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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      129 013
      A bioimaging system combining human cultured reporter cells and planar chromatography to identify novel bioactive molecules
      I. KLINGELHOEFER, L. PHAM NGOC, B. VAN DER BURG, Gertrud E. MORLOCK* (*Chair of Food Sci., Inst. of Nutrit. Sci. & TransMIT Center for Effect-Directed Anal., Justus Liebig Univ. Giessen, Heinrich-Buff-Ring26-32, 35392, Giessen, Germany, Gertrud.Morlock@uni-giessen.de)

      Anal Chim Acta 1183 (2021) 338956. Demonstration of first on-surface adherent cell assays by investigating growth and viability of modified adherent human osteosarcoma U2OS cells (Cytotox CALUX® cell system) as a biodetector for the cytotoxicity evaluation of e.g. mycotoxins, cytostatics and plant extracts, on HPTLC plates. In the planar bioassay, the cytotoxicity of sample extracts was measured with the luciferase activity in the Cytotox CALUX® reporter cells. For direct bioresponse testing on the HPTLC plate, the U2OS cell suspension was pipetted along the the sample track. RP-HPTLC-CALUX® bioassays of Saussurea costus and Ginseng extracts on HPTLC RP-18 W with n-hexane - toluene - tetrahydrofuran 25:7:15 to 6 cm. After drying, immersion in citrate phosphate buffer pH 12, and several drying and moistening steps the cytotoxic zones were detected by bioluminescence inhibition. Two different test procedures proved Cytotox CALUX® cell viability on surface and in microtiter plate assay. The isolated cytotoxic zone was analyzed by HPTLC on RP-18 W with n-hexane - toluene - tetrahydrofuran 10:1:1 and one part of the  chromatogram cut in half was derivatized with sulfuric acid reagent and evaluated under white light, while the other part was subjected to the new planar Cytotox-CALUX® bioassay. The cytotoxic zone had the same hRF as standards costunolide and dehydrocostus lactone.

      Classification: 32e
      129 012
      Nanomole-scaled high-throughput chemistry plus direct bioautography on the same chromatography plate for drug discovery
      I. YÜCE, Gertrud E. MORLOCK* (*Chair of Food Sci., Inst. of Nutrit. Sci. & TransMIT Center for Effect-Directed Anal., Justus Liebig Univ. Giessen, Heinrich-Buff-Ring26-32, 35392, Giessen, Germany, Gertrud.Morlock@uni-giessen.de)

      Anal Chim Acta 1182 (2021) 338950. On-surface synthesis and effect-directed analysis for quantitative nanomole-scaled high-throughput chemistry and biological response on a single HPTLC plate for up to 68 on-surface synthesis reactions. For generic on-surface synthesis, 1 mL or 15 nmol each of malononitrile and benzaldehyde candidates were automatically sprayed as 3-mm bands  on an HPTLC plate, dried, and oversprayed with  diketoester on the same zones, followed by drying. Development in anti-parallel with chloroform - tetrahydrofuran 6:1 up to 5 cm and detection in UV 254 nm, 366 nm and  white light. For yield calculation, four-point calibration with the preparatively synthesized standard ethyl 6-amino-5-cyano-2-methyl-4-(4-nitrophenyl)-4H-pyran-3-carboxylate, derivatization with ninhydrin collidine reagent, heating at 110°C, detection in UV 254 nm, 366 nm and white light and densitometric absorbance measurement at 430 nm. Online elution of the synthesis product zones with methanol to high-resolution mass spectroscopy and analysis by electrospray ionization MS. For the HPTLC-Bacillus subtilis bioassay 3 mm bands of ciprofloxacin solutions were developed in ethanol - water - hydrochloric acid 9:10:1, sprayed with the bacterial suspension, incubated at 37°C for 2 h, sprayed with phosphate buffered saline (PBS)-buffered thiazolyl blue tetrazolium bromide (MTT) solution, incubated at 37°C for 30 min, followed by drying. Detection of colorless (bright) inhibition zones in white light and by absorbance measurement of the blue-violet formazan background of the biodensitogram at 546 nm.

