•New HPTLC methods for major bioactive compounds in mate tea were developed.•Both aqueous and methanolic extracts could be analysed by these methods.•Methylxanthines, phenolic compounds, and saponins ...were identified by HPTLC-MS.•Mate teas of different types, ages, and geographical origins were evaluated.•PCA showed distinctive characteristics between mate teas.
New HPTLC (high-performance thin-layer chromatography) methods to evaluate major bioactive compounds in mate (Ilex paraguariensis) tea were developed. Identification of compounds was based on HPTLC-MS (high-performance thin-layer chromatography–mass spectrometry), parallel to other techniques. Quantitative evaluation of the main methylxanthines and phenolic compounds in this species was accomplished by two HPTLC methods which can be performed consecutively on one single plate. A solvent system for the separation of the identified saponins is also described for qualitative evaluations. The methods for quantification were validated in terms of: robustness; LOD and LOQ (limits of detection and quantification); repeatability; intra-day and inter-day variation; ruggedness; and accuracy. The methods have proven to be suitable for evaluation of mate tea aqueous and methanolic extracts, with the advantages of: high-throughput; reduced need of plates and application procedures; and high sensitivity – low LODs and LOQs were achieved. Aqueous extracts of nineteen mate teas of different type, geographical origin and age were evaluated in terms of methylxanthines, phenolic compounds and pH. By means of principal component analysis, distinctive characteristics between mate teas could be observed.
•HPTLC-MS techniques have the potential to be used in real time and have gained popularity in recent years.•The current work highlights pertinent and significant information regarding the hyphenated ...HPTLC-MS technique's ability to detect both food components and their adulterants.•The present work reviews the use of analytical methods in qualitative and quantitative studies of food components and their adulterants.•Study of recent advancements coupling of analytical techniques that can improve detection efficiency of food.•Hyphenated HPTLC-MS technique found to be useful in food industry for adulteration detections.
Chemometric analysis is also a powerful tool that can be used in the preliminary stages of analytical methods optimization, but is also efficient in data processing. These new approaches may be the key to the analysis of proteins, carbohydrate, sphignolipids, polyphenols in different food components. Although there are challenges in identifying and annotating those compounds due to the limited availability of analytical standards and structural diversity.
Chromatograms are a useful tool for learning more about the chemical make-up of the food being tested. This information can occasionally be implicit or presented in a subtle way. Therefore, to extract it, chemometric tools and data mining techniques are needed. A chromatogram's fingerprint allows for the possibility of performing both identity and quality testing on food. This viewpoint aims to offer a current assessment of chromatographic fingerprinting technique in the area of food authentication. Additionally, this methodology's limitations, absence from official analytical methods, and future directions are discussed. Novel techniques have been employed in the past few decades, ranging from high-pressure thin liquid chromatography (HPTLC) to mass spectrometry (MS) and other spectroscopic methods. In the last decade research developments, and the application of analytical methods in qualitative and quantitative studies of food components and their adulterants is reviewed in the present work. High-pressure Thin liquid chromatography (HPTLC) is hyphenated with high resolution mass spectrometry is particularly addressed due to its applicability in the targeted/untargeted metabolomic analysis of all food components.
In the current study, the HPTLC-MS technique was used to examine food components and food adulteration. The HPTLC-MS hyphenated techniques were found to be an extremely helpful method for the detection of food adulterants as well as the various food components, such as protein, carbohydrates, lipids, essential oils, and flavonoids, which were found to be effective as medicinally.
Application of HPTLC-MS method for the routine analysis of food vegetable analysis. The thodology have advantages to avoids various chemical steps and this techniques provides a simpler and rapid method for routine analysis of different types of samples.
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► Improved stability of carotenoids in TLC analysis. ► HPTLC method for densitometric determination of lutein. ► Screening of major dietary carotenoids in food supplements. ► Use of TLC–MS interface ...to confirm identity/structure of carotenoids.
The main problem in the densitometric determination of carotenoids is their rapid degradation during and immediately after chromatography, respectively. In this study, we show that 15ng of lutein, lycopene and β-carotene standards applied on C18 RP high-performance thin-layer chromatography (HPTLC) plates pre-developed with dichloromethane–methanol 1:1 (v/v) remained stable for 1h after the development of chromatogram using methanol–acetone 1:1 (v/v) with 0.1% of 2-tert-butylhydroquinone (TBHQ), which is a substantial improvement of their stability. An HPTLC quantification procedure for free lutein, with densitometry at 450nm based on the developed method described above, was established and validated. Repeatabilities of the chromatography expressed by the relative standard deviation (RSD) from 6 applications of lutein standard at 5, 15 and 25ng were 3.41, 1.33 and 1.65%, respectively. The best fit calibration curve from 5ng to 30ng of lutein was polynomial. Limit of detection (1.5ng) and limit of quantification (5ng) were the best achieved so far. With these chromatographic conditions dietary carotenoids lutein esters, lycopene, free lutein and β-carotene from food supplements were also well separated and were identified by visible absorption spectra scanned in situ and by mass spectra. Some additional developing solvents with the same type of chromatographic layer are proposed for the fast separation of lutein esters from free lutein in food supplements.
