Thin-layer Chromatographic Method Research Articles

Quantitative analysis of quercetin in tea mistletoe (Scurrula oortiana Dans.)

Indonesia has a variety of plants that are useful as medicine. One is the Tea Mistletoe plant (Scurrula oortiana Dans.). This plant contains flavonoid compounds that are effective as antioxidants, especially in the form of quercetin. Quercetin in the tea mistletoe plant can prevent cell damage caused by free radicals, where cells need this antioxidant to avoid the harmful effects of excessive reactive oxygen species (ROS) production and prevent damage to immune cells. This study aims to analyze the levels of quercetin in Tea Mistletoe extract using quantitative analysis methods such as thin-layer chromatography and UV-Vis spectrophotometry. Tea Mistletoe powder was extracted by maceration with 96% ethanol solvent and then concentrated using a rotary evaporator and water bath. The thick extract obtained was then tested for phytochemical screening and specific and non-specific parameters. Analysis of flavonoid quercetin compounds was carried out using a thin layer chromatography method using a mobile phase of a mixture of chloroform, ethyl acetate, and n-butanol with a ratio of 5:4:1 and a stationary phase of Silica Gel GF₂₅₄. Determination of flavonoid levels was carried out using UV-Vis spectrophotometry at a wavelength of 428 nm. The results of this study indicate that the Mistletoe Tea plant contains flavonoids, alkaloids, steroids, tannins, saponins, phenolics, and glycosides. All test parameters meet the specified requirements. The thin layer chromatography test showed the Rf value of the Mistletoe Tea extract of 0.78. The linearity test showed a correlation value (r) = 0.9988 with a total flavonoid compound content of 372.250 mg QE/g extract, or a percentage of 37.225%.

RNA Modification and Digestive Tract Tumors: A Review.

Gastrointestinal tumors, including colorectal and liver cancer, are among the most prevalent and lethal solid tumors. These malignancies are characterized by worsening prognoses and increasing incidence rates. Traditional therapeutic approaches often prove ineffective. Recent advancements in high-throughput sequencing and sophisticated RNA modification detection technologies have uncovered numerous RNA chemical alterations significantly associated with the pathogenesis of various diseases, notably cancer. These discoveries have opened new avenues for therapeutic intervention. This article delves into epigenetic modifications, with a particular emphasis on RNA alterations such as N6-methyladenosine (m6A), 5-methylcytosine (m5C), 1-methyladenosine (m1A), 7-methylguanosine (m7G), and N4-acetylcysteine (ac4C). It examines the functions and mechanisms of action of regulatory entities known as "Writers," "Readers," and "Erasers" to these modifications. Additionally, it outlines various methodologies for detecting these RNA modifications. Conventional techniques include radioactive isotope incorporation, two-dimensional thin-layer chromatography (2D-TLC), mass spectrometry, and immunological detection methods. Specialized methods such as bisulfite sequencing and reverse transcription stops are also discussed. Furthermore, the article underscores the significance of these modifications in the development, progression, and therapeutic targeting of gastrointestinal tumors, including esophageal, gastric, colorectal, liver, and pancreatic cancers. This exploration provides foundational insights for enhancing diagnostic accuracy, treatment efficacy, and prognostic assessment in gastrointestinal oncology.

Development and validation of high-performance thin-layer chromatography (HPTLC) and high-performance liquid chromatography (HPLC) methods for the simultaneous determination of myricetin and quercetin in Manilkara hexandra

Development and validation of high-performance thin-layer chromatography (HPTLC) and high-performance liquid chromatography (HPLC) methods for the simultaneous determination of myricetin and quercetin in Manilkara hexandra

Unlocking the access to nature-identical vanillin via isoeugenol ozonation: in situ ATR-IR monitoring and safety evaluation.

The transformation of renewable feedstocks into aromatic chemicals holds immense potential for advancing a green, low-carbon economy and fostering sustainable development. Herein, we present a novel approach for the conversion of isoeugenol, a renewable lignin derivative, into the valuable flavoring agent vanillin, utilizing ozone as an environmentally benign oxidant. The process optimization was significantly enhanced by the integration of in situ Attenuated Total Reflectance Infrared (ATR-IR) monitoring. The introduction of H2O not only accelerated the decay of carbonyl oxides (Criegee intermediates) but also mitigated safety hazards stemming from the vigorous decomposition and heat release of secondary ozonides. Compared to the conventional Thin Layer Chromatography (TLC) method, ATR-IR monitoring demonstrated superior sensitivity and precision in determining the reaction endpoint, leading to a remarkable vanillin yield of 96.86% upon complete conversion of isoeugenol. Additionally, a comparative assessment of the sustainability of our approach with existing methods was undertaken, and valuable recommendations for safety assessments were provided to ensure the inherent safety of chemical engineering reactions. The present study serves as a pioneering effort in facilitating the implementation of a scalable, economically feasible and environmentally sustainable strategy for biomass flavor production, while contributing to the broader adoption of in situ spectroscopic technology within the larger economy.

