TY - JOUR
T1 - HPLC method validation for measurement of sulforaphane level in broccoli by-products
AU - Campas-Baypoli, O. N.
AU - Sánchez-Machado, D. I.
AU - Bueno-Solano, C.
AU - Ramírez-Wong, B.
AU - López-Cervantes, J.
PY - 2010/4
Y1 - 2010/4
N2 - 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 C18, 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 (r2 = 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.
AB - 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 C18, 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 (r2 = 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.
KW - Broccoli
KW - By-products
KW - HPLC
KW - MSR
KW - SPE
KW - Sulforaphane
UR - http://www.scopus.com/inward/record.url?scp=77950238472&partnerID=8YFLogxK
U2 - 10.1002/bmc.1303
DO - 10.1002/bmc.1303
M3 - Artículo
SN - 0269-3879
VL - 24
SP - 387
EP - 392
JO - Biomedical Chromatography
JF - Biomedical Chromatography
IS - 4
ER -