Vitamin Analysis

Vitamins are micronutrients essential for a healthy life. These compounds, which support many functions in the human body, are taken from food or food supplements. However, it is very important to know whether the amount of vitamins stated on the label of a product is correct.

These vitamins can be found in natural or artificial (synthetic) forms. There are some key differences between the two. Natural vitamins are derived from natural sources such as plants, animals or microorganisms. For example, vitamin C can be obtained from citrus fruits, red peppers, broccoli; vitamin A from liver, carrots, sweet potatoes. They are usually more easily metabolized by the body, have better bioavailability and a lower risk of side effects. Artificial (synthetic) vitamins are produced in the laboratory by chemical processes. They mimic the structure of natural vitamins but in some cases may show isomeric differences. For example, synthetic vitamin E (dl-alpha-tocopherol) has lower bioavailability than its natural form (d-alpha-tocopherol). Synthetic vitamins are cost-effective and can be produced on a large scale, have a longer shelf life and can be prepared in specific doses. In some individuals, absorption and efficacy may be lower than natural vitamins. There may be a risk of toxicity in overdose. Synthetic vitamins can be used as supplements to address nutritional deficiencies or to manage medical conditions.

At NanoLab, water and fat-soluble vitamins are analyzed with high sensitivity.

Importance of Vitamin Analysis

The vitamin content in foods plays a critical role in determining the quality, safety and nutritional value of the product. Inadequate or excessive intake of vitamins can lead to health problems. Verifying the amount of vitamins indicated on the product label provides consumer confidence. Food codexes and regulations regulate the amounts of vitamins in certain foods and supplements. It is described in the Turkish Food Codex Regulation on the Addition of Vitamins, Minerals and Certain Other Elements to Foods.

Fat Soluble Vitamins

Vitamin A exists in two main forms. Retinoids, the active form, are found in animal foods. Carotenoids, which are provitamin A, are found in plant foods. The active form is found in liver, fish oils, milk and dairy products, egg yolks, while provitamin A is found in dark green leafy vegetables such as carrots, sweet potatoes, pumpkin, spinach, kale, red, orange and yellow peppers, and yellow-orange fruits such as melon and mango. Carotenoids from plant sources are converted to retinol in the body. This conversion can vary depending on individual bioavailability.
Vitamin D exists in two main forms. Vitamin D3 (cholecalciferol): found in animal sources and synthesized in the skin by sunlight. found in fatty fish such as salmon, mackerel, tuna, sardines, fish oil, liver, egg yolk, whole milk and dairy products.
Vitamin D2 (ergocalciferol) found in plant sources. Vitamin D2 is found in mushrooms. Vitamin D is fortified in milk and dairy products, vegetable milks (soy, almond, oat milk), breakfast cereals and orange juice.
Vitamin E is a powerful antioxidant and especially protects cell membranes from free radical damage. It occurs naturally in many foods. Vitamin E is found in vegetable oils (sunflower oil, olive oil, corn oil, wheat germ oil, soybean oil), nuts and seeds, almonds, hazelnuts, walnuts, sunflower seeds, pumpkin seeds, vegetables, green leafy vegetables such as spinach and broccoli, asparagus, berries, avocado, kiwi, mango, whole grains, eggs, oily fish.
Vitamin K is found in two main forms. Vitamin K1 (Phylloquinone): Found in plant sources. Dark green leafy vegetables: spinach, kale, broccoli, chard, lettuce, Brussels sprouts, parsley, asparagus, avocados are rich in Vitamin K1.
Vitamin K2 (Menaquinone) found in animal foods and fermented products. Vitamin K2 is found in foods of animal origin. Water Soluble Vitamins
Vitamin B1 (thiamine) plays an important role in the conversion of carbohydrates into energy and in the proper functioning of the nervous system. Cereals and cereal products, whole grains (wholemeal bread, brown rice), legumes and seeds, meat, fish, eggs, nuts, vegetables and fruits are foods rich in Vitamin B1. Breakfast cereals, bread and bakery products can be enriched with thiamine. Thiamine is sensitive to heat and water, so it can be lost when foods are overcooked.
Vitamin B2 (riboflavin) is important for energy production, skin health and the nervous system. It is also involved in the metabolism of other B vitamins. Meat and dairy products, eggs, dark green leafy vegetables, mushrooms, nuts are sources of riboflavin. Breakfast cereals, bread and baked goods are foods enriched with riboflavin. Riboflavin is water-soluble but sensitive to heat and light. It is therefore important not to keep food in direct sunlight.
Vitamin B5 (pantothenic acid) plays an important role in basic biological functions in the body, such as energy production and the synthesis of fatty acids. Meat and seafood, eggs, milk and dairy products, whole grains, legumes, nuts and seeds, mushrooms and vegetables are rich sources of vitamin B5. Vitamin B5 deficiency is rare because it is found in many foods.
Vitamin B6 (pyridoxine) is involved in many important functions in the body, such as amino acid metabolism, neurotransmitter production and hemoglobin synthesis. Food sources rich in vitamin B6 include meat, eggs, whole grains, potatoes, dark green leafy vegetables, avocados, nuts and seeds.
Vitamin B9 (folic acid or folate) plays a critical role in DNA synthesis, cell division and the production of red blood cells. It is particularly important during pregnancy because it is essential for fetal growth and development. Food sources rich in folate include dark green leafy vegetables, avocados, tomatoes, legumes, seeds, whole grains and eggs. In many countries, bread, flour and other bakery products are fortified with folic acid.
Vitamin B12 (cobalamin) is essential for DNA synthesis, red blood cell production and nervous system health. Vitamin B12 is mostly found in foods of animal origin. Meat, fish, eggs, milk and dairy products are foods rich in B12. Since natural B12 is usually found in animal sources, it is recommended that vegans use B12 supplements and consume nutritional yeast (enriched with B12). Regular consumption or supplementation from these sources is important for adequate B12 intake.
Vitamin C (ascorbic acid) plays a critical role in many biological processes such as the immune system, skin health and iron absorption. Since it cannot be produced by the body, it must be taken from outside. Fruits, vegetables, dark leafy greens, tomatoes, squash, pumpkin, potatoes, sweet potatoes are rich sources of vitamin C. Some fruit juices and processed foods are fortified with vitamin C. Since vitamin C is sensitive to heat, it is recommended to cook these foods as little as possible.

