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Detection of DNA in highly pigmented foods by membrane-based improved cetyltrimethylammonium bromide combined with real-time fluorescence polymerase chain reaction
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Yi-Chao TANG1, Wu-Yan DUAN2, Si-Yang SUN2, Chi-Yuan MA2, *, Biao XIA2, Yong-Quan DONG1
Journal of Food Safety & Quality | 2026, 17(1) : 206 - 217
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Journal of Food Safety & Quality | 2026, 17(1): 206-217
• Food Analysis and Detection •
Detection of DNA in highly pigmented foods by membrane-based improved cetyltrimethylammonium bromide combined with real-time fluorescence polymerase chain reaction
Full
Yi-Chao TANG1, Wu-Yan DUAN2, Si-Yang SUN2, Chi-Yuan MA2, *, Biao XIA2, Yong-Quan DONG1
Affiliations
  • 1.School of Environmental and Chemistry Engineering, Nanchang Hangkong University, Nanchang 330063, China
  • 2.Nanchang Customs Technical Center/National Key Laboratory of Tea Oil Testing, Nanchang 330038, China
doi: 10.19812/j.cnki.jfsq11-5956/ts.20250830001
Outline
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Objective

To establish a method for detecting DNA in highly pigmented food samples by membrane-based improved cetyltrimethylammonium bromide (CTAB) combined with real-time fluorescence polymerase chain reaction (PCR).

Methods

Black sesame, tea seed meal and rapeseed meal were selected as samples. The optimized protocol employed 0.7 μm glass fiber membrane with 70% ethanol as the rinsing solution and 3 rounds of tris(hydroxymethyl)aminomethane ethylenediaminetetraacetic acid (TE) buffer elution. Pigment removal was achieved through membrane-specific adsorption and selective permeability. The extraction efficiency of the modified CTAB method was compared with that of the traditional CTAB method, and DNA purity and amplification efficiency were evaluated via real-time fluorescence PCR targeting endogenous genes.

Results

The modified method demonstrated significantly superior DNA decolorization effects compared to the traditional CTAB method.Endogenous genes were successfully detected in all 3 kinds of samples, with DNA mass concentrations exceeding 100 ng/μL and cycle threshold (Ct) values ranging between 20 and 25. In contrast, the traditional CTAB method failed to achieve real-time fluorescence PCR amplification due to pigment residue.

Conclusion

The membrane-optimized CTAB method simplifies purification steps, enhances decolorization efficiency, and significantly improves the extraction and detection success rate of DNA from highly pigmented foods. This approach provides a reliable and efficient technical solution for food safety testing.

glass fiber membrane  /  nucleic acid purification  /  decolorization  /  real-time fluorescence polymerase chain reaction  /  highly pigmented foods
汤一超, 段吴燕, 孙思扬, 马驰远, 夏彪, 董永全. 膜法改进十六烷基三甲基溴化铵法结合实时荧光聚合酶链式反应检测高色素食品DNA. 食品安全质量检测学报, 2026 , 17 (1) : 206 -217 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250830001
Yi-Chao TANG, Wu-Yan DUAN, Si-Yang SUN, Chi-Yuan MA, Biao XIA, Yong-Quan DONG. Detection of DNA in highly pigmented foods by membrane-based improved cetyltrimethylammonium bromide combined with real-time fluorescence polymerase chain reaction[J]. Journal of Food Safety & Quality, 2026 , 17 (1) : 206 -217 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250830001
Appendix
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Year 2026 volume 17 Issue 1
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Article Info
doi: 10.19812/j.cnki.jfsq11-5956/ts.20250830001
  • Receive Date：2025-08-30
  • Online Date：2026-11-05
Article Data
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History
  • Received：2025-08-30
Funding
Affiliations
    1.School of Environmental and Chemistry Engineering, Nanchang Hangkong University, Nanchang 330063, China
    2.Nanchang Customs Technical Center/National Key Laboratory of Tea Oil Testing, Nanchang 330038, China
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https://castjournals.cast.org.cn/joweb/spaq/EN/10.19812/j.cnki.jfsq11-5956/ts.20250830001
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表12种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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