Cumulative CAMAG Bibliography Service CCBS

Our CCBS database includes more than 11,000 abstracts of publications. Perform your own detailed search of TLC/HPTLC literature and find relevant information.

The Cumulative CAMAG Bibliography Service CCBS contains all abstracts of CBS issues beginning with CBS 51. The database is updated after the publication of every other CBS edition. Currently the Cumulative CAMAG Bibliography Service includes more than 11'000 abstracts of publications between 1983 and today. With the online version you can perform your own detailed TLC/HPTLC literature search:

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      132 010
      Establishment of anti-asialo-GM1 rabbit monoclonal antibodies capable of reducing natural killer cell activity in mice
      T. KIMURA*, S. OHTA, H. MURAYAMA (*Diagnostic Division, Yamasa Corporation, Choshi, Chiba, Japan;

      PLoS ONE 18(10), e0292514 (2023). Samples were 8 glycolipids, either sialylated or not; the two main examples were monosialotetrahexosylganglioside (GM1) and gangliotetraosylceramide (= asialo-GM1 = ASGM1). TLC on silica gel with chloroform – methanol – water – acetic acid 300:200:30:1, followed by air-drying. Visualization of glycolipids by spraying 5-methylresorcinol solution (0.2 % in 2 M sulfuric acid), followed by 5 min heating at 110°C. For immunostaining, underivatized chromatograms were immersed into a blocking gel (1% gelatin, 1% polyvinylpyrrolidone and 1mM EDTA in phosphate-buffered saline solution (PBS)), followed by immersion into a solution of monoclonal (50 ng/mL) or polyclonal (1:1,000) anti-ASGM1 antibodies purposedly produced in rabbits. After 1h reaction at room temperature, the layers were washed thrice with T-PBS (0.05 % Tween 20 in PBS) and horseradish-peroxidase-conjugated anti-rabbit IgG (1:40,000) was added. After washing thrice with T-PBS, the TLC sheets were incubated in substrate solution (tetramethylbenzidine). Blue spots indicated the glycolipids bound by the antibody. In this TLC assay, each of the five monoclonal antibodies (as well as the polyclonal serum) was specific to ASGM1 (unsialylated), whereas in ELISA (enzyme-linked immunosorbent assay) at the same concentrations three of them displayed partial cross-reactivity towards GM1, GM3 or GD1b, which are sialylated glycolipids. 

      Classification: 4e, 10b, 11
      131 056
      Normal‑ and reversed‑phase high‑performance thin‑layer chromatography methods for the simultaneous determination of remogliflozin etabonate and metformin hydrochloride antidiabetic drugs in bulk and tablet formulation
      A. PATIL, A. MUNDADA, R. SHRAVANE, P. MUNDADA*, S. SURANA (*Department of Pharmaceutical Chemistry, R.C. Patel Institute of Pharmaceutical Education and Research, Shirpur, Dhule 425405, MS, India,

      J. Planar Chromatogr. 36, 89-97 (2023). HPTLC of remogliflozin etabonate (1) and metformin hydrochloride (2) in bulk and tablet formulation on silica gel with ethyl acetate - methanol - toluene - formic acid 5:2:9:4 (NP) and on RP-18 with water - methanol - glacial acetic acid 5:3:2 (RP). Quantitative determination by absorbance measurement at 226 nm for both methods. The hRF values for (1) and (2) were 56 and 10 for NP and 15 and 88 for RP, respectively. Linearity was in the range of 200-1200 ng/zone for (1) and 1000-6000 ng/zone for (2) in both NP and RP. Intermediate precisions were below 2 % (n=3). LOD and LOQ were 18 and 54 ng/zone for (1) and 105 and 318 ng/zone for (2) in NP and 17 and 51 ng/zone for (1) and 115 and 348 ng/zone for (2) in RP. Recovery was in the range of 100.3-100.8 % for (1) and 100.2-100.7 % for (2) in NP and 100.3-100.7 % for (1) and 100.3-100.8 % for (2) in RP.

