ToF-SIMS characterisation of diterpenoic acids after chromatographic separation

Microcolumn liquid chromatography (μHPLC) coupled on-line with time of flight secondary ion mass spectrometry (ToF-SIMS) was applied for mixture of diterpenoic acids (abietic, gibberellic and kaurenoic) analysis. Chromatographic effluent, with analytes separated, was carried out directly onto different, ToF-SIMS compatible surface substrates, for further ToF-SIMS analysis. Silica gel Si₆₀, aluminium backplate modified Si₆₀, monolithic silica gel and Raman spectroscopy chromatographic thin layers were used as the deposition substrates in this experiment. By ToF-SIMS surface imaging the deposition trace picture has been obtained. Effluent deposition surface area was scanned for diterpenoic acid fragment mass values based on mass spectrometric library. Measured ToF-SIMS dataset of fragment abundance and intensities were used for preliminary fragmentation schemes construction. The lowest substrate background activity has been established for monolithic silica gel thin layer and aluminium backplate modified Si₆₀ thin layer. In the case of Raman spectroscopy pre-treated thin layer or conventional chromatographic thin layer Si₆₀, the both, high background signal intensity and impossibility to construct negative ions surface image, were observed. Diterpenoic acids studied serve the similar mass spectrum but ToF-SIMS coupled with liquid chromatographic separation brings new impact to the positive identification of analytes studied.     

Thin-layer chromatographic investigation of plant pigments in selected juices and infusions of cosmetological importance and their antioxidant potential

ABSTRACT

Fruit preparations (e.g., fruit juices and nectars) are rich in plant pigments and of great demand by the alimentary and pharmaceutical industry, basically not only due to their health-enhancing properties, but also due to their attractive colors and an overall high esthetic valor. Anthocyanins and anthocyanidins are an interesting group of plant pigments, and in this study, a thin-layer chromatographic detection was carried out of two anthocyanins (cyanin and keracyanin) and two anthocyanidins (pelargonidin and delphinidin) in a selection of the commercial and homemade fruit juices and the infusions prepared of dried plants. Moreover, the same preparations were evaluated for their antioxidant properties by means of two spectrophotometric methods (based on the 2,2′-azino-di-[3-ethylbenzthiazoline] sulfonate (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) tests, respectively) and by the dot-blot test (based on DPPH). Finally, an effort was undertaken to correlate the chromatographically established occurrence of plant pigments in the investigated fruit preparations with their antioxidant properties. However, this correlation was not straightforward, and for at least two reasons. First, the four plant pigments considered are not the only secondary plant metabolites which exert an antioxidant activity. Second, different chemistries stand behind the two spectrophotometric antioxidant activity tests (ABTS vs. DPPH), and the different measuring techniques (spectrophotometry vs. dot blot) are assumed. Nevertheless, it was established that the juices and infusions with the highest detected numbers of plant pigments characterize with the high antioxidant activities also (with blueberries, chokeberries, and hibiscus flower on the top positions). In that way, confirmation of the antioxidant potential of the plant pigments was obtained. Moreover, the presence of certain plant pigments in fruit juices and plant infusions was reported for the first time.     

Coupling protocol for thin layer chromatography/matrix-assisted laser desorption ionization

Summary This paper reports the investigation and optimization of the thin layer chromatography/matrix-assisted laser desorption/ionization (TLC-MALDI) coupling protocol. The fundamental coupling parameters which influence sensitivity and lateral analyte spreading are extraction solvent selection, extraction time, and pressure. Selection of the solvent is dependent upon its extraction efficiency, which has been correlated with extraction solventR _f value. Maximum extraction efficiency occurs after two minutes for the analyte/solvent system studied. High extraction efficiency solvents cause significant lateral spreading of analyte; up to a three-fold increase in initial analyte spot size. Analyte recovery was found to be limited by the silica gel inter-partice porosity and the solvent extraction efficiency. For maintaining chromatographic resolution and maximizing signal intensity, extraction solvents withR _f values between 0.4 to 0.6 are optimal. The upper analyte recovery limit, using extraction solvents within thisR _f range, is estimated at 22%. Dedicated to Professor Werner Haerdi on the occasion of his 70^th birthday.     

A novel digitally optimized rapid quantification of carcinogenic aryl azo amines from various food matrices by HPTLC-MS

Abstract

A rapid, precise and reproducible high-performance thin-layer chromatography-mass spectroscopy (HPTLC-MS) method was developed and validated for identification and quantification of toxic aryl azo amines from chili oils, pickles and related food matrices using Merck thin-layer chromatography (TLC) silica gel F-254 plates, as a stationary phase adsorbent by CAMAG HPTLC system. Extraction of azo food colors was done using method specified as per ISO standard. The use of tert-butyl-methyl ether as final extraction solvent resulted in minimal sample clean up and high efficiency. Calibration was performed to combat matrix effect. Good linearity levels were observed for the concentrations of aniline, 2, 4-xylidine and 4 aminoazobenzene of 2–14?ppm per band. The limits of detection and quantification found for aniline, 2,4-xylidine and 4-aminoazobenzene were found to be 0.0015, 200, and 400?ppm and limit of quantification (LOQ) values were found to be 0.004, 600, and 1200?ppm, respectively, with a correlation coefficient of 99.94 %. This study thus highlights an economically viable, commercially sustainable yet highly scientific technique of HPTLC-MS methodology with structural elucidation of banned azo amines in food and related products for their identification duly detectable at trace levels in a digitized form, which can enable material integrity-related prediction capacity of suspected food matrices.