      Classification: 4e, 32
      129 007
      High-performance thin-layer chromatography hyphenated to high-performance liquid chromatography-diode array detection-mass spectrometry for characterization of coeluting isomers
      Agnes M. MORICZ*, V. LAPAT, G.E. MORLOCK, P.G. OTT (*Plant Protection Institute, Centre for Agricultural Research, Herman O. Str. 15, 1022, Budapest, Hungary, moricz.agnes@agrar.mta.hu)

      Talanta 219 (2020) 121306. Development of a workflow for compound characterization of coeluting compounds: employing an HPTLC-UV/Vis/FLD-EDA screening, followed by the characterization and identification of the most potent compounds by multi-imaging, heated electrospray ionization high-resolution mass spectrometry (HESI-HRMS) and hyphenated HPTLC-UV/Vis/FLD-HPLC-DAD-ESI-MS. HPTLC of methanolic Lemon balm leaf extract and standards oleanolic acid and ursolic acid on silica gel with n-hexane - ethyl acetate 7:3 up to 70 mm, drying for 2 min, evaluation in UV 254 nm (UV), UV 366 nm (FLD) and white light (Vis) after derivatization with a solution of vanillin (40 mg) and sulfuric acid (200 μL) in 10 mL ethanol and heating at 110°C for 5 min. Three additional chromatograms were prepared for the antibacterial assays (1) against B. subtilis, (2) A. fischeri and  (3) the α-glucosidase assay. (1) immersing in B. subtilis suspension, detectioon by immersion in aqueous 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide solution after incubation, followed by incubation until bright zones against a violet background appear, (2) immersing in Aliivibrio fischeri suspension and instantly monitoring in real-time for 30 min using 1 min exposure time in 5-min intervals and indicating the dark (or bright) active zones against the bioluminescent background, (3) immersing in α-glucosidase solution (10 units/mL) and 0.1 M sodium acetate buffer adjusted to pH 7.5 for incubation and immersing into substrate solution (1.2 mg/mL 2-naphthyl-α-D-glucopyranoside in ethanol) for further incubation at room temperature for 10 min and detection by immersion in aqueous Fast Blue Salt B solution (1 mg/mL) and drying, this revealed the enzyme inhibitors as bright zones against a violet background in white light. Online elution of zones of interest with methanol into the MS and full scan recording in the range of m/z 50–750 with a resolution of 280,000 in both negative and positive ionization modes. Expanding the HPLC-DAD-ESI-MS system by installing a TLC-MS interface enabling direct elution of HPTLC zones into the HPLC eluent.

      Classification: 4d, 32
      129 004
      New incorporation of the S9 metabolizing system into methods for detecting acetylcholinesterase inhibition
      E. AZADNIYA, J. MOLLERGUES, T. STROHEKER, K. BILLERBECK, Gertrud E. MORLOCK* (*Chair of Food Sci., Inst. of Nutrit. Sci. & TransMIT Center for Effect-Directed Anal., Justus Liebig Univ. Giessen, Heinrich-Buff-Ring 26-32, 35392, Giessen, Germany, Gertrud.Morlock@uni-giessen.de)

      Anal Chim Acta 1129 (2020) 76-84. Demonstration of a new hyphenated HPTLC-S9-AChE assay for detection of neurotoxic chemicals, including metabolic activation, at levels consistent with the threshold of toxicological concern (TTC) for organophosphates (OPs). The high sensitivity allows for the direct application of packaging migrates or extracts on the HPTLC plate without additional requirement steps. HPTLC of the ethanolic standards chlorpyrifos (CP), quinalphos (QP), eserine (ER), parathion (PT), nonylphenol (NP) and tris(nonylphenyl) phosphite (TNPP) at five different levels in the range of 0.5-10 mL/band (overall range for all six chemicals 0.1-1000 ng/band) on silica gel. After application, drying and pre-wetting the application bands by immersion up to 10 mm in water, then spraying the S9 mixture (7 mL each, 0.1 mg/mL) immediately on top of the start zones, followed by incubation at room temperature for 30 min. After drying plates for 20 min and 5 min chamber saturation, development with ethyl acetate - methanol - water 5:5:2 for ER; n-hexane - ethyl acetate - ethanol 16:3:1 for QP and PT; n-hexane - toluene - ethyl acetate 5:4:1 for CP, NP and TNPP. Evaluation in 254 nm, 366 nm and white light. Detection by immersion in AChE solution (6.6 U/mL plus 1 mg/mL bovine serum albumin in Tris-HCl buffer, 0.05 M, pH7.8) for 2 s, and incubating at 37 °C for 25 min, then immersing into the substrate-chromogenic solution (ethanolic solution of 1-naphthyl acetate  - aqueous solution of Fast Blue B salt 1:2 , 3 mg/mL each) for 1 s, drying for 10 min. Evaluation in white light, absorbance measurement at 546 nm (mercury lamp) using an inverse scan. The advantages of this straightforward workflow were demonstrated by comparison with the status quo microtiter plate assay. The method is a pragmatic new tool in risk assessment in general and can be transferred to further toxicities of interest and any other category of complex sample mixtures.