      Classification: 32a
      131 004
      Simultaneous determination of montelukast sodium and loratadine by eco-friendly densitometry and spectrophotometric methods
      Shimaa A. MAHMOUD*, A.M. EL-KOSASY, F.A. FOUAD
      (*Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt;

      J Chrom Sci, bmad025 (2023). Standards (separated and mixed) were montelukast sodium (MKT) and loratadine (LRT). Samples were methanolic solutions of commercial tablets, and purified blood plasma as biological fluid, from patients taking MKT or LRT as oral treatment. TLC on silica gel with ethyl acetate – ethanol 9:1. Visualization under UV 254nm. The hRF values were 80 for MKT and 71 for LRT. Densitometric absorbance measurement at 260 nm (20 mm/s scanning speed). System suitability was verified by resolution, selectivity, capacity and absence of tailing. The method was validated for linearity range (0.3–3.6 μg/zone for MKT, 0.2–4 µg/zone for LRT), for precision, for reproducibility, for robustness, and for accuracy expressed as average recovery values (100 % overall mean) at different concentrations. The TLC-densitometric method was also found statistically equivalent (Student’s t-test and F-test) to a previously described method (HPLC – spectrophotometry), but was better in terms of environmental and health impacts, using green analytical procedure index (GAPI) and analytical eco-scale (scores based on solvents/reagents, energy consumption, occupational hazard and waste generation). The TLC method was also compared to three (equally “green”) different analytical methods of spectrophotometry (without chromatography): response correlation, absorptivity-centering and LRT-MKT ratio derivatives. The TLC method was more sensitive (LOQ values were 82 ng/zone for MKT, 20 ng/zone for LRT).

      Classification: 5c, 7, 17c, 23d, 23e, 24, 32a, 32c
      131 005
      Green TLC-densitometric method for simultaneous determination of antazoline and tetryzoline: application to pharmaceutical formulation and rabbit aqueous humor
      (*Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Cairo, Egypt;

      J Chrom Sci, bmad042 (2023). Standards (separated and mixed) were antazoline (ANT) and tetryzoline (TET) hydrochlorides. Samples were one commercial ophthalmic solution containing both molecules (unspiked and spiked), and aqueous humour of untreated rabbits as biological fluid, spiked with various concentrations of ANT and TET. TLC on silica gel with ethyl acetate – ethanol 1:1. Visualization under UV 254 nm. Densitometric absorbance measurement at 220 nm (20mm/s scanning speed). The hRF was 47 for TET and 71 for ANT. System suitability was verified by resolution, selectivity, capacity and absence of tailing. The method was validated for linearity range (0.2 – 18 µg/band), for precision, for reproducibility, for robustness, and for accuracy expressed as average recovery values (100 % overall mean) at different concentrations. The method was also found statistically equivalent (Student’s t-test and F-test) to the official corresponding titrimetric methods of the European Pharmacopoeia. Finally, environmental and health impacts of the methods were qualitatively and quantitatively assessed better as the other described methods, using analytical greenness (AGREE), green analytical procedure index (GAPI), national environmental method index (NEMI), and analytical eco-scale (scores based on solvents/reagents, energy consumption, occupational hazard and waste generation).

      Classification: 7, 17a, 23e, 32a, 32f
      131 010
      A novel agarase, Gaa16B, isolated from the marine bacterium Gilvimarinus agarilyticus JEA5, and the moisturizing effect of its partial hydrolysis products
      Y. LEE, E. JO, Y.-J. LEE, T.-Y. EOM, Y. GANG, Y.-H. KANG, S. D. MARASINGHE, S. A. HETTIARACHCHI, D.-H. KANG, Chulhong OH* (*Jeju Marine Research Center, Korea Institute of Ocean Science and Technology, Gujwa-eup, Jeju, Korea;

      Marine Drugs 20(1), 2 (2022). Samples were the products of partial vs. complete hydrolysis of agar by rGaa16Bc, a recombinant form of agarase Gaa16B from Gilvimarinus agarilyticus (Cellvibrionaceae) overexpressed in Escherichia coli. D-galactose (G) and its oligomers (neoagarobiose (NA2), neoagarotetraose (NA4), neoagarohexaose (NA6)) were used as standards. TLC on silica gel with n-butanol – acetic acid – water 2:1:1. Visualization by spraying orcinol reagent (50 mg orcine monohydrate in 100 mL acetone and 8 mL sulfuric acid), followed by 10 min heating at 110° C. The observed patterns showed the apparition of NA6 and NA4 among the hydrolytic products already after 20 min reaction, whereas NA4 and NA2 were the main products after over-night complete hydrolysis.

