How to analyze Vitamin B6 with C18 HPLC column?

Vitamin B6 is the B group of vitamins, this product is 6-methyl-5-hydroxy-3, 4-pyridoxine dimethylol hydrochloride, also known as pyridoxine, was discovered by Paul Gyorgy in 1934, including pyridoxine, pyridoxal, and pyridoxine, mostly found in yeast, cereals, liver, eggs, dairy products.

Vitamin B6 exists in the body in the form of phosphate ester, which is a water-soluble vitamin. It is a component of some coenzymes in the human body and is involved in a variety of metabolic reactions, especially closely related to the metabolism of amino acids. It mainly ACTS on the blood, muscles, nerves, and skin of the human body. Its functions include the synthesis of antibodies, the production of gastric acid in the digestive system, the use of fat and protein (especially when losing weight), and the maintenance of sodium/potassium balance (stabilizing the nervous system). Lack of vitamin B6 general disease, general lack of appetite, the food utilization rate is low, weightlessness, vomiting, dysentery and other problems. Severe deficiency can lead to acne, anemia, arthritis, child cramps, depression, headaches, hair loss, inflammation, learning disabilities, and weakness.

Vitamin B6 Test Method (Chinese pharmacopeia 2015, Part Ⅱ)

Due to the large polarity of vitamin B6, the retention of vitamin B6 on the reversed-phase chromatography column is weak. By ion-pair chromatography, sodium pentane sulfonate is added in the mobile phase to enhance the retention of vitamin B6

Test Condition

  • Column: Galaksil C18 5um, 4.6×250mm
  • Mobile phase: 10/90 v/v MeOH; 0.04% Sodium pentane sulfonate water solvent (pH3.0)
  • Flow rate: 1 mL/min
  • Temperature: 30℃
    Injection: 10μL
  • Detection: UV 291 nm
  • Peaks: 1. Vitamin B6; 2. Vitamin B6 Impurity A; 3. Vitamin be6 Impurity B
Vitamin b6 hplc test

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