Jpc-journal of Planar Chromatography-modern Tlc最新文献

Analytical approaches that adhere to the principles of white and green chemistry are necessary to address concerns related to the environment, economy, and practicality. An analytical quality by design (AQbD)-aided ecofriendly reversed-phase thin-layer chromatography (RP-TLC) approach was utilized to quantify the flavor enhancer monosodium glutamate (E-621) in various food samples, followed by ultrasound-assisted extraction. RP-TLC layers precoated with RP-18 modified silica gel 60 F_254s were utilized as the stationary phase with green mobile phases consisting of ethanol‒water‒isopropanol (5:2.5:2.5, V/V), with densitometric detection at 570 nm with an R_F value of 0.57. Monosodium glutamate (MSG) shows linearity in 500–1500 ng/spot range. RP-TLC separation was optimized, and the most influential and interacting parameters were identified using central composite design. Validation study was performed as per the International Council for Harmonisation (ICH) recommendations, which demonstrates all the parameters are within acceptable limits. MSG shows recovery of 93.12–101.42% among various food samples. The greenness of the created method was evaluated using the green analytical procedure index (GAPI) and analytical greenness (AGREE). We utilized the blue applicability grade index (BAGI) and red–green–blue 12 (RGB12) algorithms to evaluate the concepts of blueness and whiteness. The BAGI score 80 suggested a high level of application, while the RGB12 score 85.1 proved that the approach is both cost effective and sustainable. This revolutionary RP-TLC method provides a fast and environmentally friendly alternative to traditional procedures used to analyze MSG in foods, which might help pharmaceutical industries to adopt and improve robust, environmentally friendly AQbD techniques for routine MSG assay using green solvents.

As Ayurvedic herbal medicines have grown more commercialized, it has become crucial to standardize these products using modern techniques. Mansyadi Kwatha is an Ayurvedic formulation indicated for insomnia, epilepsy, hysteria and anxiety. It consists of three potent drugs, i.e., Nardostachys jatamansi rhizome, Withania somnifera L. root and Hyoscyamus niger L. seed. In this study, a novel instrumental thin-layer chromatography (TLC) method was developed for the simultaneous estimation of atropine, rutin and vanillin. It was then employed for marker-based standardization of Mansyadi Kwatha for detection of these constituents and quantifying the same in the formulation. A pre-coated silica gel TLC plate 60 F₂₅₄ was used as the stationary phase with tetrahydrofuran‒toluene‒methanol‒formic acid (5:3.5:2:0.5, V/V) as the mobile phase for chromatographic separation with densitometric scanning at 206 nm. The developed method gave the desired separation with optimum R_F values of 0.090 ± 0.0039, 0.290 ± 0.0099 and 0.679 ± 0.0056 for atropine, rutin and vanillin, respectively. The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) Q2(R1) guidelines were followed for method validation. The method was found to be specific with comparable R_F and no interference of the mobile phase. It showed a good linear relation with correlation coefficient greater than 0.99. The method was also precise, robust with percentage relative standard deviation less than 2% and accurate with percent recovery between 95‒105%.

This paper introduces a new methodology for assessing the quality of a polyherbal preparation using thin-layer chromatography (TLC) image analysis integrated with the leave-1-out method and cross-sectional study. “Phikud Navakot”, a Thai polyherbal preparation for relieving circulatory disorders and dizziness, consisting of nine herbs, was used as a case study. The leave-1-out technique involves systematically excluding one herb at a time from the preparation to generate TLC profiles, which are then analyzed and compared with the original preparation. Another approach, based on a cross-sectional study of the TLC image, was employed by converting all track on the TLC image into red, green, and blue (RGB) profiles simultaneously for direct band-by-band comparison across chromatograms. The leave-1-out approach combined with image analysis enhances band detection clarity, allowing for a more precise assignment of each band to its corresponding herb. Regardless of whether the substance band in polyherbal preparation was distinctly noticeable, faint, closely adjacent, overlapping, or concealed, the approaches based on TLC image analysis integrated with the leave-1-out method and cross-sectional study were demonstrated to be effective analytical techniques for complex herbal preparations and could be applied to other herbal mixtures.

