TLC Comparison Research Articles

Phenolic compounds from the aerial parts of Rhodiola rosea in Japan, and their antioxidant activity and chemical adaptation to alpine environment

Twelve flavonoids (1–12) including a new flavonol glycoside, hibiscetin 3-O-glucopyranoside-8-O-glucuronopyranoside, ten known galloylglucoses (13–22) and three known phenolic acids (23–25) were isolated from the aerial parts of Rhodiola rosea growing in Japan. Their chemical structures were elucidated by UV-vis spectra, LC-MS, HR-MS, acid hydrolysis, NMR, and/or HPLC, and TLC comparisons with authentic samples. Antioxidant activity of major 14 compounds was measured by H-ORAC method, and two flavonols, herbacetin 3-O-glucoside-8-O-glucuronide, and gossypetin 3-O-glucuronide-8-O-glucoside, showed the strongest activity. Moreover, chemical adaptation to severe alpine environment of R. rosea was discussed.

New Flavonoids from the leaves and stems of Sedum formosanum (Crassulaceae)

As a series of the phytochemical research of the family Crassulaceae, the flavonoids of Sedum formosanum from two Taiwanese and one Japanese sites were isolated and characterized by UV spectra, HR-MS, LC-MS, acid hydrolysis, NMR, and HPLC and TLC comparisons with authentic samples. Eight new flavonoids (21, 22, 24-26 and 28-30) including two flavones and six isoflavones were isolated from the leaves and stems, together with 24 known compounds (1-20, 23, 27, 31 and 32), and identified as chrysoeriol 7-O-[6´´-(3-hydroxy-3-methylglutaroyl)glucoside] (21), chrysoeriol 7-O-[6´´-(5´´´-methoxy-3-hydroxy-3-methyglutaroyl)glucoside] (22), 3´-O-methylorobol 7-O-rutinoside (24), 3´-O-methylorobol 7-O-arabinosyl-(1→2)-glucoside (25), 3´-O-methylorobol 7-O-arabinosyl-(1→6)-glucoside (26), orobol 3´,4´-dimethyl ether 7-O-rutinoside (28), orobol 3´,4´-dimethyl ether 7-O-sophoroside (29) and orobol 3´,4´-dimethyl ether 7-O-arabinosyl-(1→6)-glucoside (30). The samples were collected in three sites of Taiwan and Japan, and geographic chemical variation was observed among their sites.

Flavonoids from the leaves and stems of Rhodiola ishidae (Crassulaceae)

Twenty-six flavonoids including flavonols, C-glycosylflavones, flavanone and anthocyanin were isolated from the leaves and stems of Rhodiola ishidae. Major flavonoids were flavonol glycosides based on herbacetin, gossypetin, kaempferol and quercetin, and identified by UV spectral survey, HR-MS, LC-MS, acid hydrolysis, NMR and/or HPLC and TLC comparisons with authentic samples. Of these flavonoids, three compounds, herbacetin 7-O-glucuronopyranoside-8-O-glucopyranoside, gossypetin 3-O-(2''-malonylxylopyranoside)-8-O-glucopyranoside and naringenin 7-O-sophoroside, were reported in nature for the first time. Flavonoid characters of R. ishidae were chemotaxonomically discussed.

Flavonoids and phenylethanoids from the flowers and leaves of Aeschynanthus species and cultivars (Gesneriaceae)

