花生油中黄曲霉毒素B<sub>1</sub>标准物质的研制

李鹏; 李丽; 叶金; 吴宇; 王松雪

doi:10.12338/j.issn.2096-9015.2023.0320

花生油中黄曲霉毒素B₁标准物质的研制

doi: 10.12338/j.issn.2096-9015.2023.0320

国家粮食和物资储备局科学研究院，北京 100037

基金项目: 国家重点研发计划（2022YFF0710400）。

详细信息

作者简介:
李鹏（1993-），国家粮食和物资储备局科学研究院助理研究员，研究方向：粮油质量安全与检测，邮箱：lp@ags.ac.cn

通讯作者:
王松雪（1977-），国家粮食和物资储备局科学研究院研究员，研究方向：粮油质量安全与风险防控，邮箱：wsx@ags.ac.cn

中图分类号: TB99
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出版历程
- 收稿日期: 2023-11-29
- 录用日期: 2023-12-04
- 修回日期: 2023-12-06
- 网络出版日期: 2023-12-14
- 刊出日期: 2023-10-18

Development of Aflatoxin B₁ Certified Reference Material in Peanut Oil

Academy of National Food and Strategic Reserves Administration, Beijing 100037 , China

摘要

摘要: 根据我国食品安全监测工作的需求，研制了花生油中黄曲霉毒素B₁成分分析标准物质。该标准物质由自然污染的阳性样本，经混匀、充氮、分装等多种加工工艺制备。使用高效液相色谱法（荧光检测器，HPLC-FLD）对标准物质候选物的均匀性进行评估，高效液相色谱-稳定同位素稀释质谱法（HPLC-ID-MS/MS）对稳定性进行考察，高效液相色谱法（荧光检测器）和高效液相色谱-稳定同位素稀释质谱法两种方法进行多家实验室合作定值。研究表明：该标准物质满足技术规范要求，均匀性和稳定性良好，可在−18 ℃储藏下保存16个月，黄曲霉毒素B₁的标准值为15.6 μg/kg，扩展不确定度为1.6 μg/kg （k = 2）。标准物质的不确定度评估综合考虑了均匀性、稳定性和定值过程引入的不确定度。研制的花生油中黄曲霉毒素B₁成分分析标准物质，已获得国家市场监督管理总局批准的有证标准物质证书，编号为GBW(E) 100604。该标准物质能够满足食用油中黄曲霉毒素B₁测量急需溯源标准的应用需求，为保证准确可靠的测量结果提供了有效支撑。
- 计量学 /
- 花生油 /
- 黄曲霉毒素B₁ /
- 标准物质 /
- 均匀性 /
- 稳定性 /
- 定值
Abstract: To fulfill the requirements of food safety monitoring in China, a certified reference material (CRM) for aflatoxin B₁ (AFB₁) analysis in peanut oil was developed. This CRM was derived from naturally contaminated positive samples, undergoing processes such as blending, nitrogen purging, and packaging. Homogeneity of the CRM candidate was assessed using High-Performance Liquid Chromatography with Fluorescence Detection (HPLC-FLD), while its stability was evaluated using High-Performance Liquid Chromatography - Isotope Dilution Mass Spectrometry (HPLC-ID-MS/MS). Value assignment was performed through collaborative laboratory efforts utilizing both HPLC-FLD and HPLC-ID-MS/MS. The results indicated that the CRM meets technical specifications, displaying excellent homogeneity and stability for 16 months at −18 ℃. The certified value of AFB₁ is 15.6 μg/kg with an expanded uncertainty of 1.6 μg/kg (k = 2). The uncertainty assessment encompassed factors from homogeneity, stability, and value assignment. The developed CRM for AFB₁ in peanut oil, certified as GBW(E) 100604, fulfills the urgent need for traceable standards in edible oil AFB₁ measurement and ensures accurate and reliable results.
- metrology /
- peanut oil /
- aflatoxin B₁ /
- certified reference material /
- homogeneity /
- stability /
- quantification

HTML全文

图 1 花生油中AFB₁色谱图

Figure 1. Chromatographic analysis of AFB₁ in peanut oil

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图 2 总离子流图

Figure 2. Total ion chromatogram analysis

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图 3 花生油中黄曲霉毒素B₁均匀性检测结果

Figure 3. Uniformity testing results for aflatoxin B₁ in peanut oil

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图 4 花生油中AFB₁长期稳定性监测图示

Figure 4. Trend chart illustrating long-term stability of aflatoxin B₁ in peanut oil

