High Speed Analysis of Fat-Soluble Vitamins by Ultra High-performance Liquid Chromatography with Photodiode Array Detection
August 24, 2022
Introduction
![JASCO UHPLC System](https://jascoinc.com/wp-content/uploads/2017/03/Jasco-XLC-system.jpg)
Vitamins are essential nutrients to all living organisms and are organic compounds required in small amounts for healthy physiological activity. The class of fat-soluble vitamins is insoluble in water but readily dissolve in oil and thus when consumed in excess may have a detrimental impact on the human body, as they are difficult to excrete.
This application note describes the analysis of fat-soluble vitamins using UHPLC and PDA detection with a high speed data sampling rate of 100 spectra/sec.
Experimental
Equipment
Pump: | X-LC 3185PU x 2 |
Degasser: | X-LC 3080DG |
Mixer: | X-LC 3180MX |
Column Oven: | X-LC 3067CO |
Autosampler: | X-LC 3159AS |
Detector: | X-LC 3110MD |
Conditions
Column: | ZORBAX SB-C18 (3.0 mmID x 50 mmL, 1.8 µm) |
Eluent: | Acetonitrile/Methanol (70/30) |
Flow Rate: | 0.8 mL/min |
Column Temp.: | 30 ºC |
Wave Length: | 200-500 nm |
Injection Volume: | 1 µL |
Standard Sample: | 10 fat-soluble vitamin standards |
Keywords
UHPLC, Fat-soluble vitamin, Vitamin K3, Vitamin A, Retinol acetate, Vitamin K2, d-tocopherol, Vitamin D2, Vitamin D3, a-tocopherol, Tocopherol-a-acetate, Vitamin K1, C18 Column, PDF detector
Results
A contour plot and chromatogram of the fat-soluble vitamin standard mixture are shown in Figure 1. Excellent separation of the 10 vitamins was obtained in under 4 minutes.
![](https://jascoinc.com/wp-content/uploads/2013/10/Fig-1-chromatogram-of-fat-soluble.jpg)
The on-peak spectra of fat-soluble vitamin standard mixture are shown in Fig. 2.
![](https://jascoinc.com/wp-content/uploads/2013/10/Fig-2-on-peak-spectra.jpg)
1: Vitamin K3 (Menadione) | 5: Vitamin E ( -Tocopherol) | 9: Vitamin E acetate |
2:Vitamin A (Retinol) | 6: Vitamin D2 (Ergocalciferol) | 10: Vitamin K1 (Phytonadione) (0.1 mg/mL each) |
3: Vitamin A Acetate (Retinol acetate) | 7: Vitamin D3 (Cholecalciferol) | |
4: Vitamin K2 (Menaquinone) | 8: Vitamin E ( -Tocopherol) |
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![JASCO Application Note](https://jascoinc.com/wp-content/uploads/2023/10/app-pdf-header-2.png)
High Speed Analysis of Fat-Soluble Vitamins by Ultra High-performance Liquid Chromatography with Photodiode Array Detection
Introduction
![JASCO UHPLC System](https://jascoinc.com/wp-content/uploads/2017/03/Jasco-XLC-system.jpg)
Vitamins are essential nutrients to all living organisms and are organic compounds required in small amounts for healthy physiological activity. The class of fat-soluble vitamins is insoluble in water but readily dissolve in oil and thus when consumed in excess may have a detrimental impact on the human body, as they are difficult to excrete.
This application note describes the analysis of fat-soluble vitamins using UHPLC and PDA detection with a high speed data sampling rate of 100 spectra/sec.
Experimental
Equipment
Pump: | X-LC 3185PU x 2 |
Degasser: | X-LC 3080DG |
Mixer: | X-LC 3180MX |
Column Oven: | X-LC 3067CO |
Autosampler: | X-LC 3159AS |
Detector: | X-LC 3110MD |
Conditions
Column: | ZORBAX SB-C18 (3.0 mmID x 50 mmL, 1.8 µm) |
Eluent: | Acetonitrile/Methanol (70/30) |
Flow Rate: | 0.8 mL/min |
Column Temp.: | 30 ºC |
Wave Length: | 200-500 nm |
Injection Volume: | 1 µL |
Standard Sample: | 10 fat-soluble vitamin standards |
Keywords
UHPLC, Fat-soluble vitamin, Vitamin K3, Vitamin A, Retinol acetate, Vitamin K2, d-tocopherol, Vitamin D2, Vitamin D3, a-tocopherol, Tocopherol-a-acetate, Vitamin K1, C18 Column, PDF detector
Results
A contour plot and chromatogram of the fat-soluble vitamin standard mixture are shown in Figure 1. Excellent separation of the 10 vitamins was obtained in under 4 minutes.
![](https://jascoinc.com/wp-content/uploads/2013/10/Fig-1-chromatogram-of-fat-soluble.jpg)
The on-peak spectra of fat-soluble vitamin standard mixture are shown in Fig. 2.
![](https://jascoinc.com/wp-content/uploads/2013/10/Fig-2-on-peak-spectra.jpg)
1: Vitamin K3 (Menadione) | 5: Vitamin E ( -Tocopherol) | 9: Vitamin E acetate |
2:Vitamin A (Retinol) | 6: Vitamin D2 (Ergocalciferol) | 10: Vitamin K1 (Phytonadione) (0.1 mg/mL each) |
3: Vitamin A Acetate (Retinol acetate) | 7: Vitamin D3 (Cholecalciferol) | |
4: Vitamin K2 (Menaquinone) | 8: Vitamin E ( -Tocopherol) |