Optimal conditions for reducing matrix effects when conducting chemical toxicological studies of antidepressants

The article presents an analysis of current data on the need to develop effective methods for the qualitative and quantitative assessment of antidepressants during chemical and toxicological analysis. The optimal conditions for eliminating the matrix effect by extraction and sample preparation selection have been determined. A thin-layer chromatography method for the analysis of carbamazepine and fluoxetine has been developed. The selection of the mobile phase was carried out, and the optimal conditions for the purification of antidepressants were determined using the thin-layer chromatography. A UV spectrophotometry method for the determination of carbamazepine and fluoxetine has been developed, and validation characteristics of the method have been obtained.

Stability-Indicating High-Performance Thin-Layer Chromatography Method Development and Validation for Topiroxostat in Bulk and Tablet Dosage Forms Using a Quality by Design Approach

Stability-Indicating High-Performance Thin-Layer Chromatography Method Development and Validation for Topiroxostat in Bulk and Tablet Dosage Forms Using a Quality by Design Approach

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.

Antibacterial and antioxidant activities of Eleutherine americana (Aubl.) Merr. leaves extract growing in different cultivation sites

Eleutherine americana (Aubl.) Merr. is used as a traditional remedy in Indonesia. Based on its chemical compounds, the leaves were expected to exhibit antibacterial and antioxidant activities. The aim of the study was to assess the antibacterial capacity, antioxidant capacity, total phenolic content, total flavonoid content, and phytochemical compounds in ethanolic extract of E. americana leaves collected from various cultivation sites, namely West Kotawaringin, Central Kalimantan (EW); Malang, East Java (EM); and Probolinggo, East Java (EP). The antibacterial activity assay was conducted utilizing the diffusion method, whereas the determination of antioxidant capacity was performed utilizing the DPPH method. The total phenolic content and total flavonoid content were determined using UV-Vis spectrophotometry. The phytochemical screening was conducted utilizing a tube test and thin layer chromatography (TLC) method. The growth of E. coli and S. aureus was found to be inhibited by EW, EM, and EP. Furthermore, the antibacterial activity of all samples against S. aureus was higher than that against E. coli. EP had the highest antioxidant activity (IC50 = 32.83±3.51 µg/mL), total phenolic content of 129.40±0.19 mg GAE/g extract, and total flavonoid content of 0.61±0.01 mg QE/g extract. Phytochemical screening revealed that the leaves of E. americana contained alkaloids, terpenoids, flavonoids, polyphenols, and tannins. According to the findings of this study, E. americana leaves could potentially be developed as a new natural antibacterial and antioxidant agent.

Analytical Assessment of the Quality of Dietary Supplements and Cosmetic Products Containing Xanthohumol by Thin-Layer Chromatography Along with the Estimation of Its Antioxidant Potential

Xanthohumol, a prenylated chalcone in the flavonoid group, naturally occurs in many plants and exhibits antioxidant, anticancer, anti-inflammatory, antibacterial, and antiviral effects. The growing interest in xanthohumol due to its potential therapeutic properties has led to the increase in the pool of products available on the market. The novelty of this study is the proposal of a rapid and cost-effective procedure useful for performing quality control on products containing xanthohumol in the form of dietary supplements and cosmetics as well as testing their stability. For this purpose, the thin-layer chromatography method with densitometric detection was used, which was validated in accordance with ICH (International Conference on Harmonization) guidelines. The mobile phase was toluene, 1,4-dioxane, and glacial acetic acid (37:10:1.5 v/v/v), and TLC silica gel 60 F254 plates were used as the stationary phase. The validation process assessed linearity, with a correlation coefficient (r) of 0.9987. The calculated LOD (limit of detection) and LOQ (limit of quantification) values were 3.82 and 11.57 ng/spot, respectively. Accuracy was evaluated by determining percentage recovery at three concentration levels (80, 100, and 120%), with an average recovery of 100% and RSD below 1%, confirming good accuracy. Precision was indicated by an RSD of less than 2.20%. The average content of xanthohumol in dietary supplements ranged from about 8 to 29% of the content declared by the manufacturers. The stability tests showed that XN decomposes most slowly in water (t0.5 = 10.86 h) compared with acidic (t0.5 = 10.80 h) and alkaline solutions (t0.5 = 7.39 h), as well as in the presence of an oxidizing agent (t0.5 = 18.38 h), at all tested temperatures, which is confirmed by the calculated kinetic parameters. In the tests of antioxidant capacity, xanthohumol shows significantly higher radical scavenging capacity than vitamin C in the entire range of analyzed concentrations (0.03–2.40 mmol/L).