Bauhinia vahlii Wight and Arn. is a commonly used climber in traditional medicinal practices. The present work introduces a validated high-performance thin-layer chromatography (HPTLC) method for identifying the superior B. vahlii species from various regions in India, such as Pune, Jhansi, and Ranikhet, by analyzing phytochemical markers. The current study identified and estimated lupeol and β-sitosterol in leaf extracts of B. vahlii, detecting them as magenta and violet grey bands using anisaldehyde‒sulfuric acid reagent at retention factor (R_F) ~0.68 and R_F ~0.53, respectively, at 514 nm. This was achieved using a mobile phase consisting of toluene‒ethyl acetate‒formic acid (8:2:0.2, V/V). The method was validated for parameters such as linearity, limits of detection, and quantification and precision, as per the International Council for Harmonisation (ICH) guidelines Q2R1. The calibration plots of the analyzed data exhibited a good linear relationship with R² = 0.99 ± 0.02 in the concentration range 200‒1400 ng per spot for lupeol and 600‒1300 ng per spot for β-sitosterol with respect to the peak area. Lupeol was quantified as 61.61, 37.10, and 63.14 µg in 100 mg leaf extracts of B. vahlii collected from Pune, Jhansi, and Ranikhet regions of India, respectively. Meanwhile, β-sitosterol was determined to be 116.80, 105.13, and 90.25 µg from Pune, Jhansi, and Ranikhet, respectively, in 100.0 mg of the sample. Among the three locations, Ranikhet exhibited the highest lupeol content, while Pune showed the highest β-sitosterol estimation. The validated HPTLC method employed was simple, accurate, and reproducible for the rapid screening of B. vahlii collected from diverse Indian topographies. This study may be utilized as a valuable quality assessment tool for stakeholders, aiding adherence to Good Manufacturing Practice standards.

Graphical abstract

In addressing the time-consuming and labor-intensive issues inherent in traditional marker selection mode including systematic isolation, structural elucidation, and activity screening of indicator components for a single traditional Chinese medicine (TCM), the aim of this study was to develop a method for quick simultaneous identification of multiple medicinal drugs by using high-performance thin-layer chromatography (HPTLC) technique featuring the advantages of being intuitive, easy to operate, and offering multi-dimensional information. Taking three TCMs originating from five closely related different species from Cruciferae family as an example, the study conducted parallel comparisons of optimized TLC chromatograms of seeds from the five species Lepidium apetalum, Descurainia sophia, Raphanus sativus, Sinapis alba, and Brassica juncea under different detection wavelength lights; eight characteristic components were distinctly discovered, and among them one known common constituent identified as sinapine. Subsequently, seven characteristic components were isolated on a small scale and within a short-time, and elucidated structurally. Lastly, for each seed, a combination of the common constituent and most of one of its characteristic components were chosen as identification markers for the respective seed. To sum up, a unified HPTLC method with different detection wavelengths and different marker combinations was established for the simultaneous identification of five seed-originated medicines. This research approach, characterized by simplicity, cost-effectiveness, and rapidity, presents a highly efficient alternative for establishing identification methods for chemically similar multiple TCMs from closely related species.

In recent years, green analytical chemistry (GAC) has played an important role in the development of environmentally friendly analytical methods. In the field of pharmaceutical analysis, ecofriendly analytical methods are attracting attention to ensure the safety of workers and the environment. A three-drug combination of metformin hydrochloride, sitagliptin, and dapagliflozin propanediol monohydrate was recently approved for the treatment of type II diabetes. A simple, rapid, and reproducible green thin-layer chromatography (TLC) method was developed for the simultaneous determination of metformin hydrochloride, sitagliptin, and dapagliflozin propanediol monohydrate in tablet dosage form. Chromatographic separation was carried out on precoated TLC silica gel 60F₂₅₄, using the mobile phase consisting of ethyl acetate‒acetic acid‒water (6:2:1.5, V/V). The separated bands were quantified by scanning at a detection wavelength of 205 nm. The three drugs were properly resolved (R_F values of metformin hydrochloride: 0.25 ± 0.02, sitagliptin: 0.62 ± 0.03, and dapagliflozin propanediol monohydrate: 0.86 ± 0.02) using these chromatographic conditions. The method was validated in terms of linearity, accuracy, precision, specificity, limit of detection (LOD), and limit of quantification (LOQ) in accordance with the International Council for Harmonisation (ICH) Q2(R1) guidelines. The calibration curves were found to be linear over the ranges of 5‒17.5 µg/band for metformin hydrochloride, 1‒3.5 µg/band for sitagliptin and 0.1‒0.35 µg/band for dapagliflozin propanediol monohydrate. Greenness assessment of the developed method was evaluated using three different assessment tools, viz., analytical ecoscale, green analytical procedure index (GAPI), and analytical greenness (AGREE). This is the first analytical method to be developed for the simultaneous estimation of these drugs in pharmaceutical dosage form. The developed method was applied for the simultaneous estimation of metformin hydrochloride, sitagliptin, and dapagliflozin propanediol monohydrate in tablet dosage form.

Bee propolis is a heterogeneous mixture of phytoactive metabolites with reported antioxidant, anti-inflammatory, and neuroprotective properties. The chemical composition is highly variable, complex and presents the greatest challenge for standardization. So, the study was aimed at standardization of selected phytochemicals in propolis collected from a region in India (IP). A thin-layer chromatography (TLC) method was developed for five markers with prior evidence of neuroprotective activity using orthogonal detection and subsequently validated as per the International Council for Harmonisation ICH Q2 (R1) guidelines. The methodology was assessed for greenness using different metrices and then applied to marketed samples. Furthermore, antioxidant and anticholinesterase activity were evaluated directly using TLC‒bioautography technique. The method was developed using toluene‒ethyl acetate‒formic acid (7.4:2.6:0.5, V/V) as the mobile phase, with R_F values of 0.235, 0.353, 0.552, 0.606, and 0.655 for luteolin, p-coumaric acid, chrysin, galangin, and pinocembrin, respectively. The optimized method was found to be precise (%RSD ≤ 2.0%), accurate (90‒110%), linear over the concentration ranges (R² ≥ 0.995), sensitive and robust. Pinocembrin (2.30 ± 0.12% w/w) and galangin (5.78 ± 0.30% w/w) were found in the highest concentration. TLC‒bioautography analysis confirmed the contribution of the selected phenolics in anticholinesterase and antioxidant activity. The analytical eco-score value of 76, analytical greenness calculator (AGREE) score of 0.62 and other assessed analytical pictograms indicated the methodology developed had low environmental impact. Overall, a simple, rapid, precise, and green TLC analytical methodology has been developed for IP for quality control and preliminary screening of neuroprotective activity.

