• Scopus
  • CA
  • DOAJ
  • FSTA
  • JST
  • 北大核心期刊
  • 中国科技核心期刊CSTPCD
  • 中国精品科技期刊
  • RCCSE中国核心学术期刊
  • 中国农业核心期刊
  • 中国生物医学文献服务系统SinoMed收录期刊
中国精品科技期刊2020

超高效液相色谱-荧光法测定蜂蜜中百里香酚的残留量

钱振杰 曾羲 陈敬 赵悦 游淑珠 何咏欣 冯家望

钱振杰,曾羲,陈敬,等. 超高效液相色谱-荧光法测定蜂蜜中百里香酚的残留量[J]. 食品工业科技,2022,43(23):328−333. doi:  10.13386/j.issn1002-0306.2022070302
引用本文: 钱振杰,曾羲,陈敬,等. 超高效液相色谱-荧光法测定蜂蜜中百里香酚的残留量[J]. 食品工业科技,2022,43(23):328−333. doi:  10.13386/j.issn1002-0306.2022070302
QIAN Zhenjie, ZENG Xi, CHEN Jing, et al. Determination of Thymol in Honey by Ultra Performance Liquid Chromatography with Fluorescence Detector[J]. Science and Technology of Food Industry, 2022, 43(23): 328−333. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2022070302
Citation: QIAN Zhenjie, ZENG Xi, CHEN Jing, et al. Determination of Thymol in Honey by Ultra Performance Liquid Chromatography with Fluorescence Detector[J]. Science and Technology of Food Industry, 2022, 43(23): 328−333. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2022070302

超高效液相色谱-荧光法测定蜂蜜中百里香酚的残留量

doi: 10.13386/j.issn1002-0306.2022070302
基金项目: 珠海市重点科技项目(ZH22036201210211PWC);海关总署标准制修订计划(2020B166);广东省药品监督管理局科技创新团队项目(2019TDB04)。
详细信息
    作者简介:

    钱振杰(1979−),男,硕士,高级工程师,研究方向:食品质量安全技术研究,E-mail:qianzhenjie@hotmail.com

    通讯作者:

    曾羲(1991−),女,硕士,工程师,研究方向:食品分析检测,E-mail:lovelyzeng@126.com

  • 中图分类号: O657.7+2

Determination of Thymol in Honey by Ultra Performance Liquid Chromatography with Fluorescence Detector

  • 摘要: 建立了超高效液相色谱-荧光检测法测定蜂蜜中百里香的残留量的检验方法。用40%乙腈-水溶液提取蜂蜜中的百里香酚,经离心、上清液过滤后,以0.1%甲酸水溶液-乙腈为流动相进行梯度洗脱,采用Accucore aQ色谱柱(150 mm×2.1 mm,2.6 μm)分离,荧光检测器检测(激发波长274 nm,发射波长297 nm),外标法定量。结果表明,百里香酚在0.01~3.00 μg/mL范围内线性关系良好,决定系数(R2)为0.9999,在蜂蜜基质中的平均加标回收率在87.4%~106.9%之间,变异系数在0.7%~7.3%之间,定量限为0.10 mg/kg。利用该方法测定了48份市售蜂蜜样品,仅有1份样品存在百里香酚残留。本研究建立的方法简单、迅速、可靠,适用于蜂蜜中百里香酚残留的批量筛查和定量测定。
  • 图  1  不同提取溶剂的回收率(n=3)

    Figure  1.  Recoveries of different solvents for thymol (n=3)

    图  2  不同净化方式的回收率(n=3)

    Figure  2.  Recoveries of different purification methods for thymol (n=3)

    图  3  4种色谱柱对酚类混合物(1.0 µg/mL)的分离色谱图

    Figure  3.  Chromatograms of 4 kinds of chromatographic column for mixed phenolic compounds at 1.0 µg/mL

    注:峰1:丹皮酚;峰2:丁香酚;峰3:异丁香酚;峰4:香芹酚;峰5:百里香酚;图4同。

    图  4  4种流动相对酚类混合物(1.0 µg/mL)的分离色谱图

    Figure  4.  Chromatograms of 4 kinds of mobile phase for mixed phenolic compounds at 1.0 µg/mL

    图  5  不同甲酸浓度下百里香酚(1.0 µg/mL)峰面积和保留时间的变化

    Figure  5.  Changes in peak area and retention time of thymol (1.0 µg/mL) at different concentrations of formic acid

    图  6  不同比例乙腈下香芹酚和百里香酚的分离度

    Figure  6.  Resolution of carvacrol and thymol in the presence of different concentrations of acetonitrile