Development and Validation of a High-Performance Thin-Layer Chroma-tography Method for Qualitative and Quantitative Analysis of Wogonin in S. barbata D. Don Herb

Background and Aims: Scutellaria barbata D. Don, a perennial herb with a rich history in traditional medicine, is known for its bioactive compounds, including wogonin. It possesses anti-inflammatory, antioxidant, and anticancer properties. Together with other bi-oactive compounds, wogonin contributes to the pharmacological activities associated with S. barbata D. Don, making it a subject of interest in both traditional and modern medicinal re-search. This study aimed to develop and validate a High-Performance Thin-Layer Chroma-tography (HPTLC) method for the precise and accurate quantification of wogonin in the hy-droethanolic extract of S. barbata D. Don. Methods: The process was developed and underwent rigorous validation following the ICH guidelines. Parameters such as linearity, specificity, instrument precision, precision, accura-cy, limit of detection (LOD), limit of quantification (LOQ), and robustness were systemati-cally evaluated to ensure the method's reliability. Results: The HPTLC separation was conducted on TLC aluminum plates precoated with sil-ica gel 60 F254, and optimal separation was achieved using Ethyl acetate: toluene: formic ac-id (5: 4.9: 0.1, (v/v/v/v)) as the mobile phase. Densitometric scanning at 254 nm in reflec-tion/absorbance mode revealed compact spots at Rf 0.87, corresponding to wogonin. Linear regression analysis demonstrated a robust linear relationship between peak area and the amount of wogonin in the range of 10-40 (μg/mg). The quantification of wogonin in crude extracts obtained was 3.274 μg/mg of extract. Conclusion: In summary, the developed HPTLC method emerges as a straightforward, pre-cise, and accurate tool for the rapid screening of active constituents, specifically wogonin, within the petroleum ether extracts of S. barbata D. Don. This method holds promise for routine quality control and analysis of herbal materials and formulations containing S. bar-bata D. Don, contributing to the ongoing exploration of its therapeutic potential. other: NA

TLC vs. HPLC: A green comparison for veterinary drug analysis

This work describes two analytical methods for determining lidocaine (LD) and oxytetracycline (OTC) in veterinary dosage form: a traditional Thin-Layer Chromatography (TLC) method and a more environmentally friendly Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC) method.Initially, the TLC method was used due to its speed, simplicity, and accuracy. It successfully separated LD and OTC in veterinary dosage forms with a linearity range of 0.5–11.0 µg/band for LD and 4.0–34.0 µg/band for OTC. However, TLC proved to be inadequate for resolving the binary mixture in the presence of toxic lidocaine impurity (2,6-dimethylaniline) (DMA), and was less environmentally friendly.Consequently, an RP-HPLC method was developed using a C18 column and a mobile phase consisting of ethanol and phosphate buffer (pH 7) in a 70:30 ratio, with a flow rate of 1.0 mL/min. This method effectively separated OTC, LD and DMA, using a PDA detector at wavelengths of 272, 232, and 210 nm and with retention times of 1.529, 2.77, and 3.581 min for OTC, DMA, and LD respectively. The RP-HPLC method showed linearity ranges of 270.0–972.0 µg/mL for OTC, 0.5–25.0 µg/mL for DMA, and 5.0–18.0 µg/mL for LD.Both methods were validated for linearity, precision, accuracy, robustness, detection limit, and quantitation limit. The greenness of each method was evaluated using NEMI, Eco-scale, and Agree tools, with results demonstrating the RP-HPLC method as the more environmentally sustainable method.