      Classification: 4e, 32
      128 093
      High-throughput enzyme inhibition screening of 44 Iranian medicinal plants via piezoelectric spraying of planar cholinesterase assays
      E. AZADNIYA, I. THOMÄ, J. BAAKE, Gertrud E. MORLOCK* (*Institute of Nutritional Science, and TransMIT Center for Effect-Directed Analysis, Justus Liebig University Giessen, Giessen, Germany; gertrud.morlock@uni-giessen.de)

      Journal of Chromatography B, 1184, 122956 (2021). Test for acetyl- and butyrylcholinesterase (AChE and BChE) inhibition without development of piperin (standard inhibitor of AChE and BChE) and ethanol – water (3:2) extracts of Iranian plants, on HPTLC silica gel prewashed twice with methanol – water 3:2 and dried 60 min at 120°C. After sample application the plate was immersed (speed 3.5 cm/s, time 2 s) into enzyme solution (6.6 units/mL AChE or 3.3 units/mL BChE in TRIS buffer 0.05 M, with bovine serum albumin 0.1 %, pH 7.8), incubation 25 min at 37°C and immersion (speed 3.5 cm/s, time 1 s) into chromogenic substrate solution (α-naphthyl acetate 0.1 % and Fast Blue salt B 0.2 % in ethanol – water, 1:2). Seven mobile phases were tested for the active samples. Best separation was obtained with toluene – ethyl acetate – formic acid – water 4:16:3:2 and with toluene – ethyl acetate – methanol 6:3:1. Before enzymatic assay, plates developed with acidic mobile phases were neutralized by spraying 3 mL citrate phosphate buffer (Na2HPO4 8 %, citric acid q.s. ad pH 7.5) followed by 10 min of automatic drying. Enzymatic assay was performed using a piezoelectric spraying device: a) pre-wetting by spraying 1 mL TRIS buffer (0.05 M, pH 7.8); b) spraying 3 mL of the enzyme solution; c) incubation 25 min in a humid box at 37°C; d) spraying 0.5 mL substrate solution; e) 5 min drying at room temperature, and then 10 min of automatic drying. By spraying, zone shift and zone diffusion, which occurred with plate immersion, were avoided. For development control, derivatization was done by piezoelectrically spraying 4 mL of sulfuric anisaldehyde reagent (anisaldehyde – sulfuric acid – acetic acid – methanol, 1:10:20:170), followed by heating 3 min at 110°C. For identification of zones of interest, direct elution with methanol from underivatized HPTLC plates through a TLC-MS interface directly to a MS. Identified zones were 3-O-acetyl-β-boswellic acid (triterpenoid) from Boswellia carteri gum-resin (Burseraceae), pimpinellin and psoralen (furocoumarins) from Heracleum persicum flowers (Apiaceae), oleuropein (seco-iridoid) from Olea europaea leaves (Oleaceae), harmine, harmaline, vasicine, deoxyvasine (alkaloids) from Peganum harmala seeds (Zygophyllaceae), costic acid (sesquiterpene) from Nardostachys jatamansi hypocotyl (Valerianaceae), elaidic, linoleic, palmitic, palmitoleic acids (fatty acids) from Pistacia atlantica fruits (Anacardiaceae).

      Classification: 4e, 8b, 11a, 15a, 22, 32e
      128 089
      Effect-directed profiling of 32 vanilla products, characterization of multi-potent compounds and quantification of vanillin and ethylvanillin
      Gertrud E. MORLOCK*, M. BUSSO, S. TOMEBA, A. SIGHICELLI (*Institute of Nutritional Science, and TransMIT Center for Effect-Directed Analysis, Justus Liebig University Giessen, Giessen, Germany; gertrud.morlock@uni-giessen.de)

