A high pressure liquid chromatography method for separation of prolactin forms

Area of Science:

• Biochemistry
• Analytical Chemistry
• Clinical Diagnostics

Background:

• Macroprolactin, a non-bioavailable form of prolactin, can elevate serum prolactin levels.
• Current methods like gel filtration chromatography (GFC) are labor-intensive, and polyethylene glycol (PEG) precipitation may lack accuracy.
• There is a need for a more efficient and reliable method for macroprolactin measurement.

Purpose of the Study:

• To develop and validate a high-performance liquid chromatography (HPLC) assay for the accurate quantification of macroprolactin.
• To establish an alternative method that is faster and easier to perform than existing techniques.

Main Methods:

• Serum or plasma samples were analyzed using an Agilent Zorbax GF-250 size exclusion column.
• A Tris buffer mobile phase was employed at a flow rate of 1 mL/min.
• Fractions were collected for prolactin measurement, and elution profiles were analyzed to quantify prolactin forms.

Main Results:

• The HPLC method achieved clear separation of monomeric, big, and macroprolactin forms.
• Quantification results were comparable to the gold standard GFC method with acceptable precision.
• The entire assay, from injection to fraction collection, was completed in 16 minutes.

Conclusions:

• A rapid and user-friendly HPLC method for macroprolactin quantification has been successfully developed.
• This HPLC assay is suitable for routine clinical laboratory use, offering improved efficiency.
• The method provides accurate and precise measurement of different prolactin forms.