•Rapid HPTLC-FLD quantification of coumarin in 43 cinnamon samples.•Quality control of Ceylon cinnamon and Cassia cinnamon.•Efficient high throughput method to assess seasonal food.•Specific ...fluorescence detection and compound verification by HPTLC-MS.•Effect-directed analysis for antimicrobials against A. fischeri (HPTLC-UV/FLD-EDA).
A sensitive quantitative screening of coumarin in 43 commercially available cinnamons and cinnamon-containing foods was developed via HPTLC. Complex samples like cinnamon, tea, breakfast cereals, milk rice, jam, cinnamon stars and buns were extracted with methanol only. Separation was performed on silica gel with a mixture of n-hexane, ethyl acetate and ammonia. The specific detection via derivatization with an ethanolic potassium hydroxide solution resulted in fluorescent coumarin zones, measured at 365/>400nm after stabilization. Limits of detection and quantification were 200 and 400pg/band, respectively. Over all different sample types, the contents ranged from 0.3 to 5129mg/kg with a mean repeatability and mean intermediate precision of 4% each. HPTLC-MS of selected zones, eluted via the TLC-MS Interface into MS, confirmed the identity of coumarin. Effect-directed detection as bioanalytical tool for risk assessment showed coumarin to be active against Aliivibrio fischeri bacteria down to 100ng/band.
Thin-layer chromatography (TLC) had its initial growth in the 1950s when TLC sorbents and devices for making TLC plates in the analytical laboratory became available. A resurgence in TLC use occurred ...when commercially prepared plates became available around 1965. Their advantage was greater reproducibility because of their uniformity and convenience of use. Having just passed the 50th anniversary of this date, TLC still finds wide application as a useful analytical tool throughout the world. The introduction of high-performance TLC (HPTLC)-prepared plates was also a welcome addition to the chromatography laboratory. Today, advances in TLC instrumentation that aid in sample application, plate development, qualification, and quantification continue to evolve and improve. The TLC/HPTLC plate manufacturers have continued to add new prepared plates to meet the greater demands for higher purity or special applications for these newer devices. More recently, researchers have experimented with new sorbents or preparation techniques that have resulted in special properties for thin-layer-prepared plates, particularly for use in TLC-MS applications. This article will discuss not only some classical TLC plates but also these newer thin layers, their advantages, and some of their applications.
Coupling of planar chromatography to mass spectrometry (MS) and especially ambient MS is a relatively new field of great interest. The direct sample access at ambient conditions and the feasibility ...to obtain mass spectra free of contamination within a minute or even within seconds greatly contributes to the progress of planar chromatography. Targeted recording of mass spectra on zones of interest is performed after evaluation of the chromatogram, thus providing high efficiency. Reported approaches for coupling are divided into elution-based and desorption-based techniques. Devices of both categories are commercially available. As a consequence of increasing importance, a rethink of the terminology of liquid chromatography with MS has to be considered.
The question of whether exosome lipids can be considered as potential cancer biomarkers faces our current limited knowledge of their composition. This is due to the difficulty in isolating pure ...exosomes, the variability of the biological sources from which they are extracted, and the uncertainty of the methods for lipid characterization. Here, we present a procedure to isolate exosomes and obtain a deep, repeatable, and rapid phospholipid (PL) composition of their lipid extracts, from embryonic murine fibroblasts (NIH-3T3 cell line) and none (B16-F1) and high (B16-F10) metastatic murine skin melanoma cells. The analytical method is based on High Performance Thin-Layer Chromatography with Ultraviolet and fluorescence densitometry and coupled to Electrospray (ESI)-tandem Mass Spectrometry (MS). Under the conditions described in this work, separation and determination of PL classes, (sphingomyelins, SM; phosphatidylcholines, PC; phosphatidylserines, PS; and phosphatidylethanolamines, PE) were achieved, expressed as µg PL/100 µg exosome protein, obtained by bicinchoninic acid assay (BCA). A detailed structural characterization of molecular species of each PL class was performed by simultaneous positive and negative ESI-MS and MS/MS directly from the chromatographic plate, thanks to an elution-based interface.
