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A liquid chromatography-electrospray ionization-mass spectrometry method for quantification of cyclotides in plants avoiding sorption during sample preparation
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Division of Pharmacognosy.
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2011 (English)In: Journal of Chromatography A, ISSN 0021-9673, E-ISSN 1873-3778, Vol. 1218, no 44, 7964-7970 p.Article in journal (Refereed) Published
Abstract [en]

Cyclotides are plant-produced, bioactive, cyclic mini-proteins with interesting pharmaceutical and agricultural applications. A reverse phase liquid chromatography electrospray ionization mass spectrometry (RP-LC-ESI-MS) method for analysis of cyclotides in plant materials with a minimum of sample pretreatment is presented. Three exemplary cyclotides (kalata B1, kalata B2 and cycloviolacin 02) were used as reference substances for the method development. Linearity (r(2) > 0.99) was achieved in the concentration range 0.05-10 mg/L and the limit of detection was 1.7-4.0 mu g/L. The present study is the first to demonstrate that cyclotides dissolved in water sorb to glass vials, but the addition of 15% of acetonitrile or 40 mg/L of bovine serum albumin is sufficient to keep the cyclotides in solution. Cyclotides were extracted from candied violets, violet tea, and the plants Oldenlandia affinis and Viola odorata using 70% methanol containing 0.1% formic acid (v/v). The plant content was determined to be 23.5-14,200 mu g/g (dry weight). The highest content of cyclotide was found in wild Danish V. odorata, and it is the highest content of cyclotide in a plant reported hitherto. Candied violets contained 0.00-8.66 mu g/g (dry weight), while no cyclotides were detected in commercial violet tea.

Place, publisher, year, edition, pages
2011. Vol. 1218, no 44, 7964-7970 p.
Keyword [en]
Cyclotides, LC-MS, Quantification, Sorption, Adducts, Proteins
National Category
Chemical Sciences
Identifiers
URN: urn:nbn:se:uu:diva-162450DOI: 10.1016/j.chroma.2011.08.095ISI: 000296549000008OAI: oai:DiVA.org:uu-162450DiVA: diva2:460807
Available from: 2011-12-01 Created: 2011-11-30 Last updated: 2017-12-08Bibliographically approved

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Göransson, Ulf

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