Analysis Methods Used

In accordance with the sensitivity of vitamins, the most appropriate method is determined according to the chemical properties of the vitamin to be analyzed without exposure to light, temperature and oxygen, and pre-treatments are performed. The accuracy of the results is ensured by regular calibration of the devices and quality control studies.

Different analysis methods are used according to the type of vitamins. With High Performance Liquid Chromatography (HPLC), vitamins are determined by separation in the liquid phase. It is widely used in the analysis of vitamins A, D, E, K, C and B group vitamins. Pre-treatment is carried out by applying separate extraction methods for fat-soluble and water-soluble vitamins. UV, DAD and FID detectors are used according to the type of analysis. Measurements are performed with high precision (ppm level) in our devices. Limits in vitamin analysis are determined according to the legal requirements of the relevant food and product category. These limits regulate the minimum and maximum amount of the vitamin and are usually based on the food codex, international standards or national legislation.

Vitamin Limits in Turkiye

Regulated by the Turkish Food Codex Regulation on Nutrition and Health Claims and related legislation. Vitamin amounts for dietary supplements should be less than 15% of the Nutrition Reference Values (NRV) for adults (7.5% for liquid foods). The amount of added vitamins should not pose a risk of poisoning to consumers. Vitamin additions are usually made to specific food groups. For example, vitamin B complexes are added to cereal products and vitamin D to dairy products. The purpose of fortification is to remedy the deficiency of the added vitamin in the population or to improve the quality of nutrition.

Only chemical forms specified in the regulation can be used to add vitamins to food. These forms have been scientifically evaluated and approved for safety, bioavailability (absorption and availability in the body) and stability.

The different chemical forms of vitamins are determined by the advantages of stability and bioavailability. The vitamin to be added is adapted to the type of food. For example, water-soluble vitamins are more suitable for liquid products, while fat-soluble vitamins are more suitable for fat-containing products.

Since Vitamin A is a fat-soluble vitamin, it is added to foods such as dairy products, oils and margarine. It is added in the forms of retinol, retinyl acetate, retinyl palmitate.
Vitamin D is added to fat-based products (milk, butter, margarine) and aims to meet the need for vitamin D that cannot be obtained from sunlight. It is added in the forms of cholecalciferol (vitamin D3), ergocalciferol (vitamin D2).
Vitamin E is added to vegetable oils and processed foods due to its fat-soluble nature. It is added in the form of alpha-tocopherol, alpha-tocopherol acetate.
Because Vitamin C is water-soluble, it is usually added to fruit juices, drinks and supplements. Ascorbic acid is added in the forms of sodium ascorbate, calcium ascorbate.
Group B vitamins are fortified by adding them to cereals, flour and breakfast cereals. Vitamin B1; thiamine hydrochloride, vitamin B2; riboflavin, vitamin B3; niacinamide, vitamin B6; pyridoxine hydrochloride. The added vitamin or mineral should not adversely affect the taste, color or texture of the food.

