can be grown under controlled conditions that make them a potential source of highly valuable compounds utilized in aquaculture feed and different nutraceutical industries. The main objective of this research was to optimize the culture medium of Picochlorum sp. RCC486 as a new marine isolate for improving its growth in a microplate system using the Taguchi method. The methanol extract showed maximum FRAP activity with a value increased to 455.88 μM Fe 2+ at the concentration of 1 mg/mL. Also, the antioxidant activity of all 8 fractions purified from the methanolic extract by the TLC method was evaluated. The potent activity was observed for two fractions T1 and T2 with 544.68 μM Fe 2+ and 449.08 μM Fe 2+ at the concentration of 1 mg/mL. The GC-MS analysis was performed to identify various constituents of fractions with strong reducing power. The presence of Hexadecane: Dodecane (8.97%), and Tetradecane: Nonadecane (7.01%) in T1 fraction, and ((1,2-Benzenedicarboxylic acid, bis (2-methyl propyl) ester (12.37%), Dodecane (2.84%), Hexadecane (1.03%) and Decane: Undecane (0.74%)) in T2 fraction of methanol extract of Picochlorum sp. Contribute directly to the antioxidant effect that makes the microalga as a promising feedstock for future studies in aquaculture feed or nutraceutical industries. • KNO 3 and phosphate meaningfully affect the growth of isolated microalga. • The hexane extract showed the most DPPH and ABTS radical scavenging activity. • The methanol extract showed maximum FRAP activity and fractionated using TLC. • Identify various constituents of the more active fractions using GC-MS analysis. • Determing the compounds that contribute directly to the antioxidant effect.

Identification and activity of active compound of bamboo leaves (Bambusa vulgaris Schrad ex.J.C) ethanolic extract against diabetic ulcers gram-negative bacteria from diabetic ulcer’s patient

Background: Bamboo leaves (Bambusa vulgaris Schrad ex.J.C) are plants that are known to have pharmacological activity, one of which is to have antibacterial activity. Diabetic Foot Ulcers (DFU) is one chronic complication of diabetes mellitus. DFU is open sores on the surface of the skin that can be accompanied by local tissue death. Infection that is not appropriately treated in patients can cause amputation. Improper use of antibiotics can cause resistance events. A resistance that occurs in Proteus mirabilis, Pseudomonas aeruginosa, and Escherichia coli in isolates from DFU patients encourages researchers to look for alternative antibiotics from natural resources. Objective: Determine the profile of secondary metabolites by thin-layer chromatography, foam index, total phenols, total flavonoids and the antibacterial activity of bamboo (B. vulgaris) through the antibacterial with clear zone value of bamboo leaf ethanol extract against bacteria E. coli, P. aeruginosa, and P. mirabilis. Method: Phenol, flavonoid, and triterpenoid test using TLC method. Saponin tests were carried out using foam index, fish index, and hemolytic index test methods. Total flavonoid testing using Chang method and total phenol method using the Follin Ciocalteau reagent then proceed using a spectrophotometer. Test the antibacterial activity using the agar diffusion method. Results: Chromatographic profile, phenol, flavonoid, and triterpenoid compounds were obtained; saponin index obtained foam index of 333; fish index of 200; and the hemolytic index of 1,111. The total phenol test was 16.052 ± 3.12 % and the total flavonoid was 2.42 ± 0.44 %. Antibacterial activity test obtained MIC values of 60 mg/ml for E. coli bacteria; 100 mg/ml for the bacterium P. aeruginosa; and 30 mg/ml of P. mirabilis. Conclusion: The content of secondary metabolites found in the ethanol extract of bamboo leaves were flavonoid, phenol, saponin and triterpenoids. Ethanol extract of bamboo leaves has a the potential to produce antibacterial activity against gram-negative pathogenic bacteria isolated from diabetic wound patients Keywords: Bambusa vulgaris Schrad ex.J.C, total phenol, total flavonoid, MIC

РАЗРАБОТКА СОСТАВА И СТАНДАРТИЗАЦИЯ СБОРА ДЛЯ ПРОФИЛАКТИКИ ВОСПАЛИТЕЛЬНЫХ ЗАБОЛЕВАНИЙ ПАРОДОНТА.