A novel and cost‐effective high‐performance thin‐layer chromatography (HPTLC) method, combined with densitometric quantification, was developed for the biomedical analysis of telmisartan (TEL) and ...gallic acid (GA). Recent research indicates that when used in combination, these compounds offer improved therapeutic efficacy for the treatment of cardiovascular diseases with reduced side effects. The study focused on the simultaneous quantification and pharmacokinetic analysis of drugs in rat plasma. The separation was conducted using HPTLC silica gel 60 F254 plates with dimensions of 20 × 10 cm and a thickness of 0.2 mm. The mobile phase used for separation consisted of a mixture of ethyl acetate, methanol, chloroform, and acetic acid in the ratio of 4:2:2:0.2 (v/v). GA and TEL were analyzed using ultraviolet detection at specific wavelengths, with GA at 280 nm and TEL at 296 nm. Peak purity was assessed through spectral correlation analysis using Vision CATS software. The method underwent validation following the guidelines of the US Food and Drug Administration (US FDA). Calibration plots demonstrated linearity in the concentration range of 200–1200 ng/spot, with high correlation coefficients (R2). The retention factors (Rf) were 0.67 for TEL and 0.60 for GA. The identity of the separated compounds was further confirmed using MS, with GA having a mass‐to‐charge ratio (m/z) of 168.9 in negative mode and TEL with m/z 515.2 in positive mode. In the pharmacokinetic study, the maximum peak plasma concentration (Cmax) for GA was 899.7 ng/mL, and for TEL, it was 1013 ng/mL. The time to reach maximum concentration (Tmax) was 2 h for GA and 6 h for TEL. This simultaneous qualitative and quantitative determination of the drugs in an oral pharmacokinetic study involving Wistar rats can serve as a valuable tool for future investigations into pharmacokinetic interactions, quality control, and routine analysis of these drugs, both in their pure forms and in novel formulations.
The worldwide spread coronavirus (covid-19) pandemic caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) represents a global health crisis. The world was forced to face a great ...challenge to control and overcome this health disaster through various containment measures including efficient vaccination side by side with effective medication. Remdesivir (RMD) is the first FDA approved antiviral agent for treatment of covid-19 pandemic and hence regarded as the first-in-class medication of this highly contagious respiratory disease. The current study represents the first stability indicating HPTLC method for the estimation of RMD in bulk form and pharmaceutical formulation. The method employed TLC silica gel aluminum plates 60 F254 as stationary phase and green mobile phase composed of ethyl acetate and ethanol (96: 4, v/v) with densitometric detection at 245 nm. Comprehensive validation of the adopted method was accomplished according to the ICH guidelines regarding linearity, ranges, detection and quantification limits, precision, accuracy and robustness. The developed method offered a neat separation of the drug in presence of pharmaceutical excipients as well as in presence of acidic, alkaline, neutral hydrolytic, oxidative and photolytic degradants. Additionally, structural elucidation of alkaline and hydrolytic oxidation degradation products was carried out using HPTLC-MS. Furthermore, for the first time the acidic and alkaline degradation kinetics of RMD were studied and its degradation rate constants and half-lives were calculated. Moreover, greenness appraisal of the developed method as well as comparison with previously published stability indicating HPLC methods were performed using analytical Eco-scale, GAPI and AGREE metrics.
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•For the first time, eco-friendly, fast and economic stability indicating HPTLC micro-determination of Remdesivir (RMD) was developed and validated.•Neat separation of the drug was successfully achieved either in presence of its stress degradation products or in presence of pharmaceutical formulation excipients without any interference.•For the first time, degradation kinetics of RMD were studied under acidic and alkaline degradation conditions.•Preliminary Structural elucidation of stress degradation products was carried out using HPTLC-MS.•Comprehensive greenness appraisal of the developed method as well as comparison with previously published stability indicating methods were illustrated using AES, GAPI and AGREE metrics.
The present study aims to establish a high-performance thin layer chromatography (HPTLC)-based comparative analysis, directed toward characterization of nucleobases in aqueous and alcoholic extracts ...of sea buckthorn leaves from three different varieties: Hippophae salicifolia, Hippophae rhamnoides mongolica, and Hippophae rhamnoides turkestanica. The alcoholic and aqueous leaf extracts from these sea buckthorn varieties were prepared using accelerated solvent extraction technique. A novel HPTLC method for separating and identifying six nucleobases, namely, guanosine, guanine, cytosine, adenine, uracil, and thymine were adopted. HPTLC analysis indicated the presence of one or more of these nucleobases in a total of six leaf extracts evaluated, their quantities varying from 0.23 to 7.76 µg nucleobase per mg of extract. Though a typical trend could not be observed in the values obtained, the extracts were found to be considerably rich with respect to nucleobase contents. The results acquired from HPTLC were subsequently validated by hyphenation with mass spectrometry and also by applying chemometric tools in form of heat maps, hierarchical cluster dendrograms, and principal component analysis. The presence of nucleobases in the leaf extracts was confirmed by HPLC as well but HPTLC proved to be a better approach for characterization of nucleobases in plant extracts, than high performance liquid chromatography (HPLC).
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