      J Chromatogr. A, 1652, 462377 (2021). Samples were vanilla tinctures, water − ethanol − ethyl acetate 1:1:1 extracts of vanilla-flavored food products and of natural Vanilla sp. (Orchidaceae) pods, oleoresin, paste and powders, as well as calibration standards of vanillin (1) and ethylvanillin (2). HPTLC on silica gel with n-hexane – ethyl acetate 1:1 for profiling, 3:2 for quantification. Other mobile phases were also tested and given in the supplement. Compounds (1) and (2) (hRF 68 and 82, respectively) were quantified by absorbance densitometry (at maximal wavelength 310 nm, deuterium lamp, scanning speed 10mm/s). Contents were found to be between 1 μg/g and 36 mg/g for (1) and null for (2) except in one tincture (62 µg/mL). Derivatizations performed for five assays: A) to detect radical scavengers, immersion (speed 3 cm/s, time 5 s) into DPPH• (0.5 mM in methanol), followed by drying for 90 s at room temperature and 30 s at 60 °C; B) to detect activity against Gram-negative bacteria, immersion (speed 2 cm/s, time 3 s) into Aliivibrio fischeri suspension, followed by recording the bioluminescence; C) to detect activity against Gram-positive bacteria, immersion (speed 3.5 cm/s, time 6 s) into Bacillus subtilis, followed by incubation 2 h at 37 °C, immersion in MTT solution, incubation for 30 min at 37 °C and heating for 5 min at 50 °C; D) to detect acetylcholinesterase (AChE) inhibitors, immersion (speed 2.5 cm/s, time 2 s) into AChE solution (666 units in TRIS buffer 0.05M, with bovine serum albumin 0.1 %, pH 7.8), incubation for 25 min at 37 °C and immersion into substrate solution (α-naphthyl acetate 0.1 % and Fast Blue salt B 0.18 % in ethanol – water, 1:2; E) to detect tyrosinase inhibitors, spraying with enzyme solution (400 unit/mL, in phosphate buffer 0.02 M, pH 6.8), followed by 2 min drying, immersion into substrate levodopa (18 mM in phosphate buffer, pH 6.8), 10 min incubation at room temperature and drying. For identification, zones of interest were transferred with methanol from underivatized HPTLC layer through a TLC-MS interface and a filter frit directly to a Quadrupole-Orbitrap MS (heated electrospray ionization, probe heater at 270°C, spray voltage 3.5kV, lock masses acetic acid for negative, dibutyl phthalate for positive ionization, mode full HR-MS scan in m/z range 50–750). Afterwards, the following substances assigned by MS were confirmed by using HPTLC comparison with standards: (1) and (2), vanillyl alcohol, vanillic acid, ethyl vanillyl ether, coumarin, 4-hydroxybenzoic acid, 4-methoxybenzoic acid, 4-hydroxybenzaldehyde, 4-allyl benzoic acid, oleamide, triacetin.

      Classification: 4e, 7, 8b, 32e
      128 034
      Quantification of gymnemagenin and β-sitosterol in marketed herbal formulation by validated normal phase HPTLC method
      S.E. POTAWALE, S.Y. GABHE*, K.R. MAHADIK (*Department of Pharmaceutical Chemistry, Poona College of Pharmacy, Bharati Vidyapeeth Deemed University, Pune, India; satish3619@rediffmail.com)

      Chromatography Research International 2014, 626801 (2014). HPTLC of extracts of Gymnema sylvestre (Apocynaceae) in tablets, as well as standards for calibration, on silica gel (prewashed with methanol and activated at 120°C for 15 min) with toluene – ethyl acetate – methanol 65:25:14. Derivatization by immersing into sulfuric acid (5 % in methanol) and heating at 110°C for 4 min. Densitometric evaluation within 25 min in absorbance mode at 423 nm, which was the optimal wavelength for quantifying simultaneously the triterpenoid gymnemagenin (hRF 27, linearity range 100–1200 ng/band, LOD 32 ng/band, LOQ 53 ng/band) and β-sitosterol (hRF 78, linearity range 200–1200 ng/band, LOD 97 ng/band, LOQ 159 ng/band). Interday and intra-day precisions as well as recovery rates provided relative deviation values below 1 %. This method was used to determine the analyte contents in the tablets (0.041 % gymnemagenin and 0.138 % β-sitosterol), as well as to confirm the stability of the analytes in solution at room temperature after 48h.   

      Classification: 15, 32e
      128 035
      The bacterial microbiome of the long-term aquarium cultured high microbial abundance sponge Haliclona cnidata – sustained bioactivity despite community shifts under detrimental conditions
      J. SCHELLENBERG, J. REICHERT, M. HARDT, I. KLINGELHÖFER, G. MORLOCK, P. SCHUBERT, M. BIŽIĆ, H.-P. GROSSART, P. KÄMPFER, T. WILKE, Stefanie P. GLAESER* (*Research Centre for BioSystems, Land Use and Nutrition, Institute of Applied Microbiology, Justus Liebig University Giessen, Giessen, Germany; stefanie.glaeser@umwelt.uni-giessen.de)

      Frontiers in Marine Science 7, 266 (2020). Methanol extracts from marine sponge Haliclona cnidata (Chalinidae) submitted to different stresses (antibiotics and/or darkness) were separated on HPTLC silica gel with an automated 15-step gradient based on methanol, dichloromethane and n-hexane. Bioluminescence was recorded after immersing the HPTLC plates into Aliivibrio fischeri suspension. Antibacterial activity and quorum sensing enhancement were analysed on software, and Pearson’s similarity coefficient was applied to generate similarity matrices for cluster analysis (UPGMA, Unweighted Pair Group Method with Arithmetic Mean). Only slight differences were observed, especially in QS enhanced zones in stressed vs. control cultures.

       

      Classification: 32e
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