The Hidden Hurdles in Testing Plant-Based Milks for Toxins

Exploring the challenges of detecting mycotoxins in plant-based milk alternatives using enzyme immunoassays

$29.5B

Projected market by 2031 1 4

54

PBMA products tested 1

23

Samples with mycotoxin detection 1

Introduction

Imagine reaching for a carton of oat milk, believing it to be a healthy choice, only to discover it might contain invisible toxic contaminants. This is the challenge food scientists face as plant-based milk alternatives (PBMAs) surge in popularity. With the global market projected to reach $29.5 billion by 2031, ensuring the safety of these products has never been more critical 1 4 .

Mycotoxins—toxic compounds produced by molds—can lurk in the raw ingredients used to make PBMAs. While laboratory testing methods exist, a 2022 German study revealed significant pitfalls in using rapid enzyme immunoassays (EIAs) for detecting these dangerous substances in plant-based milks 1 2 . This article explores the hidden challenges in keeping our plant-based beverages safe.

What Are Mycotoxins and Why Do They Matter?

Mycotoxins are naturally occurring toxic compounds produced by certain types of fungi that can grow on various food crops. When these contaminated crops are used to make PBMAs, the toxins can carry over into the final product 7 .

Toxin
Aflatoxins (AFB1)

Potent carcinogens linked to liver cancer

Toxin
Ochratoxin A (OTA)

Causes kidney damage and may affect fetal development

Toxin
Deoxynivalenol (DON)

Causes gastrointestinal issues and immune suppression

Toxin
Sterigmatocystin (STC)

Poses significant health risks as a carcinogen

What makes mycotoxins particularly dangerous is their stability—they often survive food processing methods that involve heat, physical, and chemical treatments 7 . The problem is significant enough that global authorities have established strict limits for mycotoxin levels in food, typically in the microgram per kilogram range 6 .

The Appeal and Limitations of Enzyme Immunoassays

Enzyme immunoassays are widely used for rapid detection of various contaminants in food products. Their appeal lies in their:

Speed

Compared to laboratory methods

Simplicity

Of use

Cost-effectiveness

For routine testing

Suitability

For on-site quality control

For decades, similar immunoassays have been successfully used to test for aflatoxin M1 in conventional cow's milk 1 . Researchers naturally wondered: could the same approach work for plant-based milks?

A Deep Dive into the Key Experiment

A comprehensive 2022 study published in Mycotoxin Research set out to answer this question, investigating the applicability of EIA methods for directly testing PBMAs without sample extraction 1 2 .

Methodology: Step-by-Step Approach

Sample Collection

Researchers acquired 54 different PBMA products from German retail stores, with the majority (34 products) labeled as organic. These represented various base ingredients including oats, rice, almonds, soy, and coconut 1 .

Mycotoxin Analysis

The study focused on detecting five mycotoxins: AFB1, STC, OTA, DON, and T-2/HT-2 toxin 1 .

Matrix Interference Testing

Since no certified toxin-free PBMA material was available, scientists assessed matrix interference by comparing toxin standard curves made in EIA buffer solution with those made with diluted PBMA 1 .

Artificial Contamination

As quality control, six PBMA materials were artificially contaminated with known mycotoxin concentrations to test recovery rates 1 .

Validation

Selected samples were additionally analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) to validate the EIA results 1 .

Key Findings and Implications

The research revealed several significant challenges:

Matrix Interference Problems

The diverse ingredients in PBMAs created substantial interference with the immunoassays 1 .

The Dilution Dilemma

Diluting samples at least 1:8 was necessary, increasing detection limits 1 .

Real-World Contamination

23 of the 54 samples showed positive results for at least one mycotoxin 1 .

Detection Limits After 1:8 Dilution
Mycotoxin Detection Limit (µg/L)
Aflatoxin B1 (AFB1) 0.4
Sterigmatocystin (STC) 2
Ochratoxin A (OTA) 0.08
Deoxynivalenol (DON) 16
T-2/HT-2 toxin 0.4

Data source: 1

Why Matrix Interference Matters

Matrix interference occurs when other components in a sample affect the detection method's ability to accurately measure the target substance. In the case of PBMAs:

  • Different ingredients (oats, nuts, legumes) each create unique interference patterns
  • Additives like stabilizers and emulsifiers further complicate detection
  • The aqueous environment of PBMAs differs from the original solid raw materials where mycotoxins typically occur 1

This variability means that a method working well for oat milk might perform poorly for almond or soy milk, creating a significant challenge for quality control laboratories.

PBMA Types and Ingredients in the Study
PBMA Base Ingredient Content Range Examples
Cereal/Pseudocereal 8.7-17% Oat, rice, spelt, millet, buckwheat
Nut-based 2.3-8.4% Hazelnut, almond, coconut, cashew
Legume-based 4-10% Soy bean, pea, lupine
Seed-based 3% Hemp
Mixed Ingredients 4.9-21% Oat + almond, rice + coconut

Data source: 1

Beyond the Core Experiment: Other Detection Methods

While EIAs face challenges with PBMAs, other technologies offer promise:

Chromatography-Based Methods

Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) provides more reliable quantification but requires sophisticated equipment and trained personnel, making it less suitable for rapid on-site testing 1 3 .

Emerging Technologies
  • Salting-out assisted liquid-liquid extraction (SALLE) combined with UHPLC-MS/MS for emerging mycotoxins 3
  • Lateral flow assays (LFAs) using various nanoparticles for enhanced sensitivity 6
  • Biosensors and portable platforms for point-of-care testing 4 7

Research Reagent Solutions for Mycotoxin Detection

Reagent/Material Function in Research
Mycotoxin standards (AFB1, OTA, STC, DON, T-2) Reference materials for calibration and quantification
Phosphate buffered saline (PBS) Dilution buffer for sample preparation
Immunoaffinity columns Selective clean-up and concentration of target mycotoxins
13C-labelled standard solutions Internal standards for precise LC-MS/MS quantification
Monoclonal antibodies Specific recognition elements for immunoassays
Organic solvents (methanol, acetonitrile) Extraction solvents for releasing mycotoxins from samples

Data source: 1 5 7

The Path Forward for Safer Plant-Based Milks

The 2022 study concluded that while EIA methods show potential for screening PBMAs, they're not yet reliable for routine analysis without further development of sample preparation methods specifically designed for these complex matrices 1 .

Future directions include:

  • Developing matrix-specific sample preparation protocols
  • Creating cleanup techniques to reduce interference while maintaining sensitivity
  • Exploring alternative rapid methods like lateral flow assays or biosensors
  • Establishing standardized reference materials for different PBMA types 1 4 6

Conclusion

The hidden challenge of detecting mycotoxins in plant-based milks reveals a broader truth about food safety: as consumer preferences evolve, our testing methods must adapt. The 2022 German study highlighted both the promise and limitations of current technologies, emphasizing that the diverse nature of PBMAs requires equally sophisticated detection approaches 1 .

While the findings might seem concerning, they represent the normal progression of scientific understanding—identifying problems is the first step toward solving them. Through continued research and method development, scientists are working to ensure that as the plant-based milk market grows, so too does our ability to verify its safety.

For consumers, the takeaway isn't to avoid plant-based milks, but to recognize the complex science behind food safety and appreciate the researchers working to develop better detection methods for these popular products.

References