Forty-one flavones, each one of flavonol, chalcone and dihydroflavonol, two flavanones, and four phenylethanoids were isolated from corollas, calyces and leaves of two Aeschynanthus species, A. fulgens and A. pulcher, and six cultivars, ‘Mahligai’, ‘Mona Lisa’, SoeKa’, ‘Redona’, ‘Freshya’ and ‘Bravera’. Flavonoids were mainly the glucuronides and/or methylglucuronides based on hispidulin, nepetin, pectolinarigenin, 6-hydroxyluteolin, scutellarein, apigenin and luteolin, and identified by UV spectra, HR-MS, LC-MS, acid hydrolysis, NMR, and/or HPLC and TLC comparisons with authentic samples. Of these flavonoids, twelve, i.e. hispidulin 7,4′-di-O-glucuronide, 7,4′-di-O-methylglucuronide, 7-O-methylglucuronide-4′-O-glucuronide, 7-O-glucuronide-4′-O-methylglucuronide, 7-O-glucosyl-(1 → 2)-glucuronide and 8-C-glucoside, nepetin 7,4′-di-O-glucuronide, 7-O-glucuronide-4′-O-methylglucuronide and 7-O-methylglucuronide-4′-O-glucuronide, pectolinarigenin 7-O-glucosyl-(1 → 2)-glucuronide and 7-O-xylosyl-(1 → 2)-(6″-malonylglucoside), and 6-hydroxyluteolin 7,4′-di-O-glucuronide, were previously undescribed.

Flavonoids from the leaves and flowers of the Himalayan Cathcartia villosa (Papaveraceae)

Five flavonols, four flavones and one C-glycosylflavone were isolated from the leaves of Cathcartia villosa which is growing in the Himalayan Mountains. They were characterized as quercetin 3-O-vicianoside (1), quercetin 7,4′-di-O-glucoside (3), quercetin 3-O-rutinoside (4), quercetin 3-O-glucoside (5), quercetin 3-O-arabinosylarabinosylglucoside (6) (flavonols), luteolin (7), luteolin 7-O-glucoside (8), apigenin (9), chrysoeriol (10) (flavones), and vicenin-2 (11) (C-glycosylflavone) by UV, LC-MS, acid hydrolysis, NMR and/or HPLC and TLC comparisons with authentic samples. On the other hand, two flavonols 1 and kaempferol 3-O-vicianoside (2) were isolated and identified from the flowers of the species. Flavonoids were reported from the genus Cathcartia in this survey for the first time. Their chemical characters were chemotaxonomically compared with those of related Papaveraceous genera, Meconopsis and Papaver.

MATERNAL CRP AS A PREDICTOR OF CHORIOAMNIONITIS IN WOMEN WITH PPROM

Introduction: Preterm prelabor rupture of the membranes (PPROM) refers to spontaneous rupture of membranes in the absence of labor pains, before 37 completed weeks of gestation. Chorioamnionitis (CAM) affects many pregnancies complicated by PPROM. Finding a serum factor that could accurately predict the presence of CAM could potentially lead to more efficient management of PPROM and improved neonatal outcomes. It has been claimed that estimation of C-reactive proteins (CRP) is helpful in the diagnosis of chorioamnionitis, and this study aims to appraise such claimsObjective: To determine the diagnostic accuracy of C-reactive protein in the detection of chorioamnionitis in women with PPROM and to test sensitivity/specificity/positive predictive valve/negative predictive value of CRP in diagnosing chorioamnionitis against histopathological examination of placenta.Method: A study conducted on total 440 antenatal women, 220 cases of PROM and 220 cases with same gestation but without PPROM used as a control. A detailed obstetrical and menstrual history was taken and systemic and local examination was done. Subjects were managed expectantly with use of tocolytics, antibiotics and steroids. Frequent vital signs monitoring and hematological investigation were done. CRP levels were determined. After delivery placenta was sent for histopathological examination for the presence of chorioamnionitis.Results: CRP appears to be the most sensitive acute phase protein; rising of less than 24 hours makes it suitable to serve as a marker for diagnosing an infection. On comparing C-reactive protein levels with other laboratory tests and indicators of infection (e.g. total leucocyte count, maternal fever, maternal tachycardia, fetal tachycardia) we found CRP level to be more sensitive (100%) but less specific (45.45%) in identifying chorioamnionitis. The positive predictive value was 31.4% and negative predictive value was 100%.Conclusion: CRP is early and reliable indicator of histopathological and clinical chorioamnionitis in comparison of TLC and clinical parameter. Thus CRP can prove useful markers in identify early and subclinical infection which could lead to premature rupture of membrane.Key-words- Preterm birth, C-reactive protein, PPROM, Chorioamnionitis.

