B Vitamin Analysis (B1–B6) in Food, Feed, Infant Formula and Premixes

B vitamins are a group of water-soluble compounds that play essential roles in energy metabolism, neurological function, and cellular processes. They act primarily as coenzymes, enabling the conversion of carbohydrates, fats, and proteins into usable energy.
Because they are not stored in significant amounts in the body, a consistent dietary supply is required to maintain normal physiological function in both humans and animals.

B vitamin analysis is used to verify nutritional composition, support formulation, and ensure compliance with product specifications and regulatory requirements.
In food and infant nutrition, B vitamins are critical for growth, energy metabolism, nervous system development, and red blood cell formation. In animal nutrition, they contribute to feed efficiency, metabolic performance, stress tolerance, and overall productivity.
Due to their low inclusion levels and high biological significance, even small deviations in concentration can impact product quality and nutritional value, making accurate measurement essential.

The analysis focuses on the most relevant and commonly supplemented B vitamins in food and feed applications, namely thiamine (B1), riboflavin (B2), niacin (B3), pantothenic acid (B5), and pyridoxine (B6).
These vitamins perform interconnected roles within metabolic pathways. Thiamine (B1) is essential for carbohydrate metabolism and energy production, particularly in high-energy diets. Riboflavin (B2) functions as a cofactor in oxidation–reduction reactions and supports cellular energy systems.
Niacin (B3), present as nicotinic acid and nicotinamide, is involved in numerous enzymatic processes related to energy metabolism and metabolic efficiency. Pantothenic acid (B5) is a precursor to coenzyme A and plays a central role in fatty acid metabolism and a wide range of biochemical reactions.
Vitamin B6 (pyridoxine and related forms) is involved in amino acid metabolism, protein utilisation, and red blood cell formation, and is also essential for normal nervous system function.

B vitamin analysis is applied across a wide range of matrices, including food products, animal feed, infant formula, and vitamin premixes.
In these applications, testing is used to verify label claims, confirm correct inclusion levels, and ensure consistency during production. Infant formula requires particularly strict control of vitamin content due to defined nutritional requirements for early development.
In feed and premixes, analysis supports formulation accuracy, stability monitoring, and overall product quality, particularly where vitamins are added at low concentrations and must remain stable throughout processing and storage.

B vitamins are sensitive compounds and can degrade under various environmental and processing conditions. Heat, light exposure, pH, and oxidation all influence their stability, with some vitamins being more susceptible than others.
Processing steps such as extrusion, pelleting, and thermal treatment can result in significant losses if not properly controlled. Storage conditions and interactions with other components in the matrix may also affect vitamin stability over time.
In addition, different sample types present unique analytical challenges. Premixes typically contain high concentrations of vitamins but may include carriers and stabilisers, while feed and infant formula are more complex matrices where vitamins are present at lower levels and may be more difficult to extract and quantify accurately.

B vitamins are analysed using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). This technique provides high sensitivity, selectivity, and accuracy, making it well-suited for the simultaneous determination of multiple vitamins in complex matrices.
LC-MS/MS allows for reliable quantification of B1–B6 at low concentration levels while minimising interference from co-extracted compounds. This is particularly important in matrices such as feed, premixes, and infant formula, where matrix effects can significantly impact analytical performance.
Careful sample preparation and extraction are required to ensure stability and accurate recovery of the vitamins prior to analysis.

B vitamin analysis is performed under SANAS-accredited ISO/IEC 17025 conditions, ensuring that methods are validated and results are reliable, traceable, and suitable for regulatory and quality control purposes.

SANAS accreditation confirms that the laboratory operates in accordance with internationally recognised standards and demonstrates technical competence in performing these analyses across a range of matrices.

B vitamin testing forms part of routine nutritional analysis in food and feed laboratories. It supports formulation, product development, and quality control by ensuring that vitamin levels meet specification throughout the product lifecycle.
When used alongside other nutritional parameters such as protein, fat, fibre, and minerals, B vitamin analysis contributes to a comprehensive understanding of product composition and performance.