    图  7  百里香酚的紫外吸收光谱

    Figure  7.  UV absorption spectrum of thymol

    图  8  百里香酚的荧光激发-发射光谱

    Figure  8.  Fluorescence excitation-emission spectrum of thymol

    图  9  阳性样品的色谱图

    Figure  9.  Chromatogram of positive sample

    表  1  百里香酚的线性范围、线性回归方程、决定系数、检出限和定量限

    Table  1.   Linear range, linear equation, correlation coefficients, limit of detection (LOD) and limit of quantitation (LOQ) of thymol

    线性范围
    (µg/mL)
    线性方程决定系数
    (R2)
    检出限
    (mg/kg)
    定量限
    (mg/kg)
    0.01~3.00y=1.537×107x−31960.99990.030.10
    下载: 导出CSV

    表  2  百里香酚的加标回收率和精密度(n=6)

    Table  2.   Spiked recoveries and relative standard deviations (RSDs) of thymol (n=6)

    基质种类添加浓度(mg/kg)平均加标回收率(%)精密度(%)
    雪脂莲蜜0.198.15.7
    0.898.33.2
    3096.90.8
    椴树蜜0.189.54.3
    0.8100.32.1
    3087.40.9
    玄参蜜0.1106.96.0
    0.896.42.1
    3091.11.8
    洋槐蜜0.195.25.8
    0.8103.21.3
    3089.50.8
    百花蜜0.198.87.3
    0.896.81.5
    3096.70.7
    下载: 导出CSV
  • [1] 刘欢, 赵巨堂, 罗海涛, 等. 提取方法对百里香精油化学成分和抗氧化活性的影响[J]. 食品工业科技,2022,43(19):333−341. [LIU H, ZHAO J T, LUO H T, et al. Effect of different extraction methods on chemical constituents and antioxidant activity of essential oil from Thymus vulgaris L

    J]. Science and Technology of Food Industry,2022,43(19):333−341.
    [2] 李旭领, 郭月婷, 王一佳, 等. 药用植物挥发油的抗菌作用及机制研究进展[J]. 动物医学进展,2022,43(5):97−101. [LI X L, GUO Y T, WANG Y J, et al. Progress on antimicrobial effects and mechanisms of volatile oils from medicinal plants[J]. Progress in Veterinary Medicine,2022,43(5):97−101. doi:  10.3969/j.issn.1007-5038.2022.05.019
    [3] 崔醒, 朱秋劲, 侯瑞, 等. 丁香酚、香芹酚和百里香酚对禾谷镰刀菌的抑菌活性及机制[J/OL]. 食品科学: 1−14 [2022-07-24]. http://kns.cnki.net/kcms/detail/11.2206.TS.20220414.1359.094.html

    CUI X, ZHU J Q, HOU R, et al. Antibacterial activity and mechanism of eugenol,carvacrol and thymol against Fusarium graminearum[J/OL]. Food Science:1−14. [2022-07-24]. http://kns.cnki.net/kcms/detail/11.2206.TS.
    [4] 靳盼盼, 刘亚文, 刘旖旎, 等. 百里香酚对食源阴沟肠杆菌生物膜形成的抑制作用[J]. 食品工业科技,2019,40(22):104−109, 114. [JIN P P, LIU Y W, LIU Y N, et al. Inhibition of biofilm formation of Foodborne-enterobacter cloacae by thymol[J]. Science and Technology of Food Industry,2019,40(22):104−109, 114.
    [5] KACHUR K, SUNTRES Z. The antibacterial properties of phenolic isomers, carvacrol and thymol[J]. Critical Reviews in Food Science and Nutrition,2020,60(18):3042−3053. doi:  10.1080/10408398.2019.1675585
    [6] 胡福良, 朱威, 李英华. 香精油的抗蜂螨作用及其在蜂群中的应用[J]. 昆虫知识,2005(4):375−378. [HU F L, ZHU W, LI Y H. Use of essential oils for controlling Varroa destructor in honey bee colonies[J]. Entomological Knowledge,2005(4):375−378.
    [7] REGULATION H. Council Regulation (EEC) No 2377/90 of 26 June 1990 laying down a community procedure for the establishment of maximum residue limits of veterinary medicinal products in foodstuffs of animal origin[J]. Official JL,1990,224:0001−0008.
    [8] 日本厚生劳动省. 生食発0822第5号[S]. 食药事务和卫生委员会, 平成30年8月22日