Assessment of Nutritional Status of Patients Suffering from Asthma

Background and aim: Several studies have suggested that increasing prevalence of asthma could be associated with nutrients intake. These nutrients can play beneficial as well as detrimental role in asthma and may affect the nutritional status of asthma patients. This case-control study was undertaken to assess the nutritional status and differences in dietary intake between a population with and without asthma. Methods: A case-control study was carried out in clinically and spirometrically diagnosed asthma patients and (n=143, average age 29.35 ± 8.64 years) and age, sex and socioeconomic status matched healthy volunteers (n=143, average age 29.54 ± 8.61 years) to assess and evaluate the nutritional status of asthmatics. Anthropometric measurements(BMI, MUAC, and TSF), biochemical investigations (Hb, total and differential leucocyte counts and serum calcium and protein and serum IgE), pulmonary function test, nutrient intake (3 day diary method and food frequency questionnaire) and subjective global assessment were used asstandard nutritional assessment tools. Results were considered statistically significant if the observed two-sided significance level (P-value) was not greater than 0.05. Results: Thepresentstudy resultsfoundthatthemajorityof casesweresignificantly overweight or obese on the basis of body mass index (p=0.01), mid upper arm circumference (p=0.01) and triceps skin fold thickness (p=0.01) than controls. Hb, serum protein and serum calcium, levels were significantly lower in asthma cases in comparison of non-asthmatic controls (p=0.01, 0.01, 0.01 respectively) and TLC, eosinophils, monocytes and serum Ig E levels were significantly higher in asthmatics in comparison to nonasthmatics (p=0.01, 0.01, 0.01 and 0.00 respectively). Daily intake of fat was significantly higher whereas protein intake was significantly lower in asthmatics than in non-asthmatic subjects (p=0.01 and p=0.01 respectively). Significant decrease in daily intakes of thiamine, riboflavin, vitamin C, calcium and iron was noticed in asthmatic than non-asthmatic subjects (p=0.01, p=0.02, p=0.01, p=0.01, p=0.01, p=0.00). Based on subjective global assessment rating scale, significant difference was observed between the groups (p=0.04) as 48.9% and 21.7% of the asthmatics and 23.6% and 47% of the nonasthmatics were moderately malnourished and well-nourished respectively. Conclusion: It can be concluded that overall nutritional status of asthma patients was poor based on the standard tools used. Most of them were obese, had unbalanced nutrient intake and were not well nourished. Thus the time may be close for consideration of dietary advice as per recommended and modified dietary allowances followed by intervention trials.

Bioassay-guided fractionation and identification of α-amylase inhibitors from Syzygium cumini leaves

Context: Pancreatic α-amylase and α-glucosidase inhibitors serve as important strategies in the management of blood glucose. Even though Syzygium cumini (L.) Skeels (Myrtaceae) (SC) is used extensively to treat diabetes; scientific evidence on antidiabetic effects of SC leaves is scarce. Objective: SC leaf extract was investigated for α-amylase inhibitory effect and continued with isolation and identification of α-amylase inhibitors. Materials and methods: Bioassay-guided fractionation was conducted using in vitro α-amylase inhibitory assay (with 20–1000 μg/mL test material) to isolate the inhibitory compounds from ethyl acetate extract of SC leaves. Structures of the isolated inhibitory compounds were elucidated using 1H NMR and 13C NMR spectroscopic analysis and direct TLC and HPLC comparison with authentic samples. Study period was from October 2013 to October 2015. Results: An active fraction obtained with chromatographic separation of the extract inhibited porcine pancreatic α-amylase with an IC50 of 39.9 μg/mL. Furthermore, it showed a strong inhibition on α-glucosidase with an IC50 of 28.2 μg/mL. The active fraction was determined to be a 3:1 mixture of ursolic acid and oleanolic acid. Pure ursolic acid and oleanolic acid showed IC50 values of 6.7 and 57.4 μg/mL, respectively, against α-amylase and 3.1 and 44.1 μg/mL respectively, against α-glucosidase. Discussion and conclusions: The present study revealed strong α-amylase and α-glucosidase inhibitory effects of ursolic acid and oleanolic acid isolated from SC leaves for the first time validating the use of SC leaves in antidiabetic therapy.

