Estimation of amniotic fluid phospholipids by high-performance liquid chromatography

Area of Science:

• Clinical Chemistry
• Biochemistry
• Analytical Chemistry

Background:

• Amniotic fluid phospholipids are crucial biomarkers for predicting fetal lung maturity.
• Accurate and efficient analytical methods are needed for routine clinical assessment.

Purpose of the Study:

• To develop and validate a high-performance liquid chromatography (HPLC) method for analyzing key amniotic fluid phospholipids.
• To compare the performance of the developed HPLC method against the standard thin-layer chromatography (TLC) technique.

Main Methods:

• A novel HPLC method was established using a DIOL or CN column and a variable-wavelength detector.
• The method involved extracting phospholipids from amniotic fluid with an internal standard.
• Analysis was performed on 40 amniotic fluid samples.

Main Results:

• The HPLC method demonstrated good agreement with TLC for lecithin/sphingomyelin ratio, phosphatidyl glycerol, and phosphatidyl inositol.
• High correlation coefficients were observed: rPG=0.94, rPI=0.95, rL/S=0.97.
• The HPLC procedure provided selective separation of multiple phospholipids simultaneously.

Conclusions:

• The developed HPLC method is a rapid, selective, and accurate tool for quantifying amniotic fluid phospholipids.
• This HPLC technique facilitates precise estimation of individual phospholipid concentrations, aiding in fetal lung maturity assessment.
• HPLC offers significant advantages over TLC in terms of speed and analytical capability.