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Determination of 4 Vitamines Using UHPLC/MS/MS Method

Method: HPLC-MS
Matrix: Medicine
Application No.: 101931
Componds: L-Ascorbic acid, Pyridoxal hydrochloride, Folic acid, Riboflavin
Columns: Spursil C18 3 μm 100 x 2.1 mm
Cat No.: 82012
Sample Pretreatment: Standards: Individual stock solutions of tested compounds were prepared in different solutions at a concentration of 1 mg/mL.L-Ascorbic acid and pyridoxal hydrochloride stock standard solutions were prepared by water. Riboflavin stock standard solution was prepared by 3 M hydrochloric acid solution. Folic acid was dissolved by a mixed solution of 0.02 M sodium hydroxide solution and ethanol (1:1 v/v). A working solution of the mixture of all 4 compounds was obtained by fresh dilution of the stock solution with mobile phase.
Conditions: Mobile Phase:
A: 10 mM ammonium formate, pH 2.2
B: methanol:acetonitrile = 4:6
Gradient: hold 100% A for 3.5 min, 100 to 20% A in 3.5 min, hold 20% A for 1 min, 20 to 100% A in 1 min, equilibrate at 100% A for 6 min.
Flow Rate: 0.25 mL/min;
Injection: 2 µL;
Temperature: 25 ℃
Source: ESI; Ionization Mode: Positive
Publisher: Dikma Technologies Inc.
Keyword: L-Ascorbic acid, Pyridoxal hydrochloride, Folic acid, Riboflavin, Vitamines
Abstract: These vitamins were well separated and eluted from the column within 7.5 min. This method is simple, effective, and can be applied to vitamin detection in a broad set of fields.

 

No.

Compounds

Concentration (µg/mL)

Retention Time (min)

MS→MS1 Ions

(Precursor→Product 1)

MS→MS2 Ions

(Precursor→Product 2)

1

L-Ascorbic acid

17

1.33

177.1→141.0

177.1→95.1

2

Pyridoxal hydrochloride

0.1

2.43

168.1→150.0

168.1→122.2

3

Folic acid

3.0

6.76

442.2→295.1

442.2→176.1

4

Riboflavin

1.0

7.13

377.2→243.0

377.2→172.1