Determination of sulforaphane in cruciferous vegetables by HPLC
KONG Fanhua1, YANG Chunxue1, FANG Congrong2, QIU Nannan2, XU Jiajia1, BAI Shasha1, LI Dong1, CUI Yajuan1*
1(Beijing Institute of Nutrition Resources, Beijing 100069, China) 2 (NHC Key Laboratory of Food Safety Risk Assessment, Food Safety Research Unit of Chinese Academy of Medical Science, China National Center for Food Safety Risk Assessment, Beijing 100021, China)
Abstract: In this study, an analytical method was developed for determining sulforaphane in cruciferous vegetables. The extraction and enzymolysis conditions of sulforaphane in broccoli were selected by a single factor experiment. The conditions that pH=3.0 hydrochloric acid solution hydrolyzed at 25 ℃ for 0.5 h, extracted with ethyl acetate were used for the pretreatment. The separation of sulforaphane was achieved on C18 column(150 mm×4.6 mm, 5μm)using acetonitrile-water(20∶80,V/V)as the mobile phase at a flow rate of 1.0 mL/min. The detection wavelength and column temperature were 201 nm and 30 ℃, respectively. The results showed that the linear range of sulforaphane was 2.00-100.00 μg/mL. with a relative standard deviation (RSD) of sulforaphane in broccoli and turnip less than 5%. The average recovery rate of broccoli and turnip was 100.09% and 96.16%, and RSD was less than 5%. Therefore, this developed method is high reproducibility and accuracy. It can be applied to accurately determine sulforaphane in cruciferous vegetables.
RONG Y, HUANG L X, YI K Z, et al.Co-administration of sulforaphane and doxorubicin attenuates breast cancer growth by preventing the accumulation of myeloid-derived suppressor cells[J].Cancer Letters, 2020, 493:189-196.
[2]
HUANG B, LEI S X, WANG D, et al.Sulforaphane exerts anticancer effects on human liver cancer cells via induction of apoptosis and inhibition of migration and invasion by targeting MAPK7 signalling pathway[J].Journal of B.U.ON.:Official Journal of the Balkan Union of Oncology, 2020, 25(2):959-964.
[3]
韩玉芳, 赵延兵, 李俊, 等.萝卜硫素诱导肝癌Huh-6细胞增殖和凋亡[J].河南科技大学学报(医学版), 2017, 35(4):256-259. HAN Y F, ZHAO Y B, LI J, et al.Proliferation and apoptosis induced by sulforaphane in cultured human hepatoblastoma Huh-6 Cell[J].Henan Univ Sci Tech(Med Sci), 2017, 35(4):256-259.
[4]
GE M, ZHANG L, CAO L, et al.Sulforaphane inhibits gastric cancer stem cells via suppressing sonic hedgehog pathway[J].International Journal of Food Sciences & Nutrition, 2019, 70(5):1-9.
[5]
ZHENG Z, LIN K, HU Y, et al.Sulforaphane metabolites inhibit migration and invasion via microtubule-mediated Claudins dysfunction or inhibition of autolysosome formation in human non-small cell lung cancer cells[J].Cell Death and Disease, 2019, 10(4):1 755-1 433.
[6]
LUO L, ZHAO H, LUO Q.Swerchirin exerts anticancer activity on SKOV3 human ovarian cancer cells via induction of mitochondrial apoptosis, G2/M cell cycle arrest and inhibition of Raf/MEK/ERK cascade[J].Journal of Buon, 2018, 23(1):111-116.
[7]
张亚芳, 赵晓红, 王德珠, 等.萝卜硫素对宫颈癌HeLa细胞凋亡及放射敏感性的影响[J].中国药业, 2020, 29(15):22-26. ZHANG Y F,ZHAO X H,WANG D Z,et al.Effect of sulforaphane on apoptosis and radiosensitivity of hela cells of cervical cancer[J].China Pharmaceuticals, 2020, 29(15):22-26.
[8]
MA T, ZHENG Z, GUO H, et al.4-O-methylhonokiol ameliorates type 2 diabetes-induced nephropathy in mice likely by activation of AMPK-mediated fatty acid oxidation and Nrf2-mediated anti-oxidative stress[J].Toxicology and Applied Pharmacology, 2019, 370:93-105.
