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Home / Applications / High Speed Analysis of Fat-Soluble Vitamins by Ultra High-performance Liquid Chromatography with Photodiode Array Detection

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High Speed Analysis of Fat-Soluble Vitamins by Ultra High-performance Liquid Chromatography with Photodiode Array Detection

By Heather Haffner

PDF IconDownload This Application

August 24, 2022

Introduction

JASCO UHPLC System
JASCO UHPLC System

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.

Fig. 1. Chromatogram of Fat-soluble Vtamin Standard Mixture

The on-peak spectra of fat-soluble vitamin standard mixture are shown in Fig. 2.

Fig. 2. On-peak Spectra of Fat-soluble Vitamin Standard Mixture
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)
This document has been prepared based on information available at the time of publication and is subject to revision without notice. Although the contents are checked with the utmost care, we do not guarantee their accuracy or completeness. JASCO Corporation assumes no responsibility or liability for any loss or damage incurred as a result of the use of any information contained in this document. Copyright and other intellectual property rights in this document remain the property of JASCO Corporation. Please do not attempt to copy, modify, redistribute, or sell etc. in whole or in part without prior written permission.

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About the Author

JASCO Application Note

High Speed Analysis of Fat-Soluble Vitamins by Ultra High-performance Liquid Chromatography with Photodiode Array Detection

Introduction

JASCO UHPLC System
JASCO UHPLC System

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

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.

Fig. 1. Chromatogram of Fat-soluble Vtamin Standard Mixture

The on-peak spectra of fat-soluble vitamin standard mixture are shown in Fig. 2.

Fig. 2. On-peak Spectra of Fat-soluble Vitamin Standard Mixture
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)

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

This document has been prepared based on information available at the time of publication and is subject to revision without notice. Although the contents are checked with the utmost care, we do not guarantee their accuracy or completeness. JASCO Corporation assumes no responsibility or liability for any loss or damage incurred as a result of the use of any information contained in this document. Copyright and other intellectual property rights in this document remain the property of JASCO Corporation. Please do not attempt to copy, modify, redistribute, or sell etc. in whole or in part without prior written permission.
28600 Mary’s Court, Easton, MD 21601 USA • (800) 333-5272 • Fax: (410) 822-7526 • jascoinc.com/applications

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