TESTING FOR NITROSAMINES IN FOOD AND WATER

Nitrates (NO₃^–) occur naturally in Soil as Nitrogen Compounds and are also a component of Synthetic Nitrogenous Fertilizers. Other than Rice, Plants absorb Nitrogen in Nitrate form. Leafy greens like Spinach are relatively higher in Nitrate content. Others are Celery, Carrot, and Beetroot. Organic Vegetables are likely to have lower Nitrate content compared to Non-organic ones due to the non-use of Synthetic Nitrate Fertilizers.

Nitrosamines are Reaction Products of Nitrites or Nitrogen Oxides with Secondary (NH-R₂) or Tertiary (NH-R3) Amines or Substances containing Amino Groups (NH₂). Nitrite (NO₂^–) is an intermediate in the Nitrate (NO₃^–) conversion process formed with the help of Soil (Nitrobacteraceae) and Oral (Veillonella, Neisseria) Bacteria and Enzymes.

Nitrosamines form when Nitrite or other Nitrogenous Agents like Dinitrogen Trioxide (N₂O₃)_, and Nitrogen Tetroxide (N₂O₄) react with Secondary and Tertiary Amines, Amides, Carbamates, Urea, and Guanidines.

The International Agency for Research on Cancer (IARC) categorizes NOCs as Class 2a and Class 2b i.e. as “Probable and Possible Carcinogens”, while the International Council for Harmonisation (ICH) M7 considers them to be “Cohort of Concern”.

N-nitrosamines, the Amine Derivatives exhibit their Mutagenicity or Carcinogenicity on being Activated Metabolically while the N-nitrosamides are Direct Acting Mutagens.

NO₂–                 —————>                N₂O₃ (Nitrous Anhydride)

Nitrite Ion         Acidic Aq. Soln             Nitrosating Agent

Secondary/     + Nitrosating Agent—–> Nitrosamines

Tertiary Amine

Volatile Nitrosamines Nitrosodimethylamine (majorly) and Nitrosopyrrolidine occur most commonly in Foods like Cured Meats, Primarily Cooked Bacon; Beer; few Cheeses; Nonfat Dry Milk; and sometimes Fish. Nitrosamines can also be created directly in foods through high-heat cooking, as with fried bacon.

Direct-flame Heated Air Drying of Certain Foods

N₂                Nitrosates      Amines in Certain Foods

in Drying Air

Nitrosamine risks from leafy greens and other vegetables in less than from Processed Meats as they are not Protein-rich and also contain protective components like Vitamin C, Polyphenols, and Fibre all of which Reduce Nitrosamine Formation. Research suggests that the Bacterial Catalyzation in Nitrosation of Secondary Amines may be important for People with Urinary Tract Infections in locations having high levels of Nitrates in Drinking Water.

Nitrosamine Regulations

The Annex to Commission Regulation (EU) 1333/2008 of the European Parliament and Council, and Commission Regulation (EU) 231/2012 have been amended and Set New Limits for Nitrite (E 249-250) and Nitrate (E 251-252) Additives in Food in the E.U. from 29-10-2023. The US FDA has stated limits for NDMA or total Nitrosamines in Bacon, Barley Malt, Ham, and Malt Beverages.

Nitrosamine Analysis in Food and Feed

Matrices involving Quantification of Extremely Minute Levels of Contaminants demands

Distinct Experiential Scientific, Technical, and Analytical Rigour.

Food Types Tested

Fruits, Vegetables, Processed and Cooked Meat, Marine Foods – Dried Fish and Edible

Molluscs, Alcoholic Beverages, etc.

         Parameters

N-Nitrosopyrrolidine(NPYR), N-Nitrosodibutylamine(NDBA), N-Nitrosodiethanolamine

(NDELA), N-Nitrosodiethylamine (NDEA), N-Nitrosodimethylamine (NDMA), N-

Nitrosodipropylamine (NDPA), N-Nitrosomethylethylamine (NMEA), N-Nitrosomorpholine

(NMOR), N-Nitrosopiperidine (NPIP)

Methods

IS, EU, USFDA

Instruments

ICP-MS, LC-MS/MS, HR-MS, GC, GC-MS, GCMSMS, HPLC, FTIR, ELISA, RT-PCR, NMR

Probiotics: At the Gut of the Health & Wellness Revolution

Around 60 tons of food passes through the human gastrointestinal tract (GIT) over an average person’s lifetime. The GIT represents a biotic ecosystem within an abiotic environment, influenced by factors such as temperature, pH, and water. The term “biotic” derives from the Greek word “biotikos,” meaning “relating to life.” The GIT is home to an estimated 39 trillion microorganisms, collectively referred to as the ‘Gut Microbiota,’ which includes bacteria, archaea (anaerobes living in low-oxygen environments), and eukarya (fungi, simple algae, protozoa).

