LC MS

Ibotenic acid, muscimol and muscarine were recognized as responsible for psychotropic effects of A. muscaria and A. pantherina. Demand for their specific and sensitive identification and quantitation in biological material lead to effort to develop reliable analytical method, but satisfactory solution is still lacking. Presented article describes liquid chromatography-mass spectrometry method suitable for isolation and identification of principal toxins of A. muscaria and A. pantherina in urine. Dedicated liquid chromatography-mass spectrometry method is reported. Technique consists of an extraction of analytes on Strata X-CW and Discovery SCX SPE cartridges and separation is achieved using a Gemini C18 column (150 mm x 2.0 mm, 5 micron) and 8 mM heptafluorobutyric acid as mobile phase. Detection at m/z 159 for ibotenic acid, m/z 115 for muscimol and m/z 174 for muscarine was used. Retention times and LODs are 2.6 min and 50 ng.ml-1 for ibotenic acid, 4.6 min and 40 ng.ml-1 for musc...

Withania somnifera (Ashwagandha) root extract is an important herbal medicine. The current study was designed to investigate the impact of Consciousness Energy Healing Treatment (The Trivedi Effect®) on the characteristic properties of the phytoconstituents present in the ashwagandha root extract using LC-MS, GC-MS, and NMR spectroscopy. Ashwagandha root extract was divided into two parts – one part was control (without treatment), while another part was treated with the Consciousness Energy Healing Treatment remotely by seven renowned Biofield Energy Healers and defined as the Biofield Energy Treated sample. The LC-MS analysis revealed that the retention time (Rt) of the phytoconstituents remained same in both the control and treated samples, whereas the peak area% was significantly altered. The peak area% of the treated sample at Rt of 5.06, 5.24, 5.49, 5.68, 6.78, 6.87, 8.12, 8.22, 8.36, 8.55, 9.02, and 10.13 minutes were significantly increased in the range of 0.27% to 146.94% compared to the control sample. In addition, the peak area% of the treated sample at Rt of 5.29, 5.36, 6.40, 6.49, 6.56, 6.67, 7.08, 7.37, 7.76, 7.87, 8.02, 8.43, 8.85, 8.97, and 9.12 minutes were significantly decreased in the range of 1.83% to 23.53% compared with the control sample. A total of 18 withanolides such as sitoindoside IX, viscosa lactone B, 24, 25-dihydrowithanolide D, withanolide A, withanone, withaferin A, withanolide D, ixocarpalactone A, withanolide S, withanolide sulfoxide, etc. were proposed with their structure from the molecular mass at m/z 650, 489, 473, 471, 505, and 992 at retention times of 6.87, 7.10, 7.87, 8.22, 8.54, and 9.11 minutes with the help of GC-MS and NMR data of both the control and Biofield Energy Treated samples. The mass peak intensity values of the Biofield Energy Treated sample at the same retention time in was significantly decreased in the range of 3.99% to 52.28% compared with the control sample. These findings suggest that Energy of Consciousness Healing Treatment could be beneficial for altering the concentration of the phytoconstituents in the ashwagandha root extract by modifying their intrinsic physicochemical properties, which might be helpful to improve the bioavailability of active constituents of ashwagandha root extract that might provide better therapeutic response against inflammatory diseases, immunological disorders, arthritis, cancer, diabetes, sexual disorders, aging and other chronic infections.

Aerial parts of some Veronica species are used as traditional medicine for treatment of various inflammatory conditions, including rheumatism. In the present study, phenolic content and anti-inflammatory activity of three Veronica species (Plantaginaceae), Veronica jacquinii Baumg., Veronica teucrium L. and Veronica urticifolia Jacq was studied. The identification and quantification of 30 phenolic compounds in aqueous-acetone extracts were performed using LC–MS/MS technique, and the main components were baicalin, hyperoside, isoquercetine, chlorogenic acid and quinic acid. Genistein and baicalein were detected for the first time in Plantaginaceae family and Veronica genus, respectively.The anti-inflammatory potential of tested extracts was determined using calcium ionophore-stimulated human platelets as a source of cyclooxygenase-1 (COX-1) and 12-lipoxygenase (12-LOX) enzymes, and highly sensitive and specific LC–MS/MS technique was performed for detection of the main arachi-donic acid metabolites formed by them. V. urticifolia extract exerted the strongest COX-1 inhibition(IC50< 0.4 mg/mL), while V. jacquinii extract exhibited the highest ability to inhibit 12-LOX activity(IC50= 1.072 mg/mL). The results obtained in this study may be used to promote better use of the genus Veronica as anti-inflammatory agents.

The colorant behaviour of cochineal and kermes insect dyes in 141 experimentally-dyed and 28 artificially-aged samples of silk and wool was investigated using ultra-high performance liquid chromatography coupled to photodiode array detector (UHPLC-PDA), liquid chromatography electrospray ionisation mass spectrometry (LC-ESI-MS) and image scanning electron microscopy – energy dispersive X-ray spectroscopy (SEM-EDX). Partial-least squares discriminant analysis (PLS-DA) was then used to model the acquired UHPLC-PDA data and assess the possibility of discriminating cochineal insect species, as well as their correspondent dyed and aged reference fibres. The resulting models helped to characterize a set of 117 red samples from 95 historical textiles, in which UHPLC-PDA analyses have reported the presence of cochineal and kermes insect dyes.

Analytical investigation of the experimentally-dyed and artificially-aged fibres has demonstrated that the ratio of compounds in the insects dye composition can change, depending on the dyeing conditions applied and the type of fibres used. Similarities were observed when comparing the UHPLC-MS and SEM-EDX results from the dyed and aged references with the historical samples. This was verified with PLS-DA models of the chromatographic data, facilitating the classification of the cochineal species present in the historical samples. The majority of these samples were identified to contain American cochineal, which is in agreement with historical and dye identification literature that describe the impact of this dyestuff into European and Asian dyeing practices, after the Iberian Expansion in the 16th century.

The analytical results emphasize the importance of using statistical data interpretation for the discrimination of cochineal dyes, besides qualitative and quantitative evaluation of chromatograms. Hence, the combination of UHPLC-PDA with a statistical classification method, such as PLS-DA, has been demonstrated to be an advisable approach in future investigations to assess closely related species of natural dyes in historical textile samples. This is particularly important when aiming to achieve more accurate interpretations about the history of works of art, or the application of natural dyes in old textile production.