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  • Phenolic compounds involved in grafting incompatibility of Vitis spp: development and validation of an analytical method for their quantification.

Phenolic compounds involved in grafting incompatibility of Vitis spp: development and validation of an analytical method for their quantification.

Phytochemical analysis : PCA (2014-06-04)
Sara Canas, Maria Assunção, João Brazão, Geni Zanol, José Eduardo Eiras-Dias
ABSTRACT

Graft incompatibility of Vitis spp is an unresolved worldwide problem with important economic consequences. Grafting comprises a complex set of morphological and physiological alterations, in which the phenolic compounds seem to be strongly involved. Therefore, a detailed analysis and recognition of structural phenolic compounds diversity in the two partners of a Vitis graft is of great importance to evaluate their role as markers of graft establishment. To optimise a sample extraction method, and to develop and validate a high-performance liquid chromatography (HPLC) method for the simultaneous determination of phenolic acids and flavonols in the graft union so as to understand their behaviour in the metabolism of the scion-rootstock system, using compatible and incompatible combinations of a Syrah cultivar and two rootstocks (R110 and SO4). Sixty extracts of Vitis grafting tissues were prepared and analysed by HPLC for the qualitative and quantitative determination of their phenolic profile. Among the phenolic compounds identified in the samples, one benzoic acid (gallic acid), three cinnamic acids (caffeic acid, ferulic acid and sinapic acid) and two flavonols (catechin and epicatechin) are potentially suitable as markers of graft incompatibility. The method developed presents good performance and lends itself readily for application in routine analysis of the phenolic composition of Vitis grafting tissues to distinguish compatible and incompatible combinations in the graft callusing stage.

MATERIALS
Product Number
Brand
Product Description

Supelco
(+)-Catechin, analytical standard
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Methanol solution, contains 0.10 % (v/v) formic acid, UHPLC, suitable for mass spectrometry (MS), ≥99.5%
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Methanol solution, NMR reference standard, 4% in methanol-d4 (99.8 atom % D), NMR tube size 3 mm × 8 in.
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Methanol-12C, 99.95 atom % 12C
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trans-Ferulic acid, ≥99%
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trans-Sinapic acid, analytical standard
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Caffeic acid, matrix substance for MALDI-MS, ≥99.0% (HPLC)
Sigma-Aldrich
trans-Ferulic acid, 99%
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Caffeic acid, ≥98.0% (HPLC)
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Triton X-100, laboratory grade
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β-D-Allose, rare aldohexose sugar
USP
Methyl alcohol, United States Pharmacopeia (USP) Reference Standard
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trans-Ferulic acid, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Ferulic acid, European Pharmacopoeia (EP) Reference Standard
Supelco
trans-Ferulic acid, matrix substance for MALDI-MS, ≥99.0% (HPLC)
Supelco
Methanol, analytical standard
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Sinapic acid, matrix substance for MALDI-MS, ≥99.0% (T)
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Acetone, ≥99%, meets FCC analytical specifications
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Acetone, natural, ≥97%
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Gallic acid monohydrate, ACS reagent, ≥98.0%
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Methanol, suitable for HPLC, gradient grade, 99.93%
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Acetone, suitable for HPLC, ≥99.9%
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Methanol, anhydrous, 99.8%
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Sinapic acid, ≥98%, powder
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Chlorogenic acid, ≥95% (titration)
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Acetone, for residue analysis, ≥99.5%
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Acetone, SAJ first grade, ≥99.0%
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Methanol, SAJ special grade
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Acetone, for residue analysis, JIS 5000
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Acetone, analytical standard