The bacteria in the gut microbiome can be categorized as “good” or “bad” and undergo a synbiotic, or mutualistic, co-evolution with their host. This complex and dynamic microbial population and its interactions are vital for maintaining the health of the ecosystem. Dysbiosis, or an imbalance within this microorganism community, can disrupt these crucial mechanisms.

The first probiotic, Lactobacillus bulgaricus, was identified by Bulgarian physician and microbiologist S.G. Grigorov in 1905. Subsequently, zoologist and the father of natural immunity, Élie Metchnikoff, suggested that this bacterium contributed to the extended longevity of Bulgarians.

• Strengthened Gut Integrity: Enhancing the intestinal epithelium.
• Harvesting Energy: Improving energy extraction from the diet.
• Protection Against Pathogens: Acting as a barrier to harmful microorganisms.
• Regulating Host Immunity: Potentially aiding in conditions like hay fever, eczema, and the common cold.

The Indian and global probiotics markets are projected to reach $2 billion and $78 billion by 2028 and 2026, respectively, according to Knowledge Source Intelligence (KSI), Research Markets (RM), and Coherent Market Insights (CMI). As these markets expand, it becomes increasingly important to understand the different types of biotics, their safety aspects, the regulations governing them, and the scientific tests that ensure their safety and efficacy.

The Role of Food Testing and EFRAC Labs

Food testing is essential in the probiotics sector to ensure the safety, efficacy, and quality of probiotic products. Here’s how food testing, conducted by laboratories such as EFRAC (Edward Food Research and Analysis Centre), contributes:

• Verification of Probiotic Strains: Food testing verifies that probiotic products contain the strains and quantities listed on their labels, which is critical for their effectiveness.
• Effectiveness Evaluation: Testing ensures that probiotics can survive the digestive process and reach the intestines in adequate numbers to provide health benefits.
• Regulatory Compliance: Food testing supports compliance with regulations and standards, such as those set by EFRAC labs, ensuring products meet safety and quality benchmarks.

As the popularity of probiotics continues to rise, understanding the importance of food testing and staying informed about regulatory standards and scientific assessments, such as those provided by EFRAC, can help consumers make educated choices about their health.

PFAs in Food & Water

Perfluoroalkyl and polyfluoroalkyl substances (PFAs) are a diverse group of thousands of synthetic chemicals categorized as and belonging to the Persistent Organic Pollutants (POPs) Class of Chemicals.

Persistent because they “Resist Degradation through Chemical, Biological, and Photolytic Processes” Organic, as they have “Carbon atoms” covalently linked to Fluorine atoms, and Pollutant as they are “Extremely Toxic, and thus Detrimental to Human Health & Global Environment”. Some of the PFAS have been in Commercial use since the 1940s.

Since the 1960s, the FDA authorized Oil and liquid-resistant PFAS for limited use in Non-stick Cookware, Food Packaging, and Food Processing Equipment. However, these damage the Immune & Reproductive, Neurological Systems, and are Cancer and Birth Defect-causative.

Their Widespread use, Environmental Persistence, and consequently elevated levels in Air, Water, and Soil, have been contaminating Humans. Certain Vegetables and Drinking Water are important sources.  Seafood including Fish, Meat, and its Products, Eggs, Milk, and Dairy Products are strong pathways for their bioaccumulation in Humans.

EU Stipulated Maximum levels for Selected PFAS’ in Certain Foods with effect from 1^st January 2023.  USEPA’s limits on 02 common types of PFAS – PFOA and PFOS are extremely strict i.e. at 04 parts/ trillion. This is phenomenal considering that less than a decade back, the USEPA, had suggested a 70 parts/trillion upper limit on combined PFOS and PFOA exposure.

The increasing precision of cutting-edge Instruments has made PFAS Detection and Quantitation, a reality affording us the chance to make Food, Scientifically Safe.

Let’s do it!