Standardisation and Antimicrobial Activity of <i>Hedychium spicatum</i> (<i>Shati</i>)

Background: Hedychium spicatum Buch. Ham. (Zingiberaceae), also known as the spiked ginger lily, is a versatile herbal medicine used in traditional systems of medicine for its therapeutic properties. It has been used as an analgesic, antifungal, anti-inflammatory, antimicrobial, antioxidant, antispasmodic, Central Nerves System (CNS) depressant and for cytotoxic activities, fevers, pediculicidal and respiratory disorders. Aim: To standardise and evaluate the antimicrobial activity of H. spicatum (Shati). Method: A study was conducted to standardise the anti-microbial activity of H. spicatum (Shati). An extract was prepared using hydroalcoholic and organoleptic methods and physico-chemical analyses were performed. Results: It was discovered that High-Performance Thin Layer Chromatography (HPTLC) was more flexible than traditional Thin Layer Chromatography (TLC) methods. The hydroalcoholic extracts inhibited good antimicrobial activity against all tested fungi and bacteria. The study concludes that the standardisation and potential antimicrobial activity of H. spicatum extracts against tested bacteria and fungi have been evaluated. Conclusion: Further investigation is needed to explore polyherbal formulations for infectious skin disorders.

Method Development and Validation of HPTLC Method for Simultaneous Estimation of Atropine, Piperine and Vasicine in <i>Ayurvedic</i> Formulation

Background: Dhumayoga, an Ayurvedic formulation is used for the management of respiratory disorders, including dyspnea and asthma. Aim: To develop and validate a High-Performance Thin-Layer Chromatography (HPTLC) method for the simultaneous estimation of atropine, vaccine and piperine in Dhumayoga Ayurvedic formulation. Methods: The three biomarkers were resolved using precoated silica gel 60F254 as stationary phase, dioxane: toluene: methanol: ethyl acetate: Aqueous ammonia (4:7:3:5:1v/v/v/v/v) as mobile phase and detected and quantified at 254nm. Results: The Rf values of atropine, vasicine, and piperine were found to be 0.406 ± 0.02, 0.56 ± 0.02, 0.87 ± 0.02, respectively. The calibration plot was linear in the range of 500-2500ng per zone with a correlation coefficient (r2) of atropine, vasicine and piperine was 0.994, 0.9986 and 0.9935, respectively. Conclusion: The developed method was applied for the quantitation of biomarkers in Dhumayoga. The method was validated in terms of linearity, precision, specificity, accuracy and robustness as per International Council for Harmonisation (ICH) guidelines.

Sustainable and efficient monitoring of tryptophan and tyrosine serum levels: a green HPTLC method as a biomarker for type 2 diabetes

In recent years, there has been considerable interest in using amino acids like tryptophan (Trp) and tyrosine (Tyr) as biomarkers for various diseases, including type 2 diabetes mellitus (T2D). In diseases like T2D, the metabolism of Trp and Tyr is altered. The activity of enzymes involved in Trp metabolism increases, leading to a decrease in its serum level. On the other hand, the serum level of Tyr increases due to the suppressed activity of its metabolizing enzymes. These observations suggest that Trp and Tyr metabolism may play a crucial role in the pathophysiology of type 2 diabetes. Our study highlights the potential utility of Trp and Tyr as biomarkers for the early detection, prognosis, and monitoring of this metabolic disorder. Given these observations, we aimed to develop a high-performance thin-layer chromatographic (HPTLC) method that is sensitive, selective, rapid, and environmentally friendly for estimating the concentrations of Trp and Tyr in biological fluids, particularly serum samples. To evaluate the method, we performed analysis using serum samples from controlled and streptozotocin-induced diabetic rats. Our main objective was to develop a method that is sensitive and selective for precisely determining Trp and Tyr serum levels, which could serve as potential biomarkers for T2D. Fluorescence and absorption modes were employed for densitometry scanning. We assessed the precision and high separation efficiency of the chromatographic system by calculating parameters such as separation and resolution factors, number of theoretical plates, and height equivalent to theoretical plates. To evaluate the environmental impact of our proposed method, we employed the AGREE (Analytical GREEnness metric) and GAPI (Green Analytical Procedure Index) greenness assessment tools. The results confirmed that our method is environmentally friendly and exhibits superior eco-friendliness and greenness compared to other reported methods.Graphical

A high-performance thin-layer chromatography densitometric method for the separation of isomeric ceftriaxone in powder for injection formulation