[9]
LI Y Y, LIU Y T, YAN X, et al.Pharmacological effects and mechanisms of chinese medicines modulating NLRP3 inflammasomes in ischemic cardio/cerebral vascular disease[J].The American Journal of Chinese Medicine, 2018, 46(8):1 727-1 741.
[10]
SIVAKUMAR G, ALIBONI A, BACCHETTA L.HPLC screening of anti-cancer sulforaphane from important European Brassica species[J].Food Chemistry, 2007, 104(4):1 761-1 764.
[11]
RENATO I, PATRICK R.HPLC screening of anti-cancer sulforaphane from important European Brassica species[J].Food Chemistry, 2008,108(1):407-408.
[12]
NATHAN V M, JOHN A J, ELIZABETH H J.Heating decreases epithiospecifier protein activity and increases sulforaphane formation in broccoli[J].Phytochemistry, 2004, 65(9):1 273-1 281.
[13]
ZHANG Z S, WANG L J, LI D, et al.Ultrasound-assisted extraction of oil from flaxseed[J].Separation and Purification Technology, 2008, 62(1):192-198.
[14]
VANDUCHOVA A, TOMANKOVA V, ANZENBACHER P, et al.Influence of sulforaphane metabolites on activities of human drug-metabolizing cytochrome P450 and determination of sulforaphane in human liver cells[J].Journal of Medicinal Food, 2016, 19(12):1 141-1 146.
[15]
KOKOTOU M G, REVELOU P K, PAPPAS C, et al.High resolution mass spectrometry studies of sulforaphane and indole-3-carbinol in broccoli[J].Food Chemistry, 2017, 237:566-573.
[16]
CHAU O,FAWZY E.A new validated HPLC method for the determination of sulforaphane:application to study pharmacokinetics of sulforaphane in rats[J].Biomedical Chromatography, 2016, 30(7):1 016-1 021.
[17]
何珺, 谢述琼, 杨佳年, 等.12种十字花科蔬菜种子中萝卜硫素含量研究[J].食品研究与开发, 2015, 36(4):11-13. HE J, XIE S Q, YANG JN, et al.Determining the sulforaphane content of twelve cruciferous plants'seeds[J].Food Research And Development, 2015, 36(4):11-13.
[18]
董俊杰, 施琪浩, 张英杰, 等.拓展实验萝卜硫素质谱分析与指认[J].当代化工, 2020, 49(8):1 643-1 646;1 663. DONG J J, SHI Q H, ZHANG Y J, et al.Mass spectroscopic analysis and identification of sulforaphane for materialsand pharmaceutical professional expansion experiments[J].Contemporary Chemical Industry, 2020, 49(8):1 643-1 646;1 663.
[19]
赵登奇, 孙亚天, 黄建颖, 等.西兰花叶中生物活性成分的测定[J].核农学报, 2020, 34(6):1 266-1 271. ZHAO D Q, SUN Y T, HUANG J Y, et al.Determination of bioactive components in broccoli leaves[J].Journal of Nuclear Agricultural Sciences, 2020, 34(6):1 266-1 271.
[20]
中华人民共和国国家质量监督检验检疫总局, 中国国家标准化管理委员会.GB/T 27404—2008食品安全国家标准 实验室质量控制规范 食品理化检测[S].北京:中国标准出版社, 2008. General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China.GB/T 27404—2008 Standardization Administration.Criterion on quality control of laboratories-chemical testing of food[S].Beijing:Standards Press of China, 2008.
[21]
BERTELLI D, PLESSI M, BRGHIROLI D, et al.Separation by solid phase extraction and quantification by reverse phase HPLC of sulforaphane in broccoli[J].Food Chemistry, 1998, 63(3):417-421.
[22]
张品南, 马绍英, 杨海荣, 等.西兰花种子及其愈伤组织中萝卜硫素含量比较[J].天然产物研究与开发, 2013, 25(6):796-798;811. ZHANG P N, MA SY, YANG H R, et al.Comparison of sulforaphen content in seeds and callus induced from different organs of broccoli species[J].Nat Prod Res Dev, 2013, 25(6):796-798;811.
2021, Vol. 47 
