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4 results on '"Marrubini G"'

2. CE analysis and molecular characterisation of depurinated DNA samples

Author

Solange Sorçaburu-Cigliero, Giorgio Marrubini, Paola Pitacco, Paolo Fattorini, Carlo Previderè, Pierangela Grignani, Fattorini, Paolo, Marrubini, G., SORCABURU CIGLIERO, Solange, Pitacco, P., and Grignani, P. Previderè C.

Subjects
Male, Guanine, Biochemical Phenomena, Clinical Biochemistry, Biology, STR, Polymerase Chain Reaction, Biochemistry, Analytical Chemistry, law.invention, chemistry.chemical_compound, law, Humans, A-DNA, Typing, Polymerase chain reaction, Electrophoresis, Agar Gel, Adenine, Electrophoresis, Capillary, DNA, Middle Aged, CE, Depurination, DNA primary structure, QPCR, STR, Molecular biology, CE, DNA primary structure, QPCR, Molecular Weight, Real-time polymerase chain reaction, chemistry, Depurination, Agarose gel electrophoresis, Linear Models, Nucleic Acid Conformation, DNA Damage, Microsatellite Repeats
Abstract

A DNA sample was partially degraded by scalar heat-acid treatments to study the extent of apurinic-apyrimidinic (A-P) lesions produced along the molecule. A CE-UV method allowed us to measure the rate of depurination at pH 5.0 and 70°C which was calculated to be 5.41×10(-6) s(-1) for adenine and 6.27×10(-6) s(-1) for guanine. CE identified depurination on treated samples when it occurred with a loss of >4% of the basic moieties. The molecular features of the A-P enriched samples were investigated by using molecular assays (agarose gel electrophoresis, UV spectrophotometry and quantitative PCR) and the consistency of the results of the STR typing were compared with the degree of depurination of the PCR template. The treated DNA samples showed molecular features such as fragmentation, altered OD(260) /OD(280) ratios and decreased ability of the quantitative PCR to synthesise the human target, related to the severity of depurination. A satisfactory correlation between the degree of damage and the amount of residual PCR-sensitive target sequences was also demonstrated (r(2) =0.9717). The conventional and mini-STR typing of the samples showed that the genetic outcome was influenced by a depurination damage that exceeded 4% when locus drop-outs and artefactual PCR results were evident. As the success of STR typing depends on the integrity of the DNA recovered from the samples, the CE-UV, physical and molecular assays described here are proposed as a set of useful methods in the analysis of certain forensic and clinical samples, for a critical evaluation of the outcome of the genetic testing.

Published
2011
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3. The use of a microwave-assisted solvent extraction coupled with HPLC-UV/PAD to assess the quality of Marrubium vulgare L. (white horehound) herbal raw material.

Author

Martino E, Della Volpe S, Cavalloro V, Amri B, Kaab LBB, Marrubini G, Rossi D, and Collina S

Subjects
Biomarkers analysis, Chemical Fractionation, Chromatography, High Pressure Liquid, Marrubium physiology, Microwaves, Plant Extracts chemistry, Plant Leaves chemistry, Research Design, Stress, Physiological, Diterpenes analysis, Marrubium chemistry, Phytochemicals analysis, Plant Extracts isolation & purification
Abstract

Introduction: Marrubium vulgare is a herbal remedy presents in several European Pharmacopoeias and commonly marketed as white horehound. The chemotaxonomic marker of Marrubium genus is marrubiin and its content may change in response to biotic and abiotic stress., Objective: Development of a microwave-assisted solvent extraction (MASE) methodology suitable for exhaustively extracting marrubiin from M. vulgare leaves, easily applicable to large sets of samples. Evaluation of the influence of copper(II) on marrubiin production., Material and Methods: M. vulgare leaves were dried, extracted exploiting MASE and analysed via high-performance liquid chromatography ultraviolet photodiode array detection (HPLC-UV/PAD) system. A design of experiments approach was adopted to select the best extraction conditions. Extraction parameters (solvent composition, extraction time and temperature), were studied applying two full factorial experimental designs in a sequential approach. To analyse samples, a rapid HPLC-UV/PAD method was set up., Results: The best results in terms of marrubiin extraction yield were obtained extracting samples at 120°C with 100% ethanol, for 15 min (3 × 5 min microwave cycles). The developed methodology was successfully applied to matrices grown in Greenhouse conditions and under stress induced by copper(II), selected as model agent for abiotic stress. Progressively decreasing production of marrubiin was evidenced in connection with treatment with 80, 200 and 300 mg/L copper sulphate., Conclusion: An efficient methodology for the extraction and determination of the amount of marrubiin in large sets of samples of M. vulgare plants was developed. Results demonstrated that marrubiin is an easily detectable marker useful for evaluating M. vulgare reaction to stress., (© 2019 John Wiley & Sons, Ltd.)

Published
2019
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