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Article

An UFLC-DAD Method for the Quantification of Menaquinone-4 in Spiked Rabbit Plasma

Karthika Anoop1, Gullapalli Kowmudi1, Mukkamala Sailaja1, Krishna Veni Nagappan1★, Nagarajan Janaki Sankarachari Krishnan1, Ramalingam Peraman2★ Corresponding author

  1. 1Department of Pharmaceutical Analysis JSS College of Pharmacy, Ooty [A Constituent College – JSS Academy of Higher Education & Research], Rocklands, Ooty, Tamil Nadu, INDIA.
  2. 2Department of Medical Chemistry, Raghavendra Institute of Pharmaceutical Education and Research, Chiyyedu, Anantapur, Andhra Pradesh, INDIA.

CORRESPONDENCE

Krishna Veni Nagappan

Department of Pharmaceutical Analysis JSS College of Pharmacy, Ooty [A Constituent College – JSS Academy of Higher Education & Research], Rocklands, Ooty, Tamil Nadu, INDIA.

krisath@jssuni.edu.in

Received: 04-12-2018; Revised: 21-02-2019; Accepted: 01-03-2019.

Volume 11, Issue 2 · pp. 149–154 · PUBLISHED · DOI: JYoungPharm-11-2-149

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ABSTRACT

Objective:The present study is aimed to develop and validate an Ultra-Fast Liquid Chromatography-Diode Array Detector (UFLC-DAD) method for the quantification of vitamin K2 as Menaquinone-4 (MK-4) in rabbit plasma. Method: Standard solutions and spiked plasma samples of MK-4 and Internal Standard (IS) were prepared from primary stock solutions of 1mg/ ml concentration in ethanol each. Protein precipitation was carried out for the MK-4 and IS extraction from plasma spiked samples. Chromatographic separation was employed using Isopropyl Alcohol and Acetonitrile (50:50 v/v) as mobile phase and a C-18 column with 1ml/min flow rate and a run time of 10 mins. Detection was carried out in the range 190-600 nm with 269 nm set as a reference wavelength. Result:The retention times of MK-4 and IS were at 5.5 ± 0.5 mins and 8 ± 0.5 mins respectively. Calibration curve for MK-4 was found to be linear in the range of 0.374 to 6 µg/ ml with an R2 value of 0.9934. The % RSD for accuracy was <15%, inter and intraday precisions were <10%. The samples were found to be stable throughout the study. Conclusion:This method can be applied to the estimation of MK-4 in rabbit plasma using UFLC-DAD.

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    Anoop, K., Kowmudi, G., Sailaja, M., Nagappan, K. V., Krishnan, N. J. S., & Peraman, R. (). An UFLC-DAD Method for the Quantification of Menaquinone-4 in Spiked Rabbit Plasma. Journal of Young Pharmacists, 11(2), 149–154. https://doi.org/JYoungPharm-11-2-149