Determination of 25-Hydroxycholecalciferol Content in Egg Yolk by HPLC

✍ Scribed by Pirjo Mattila; Vieno Piironen; Esko Uusi-Rauva; Pekka Koivistoinen

• Publisher: Elsevier Science
• Year: 1993
• Tongue: English
• Weight: 295 KB
• Volume: 6
• Category: Article
• ISSN: 0889-1575
• DOI: 10.1006/jfca.1993.1027

✦ Synopsis

A high-performance liquid chromatographic (HPLC) procedure for determining 25 -hydroxycholecalciferol contents in chicken egg yolk has been developed. The procedure consists of the following steps: saponification. extraction of the unsaponifiable fraction, purification using solidphase extraction, clean-up on straight-phase HPLC, and quantification with reverse-phase HPLC using an internal standard ( (25-\mathrm{OH}-\mathrm{D}{2}) ) method and diode array detection. The suitability of the method was tested on a pooled egg-yolk sample. The overall mean recovery for (25-\mathrm{OH}-\mathrm{D}{2}) was (71 \pm 5.4 %) and for (25-\mathrm{OH}-\mathrm{D}{3}) was (71 \pm 3.2 %). The recovery of (25-\mathrm{OH}-\mathrm{D}{3}), calculated on the basis of the internal standard, was (98 \pm 4.6 %). The detection and determination limits of the method were 2 and (8 \mathrm{ng} /) injection, respectively. The pooled egg yolk contained (0.98 \pm 0.069 \mu \mathrm{g}) of (25-) (\mathrm{OH}-\mathrm{D}{3} / 100 \mathrm{~g}) of egg yolk. A simultaneous determination of the vitamin (D{3}) contents in yolks is also possible with the method described. (c 1993) Academic Press. Inc.