The anti-obesity activity of the bioactive fractions of Haldina cordifolia (HC) was investigated in pursuit of a novel herbal plant targeting obesity, as its anti-obesity potential has not yet been investigated. This innovative study examined the HC bioactive fraction to select the most significant fraction using an array of studies targeting obesity. Initially, a quantitative phytochemical assessment including total phenolic, flavonoid, and steroidal contents was performed. Additionally, a series of in vitro antioxidant ABTS, DPPH, nitric oxide, and hydrogen peroxide scavenging assays were implemented, whereas, alpha-glucosidase, alpha-amylase, and pancreatic lipase assays were conducted for the inhibition of enzyme capabilities. The findings demonstrated that fractions F2 and F3 have better efficacy, based on IC₅₀ values. Subsequently, the anti-obesity potential of fractions was assessed utilizing cultured 3T3-L1 adipocytes, oil red O staining, and triglyceride content, as markers of accumulation of lipid which showed significant results. Furthermore, high-performance thin-layer chromatography‒multistage mass spectrometry (HPTLC‒MSⁿ) was employed to identify the phytoconstituents within these fractions, particularly the anti-obesity agent, β-sitosterol. HPTLC‒MSⁿ analysis revealed the presence of five major compounds in fraction F2: caffeine, quercetin, β-sitosterol naringin, and ascorbic acid along with retro Diels–Alder fragmentation. Finally, an in silico study was designed which subsequently established the optimal binding activity of the obtained compounds towards the particular receptors targeting obesity. Overall, fraction F2 showed the most potent activity. The study unveiled the promising anti-obesity potential of bioactive fractions from HC and a potential treatment strategy has been established to combat obesity.

This study presents the enantiomeric resolution of three profen drugs, viz., flurbiprofen (FLB), fenoprofen, and zaltoprofen by employing sitafloxacin (SIT) as a chiral selector within a direct thin-layer chromatography (TLC) method. Chromatographic resolution of three drugs was carried out using a mobile phase consisting of acetonitrile‒methanol‒trimethylamine (6:3:1, V/V, pH 8.5), with a stationary phase maintained at pH 5. Analytical validation of the FLB enantiomeric resolution exhibited consistent recovery rates, with coefficient of variation (%CV) values consistently below 2%. Furthermore, the method demonstrated sensitivity, with low limits of detection and quantification values of 0.058 and 0.172 µg per spot, respectively, for FLB enantiomers, indicating reliable detection at low concentrations. The analytical data validate the effectiveness and reliability of the developed TLC method for the enantiomeric resolution of profen drugs, offering significant implications for pharmaceutical research and development.

Traditional formulations owing to their holistic approach to health and wellness with minimum side effects are widely used all over the globe. Itrifal Muqawwi Dimagh (IMD) is a traditional polyherbal formulation used in the Unani system of medicine conventionally claimed to support and enhance cognitive function. The formulation is widely used by a large population due to its alleged therapeutic efficacy. Despite being widely used, the formulation lacks scientific validation and justification in establishing its therapeutic potential. The present study was designed to detect and identify bioactives responsible for acetylcholinesterase inhibitory activity by thin-layer chromatography (TLC)‒bioautography‒mass spectroscopy (MS) and its validation using in silico molecular approach. Quality control was done using ultraperformance liquid chromatography (UPLC)‒MS fingerprint analysis. The authentication of individual constituents of the formulation was done by using powder microscopy. Ethyl acetate and hydrolyzed ethyl acetate fraction of hydroalcoholic extract of IMD were prepared, and in vitro acetylcholinesterase (AChE) inhibitory activity by Ellman’s method was performed. The TLC‒MS bioautography revealed the presence of scopoletin, tannic acid, ellagic acid, and catechin as potential bioactive anticholinesterase metabolites. In silico analysis of the identified metabolites from IMD showed AChE activity of ten identified metabolites, moreover, catechin and naringenin showed the best of potential activity. UPLC‒MS analysis demonstrated separation of 33 phytocompounds in the best active fraction of formulation. Hence, based on our findings, it can be concluded that IMD has a great potential to overcome cholinergic deficiency and can be used for neuroprotection and other neurological disorders after successful in vivo pharmacokinetic and toxicity validation.