Isolation and characterization of three benzylisoquinoline alkaloids from Thalictrum minusL. and their antibacterial activity against bovine mastitis

Journal of Ethnopharmacology

Available online 16 July 2016

In Press, Accepted Manuscript — Note to users

• Saleem Mushtaq^{a, d},
• Muzaffar Ahmad Rather^b,
• Parvaiz H Qazi^{a, , ,} ,
• Mushtaq A Aga^c,
• Aabid Manzoor Shah^a,
• Aiyatullah Shah^a,
• Md. Niamat Ali^{d, ,}

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doi:10.1016/j.jep.2016.07.040

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Abstract

Ethno-pharmacological relevance

The roots of Thalictrum minus are traditionally used in the treatment of inflammation and infectious diseases such as bovine mastitis. However, there are no reports available in literature till date regarding the antibacterial studies of T. minus against bovine mastitis.

Aim of the study

The present study was undertaken to evaluate the antibacterial potential of crude extract of T. minus (root) and some of its isolated constituents against bovine mastitis in order to scientifically validate its traditional use.

Materials and methods

A total of three alkaloid compounds were isolated from the DCM: MeOH extract of roots of T. minus using silica gel column chromatography. Structural elucidation of the isolated compounds was done by using spectroscopic techniques like mass spectrometry and NMR spectroscopy. Pathogens were isolated from cases of bovine mastitis and identified by using 16 S rRNA gene sequencing. The broth micro-dilution method was used to evaluate the antibacterial activities of DCM: MeOH extract and isolated compounds against mastitis pathogens.

Results

The three isolated compounds were identified as benzylisoquinoline alkaloids (1) 5’-Hydroxythalidasine, (2) Thalrugosaminine and (3) O-Methylthalicberine. Compounds (2) and (3) are reported for the first time from the roots of T. minus. Five mastitis pathogens viz., Staphylococcus xylosus, Staphylococcus lentus, Staphylococcus equorum, Enterococcus faecalis and Pantoea agglomerans were identified on the basis of sequence analysis of isolates using the nucleotide BLAST algorithm. This study reports for the first time the isolation and molecular characterization of mastitis pathogens from Kashmir valley, India. The DCM: MeOH extract exhibited broad spectrum antibacterial activities that varied between the bacterial species (MIC = 250-500 µg/ml). 5’-Hydroxythalidasine and Thalrugosaminine showed promising antibacterial activity with MIC values of 64-128 µg/ml while Staphylococcus species were found to be the most sensitive strains.

Conclusions

The antibacterial activities of the DCM: MeOH extract and isolated compounds support the traditional use of T. minus in the treatment of bovine mastitis.

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Graphical abstract

Chemical compounds studied in the article

• Thalrugosaminine (PubChem CID: 321938);
• O-Methylthalicberine (PubChem CID: 100230)

Keywords

• Bovine mastitis;
• Thalictrum minus;
• 5’-Hydroxythalidasine;
• Thalrugosaminine;
• antibacterial activity;
• minimum inhibitory concentration (MIC)

Corresponding authors. Cytogenetics and Molecular Biology Research Laboratory, Centre of Research for Development (CORD), University of Kashmir, Hazratbal–190006, Srinagar, Jammu & Kashmir, India. Tel.: +91 9906717120; fax: +91 194 227 2096.

Corresponding authors. Biotechnology Division, CSIR - Indian Institute of Integrative Medicine, Sanatnagar-190005, Srinagar, Jammu & Kashmir, India. Tel.: +0194 2431253, 0194 2431255; fax: +91 01942441331.

© 2016 Published by Elsevier Ireland Ltd.