      Classification: 4e, 10a
      131 003
      Development of a high-performance thin-layer chromatography method for the quantification of alkyl glycerolipids and alkenyl glycerolipids from shark and chimera oils and tissues
      M. PAPIN, C. GUIMARAES, B. PIERRE-AUE, D. FONTAINE, J. PARDESSUS, H. COUTHON, G. FROMONT, K. MAHÉO, A. CHANTÔME, C. VANDIER*, M. PINAULT (*Nutrition, Growth and Cancer INSERM UMR 1069, University of Tours, Tours, France;

      Marine Drugs 20(4), 270 (2022). Samples were ether glycerols (EG) purified: (A) from Chimaera monstrosa liver oil (Chimaeridae); (B) from mixed liver oil of sharks Centrophorus squamosus (Centrophoridae) and Somniosus microcephalus (Somniosidae); (C) from Macaca fascicularis hearts (Cercopithecidae); (D) from tumors obtained by grafting in mice the human melanoma cell line MDA-MB-435s, and (E) from periprostatic adipose tissue of men with prostate cancer. Reduction of (phospho)ester glycerolipids into EG and fatty alcohols was part of the purification process. Octadecyl-glycerol and octadecenyl-glycerol were used as standards of alkyl- and alkenyl-glycerols, respectively. HPTLC on silica gel previously developed with chloroform – methanol 1:1, air-dried and activated for 30 min at 110° C. Application under nitrogen stream (6 bar). Development with petroleum ether – diethyl ether – acetic acid 60:140:1. After 2 h drying at room temperature under ventilation hood, visualization by 50 s immersing into sulfuric acid (7 % in ethanol), followed by 2 h drying under air-stream, and 14 min heating at 140° C. Plates were documented under white light illumination and densitometry was performed by computered scanning of the pictures. Alkyl-glycerols (mean hRF 34, LOQ 1235 ng/band) and alkenyl-glycerols (mean hRF 44, LOQ 2352 ng/band), present in all samples (except alkenyl-glycerols in shark oil), were quantified after method validation for specificity, sensitivity, accuracy, precision and repeatability. Linearity range was 1000 ng – 7000 ng for both EG types. To confirm the band identification, samples and standards were also submitted to acidic hydrolysis before HPTLC application. In this case, the bands of alkenyl glycerols did not appear, because chlorhydric acid reacted with the vinyl ether bonds to form glycerol and aldehydes.

      Classification: 4d, 4e, 9, 11c, 32f
      130 141
      Two-dimensional high-performance thin-layer chromatography for the characterization of milk peptide properties and a prediction of the retention behavior – a proof-of-principle study
      M. TREBLIN, T. VON OESEN, L.-C. CLASS, G. KUHNEN, I. CLAWIN-RÄDECKER, D. MARTIN, J. FRITSCHE, S. ROHN* (*Department of Food Chemistry and Analysis, Institute of Food Technology and Food Chemistry, Technical University of Berlin, Berlin, Germany;

      J Chromatogr A 1653, 462442 (2021). Samples were peptides obtained through tryptic hydrolysis of the 5 most abundant milk proteins: α-lactalbumin (α-LA), β-lactoglobulin (β-LG), α-, β- and κ-casein (CA). As standards, synthetic whey and pea (Pisum sativum, Fabaceae) peptides (selected based on the in silico tryptic digest of α-LA, β-LG, legumin A, and vicilin with one or zero miscleavages) were only used in the last assay for prediction of the RF values of peptides with known amino-acid (AA) sequences. Two-dimensional HPTLC on silica gel (pre-washed with methanol and activated 10 min at 100°), first with basic mobile phase sec-butanol – pyridine – ammonia – water 39:34:10:26, and (after 12h drying) in the orthogonal direction with acidic mobile phase sec-butanol – pyridine – acetic acid – water 11:8:2:5. Derivatization for peptides and proteins by immersion into fluorescamine (0.05 % in acetone); visualization under UV 254 nm and 365 nm. Computer-assisted determination of the x- and y-coordinates of the derivatized zones. Repeatability (n=8) of the 2D-HPTLC was statistically tested with the Kolmogorov-Smirnov test for normal distribution and with Dixon’s Q test for outliers. Relative standard deviation (RSD) for the RF values was 12.9 % for the first dimension (y-coordinates) and 16.5 % for the second dimension (x-coordinates). According to their higher intensity and sharpness, 15 – 20 detected zones from each protein hydrolyzate were selected, manually scraped from the derivatized layer, dissolved in formic acid solution (0.1 % in acetonitrile – water 3:2), mixed with an equal volume of matrix (dihydroxybenzoic acid 2 % in acetonitrile – water 3:7), crystallized on air on a ground steel target, before being desorbed by the laser beam of the MALDI-TOF-MS/MS (matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry). Direct hyphenation of HPTLC to MS was not performed, to avoid zone diffusion during plate coating with the matrix and to circumvent the stronger binding of polar peptides on the layer.  The MS spectra were acquired in positive reflector mode in m/z range 340 – 4000 (10 – 2500 for fragments), using an external peptide as calibration standard. Identification of 51 from the 85 selected peptides according to AA sequences was performed, using software programs allowing m/z calculation of protein fragments and estimation of cleavage sites. Correlation of the retention behaviour of the peptides with their properties (molecular weight MW, isoelectric point IEP, charges, polarity) was tested with Student’s two-sided t-test after calculation of Pearson’s correlation coefficients. The correlation was significant with IEP, percentages of anionic AA and of non-polar AA; but not with the following properties: MW, percentages of cationic AA and of uncharged polar AA. Finally, based on the correlation results, regression formulas were found to calculate the x- and y-coordinates of any known peptide from the percentage of non-polar AA (or vice-versa). The prediction power of these formulas was verified by repeating the complete 2D-HPTLC-MS experiment with the standard peptides of whey and of peas, and measuring the absolute and relative deviations between the actual x- and y-coordinates and the predicted values. The absolute deviations were higher in the lower RF zones. The average, relative RF value deviations (range 22.1 – 25.7 %) were not different between whey and pea peptides.

