Premium
HPLC method validation for measurement of sulforaphane level in broccoli by‐products
Author(s) -
CampasBaypoli O. N.,
SánchezMachado D. I.,
BuenoSolano C.,
RamírezWong B.,
LópezCervantes J.
Publication year - 2010
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1303
Subject(s) - sulforaphane , chemistry , chromatography , glucoraphanin , repeatability , high performance liquid chromatography , extraction (chemistry) , acetonitrile , relative standard deviation , detection limit , glucosinolate , brassica , botany , biochemistry , biology
A simple and specific analytical method was developed and tested for the determination of sulforaphane in broccoli by‐products. The method includes the optimization of the conversion of glucoraphanin to sulforaphane, followed by purification of extracts using solid‐phase extraction and high‐performance liquid chromatography (HPLC) analysis. The response surface methodology was used to find optimum conditions for the preparation and purification procedure. Chromatographic conditions for reversed‐phase HPLC with UV photodiode array detection were as follows: column, Exil ODS C 18 , 25 × 0.46 cm, 5 μm; column temperature, 36°C; mobile phase, a 30 : 70 (v/v) mixture of acetonitrile:water; flow rate, 0.6 mL/min. The detection wavelength was UV 202 nm. Under these conditions, excellent linearity was obtained ( r 2 = 1), and the overall recovery was 97.5 and 98.1% for fresh florets and lyophilized florets, respectively. The precision results showed that the relative standard deviation of the repeatability for florets fresh and lyophilized was 3.0 and 4.0%, respectively. Sulforaphane contents were determined in the edible portion of fresh broccoli, and broccoli crop remains. Copyright © 2009 John Wiley & Sons, Ltd.