HPLC method validation for measurement of sulforaphane level in broccoli by-products

O. N. Campas-Baypoli, D. I. Sánchez-Machado, C. Bueno-Solano, B. Ramírez-Wong, J. López-Cervantes*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

66 Scopus citations


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.

Original languageEnglish
Pages (from-to)387-392
Number of pages6
JournalBiomedical Chromatography
Issue number4
StatePublished - Apr 2010


  • Broccoli
  • By-products
  • HPLC
  • MSR
  • SPE
  • Sulforaphane


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