Taking into account the literature data, as well as the research conducted by the authors, the composition of the collection for the prevention of inflammatory periodontal diseases has been developed, which includes medicinal plant raw materials:medicinal pimenta fruits, medicinal sage leaves, yarrow grass, badan rhizomes in a ratio of 1:1:1: 1 by weight. The quantitative ratio of the components is determined in accordance with the results of the evaluation of the antimicrobial effect of the extract from the proposed collection. To confirm the composition of biologically active substances of the collection, pharmacopoeial qualitative reactions to flavonoids and tannins, as well as the TLC method, which allows identifying substances of a polyphenolic nature, are proposed. The quantitative determination of extractive substances, essential oil, tannins, and flavonoids in the studied collection was carried out and the norms of their content were proposed. The definition of commodity characteristics is carried out.

Mini scale production of Brazilian Cherry Fruits (Eugenia uniflora L.) for the first time in NSTU campus in Bangladesh &amp; the determination of antioxidant activity of different solvent extracts of ripe and non-ripe Brazilian Cherry Fruits

The aim of this research is to cultivate Brazilian Cherry Fruits (Eugenia uniflora L.) Plants in NSTU (Noakhali Science and Technology University) Campus, Noakhali, Bangladesh for fruits production and to determine the qualitative antioxidant test, antioxidant activities and total phenolic content of different solvent extracts (Water, Methanol and Ethanol) of Ripe Brazilian Cherry Fruits in Red Color and compare those activities with non-ripe Brazilian Cherry fruits in Green color. The qualitative antioxidant test was determined by simple TLC method and antioxidant activity was determined by DPPH radical inhibition method while the total phenolic content was determined by Folin-Ciocalteu method. The result of the qualitative determination of Antioxidant test indicated that all extracts of Brazilian Cherry Fruits contain high antioxidant compounds. Among the analyzed fruits of all extracts, in both Ripe and Non-Ripe stage, the water extract of non-ripe Brazilian Cherry Fruits (BC-GC-W) showed the highest DPPH radical scavenging activity (73.66%) and showed the lowest IC50 of DPPH scavenging activity (1.01 µg/ml) which may be due to the presence of high content of Vitamin C. Among all extracts of Ripe and non-ripe, the methanol extract of non-ripe Brazilian Cherry Fruits in Green Color (BC-GC-M) was found to be highest total phenolic content 111.52 mg GAE/100g, and the ethanol extracts of Ripe Brazilian Cherry Fruits in Red Color (BC-RC-E) was found to be lowest total phenolic content, 106.004 mg GAE/100g.The overall results that were obtained in this study will be one of the important key strategies by adding this fruits in the diet chart to prevent and fight against infection recently caused by COVID-19 in worldwide and other infectious diseases by increasing our body immunity in the long term and this fruits also will be useful as a raw materials in the pharmaceutical and food industries to prepare drugs and food products.

Eco-Friendly Chromatographic Methods for Determination of Acemetacin and Indomethacin; Greenness Profile Assessment.

