HPLC Determination of Vitamin K1in Neonatal Plasma Following Oral or Parenteral Supplementation with Konakion

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Abstract
A rapid high-performance liquid chromatographic method for the determination of Vitamin K1(20) in plasma after a single hexane extraction is described. A ternary mobile phase consisting of acetonitrile/propan-2-o1/dichloromethane (68.5/22.2/9.3, v/v) with a flow rate of 1.8 m1/min was used in combination with a Waters Associates Z Module RCSS containing a Nova-Pak C18 Radial-Pak cartridge to provide separation from co-extracted UV absorbing contaminants. The analytical column was protected by a Waters Associates Guard-Pak precolumn module with a Guard-Pak μBondapak Cig cartridge. Using only 250μl of sample, plasma levels in trie region of 15–20 ng/ml for Vitamin Kw1(25), can be determined using UV detection at 270 nm. Vitamin K1 (25), a synthetic homologue of K1 (20) was used as internal standard. The method has been developed for measuring plasma levels in neonates after supplementation with Vitamin K1 (20)