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图 5 花生油中AFB₁短期稳定性监测图示

Figure 5. Trend chart depicting short-term stability of aflatoxin B₁ in peanut oil

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表 1 两种不同原理方法结果比较（μg/kg）

Table 1. Comparative results from two distinct analytical methods for AFB₁ (μg/kg)

序号	液相法	质谱法
1	15.2	15.1
2	15.4	15.4
3	15.6	15.2
4	15.1	15.6
5	15.8	15.1
6	15.3	15.7
7	15.7	15.2
8	15.8	15.0
9	15.0	15.6
10	15.9	15.3
平均值	15.48	15.32
SD	0.32	0.24
RSD	2.08%	1.59%

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表 2 方差分析结果

Table 2. Statistical analysis of variance results

差异源	差方和	自由度	方差	F_统计量	F_临界值
组间	10.984	24	0.458	1.226	1.737
组内	18.660	50	0.373
总计	29.644	74

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表 3 稳定性研究线性回归分析

Table 3. Linear regression analysis of long-term stability

类型	时间/月	平均测定值（μg/kg）	类型	时间/天	平均测定值（μg/kg）
长期稳定性	0	15.8	短期稳定性	0	16.2
	0.5	14.9		1	16.9
	1	16.1		2	16.8
	2	15.2		3	16.2
	4	15.8		5	16.2
	8	15.5		6	16.1
	12	15.3		7	15.9
	16	15.2		−	−
	平均值	15.5		平均值	16.3
	β₁	−0.021		β₁	0.0932
	β₀	15.590		β₀	16.648
	s²	0.168		s	0.0945
	s(β₁)	0.0260		s(β₁)	0.0476
	t _{0.95, n-2}	2.45		t _{0.95, n-2}	2.45

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表 4 花生油中AFB₁ 标准物质定值结果

Table 4. Certified value assignment results for aflatoxin B₁ in peanut oil

实验室	测定值（μg/kg）	平均值（μg/kg）	标准偏差	定值方法
A	15.6, 14.9, 15.7, 16.9, 15.2, 16.2	15.8	0.72	HPLC-ID-MS/MS
B	14.9, 15.3, 14.6, 15.0, 16.6, 15.3	15.3	0.70	HPLC-ID-MS/MS
C	15.9, 15.4, 15.4, 16.0, 15.3, 15.7	15.6	0.29	HPLC-ID-MS/MS
D	15.6, 16.3, 15.4, 15.2, 15.4, 15.6	15.6	0.38	HPLC-FLD
E	15.6, 15.7, 15.9, 15.5, 16.6, 16.0	15.9	0.40	HPLC-FLD
F	14.9, 15.7, 15.5, 15.5, 15.5, 15.8	15.5	0.31	HPLC-FLD
G	15.8, 15.9, 16.1, 15.7, 15.5, 16.0	15.8	0.22	HPLC-FLD
总平均值		15.6	0.20

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表 5 花生油中AFB₁标准物质研制过程的不确定度来源

Table 5. Uncertainty sources identified during the development of aflatoxin B₁ CRM in peanut oil

不确定度来源	计算公式	相对不确定度 %
均匀性引入	$ {u}_{\mathrm{H}}=\dfrac{{S}_{\mathrm{H}}}{\overline{X}} $	1.06
稳定性引入	$ {u}_{\text{S}}=\sqrt{{{u}_{\mathrm{l}\mathrm{S}}}^{2}+{{u}_{\mathrm{S}\mathrm{S}}}^{2}} $	3.42
定值引入	$ {u}_{\mathrm{A}}=\dfrac{{S}_{\overline{X}}}{\overline{X}}=\dfrac{\sqrt{{\displaystyle\sum }_{i=1}^{m}(\overline{{X}_{i}}-\overline{X}{)}^{2}/m(m-1)}}{\overline{X}} $	1.11
定值引入	$ {u}_{\mathrm{B}} $	3.32
合成相对不确定度		5.1
k = 2（95%置信区间），相对扩展不确定度		10.2

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