3.3 Vitamin Analysis

These are low molecular weight, biologically active organic compounds essential for an organism’s normal health and growth.

Vitamins

What are the fat-soluble vitamins?

ADEK

These vitamins are absorbed with dietary fat and stored in the liver and adipose tissues.

Fat-soluble vitamins (ADEK)

These vitamins have more risk of toxicity because they accumulate in the body.

Fat-soluble vitamins (ADEK)

This vitamin functions for vision, immune system, and skin health.

Vitamin A

This vitamin can be sourced from liver, carrots, sweet potatoes, and leafy greens.

Vitamin A

Which foods can provide vitamin A?

• Liver

• Carrots

• Sweet potatoes

• Leafy greens

Deficiency of this vitamin leads to night blindness and xeropthalmia.

Vitamin A

Toxicity of this vitamin leads to liver damage and birth defects.

Vitamin A

This vitamin is helpful in calcium absorption, bone health, and immune system.

Vitamin D

Where can we source vitamin D?

• sunlight

• fatty fish

• fortified milk

Deficiency in this vitamin leads to rickets (children) and osteomalacia (adults).

Vitamin D

Toxicity of this vitamin leads to hypercalcemia.

Vitamin D

This vitamin serves as an antioxidant and protects cell membranes.

Vitamin E

What foods provide vitamin E?

• vegetable oils

• nuts

• seeds

Deficiency in this vitamin leads to hemolytic anemia and neurological problems.

Vitamin E

Toxicity of this vitamin leads to bleeding disorders as it interferes with vitamin K.

Vitamin E

This vitamin helps in blood clotting and bone metabolism.

Vitamin K

Where can we source vitamin K?

• leafy greens

• gut bacteria synthesis

Deficiency in this vitamin leads to impaired blood clotting.

Vitamin K

These vitamins are not stored extensively and are excreted in urine.

Water-soluble vitamins

True or false: Water-soluble vitamins are less toxic than fat-soluble vitamins but must be consumed regularly.

True

It acts as a powerful antioxidant, essential for collagen synthesis, supports immune function, and significantly enhances iron absorption.

Vitamin C

What foods provide vitamin C?

• citrus fruits

• bell peppers

• tomatoes

Deficiency of this vitamin leads to scurvy, which is characterized by bleeding gums, joint pain, and poor wound healing due to failed collagen synthesis

Vitamin C

Vitamin C deficiency leads to this disease, which is characterized by bleeding gums, joint pain, and poor wound healing due to failed collagen synthesis.

Scurvy

High doses of this vitamin can cause gastrointestinal distress and increase the risk of kidney stones.

Vitamin C

These vitamins act as oenzymes that drive energy metabolism (helping the body extract energy from carbs, fats, and proteins), support DNA synthesis, and maintain proper nerve function.

Vitamin B Complex

What foods provide the vitamin B complex?

• whole grains

• meats

• eggs

• leafy greens

This vitamin functions in energy metabolism (coenzyme for carbohydrate breakdown); nerve signaling.

B1 (Thiamin)

Deficiency in this vitamin leads to beriberi (Wet = cardiovascular issues; Dry = muscle wasting/neurological).

B1 (Thiamin)

This vitamin functions in energy production (electron transport via FAD/FMN); cellular growth.

B2 (Riboflavin)

Deficiency in vitamin B2 leads to a group of diseases called __.

Ariboflavinosis

Deficiency in this vitamin leads to cheliosis (cracked lips), glossitis (magenta tongue), and angular stomatitis.

B2 (Riboflabin)

This vitamin functions in energy metabolism (redox reactions via NAD/NADP); DNA repair.

B3 (Niacin)

Deficiency in this vitamin leads to pellagra.

B3 (Niacin)

What are the 4D’s of pellagra?

• Diarrhea

• Dermatitis

• Dementia

• Death

This vitamin functions as a structural component of Coenzyme A (CoA); vital for fatty acid synthesis and energy extraction.

B5 (Pantothenic Acid)

Deficiency in this vitamin causes the burning feet syndrome, fatigue, and GI distress.

B5 (Pantothenic Acid)

This vitamin functions as a coenzyme for carboxylation reactions (synthesis of fats and glucose).

B7 (Biotin)

This vitamin functions in DNA synthesis and cell division; vital for rapid growth phases.

B9 (Folate)

Deficiency in these vitamins leads to megaloblastic anemia (enlarged blood cells).

B9 and B12

Deficiency in this vitamin leads to neural tube defects (e.g., spina bifida) in fetal development.

