Lab Glassware Division 中文(Chinese)

Simultaneous Analysis of 6 Water-soluble Vitamins on a HILIC Column

Method: HPLC
Matrix: Medicine
Application No.: 101970
Componds: Ascorbic acid, Pyridoxine hydrochloride, Nicotinamide, Thiamine hydrochloride, Folic acid, Riboflavin sodium phosphate
Columns: Inspire™ HILIC 3 μm 150 x 4.6 mm
Cat No.: 81418
Sample Pretreatment: Stock solution: Most of vitamins are prepared in water at the concentration of 10mg/mL. Stock solutions of folic acid and riboflavin are prepared in 0.1mol/L NaOH at the concentration of 10mg/mL.
Working solution: Working solution are prepared by diluting stock solution with water: acetonitrile (1:4).
Conditions: Mobile Phase:
A: 20 mM ammonium dihydrogen phosphate, pH 3.0
B: acetonitrile
Gradient: hold 17% A for 3.1 min, 17 to 60% A in 1.9 min, 60 to 80% A in 6 min, 80 to 17% A in 1 min, equilibrate at 17% A for 5 min.
Flow Rate: 1.0 mL/min
Injection: 2 µL
Temperature: 35 ℃
Wavelength: 260 nm
Publisher: Dikma Technologies Inc.
Keyword: Ascorbic acid, Pyridoxine hydrochloride, Nicotinamide, Thiamine hydrochloride, Folic acid, Riboflavin sodium phosphate, HPLC, Inspire HILIC C18
Abstract: In this application note, a DIKMA Inspire HILIC 3 µm C18 column was used for establishment of an analysis method for simultaneous determination of 6 vitamins. These vitamins were well separated and eluted from the column within 10 min.

 

No.

Compounds

Concentration 

(µg/mL)

Retention Time

(min)

Resolution

Asymmetry

1

Ascorbic acid

40

2.765

-

1.13

2

Pyridoxine hydrochloride

40

3.093

3.17

1.11

3

Nicotinamide

40

3.450

3.21

1.15

4

Thiamine hydrochloride

40

4.181

5.46

1.20

5

Folic acid

40

6.740

20.82

1.05

6

Riboflavin sodium phosphate

40

9.334

22.46

1.38