The aim of this study was to develop and validate a High-Performance Thin Layer Chromatographic (HPTLC) method for simultaneous determination of ceftriaxone and ceftriaxone e-isomer in powder for injection formulation. Ceftriaxone sodium injection is an antibiotic that used globally. It has Z/E geometrical conformation, in which ceftriaxone sodium and 3 ene-isomer have Z- conformation while (E)-isomer has E- conformation and the potential toxicity of ceftriaxone (E)-isomer has been reported. Thus, to safeguard the public health, a simple and easy to use, rapid and reliable method was developed for qualitative and quantitative determination of ceftriaxone sodium and its (E)–isomer. Samples were applied on HPTLC glass plates precoated with silica gel 60F254 by using Linomat semi-auto sampler. Separation was carried out using acetone, triethyl amine, water, chloroform and ethyl acetate as a mobile phase in different ratios. The Rf values of separated compounds were 0.51 ± 0.01 and 0.62 ± 0.01 for ceftriaxone sodium and ceftriaxone (E)-isomer respectively. The method was validated by studying Specificity, Linearity, Accuracy, Precision, Robustness, Limit of Detection (LOD) and Limit of Quantification (LOQ) and Solution stability. The developed method was successfully, sensitive, simple, precise, accurate, robust and applicable for the simultaneous determination of ceftriaxone sodium and ceftriaxone (E)-isomer in powder for injection formulation.

Enzymatic Production of Chitooligosaccharide Using a GH Family 46 Chitosanase from Paenibacillus elgii and Its Antioxidant Activity

Chitooligosaccharide (COS), a natural antioxidant, is a hydrolysis product of chitosan created using enzymatic or chemical methods. COS has received considerable attention recently, making its efficient bioproduction of great value. This study investigated the optimal conditions for the enzymatic method using a GH family 46 chitosanase from Paenibacillus elgii TKU051 to prepare COS based on the response surface methodology (RSM). The results showed optimal values for chitosan hydrolysis, such as a pH of 5.5, an incubation temperature of 58.3 °C, an [E]/[S] ratio of 118.494 (U/g), and an incubation time of 6.821 h. Under the optimal conditions, the highest reducing sugar level (per substrate, w/w) of the chitosan hydrolysis process that could be reached was 690.587 mg/g. The composition of the obtained COS was analyzed using the thin-layer chromatography (TLC) method, yielding (GlcN)2 and (GlcN)3 as the products. The ascorbic acid equivalent antioxidant capacity (AEAC) of the obtained COS was found to be 1246 mg/100 g (via a DPPH (2,2-diphenyl-1-picrylhydrazyl) radical-scavenging assay) and 3673 mg/100 g (via an ABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) radical-scavenging assay). This green and efficient bioproduction method may possess excellent potential for application in bioactive COS preparation.

Separation of Gallic acid and Quercetin from the Cassia tora extract Using Thin Layer Chromatography: Simple and Cost-Effective Approach

Gallic acid and quercetin are the flavonoids present in the Cassia tora and play significant role in healing number of ailments. No method has yet been developed to separate and identify gallic acid and quercetin from Cassia tora extract using thin layer chromatography (TLC). The TLC method is simple and affordable for the identification of flavonoids compared to advanced chromatography tools. Therefore, the aim was to create a TLC-based method for identifying and isolating gallic acid and quercetin from the Cassia tora extract. The petroleum ether and hydroalcoholic extract were prepared form the aerial part of the Cassia tora. The petroleum ether and hydroalcoholic extract were subjected to the phytochemical analysis. The TLC was performed with hydroalcoholic extract to separate and identify gallic acid and also quercetin. Mobile phase made in ratio (4:5:1) and (5:7:1), from reagents i.e. ethyl acetate, toluene, and formic acid, was used for the separation of quercetin and gallic acid, respectively. The findings of phytochemical studies expressed existence of polyphenol and flavonoids in extract with hydro-alcohol, so that hydroalcoholic extract was used for TLC study. The extract and pure quercetin exhibited 0.62 value of Rf, while extract and gallic acid revealed 0.34 Rf value, authorizing existence of quercetin and also gallic acid in Cassia tora extract. This analysis thus concluded that TLC is most effective approach for identifying and isolating components i.e. gallic acid and also quercetin from extract of Cassia tora.