Determination of Botanical Origin of Propolis from Monte Region of Argentina by Histological and Chemical Methods

Propolis production by honey bees is the result of a selective harvest of exudates from plants in the neighborhood of the hive. This product is used in Argentina as a food supplement and alternative medicine. The aim of this study was to determine the botanical origin of propolis from the arid regions of Monte of Argentina using rapid histochemical techniques and by comparison of TLC and HPLC-DAD chromatographic profiles with extract profiles obtained from Zuccagnia punctata, Larrea divaricata and Larrea cuneifolia, plant species that grow in the study area as a natural community named "jarillal". Microscopical analysis revealed the presence of several Z. punctata structures, such as multicellular trichomes, leaflets, stems and young leaves. Remarkable was the richness of the propolis in two bioactive chalcones, also present in Z. punctata resin; these compounds can be regarded as possible markers for propolis identification and justify its use as a dietary supplement, functional food and medicinal product. This study indicates that the source of resin used by honey bees to produce propolis in the Monte region of Argentina is only Z. punctata, a native shrub widespread in this phytogeographical region, while other more abundant species (L. divaricata and L. cuneifolia) in the region were not found, indicating that this propolis could be defined as a mono-resin, type-Zuccagnia.

C14-Polyacetylene glucosides from Codonopsis pilosula

Seven new C14-polyacetylene glucosides codonopilodiynosides A–G (1–7) were isolated from an aqueous extract of the Codonopsis pilosula roots. Their structures were determined by spectroscopic and chemical methods as (–)-(5S,6E,12E)-tetradeca-6,12-dien-8,10-diyn-1,5,14-triol 5-O-β-d-glucopyranoside (1), (–)-(5S,6E,12E)-tetradeca-6,12-dien-8,10-diyn-1,5,14-triol 5-O-β-d-glucopyranosyl-(1″ → 2′)-β-d-glucopyranoside (2), (–)-(5S,6E,12E)-tetradeca-6,12-dien-8,10-diyn-1,5,14-triol 5,14-di-O-β-d-glucopyranoside (3), (–)-(5S,6E)-tetradeca-6-en-8,10-diyn-1,5,14-triol 5-O-β-d-glucopyranoside (4), (–)-(5S,6E,12E)-tetradeca-6,12-dien-8,10-diyn-1,5-diol 5-O-β-d-glucopyranosyl-(1″ → 2′)-β-d-glucopyranoside (5), (–)-(6S,4E,12E)-tetradeca-4,12-dien-8,10-diyn-1,6-diol 6-O-β-d-glucopyranosyl-(1″ → 2′)-β-d-glucopyranoside (6), and (–)-(5S,6E)-tetradeca-6-en-1,5-epoxy-8,10-diyn-14-ol 14-O-β-d-glucopyranosyl-(1″ → 2′)-β-d-glucopyranoside (7), respectively. The absolute configurations of 1–7 were assigned by enzymatic hydrolysis followed by isolation of glucose and aglycones (1a and 4a–7a), and subsequent comparison of specific rotation, TLC, and 1H NMR data of the glucose with an authentic sugar sample and application of modified Mosher's method based on the MPA determination rule of ΔδRS values for 1a and 4a, and ΔδS values for 6a. The configuration of 7 was assigned by electronic circular dichroism calculations based on the quantum-mechanical time-dependent density functional theory.

