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

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

钱振杰^1,,
曾羲^1, ,,
陈敬²,
赵悦²,
游淑珠²,
何咏欣¹,
冯家望²

1.
广州市食品检验所，广东广州 511400
2.
拱北海关技术中心，广东珠海 519000

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
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- 文章访问数: 135
- HTML全文浏览量: 15
- PDF下载量: 16
- 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

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

1.
Guangzhou Institute for Food Inspection, Guangzhou 511400, China
2.
Gongbei Customs Technology Center, Zhuhai 519000, China

摘要

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。
- 蜂蜜 /
- 百里香酚 /
- 超高效液相色谱 /
- 荧光法 /
- 残留量
Abstract: The presented method was developed to determine the concentration of thymol residues in honey by ultra performance liquid chromatography with a fluorescence detector. The thymol in honey was extracted by 40% acetonitrile-water solution. After centrifugation and filtration, the separation of thymol was performed on an Accucore aQ liquid chromatography column (150 mm×2.1 mm, 2.6 μm) with the gradient elution of acetonitrile and water (containing 0.1% formic acid) as mobile phases, and quantitated by external standard method with fluoresce detector (excitation wavelength 274 nm, emission wavelength 297 nm). A good linear relationship (R²=0.9999) was observed with concentration ranging from 0.01 μg/mL to 3.00 μg/mL. The average recovery of thymol was from 87.4% to 106.9%, the coefficient of variation was between 0.7% and 7.3%, and the limit of quantification was 0.10 mg/kg. The developed method was applied to determine residual thymol in 48 commercial honey samples, and the thylmol was only detected in one sample. The presented method is simple, quick, reliable, and suitable to be used for batch screening and quantitative determination of residual thymol in honey.
- honey /
- thymol /
- ultra performance liquid chromatography /
- fluorescence method /
- residue

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图 1 不同提取溶剂的回收率（n=3）

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

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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)

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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。

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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

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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

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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

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图 7 百里香酚的紫外吸收光谱

Figure 7. UV absorption spectrum of thymol

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图 8 百里香酚的荧光激发-发射光谱

Figure 8. Fluorescence excitation-emission spectrum of thymol

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图 9 阳性样品的色谱图

Figure 9. Chromatogram of positive sample

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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)
0.01~3.00	y=1.537×10⁷x−3196	0.9999	0.03	0.10

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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	0.1	98.1	5.7
	0.8	98.3	3.2
	30	96.9	0.8
椴树蜜	0.1	89.5	4.3
	0.8	100.3	2.1
	30	87.4	0.9
玄参蜜	0.1	106.9	6.0
	0.8	96.4	2.1
	30	91.1	1.8
洋槐蜜	0.1	95.2	5.8
	0.8	103.2	1.3
	30	89.5	0.8
百花蜜	0.1	98.8	7.3
	0.8	96.8	1.5
	30	96.7	0.7

下载: 导出CSV

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