Vitamins that can be added to feed in Turkiye and their maximum limits

It is regulated by the "Regulation on the Market Supply and Use of Feed" published by the Ministry of Agriculture and Forestry. Vitamins are considered in the category of nutritional additives within feed additives. These include fat-soluble vitamins (A, D, E, K), water-soluble vitamins (B1, B2, B3, B5, B6, B7, B9, B12, C).

Each vitamin has different limits of use and dosages according to animal species.

Vitamin A varies for specific animal species, but generally up to 10,000 IU/kg can be added.
Vitamin D is 5,000 IU/kg for poultry, with different limits for other animal species.
Vitamin E is generally used between 100-500 mg/kg in feeds.
Vitamin K maximum limit may vary between 2-10 mg/kg.
Vitamin C is limited to 100 mg/kg.
Vitamin B1 (Thiamine) maximum: 10-15 mg/kg,
Vitamin B2 (Riboflavin) 4-10 mg/kg for poultry, 2-8 mg/kg for other animal species,
Vitamin B3 (Niacin) 30-60 mg/kg for poultry, 20-40 mg/kg for pigs,
Vitamin B5 (Pantothenic Acid) 10-20 mg/kg for poultry, 8-15 mg/kg for pigs,
Vitamin B6 (Pyridoxine) maximum 3-6 mg/kg,
Vitamin B7 (Biotin) 0.1-0.3 mg/kg for poultry, 0.05-0.2 mg/kg for other animal species,
Vitamin B9 (Folic Acid) maximum 1-5 mg/kg,
Vitamin B12 (Cobalamin) is added at levels of 10-20 µg/kg for poultry and 5-15 µg/kg for other animal species.

It is essential that vitamins are used at a level that meets the needs of the animals. This is formulated in feed additive premixes. The specified limits should not be exceeded as overdose of B vitamins can lead to economic losses and environmental problems. Product labels should indicate the type, amount and premix form of vitamin used.

These analyzes guarantee the quality and reliability of food products. While consumers pay attention to the amount of vitamins in the products they buy, it is of great importance for producers to have regular laboratory analyzes because a healthy life requires access to accurate information and reliable products.

Why Should We Have a Vitamin Analysis?

Vitamin analyzes are required to fulfill the legal requirements we have mentioned and to create brand trust in the face of the consumer.
In order to bring your products to the forefront in the market, you can catch trends with the production of innovative products with vitamin additives.
Since the amount of vitamins added in vitamin-added products is proportionally small, it must be checked whether a correct homogenization is made.
In some country standards such as European regulation and FDA, it is an obligation to include Vitamin analysis in nutrient tables. Vitamin tests may be indispensable for exported products.

Why Nanolab?

Vitamin analysis is a process that requires scientific precision and technical equipment. By providing the most reliable results with these analyzes, consumer health and the sustainability of the sector are supported. These analyzes will be carried out in our laboratories with our high-precision equipment and devices and by our expert staff and will provide you with the most accurate results. By having your vitamin-containing food and supplement products analyzed in our laboratories, you can both ensure compliance with legal requirements and gain consumer confidence!

Food, Feed	Vitamin A (Retinol) Vitamin E (DL-Alpha Tocopherol) Vitamin D2 (Ergocholecalciferol) Vitamin D3 (Cholecalciferol)	R.G. 29955, 2017
Food, Baby Food, Infant and Young Child Supplementary Food, Follow-Up Formulas	Vitamin B1 (Thiamine) Vitamin B2 (Riboflavin) Vitamin B5 (Panthetonic Acid) Vitamin B6 ((Pyridoxine) Vitamin B8 Vitamin B9 (Folic Acid) Vitamin B12 (Cobalamin)	Development Of HPLC Methods For The Determination Of Watersoluble Vitamins In Pharmaceutıcals And Fortified Food Products Clemson University
Food, Feed	Vitamin C (L-Ascorbic Acid)	Rückemann H. 1980 (Z. Lebensm. Unters. Forsch, 171, 446-448, 1980 ) - HPLC-DAD
Food, Feed, Baby Food, Infant and Young Child Supplementary Food, Follow-Up Formulas	Vitamin K (K1 Pthylloquinone)	AOAC 999.15 - HPLC-DAD