The green chemistry approach was developed for the purpose of saving the environment by using green solvents. Applying green analytical chemistry principles to traditional methods is considered a challenge. Acemetacin is a commonly used analgesic prodrug that bioactivates to indomethacin. Developing two simple, eco-friendly chromatographic methods for simultaneous determination of acemetacin and indomethacin. The first method is HPLC-DAD. Separation was performed on a Waters XBridge® Shield RP18 (250 × 4.6 mm, 5 µm) analytical column with ethanol-ammonium acetate buffer (50 mM, pH 3.5 ± 0.1; 60:40, v/v) as a mobile phase at a flow rate of 1 mL/min at 25 ± 0.5°C and UV detection at 254 nm. The other method is TLC coupled with densitometric quantification using pre-coated silica TLC plates and butanol-ethyl acetate (70:30, v/v) elution system. The plates were scanned at 254 nm. Both methods were validated according to International Conference on Harmonization guidelines. Linearity was confirmed for both over a concentration range of 1-100 µg/mL for the HPLC method and 0.2-7 µg/band for TLC-densitometric method. The methods' greenness was evaluated by the National Environmental Methods Index, Eco-Scale, Green Analytical Procedure Index metrics and Analytical GREEnness Metric Approach. The proposed methods were found to be suitable for determination of studied drugs in their marketed formulations and is suggested for routine analysis in quality control laboratories. The developed HPLC method shortened the elution time of the analyzed drugs saving more time and money and the TLC method lowered the drugs' detection limit. HPLC and TLC methods were validated for the assay of acemetacin and indomethacin. The methods' greenness was evaluated and compared to published methods. The developed HPLC method shortened the elution time of the analyzed drugs, saving time and money and the TLC method lowered the drugs' detection limit.

Acetylcholinesterase inhibitory activity of extract and fractions from the root of Rauvolfia serpentina(L.) Bth.ex Kurz.

Alzheimer's disease (AD) is a degenerative brain disease characterized by confusion, behavior changes, decline in memory and cognitive skills. One of the strategies in the treatment of AD is to use acetylcholinesterase (AChE) inhibitors. The current study aims to determine the AChE inhibitory activities of the extract and fractions of the root of Rauvolfia serpentina. Extraction was carried out by maceration method using ethanol, followed by liquid-liquid partition using n-hexane, ethyl acetate and n-butanol. Further fractionation was conducted by using vacuum liquid chromatography (VLC). The AChE inhibitory assays were performed by using Ellmann's method. Phytochemical screening was carried out by TLC method. The ethanolic extract of R.serpentina showed inhibition against AChE enzyme with an IC50 value of 7.46μg/mL. The extract and fractions showed higher inhibition against butyrylcholinesterase (BChE) compared to AChE. Amongst three fractions obtained, the n-butanol fraction showed the strongest inhibition with an IC50 value of 5.99μg/mL against AChE. VLC fractionation of the n-butanol fraction yielded 13 subfractions (VLC 1-VLC 13). Four out of 13 subfractions gave more than 80% inhibition against AChE, namely subfractions 4-7, with IC50 values ranging from 4.87 to 47.22μg/mL. The phytochemical screening of these subfractions suggested the presence of alkaloids. The ethanolic extract, as well as fractions of R.serpentina root, are potential for AChE inhibitor. The alkaloid compound may be responsible for this activity.

A pattern recognition method on smartphones for planar chromatography and verification on chromatograms of four herbal medicines from citrus fruits

Image retrieval techniques in combination with support vector machine (SVM) on smartphones were studied to make pattern recognition of planar chromatograms. A micellar TLC method employed less toxic and flammable solvents was studied to discriminate four herbal medicines (HMs) from citrus fruits. And the chromatograms were used to test the pattern recognition method. The TLC method employed O/W microemulsion–formic acid (98:2) as the mobile phase, polyamide as the solid phase, and 1% aluminum trichloride aqueous solution as the visualization reagent. TLC of samples of dried lemon peel, dried sweet-orange peel, pummelo peel, and mandarin orange peel of varying origins were performed in three separate runs. Image retrieval techniques combined with SVM directly classified the chromatograms based on migration path images. A mobile application running on an Android smartphone was developed to perform the pattern recognition. The micellar TLC method satisfactorily characterized the chemical profile of the four HMs from citrus fruits. The pattern recognition method combining color layout descriptor with SVM correctly classified all the chromatograms. A smartphone with an application combining color layout descriptor with SVM would be a convenient tool for pattern recognition of planar chromatograms.