    Ministry of Health, Labor and Welfare of Japan, Life Hygiene and Food Safety Regulations 0822 No. 5 [S]. Pharmaceutical Affairs and Food Sanitation Council, August 22, 2018.
    [9] BOGDANOV S, KILCHENMANN V, FLURI P, et al. Influence of organic acids and components of essential oils on honey taste[J]. Am Bee J,1999,139(1):61−63.
    [10] BOGDANOV S, KILCHENMANN V, IMDORF A, et al. Residues in honey after application of thymol against Varroa using the Frakno Thymol Frame[J]. Am Bee J,1998,138(8):610−611.
    [11] 李瑞珍, 方兵兵, 刘世丽. 世界蜂蜜生产、贸易与消费发展态势分析[J]. 中国食物与营养,2021,27(8):15−20. [LI R Z, FANG B B, LIU S L. Analysis on the development trend of world honey production, trade and consumption[J]. Food and Nutrition in China,2021,27(8):15−20. doi:  10.3969/j.issn.1006-9577.2021.08.003
    [12] TONELLO N, MORESSI M B, ROBLEDO S N, et al. Square wave voltammetry with multivariate calibration tools for determination of eugenol, carvacrol and thymol in honey[J]. Talanta,2016,158:306−314. doi:  10.1016/j.talanta.2016.05.071
    [13] AGHAMOHSENI B, HASSANINEJAD-DARZI S K, ASADOLLAHI-BABOLI M. A new sensitive voltammetric determination of thymol based on MnY nanozeolite modified carbon paste electrode using response surface methodology[J]. Microchemical Journal,2019,145:819−832. doi:  10.1016/j.microc.2018.11.045
    [14] ZIYATDINOVA G, ZIGANSHINA E, CONG P N, et al. Voltammetric determination of thymol in oregano using CeO2-modified electrode in Brij® 35 micellar medium[J]. Food Analytical Methods,2017,10(1):129−136. doi:  10.1007/s12161-016-0562-y
    [15] 董茂锋, 韩薇, 白冰, 等. 气相色谱法测定饲料香味剂中香芹酚和百里香酚的含量[J]. 上海农业学报,2015,31(3):79−81. [DONG M F, HAN W, BAI B, et al. Determination of carvacrol and thymol contents in feed aromatizer by gas chromatography[J]. Acta Agriculturae Shanghai,2015,31(3):79−81.
    [16] 宫玲玲, 李会荣. 气相色谱法同步测定饲料添加剂中百里香酚、香芹酚的研究[J]. 中国饲料,2014(2):38−40. [GONG L L, LI H R. Determination of thymol and carvacrol in feeds by gas chromatograpahy[J]. China Feed,2014(2):38−40. doi:  10.3969/j.issn.1004-3314.2014.02.021
    [17] 罗成江, 周炜, 侯轩, 等. 气相色谱法测定香味剂中桉油精肉桂醛百里香酚及香芹酚的含量[J]. 中国兽医杂志,2019,55(2):113−115. [LUO C J, ZHOU W, HOU X, et al. Determination of eucalyptol, cinnamaldehyde, thymol and carvacrol contents in fodder flavouring matter by gas chromatography[J]. Chinese Journal of Veterinary Medicine,2019,55(2):113−115.
    [18] ANTIH J, HOUDKOVA M, URBANOVA K, et al. Antibacterial activity of Thymus vulgaris L. essential oil vapours and their GC/MS analysis using solid-phase microextraction and syringe headspace sampling techniques[J]. Molecules,2021,26(21):6553. doi:  10.3390/molecules26216553
    [19] 王晓峰. 高效液相色谱法测定食品包装薄膜中的香芹酚和百里香酚[J]. 食品安全导刊,2021(24):140−141. [WANG X F. High performance liquid chromatography method for rapid detection of carvacrol and thymol in active food packaging film[J]. China Food Safety Magazine,2021(24):140−141.
    [20] 张玮, 李会荣, 宫玲玲. 高效液相色谱法测定混合型饲料添加剂中肉桂醛、丁香酚、香芹酚和百里香酚含量[J]. 饲料研究,2020,43(2):64−69. [ZHANG W, LI H R, GONG L L. Study on the determination of cinnamaldehyde, eugenol, carvacrol and thymol in mixed feed additives by liquid chromatograph[J]. Feed Research,2020,43(2):64−69.
    [21] 窦茂鑫, 赵迪, 侯永清, 等. 饲料添加剂百里香油中百里香酚和香芹酚含量的测定[J]. 中国饲料,2013(7):39−40. [DOU M X, ZHAO D, HOU Y Q, et al. Determination of the content of thymol and carvacrol in feed additives[J]. China Feed,2013(7):39−40.
    [22] TSIGOURI A, PASSALOGLOU-KATRALI M, SABATAKOU O. Determination of eucalyptol camphor menthol and thymol in greek thyme honey by GC-FID[J]. Acta Alimentaria,2008,37(2): 181−189. doi:  10.1556/AAlim.2007.0032
    [23] NOZAL M J, BERNAL J L, JIMENEZ J J, et al. Extraction of thymol, eucalyptol, menthol, and camphor residues from honey and beeswax: Determination by gas chromatography with flame ionization detection[J]. Journal of Chromatography A,2002,954(1−2):207−215. doi:  10.1016/S0021-9673(02)00153-X
    [24] ARES A M, NOZAL M J, BERNAL J L, et al. Simultaneous determination of carvacrol and thymol in bee pollen by using a simple and efficient solvent extraction method and gas chromatography-mass spectrometry[J]. Journal of Pharmaceutical and Biomedical Analysis,2020,181:113124. doi:  10.1016/j.jpba.2020.113124
    [25] BADERTSCHER R, KILCHENMANN V, LINIGER A, et al. Determination of 1,4-dichlorobenzene, naphthalene and thymol residues in honey using static headspace coupled with GC-MS[J]. Journal of Api Product and Api Medical Science,2010,2:87−92. doi:  10.3896/IBRA.4.02.3.01
    [26] 中华人民共和国卫生部. GB 14963-2011 食品安全国家标准 蜂蜜[S]. 北京: 中国标准出版社

