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<p>Milk and milk products are important sources of nutrients for humans. However, milk quality and safety remain major concerns due to adulteration. Nitrates and nitrites in milk indicate contamination with pond water containing fertilizers; however, rapid, field-deployable detection methods are lacking. A paper-based strip assay for the simultaneous detection of both nitrites and nitrates in milk was developed. The strips contain two detection zones and are applied by dipping into milk and observing the color change. Adulterated milk turns the strip pink, while pure milk shows no color change. The limit of detection was 0.0005% (5&#xa0;mg/L) for nitrites and 0.005% (50&#xa0;mg/L) for nitrates, with a response time of 1–2&#xa0;min. The method showed excellent sensitivity and specificity (100%), with no false positive or false negative results during blind testing. Variations in pH, temperature, and processing did not affect strip performance. The strips demonstrated excellent repeatability (RSD &lt; 3%) and reproducibility (RSD &lt; 5%). Other adulterants (glucose, maltodextrin etc.) showed no interference. The strips remained stable for 3 months at 0–4&#xa0;°C. Validation against the spectrophotometric method showed strong correlation (R² = 0.998 for nitrite, R² = 0.997 for nitrate) with no significant difference (<i>p</i> &gt; 0.05). The strip assay offers a rapid, user-friendly tool for detecting milk adulteration in industrial, field and household settings.</p> Graphical Abstract <p></p>

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Development of a paper strip assay for the simultaneous detection of nitrites and nitrates in milk

  • 期刊论文

  • 发表日期：

    2026年4月17日

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  • Saya Vilas Deshmukh,
  • Kamal Gandhi,
  • Rajan Sharma,
  • Sonu K. Shivanna,
  • Prince Dhanda,
  • Jessica Pahsyntiew

摘要

Milk and milk products are important sources of nutrients for humans. However, milk quality and safety remain major concerns due to adulteration. Nitrates and nitrites in milk indicate contamination with pond water containing fertilizers; however, rapid, field-deployable detection methods are lacking. A paper-based strip assay for the simultaneous detection of both nitrites and nitrates in milk was developed. The strips contain two detection zones and are applied by dipping into milk and observing the color change. Adulterated milk turns the strip pink, while pure milk shows no color change. The limit of detection was 0.0005% (5 mg/L) for nitrites and 0.005% (50 mg/L) for nitrates, with a response time of 1–2 min. The method showed excellent sensitivity and specificity (100%), with no false positive or false negative results during blind testing. Variations in pH, temperature, and processing did not affect strip performance. The strips demonstrated excellent repeatability (RSD < 3%) and reproducibility (RSD < 5%). Other adulterants (glucose, maltodextrin etc.) showed no interference. The strips remained stable for 3 months at 0–4 °C. Validation against the spectrophotometric method showed strong correlation (R² = 0.998 for nitrite, R² = 0.997 for nitrate) with no significant difference (p > 0.05). The strip assay offers a rapid, user-friendly tool for detecting milk adulteration in industrial, field and household settings.

Graphical Abstract