      Classification: 2c, 2d, 4e, 18b, 19, 32e
      130 081
      Applicability of the Universal Mixture for describing system suitability and quality of analytical data in routine normal phase High Performance Thin Layer Chromatography methods
      M. SCHMID, T.K. Tiên Do*, I. TRETTIN, E. REICH (*CAMAG, Muttenz, Switzerland;

      J Chromatogr A 1666, 462863 (2022). Theoretical discussion on the factors determining the RF value of a given substance in a chromatographic system: A) the stationary phase (SP); B) the mobile phase (MP), the composition of which can be different from the solvent mixture prepared because of evaporation, saturation and liquid or gas adsorption effects over migration time; C) the difference of the free energies for the analyte transfer from SP to MP; D) external parameters like temperature and humidity. The universal HPTLC mixture (UHM) is a mixture of reference compounds that can be used for the system suitability test (SST) for the full RF range in all HPTLC experiments. Its composition is: thioxanthen-9-one (0.001 %), guanosine (0.05 %), phthalimide (0.2 %), 9-hydroxyfluorene, octrizole, paracetamol, sulisobenzone and thymidine (each 0.1 %), in methanol. The purpose was to study the potential of UHM to replace SST (described with specific markers in European Pharmacopoeia monographs) and to assess the quality of HPTLC results. TLC and HPTLC silica gel on different support (aluminium, glass) or with different granulometries and binders (classic, Durasil, Adamant), of the UHM, an acetonitrile extract of Abelmoschus manihot flowers (Malvaceae), a methanol extract of Sambucus canadensis flowers (Adoxaceae), and essential oils of Lavandula angustifolia, of Mentha × piperita (Lamiaceae) and of Myristica fragrans (Myristicaceae), as well as the following specific markers (standards): borneol, bornyl acetate, linalool, linalyl acetate (terpenoids), isoeugenol, isoeugenol acetate, chlorogenic acid (phenylpropanoids), gossypin (flavone), gossypetin-glucuronide, hyperoside (flavonol heterosides). Development (after 20 min plate conditioning with a saturated MgCl2 solution) with one of the following mobile phases: (MP1) toluene – ethyl acetate 19:1, especially for essential oils; (MP2) ethyl acetate – butanone – formic acid – water 5:3:1:1, especially for S. canadensis; (MP3) ethyl acetate – acetic acid – formic acid – water 100:11:11:26, especially for A. manihot. Documentation in UV 254 nm and 350 nm, and with white light (reflection + transmission), before and after derivatization. RF values were determined by scanning densitometry at 254 nm in absorption mode (for octrizole, at 366 nm in fluorescence mode with mercury lamp and optical filter K400 nm). For each HPTLC condition, intra-laboratory precision assay of UHM separation was performed (at least 5 analyses) with average RF values and 95 % prediction intervals, and calculating RF differences between pairs of UHM constituents and 95 % confidence intervals, which were max. +/-0.012 of the RF values for all UHM and markers. The sensitivity of UHM, and thus its usefulness as generic SST was demonstrated by repeating the HPTLC experiments with modifying by 10 % the quantity of one of the solvent each time. There were always significant changes in RF values of UHM components and/or in RF differences between pairs of UHM bands; it was often but no always the case with the official specific markers. UHM underwent also significant changes (although less than A. manihot extract) when several silica gel phases were compared under the same HPTLC conditions. This property is crucial to verify the right stationary phase before doing any RF correlations, and could make UHM a universal tool to identify discrepancies between different analyses. Finally, the use of UHM for a computer-supported evaluation of HPTLC results was discussed, either for zone identification and RF corrections (within confidence intervals), or for correlations of entire fingerprints as first step to implement machine learning algorithms.

      Classification: 2a, 2f, 3g, 7, 8a, 15a, 15b, 32e