Improvement and validation of ¹²⁵I-high-performance liquid chromatography method for determination of total human serum choline and ethanolamine plasmalogens.

Abstract

BACKGROUND:

Serum plasmalogens (Pls) have gained interest in several clinical symptoms such as metabolic syndrome/atherosclerosis or Alzheimer's disease possibly because of their antioxidant properties. We have developed a highly sensitive and simple method to determine plasmenylcholine (PlsCho; choline plasmalogen) and plasmenylethanolamine (PlsEtn; ethanolamine plasmalogen) separately, using a radioactive iodine and high-performance liquid chromatography ((125)I-HPLC method). The present study reports the improvement and validation of (125)I-HPLC method by introducing a quantitative standard (QS) and online detection with a flow γ-counter.

METHODS:

1-Alkenyl 2,3-cyclic glycerophosphate was prepared as QS from l-α-lyso plasmenylcholine by enzymatic treatment with phospholipase D. Online detection with a flow γ-counter was investigated to be available to quantify Pls. The method validation was carried out in terms of selectivity, sensitivity, linearity, precision, accuracy and recovery.

RESULTS:

Linearity was established over the concentration range 5-300 μmol/L for Pls and QS with regression coefficients >0.99. The accuracy and reliability were satisfactory. The method has been applied to the determination of human serum Pls from healthy subjects and the elderly with dementia or artery stenoses.

CONCLUSIONS:

The improved (125)I-HPLC method is useful as an autoanalytical system for a routine diagnostic test of human serum Pls.

• Authors: Ryouta Maeba, Yuya Yamazaki, Toru Nezu, Tomoki Okazaki
• Journal: Annals of clinical biochemistry (Ann Clin Biochem) Vol. 49 Issue Pt 1 Pg. 86-93 (Jan 2012) ISSN: 1758-1001 [Electronic] England
• PMID: 22042981 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Validation Study)

Chemical References

• Glycerophosphates
• Iodine Radioisotopes
• Plasmalogens
• choline plasmalogens
• phosphatidal ethanolamines
• Phospholipase D

Topics

• Aged
• Aged, 80 and over
• Animals
• Calibration
• Case-Control Studies
• Cattle
• Chromatography, High Pressure Liquid (methods)
• Coronary Stenosis (blood)
• Dementia (blood)
• Female
• Gamma Rays
• Glycerophosphates (biosynthesis)
• Humans
• Iodine Radioisotopes
• Male
• Phospholipase D (metabolism)
• Plasmalogens (blood)
• Reference Standards
• Reproducibility of Results
• Sensitivity and Specificity