B9 (Folate)

Deficiency in this vitamin leads to pernicious anemia.

B12 (Cobalamin)

This vitamin functions in myelin sheath maintenance (nerve protection); works with Folate for RBC formation and DNA synthesis.

B12 (Cobalamin)

What are the classical methods of vitamin analysis?

• Chemical assay

• Biological assay

• Titrimetry (Redox with DCPIP)

This vitamin analysis is suitable for vitamin C analysis.

Redox titration with DCPIP

Redox titration with DCPIP is suitable for the analysis of which vitamin?

Vitamin C

This vitamin reduces DCPIP from blue to colorless.

Vitamin C

This vitamin analysis method is simple and inexpensive but it is not specific (other reducing agents may interfere).

Titrimetric Method

Match the specific microorganism to the vitamin it is used to assay:

1. Lactobacillus casei

2. Lactobacillus leichmannii

1. Lactobacillus casei → B9 (Folic acid)

2. Lactobacillus leichmannii → B12 (Cobalamin)

This vitamin analysis method is sensitive, reflects biological availability, but time-consuming and less precise than modern methods.

Microbiological assays

It is used to quantify certain vitamins by measuring their characteristic absorption of light within the ultraviolet or visible spectrum.

UV-Vis Spectrophotometer

At what specific wavelength is Vitamin A typically measured using UV-Vis spectrophotometry?

325 nm

Beyond standard absorption, fluorescence of this vitamin is used for its spectrophotometric quantification.

B2 (Riboflavin)

UV-Vis Spectrophometry is applicable for these vitamins.

• A → 325 nm absorption

• B2 (RIboflavin) → Fluorescence

• C

This chromatographic method is widely used for most vitamins, both fat- and water-soluble.

HPLC

This separates vitamins using different stationary phases and its detection is through UV, fluorescence, or MS.

HPLC

What specific type of HPLC is used for the analysis of B-complex vitamins?

Reversed-phase HPLC

What is the standard detection method and specific wavelength used for Vitamin C in HPLC?

UV detection (245 nm)

Identify the detection type for each:

1. Vitamin C

2. B-complex

3. Fat-soluble (A,D,E,K)

1. Vitamin C: UV at 245 nm.

2. B-complex: Reversed-phase.

3. Fat-soluble: Fluorescence or MS.

This vitamin analysis is highly sensitive, specific, and QUANTITATIVE, but it requires expensive equipment and skilled operation.

HPLC

This vitamin analysis is suitable for volatile and thermally stable derivatives of vitamins.

Gas Chromatography

This vitamin analysis is not widely used due to the non-volatile nature of many vitamins.

Gas Chromatography

This vitamin analysis is suitable for multivitamin analysis (mainly water-soluble).

LC-MC/MS (Liquid Chromatography-Tandem Mass Spectrometry)

This vitamin analysis is ultra-sensitive and has simultaneous multivitamin detection.

LC-MC/MS (Liquid Chromatography-Tandem Mass Spectrometry)

This vitamin analysis is very costly and has complex sample preparation.

LC-MC/MS (Liquid Chromatography-Tandem Mass Spectrometry)

This vitamin analysis is mainly suitable for water-soluble vitamins (B-complex).

Capillary electrophoresis

This vitamin analysis is based on the separation by charge and size in an electric field.

Capillary electrophoresis

This vitamin analysis is fast and has low solvent usage, but it is less robust and has lower reproducibility than HPLC.

Capillary electrophoresis

This vitamin analysis method is suitable for vitamin C and some B vitamins.

Biosensors or immunoassays

This vitamin analysis is based on the enzyme or antibody detection.

Biosensors or immunoassays

This vitamin method is rapid, portable, and has low cost potential, but it is still developing and has limited availability.

Biosensors or immunoassays

This physically blocks UV and visible light and prevents photodegradation (light destruction), which is strictly necessary for highly light-sensitive vitamins like Riboflavin (B2) and Vitamin A.

amber bottles

This vitamin is extracted through cold extraction using metaphosphoric/acetic acid.

Vitamin C

Vitamin C is extracted through __.

Cold extraction using metaphosphoric/acetic acid.

These vitamins are extracted through boiling/autoclaving in acid, followed by further acid treatment.

• B1 (Thiamin)

• B2 (Riboflavin)

Vitamins B1 (Thiamin) and B2 (Riboflavin) are extracted through _.

boiling/autoclaving in acid, followed by further acid treatment

This vitamin is extracted through autoclaving in acid (for non-cereals) or autoclaving in base (for cereals).

B3 (Niacin)