    Ministry of Health of the People's Republic of China. GB 14963-2011 National food safety standard honey[S]. Beijing: China Standard Press.
    [27] MARTEL A C, ZEGGANE S. Determination of acaricides in honey by high-performance liquid chromatography with photodiode array detection[J]. Journal of Chromatography A,2002,954(1−2):173−180. doi:  10.1016/S0021-9673(02)00126-7
    [28] SÁNCHEZ L M, RAMOS M J G, DEL MAR GÓMEZ-RAMOS M, et al. Presence, persistence and distribution of thymol in honeybees and beehive compartments by high resolution mass spectrometry[J]. Environmental Advances,2021,5:100085. doi:  10.1016/j.envadv.2021.100085
    [29] GHOBADLOO P A, HAMIDI S, NEMATI M, et al. Ultrasound assisted dispersive solid phase microextraction of thymol and carvacrol in pharmaceutical products using graphene oxide as an adsorbent prior to analysis by high performance liquid chromatography[J]. Current Pharmaceutical Analysis,2020,16(5):578−584. doi:  10.2174/1573412915666190220102628
    [30] 黄学者, 贾光群, 魏红跃, 等. 超高效液相色谱-紫外/荧光法检测蜂蜜中百里香酚含量[J]. 食品安全质量检测学报,2020,11(20):7220−7226. [HUANG X Z, JIA G Q, WEI H Y, et al. Determination of thymol content in honey by ultra performance liquid chromatography-ultraviolet/fluorescence method[J]. Journal of Food Safety & Quality,2020,11(20):7220−7226.
    [31] 张晓华, 吕敏明, 郑晶晶, 等. 化学计量学二阶校正方法结合高效液相色谱用于蜂蜜中10种酚酸类物质的快速定量分析[J]. 色谱,2020,38(5):572−580. [ZHANG X H, LÜ M M, ZHENG J J, et al. Fast high-performance liquid chromatography quantification of ten phenolic acids in honey using chemometric second-order calibration method[J]. Chinese Journal of Chromatography,2020,38(5):572−580.
    [32] 吴升德, 谷群远, 沈校校, 等. 蜂蜜中农药残留检测前处理和分析方法研究进展[J]. 中国蜂业,2021,72(12):58−60. [WU D S, GU Y Q, SHEN X X, et al. Research progress of pretreatment and analytical methods on pesticide residues in honey[J]. Apiculture of China,2021,72(12):58−60. doi:  10.3969/j.issn.0412-4367.2021.12.032
    [33] 胡福良, 朱威, 李英华. 香精油抗蜂螨作用的研究进展[J]. 养蜂科技,2004(5):21−23, 25. [HU F L, ZHU W, LI Y H. The recent advances in essential oils for the control of Varroa jacobsoni[J]. Apicultural Science and Technology,2004(5):21−23, 25.
  • 加载中
图(9) / 表(2)
计量
  • 文章访问数:  63
  • HTML全文浏览量:  7
  • PDF下载量:  12
  • 被引次数: 0
出版历程
  • 收稿日期:  2022-08-01
  • 网络出版日期:  2022-10-21
  • 刊出日期:  2022-11-23

目录

    /

    返回文章
    返回

    重要通知

    1、快速见刊:客座主编专栏征稿-食源性功能物质挖掘及评价
           2、喜讯 :《食品工业科技》被DOAJ数据库收录!
           3喜报:《食品工业科技》世界期刊影响力稳居Q2区
           4、祝贺:《食品工业科技》中国期刊影响力稳居Q1第二名