A high‐throughput monolithic HPLC method for rapid Vitamin C phenotyping of berry fruit

Phytochemical Analysis - Tập 17 Số 5 - Trang 284-290 - 2006
Paul G. Walker1, Sandra Gordon1, Rex Brennan1, Robert D. Hancock1
1Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, UK

Tóm tắt

AbstractA rapid method for the quantification of łascorbic acid (1) in berry fruit by HPLC with photodiode array detection is presented. łAscorbic acid was resolved on a C18 monolithic column with aqueous buffer, after which the column was washed with acetonitrile to remove lipophilic compounds prior to re‐equilibration for analysis of the next sample. Using the monolithic column format with high mobile phase flow rates, the entire separation, wash and re‐equilibration were achieved in 3 min. With the exception of gooseberry (Ribes uva‐crispa), for which an interfering compound co‐eluted, concentrations of 1 could be determined in a wide range of berry fruits after extraction in metaphosphoric acid without further sample preparation. Using this extraction method, recoveries of 1 in excess of 85% were achieved. Fruit or juice extracts were stable in 5% metaphosphoric acid for at least 4 h and stability could be extended to longer than 150 h by the addition of the reducing agent tris(2‐carboxethyl)phosphine hydrochloride. Following validation, the method was utilised for the phenotyping of fruit in a Scottish Crop Research Institute (SCRI) Ribes nigrum L. breeding population of 300 individuals. An improved extraction method allowed extraction, quantification of 1 and data analysis to be undertaken in less than one working week. Copyright © 2006 John Wiley & Sons, Ltd.

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Phytochemical Analysis

Tập 17 Số 5

284-290

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