Validation of chromatographic methods, TLC and HPLC, to quantify known radiochemical impurities and the radiochemical purity the radiopharmaceutical [177Lu]Lu-PSMA I&T

The Energy and Nuclear Research Institute (IPEN) is studying the production of the radiopharmaceutical [177Lu]Lu-PSMA I&T, in accordance with the good manufacturing practices recommended by the National Health Surveillance Agency (ANVISA), to be used in the therapy of prostate cancer. This work aims to validate chromatographic methods, Thin-Layer Chromatographic (TLC) and high-performance liquid chromatography (HPLC), to determine the radiochemical purity (RCP) of the product. The entire validation process of this work was based on ANVISA's RDC 166, 2017 and the Guide 10, version 1, 2017, guided the statistical treatments adopted. With the selectivity study we can observe that the presence of impurities or excess excipients in the sample does not interfere with the quantification of the product. The proposed methods were linear with linear correlation coefficients (r) above 0.99. The precision and repeatability presented relative standard deviation values lower than specified (RSD <5 %). The small controlled variations in the method suggested for the robustness test also did not affect the radiochemical purity of the product. In view of the results and in accordance with the criteria established by the National Health Surveillance Agency (ANVISA), the two chromatographic methods were validated in accordance with RDC 166, 2017, proving to be selective, precise, linear and robust. The validation of TLC and HPLC methods enables their application in the batch release routine of the new radiopharmaceutical at Radiopharmacy Center of IPEN.

Quantification of Metronidazole in Tablets: Combining Thin-Layer Chromatography in the GPHF-Minilab™ with Image Processing Using Open-Source ImageJ Software

The GPHF-Minilab™ is a portable toolkit for performing qualitative methods such as thin-layer chromatography (TLC) on common pharmaceuticals. It is particularly useful in resource-limited locations where it is more challenging to monitor for substandard and falsified (SF) medicines. However, the GPHF-Minilab™ TLC methods are only semi-quantitative at best and thus have issues monitoring product quality effectively. We have improved on the GPHF-Minilab™ TLC method for metronidazole, a common antibiotic, by making it fully quantitative. Sample solutions were spotted on TLC plates alongside three metronidazole standards at different concentrations. After development, plates were imaged in a lightbox with two different smartphone cameras. Images were processed through the open-source program ImageJ and resulting pixel data from the standard spots were used to create a calibration curve, enabling quantitation of the sample. The USP Metronidazole Tablet high-performance liquid chromatography (HPLC) assay was used as the reference method. We validated this TLC method using 250 and 500 mg metronidazole tablets from different manufacturers and assessed linearity, range, accuracy, precision, intermediate precision, specificity, and robustness. These improvements should enhance the GPHF-Minilab™ TLC methods for metronidazole product screening. Additionally, the procedure is extensible to other analytes, although further validation would be required for each Minilab method.

Енергетичний обмін у печінці щурів із паркінсонічним синдромом, індукованим ротеноном, при дії метаніндіазенону

Parkinson’s disease (PD) is symptomatically characterized by motor disorders in the human body, which are associated with the degeneration of dopamine neurons in the substantia nigra of the midbrain. It is widely recognized that mitochondrial dysfunction occurs in dopaminergic neurons, leading to their death and the development of this pathology. Recently, benzodiazepine derivatives have been found to have a neuroprotective effect against damage of dopaminergic neurons in an experimental model of Рarkinsonian syndrome (PS). The aim of this study was to investigate the effect of a novel molecule, named methanіdiazenone, on energy metabolism in the liver of rats induced by rotenone. To achieve the set goal, the concentration of ATP, ADP, AMP, hypoxanthine аnd xanthine were determined in the bile samples of rats with PS by the method of thin-layer chromatography. The obtained data indicate that under the influence of rotenone compared to the control values the content of ATP in bile decreased by 40%, while the levels of AMP, xanthine and hypoxanthine, on the contrary, increased by 87,5%, 55,6%, and 25%, respectively. Our findings suggested that the ratio of AMP/ATP, which is an important indicator of the functional state of mitochondria, under the influence of rotenone increased by 200% compared to the control. Metanindiazenone in a dose of 1,0 and 2,0 mg/kg normalized all studied parameters of purine metabolism. The presented results show that methanindiazenone significantly improves purine metabolism in the liver of rats with PS induced by rotenone, indicating the normalization of mitochondrial function in hepatocytes. Thus, methanindiazenone may be recommended for clinical trials regarding its use in combination with other drugs to treat Parkinson’s disease and possibly other neurodegenerative diseases, in which mitochondrial dysfunction occurs.