Application of direct HPTLC‐MALDI for the qualitative and quantitative profiling of neutral and acidic glycosphingolipids: The case of NEU3 overexpressing C2C12 murine myoblasts

Glycosphingolipids (GSLs) are a class of ubiquitous lipids characterized by a wide structural repertoire and a variety of functional implications. Importantly, altered levels have been correlated with different diseases, suggesting their crucial role in health. Conventional methods for the characterization and quantification are based on high-performance TLC (HPTLC) separation and comparison with the migration distance of standard samples or on MS. We set up and herein report the application of an ImagePrep method for glycosphingolipids qualitative and quantitative profiling through direct HPTLC-MALDI with particular application to wild-type and NEU3 sialidase-overexpressing C2C12 myoblasts. Lipids were analyzed by HPTLC, coupled with MALDI-TOF, and the resulting GSLs profiles were compared to the [³H]sphingolipids HPTLC patterns obtained after metabolic radiolabeling. GSLs detection by HPTLC-MALDI was optimized by testing different methods for matrix delivery and by performing quantitative analyses using serial dilutions of GSLs standards. Through this approach an accurate analysis of each variant of neutral and acidic GSLs, including the detection of different fatty-acid chain variants for each GSL, was provided and these results demonstrated that HPTLC-MALDI is an easy and high-throughput analytical method for GSLs profiling, suggesting its use for an early detection of markers in different diseases, including cancer and heart ischemia.

Comparison of TLC and Different Micro TLC Techniques in Analysis of Tropane Alkaloids and Their Derivatives Mixture from Datura Inoxia Mill. Extract.

Planar chromatography is a very useful tool for analysis of wide range of different mixtures. Thanks to its possibility for rapid separation of large number of samples simultaneously, low solvent consumption and ability to analyse rough material allow to receive precise and reliable results in short time and low cost. Miniaturization of planar techniques brings a lot of advantages, such as shortening distance and time of chromatogram development, and further lowering of solvent consumption. Besides, it often allows to improve separation parameters and raise efficiency of chromatographic system. In this paper, ability of analysis of tropane alkaloids mixture from Datura Inoxia Mill. extract using conventional TLC technique with five micro TLC techniques (short distance TLC, HPTLC, UTLC, OPLC and ETLC) in maximally closed chromatographic conditions was compared in order to present abilities of micro TLC techniques in plant material analysis.

Chemometric Processing of Pharmaceutical Essential Oil Fingerprints—Comparison of GC, HPLC, TLC, IR Spectroscopy, and Differential Scanning Calorimetry

Fifteen essential oils of pharmaceutical grade were fingerprinted by five techniques: TLC, GC, HPLC, attenuated total reflectance FTIR spectroscopy, and differential scanning calorimetry (DSC). Denoising and baseline removal was found to be a crucial step for correct comparative analysis. Standardization of the signal was not necessary in the presented case; however, it should be considered and checked in each case. Due to small variance explained by first two principal components (below 50%) and outlying observations, the main analysis was performed by Euclidean dendrograms. It was found that almost all techniques besides DSC find real chemical similarities; however, DSC can be used as an additional tool. The similarities among the five techniques were also compared and discussed.

Comparison of TLC and HPLC methods used for analysis of (−)-epicatechin and its dimer procyanidin B2 in chocolate

TLC test solutions of baking chocolate (Standard Reference Material (SRM) 2384, National Institute of Standards and Technology, USA) were prepared by extraction of 0.0200 g pulverized defatted chocolate (the amount [%] of fat was determined separately) with 2 mL 70% acetone (aq.) followed by centrifugation and filtration (0.45 μm PVDF filter; Millipore, USA). Liquid–liquid partitioning (1 mL TLC test solution, 1 mL 20% aqueous sodium chloride, and 1.5 mL ethyl acetate) was needed to obtain sufficiently pure test solutions for the HPLC analysis.

Fusarium verticillioides as a new pathogen of the parasitic weed Orobanche spp.

A strain of Fusarium verticillioides was isolated from the tubercles of the parasitic weed Orobanche cumana in Israel. The pathogenicity was tested in polyethylene bags on O. cumana, O. crenata, O. aegyptiaca and O. ramosa and in pots on O. cumana and O. crenata. F. verticillioides was highly pathogenic to O. aegyptiaca, O. ramosa and O. cumana in the polyethylene bags. In pots, the fungus caused wilting and necrotic areas on flowering shoots of O. cumana, but did not cause disease symptoms on O. crenata. F. verticillioides grown on liquid Czapek growth medium produced a phytotoxic metabolite, which in leaf-puncture bioassay caused large necrotic areas on Orobanche shoots and on leaves of various crops. An extract of the fungal growth medium caused complete mortality of O. cumana and O. aegyptiaca seedlings in vitro. The toxic metabolite was isolated and identified by spectroscopic methods as fusaric acid. The identity of the compound was confirmed by conversion into the corresponding methyl ester, and by TLC comparison against authentic fusaric acid. No other phytotoxic metabolites could be detected in the growth medium extracts.

