HPLC method validation for measurement of sulforaphane level in broccoli by-products.
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 x 0.46 cm, 5 microm; column temperature, 36 degrees 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.
Campas-Baypoli
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ON
Departamento de Biotecnología y Ciencias Alimentarias, Instituto Tecnológico de Sonora, 5 de febrero 818 Sur, Cd. Obregón, Sonora, México.
Sánchez-Machado
D I
DI
Bueno-Solano
C
C
Ramírez-Wong
B
B
López-Cervantes
J
J
D001937:Brassica / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005961:Glucosinolates / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007096:Imidoesters / Q000737:chemistry; D017879:Isothiocyanates; D010091:Oximes; D015203:Reproducibility of Results; D052616:Solid Phase Extraction; D013454:Sulfoxides; D013861:Thiocyanates / Q000032:analysis* / Q000737:chemistry
D005961:Glucosinolates; D007096:Imidoesters; D017879:Isothiocyanates; D010091:Oximes; D013454:Sulfoxides; D013861:Thiocyanates; C016766:sulforaphane; C119494:glucoraphanin