Journal of Food Measurement and Characterization

, Volume 13, Issue 3, pp 1947–1957 | Cite as

Simultaneous analysis of vitamin D and K in processed food products via ultra high- performance liquid chromatography (UHPLC)

  • Devi-Nair RathiEmail author
  • Mohd Fairulnizal Md Noh
  • Aswir Abd Rashed
  • Isameyah Dasuki
Original Paper


This study highlights a simplified extraction and method development of Ultra High-Performance Liquid Chromatography for simultaneous determination of vitamin D and K in selected processed food products. Simultaneous analysis of vitamin D and K was achieved via gradient elution of methanol with ultra-pure water (95:5–100:0–95:5), at programmed flow rate of (1.0–0.2–1.0 mL/min) for left pump and constant 1 mL/min for right pump within 25 min on HPLC Dionex Ultimate 3000 Series system using two Dionex Acclaim Polar Advantage II C18 columns at 30 °C and acquisition wavelength of 265 nm. The limits of detection and quantification ranged between 0.05 and 0.12 µg/mL and 0.16–0.41 µg/mL; for both vitamins. The method showed good linearity (R2: 0.996–0.999) and high accuracy (85.35–106.36%) for both vitamin D and K. Recovery above 70% was achieved for vitamin D, whereas lower recovery of 60–70% was obtained for vitamin K. The proposed analytical and extraction procedure was advantageous for sample purification and improved detection of trace vitamin levels from selected cereal and flour products.


Vitamin D Vitamin K On-Line solid phase extraction Ultra High Performance Liquid Chromatography 



The authors wish to thank the Director General of Health Malaysia and the Director of Institute for Medical Research (IMR) for granting the permission to publish this study. We would also like to express our gratitude to the staff of Nutrition Unit for their valuable support. This project was funded by a grant from the Ministry of Health Research Grant.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Nutrition Unit, Cardiovascular, Diabetes and Nutrition Research Centre, Institute for Medical ResearchMinistry of HealthKuala LumpurMalaysia

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