Regioselective acylation of 3-O-angeloylingenol by Candida antarctica Lipase B

Acylation of 3-O-angeloylingenol (1) with vinyl acetate, vinyl decanoate and vinyl cinnamate, catalyzed by Candida antarctica Lipase B, was investigated. In each case, compound 1 was quantitatively and regioselectively acylated to afford a single product, 3-O-angeloyl-20-O-acetylingenol (1a), 3-O-angeloyl-20-O-decanoylingenol (1b) and 3-O-angeloyl-20-O-cinnamoylingenol (1c), respectively. The structures of the novel compounds 1b-1c were determined by MS and NMR, and product 1a by comparison of RP-HPLC and TLC with a standard. Compounds 1b-1c induced a bipolar morphology of MM96L melanoma cells at a similar concentration as compound 1, as well as having activity in inhibiting the growth of MM96L melanoma cells.

HPLC Estimation of berberine in Tinospora cordifolia and Tinospora sinensis.

A high-performance liquid chromatographic method for the estimation of berberine in the stem of Tinospora cordifolia (Willd.) Miers. ex Hook.f. and Thoms. and Tinospora sinensis (Lour.) Merrill is described. The dried stems of T. cordifolia and T. sinensis were defatted with petroleum ether (60-80°). The marc was dried and further extracted with methanol. The concentration of berberine in methanol extract was determined using a C-18 reverse phase column with a mobile phase of acetonitrile:water (10:90 v/v) at a flow rate of 0.6 ml/min and with UV detection at 266 nm. TLC and HPLC comparison of both the species revealed significant variation in the chemical constitution of the two species. This observation becomes important in the context of the use of T. sinensis in place of the genuine drug T. cordifolia.

Comparison of normal and reversed-phase TLC for separation of selected pesticides

Fifteen urea pesticides have been separated on RP-18WF 254 plates with methanol-water and mixed organic (acetonitrile-methanol, 1:1 v/v )-0.1% aqueous orthophosphoric acid (H 3 PO 4 ) mobile phases (RP-HPTLC), and on silica gel 60F 254 plates with benzene-methanol and benzene-ethanol mobile phases (NP-TLC). The pesticides could be classified into three groups: monolinuron (1), chlorotoluron (2), diuron (3), isoproturon (4), and linuron (5) dimefuron (6), diflubenzuron (7), teflubenzuron (8), and lufenuron (9); and thifensulfuron methyl (10), triasulfuron (11), chlorsulfuron (12), rimsulfuron (13), amidosulfuron (14), and tribenuron methyl (15). Relationships between R F values and mobile phase composition were determined for the pesticides. Δ R F values, separation factors ( α ), and constants for separation of pairs of compounds ( R F α ) were used to determine optimum chromatographic conditions for the separations. Comparison of results from normal and reversed-phase TLC revealed the pesticides in the f...

Determination of the plasma input function of [18F]altanserin by selective liquid–liquid extraction. Validation of a TLC method and comparison to HPLC

Determination of the plasma input function of [18F]altanserin by selective liquid–liquid extraction. Validation of a TLC method and comparison to HPLC

Determination of the Complete Absolute Configuration of Petriellin A

We report the full structural determination of the depsipeptide petriellin A. The absolute configuration of the amino acid residues, N-methyl isoleucine and N-methyl threonine, have been determined by a combination of HPLC and TLC comparison of synthetic Marfey’s derivatives and Marfey’s derivatives of the natural product hydrolysate. The configuration of the chiral centres in these two N-methylated residues was found to be the same as those of the common unmethylated l-amino acids.