A high pressure liquid chromatography method for separation of prolactin forms

Damon A. Bell, Kirsten Hoad, Lillian Leong, Juwaini Abu Bakar, Paul Sheehan, Samuel D. Vasikaran

    Research output: Contribution to journalArticle

    3 Citations (Scopus)

    Abstract

    Background: Prolactin has multiple forms and macroprolactin, which is thought not to be bioavailable, can cause a raised serum prolactin concentration. Gel filtration chromatography (GFC) is currently the gold standard method for separating macroprolactin, but is labour-intensive. Polyethylene glycol (PEG) precipitation is suitable for routine use but may not always be accurate. We developed a high pressure liquid chromatography (HPLC) assay for macroprolactin measurement. Methods: Chromatography was carried out using an Agilent Zorbax GF-250 (9.4 × 250 mm, 4 μ1/2m) size exclusion column and 50 mmol/L Tris buffer with 0.15 mmol/L NaCl at pH 7.2 as mobile phase, with a flow rate of 1 mL/min. Serum or plasma was diluted 1:1 with mobile phase and filtered and 100 μ1/2L injected. Fractions of 155 μ1/2L were collected for prolactin measurement and elution profile plotted. The area under the curve of each prolactin peak was calculated to quantify each prolactin form, and compared with GFC. Results: Clear separation of monomeric-, big- and macroprolactin forms was achieved. Quantification was comparable to GFC and precision was acceptable. Total time from injection to collection of the final fraction was 16 min. Conclusions: We have developed an HPLC method for quantification of macroprolactin, which is rapid and easy to perform and therefore can be used for routine measurement.

    Original languageEnglish
    Pages (from-to)285-288
    Number of pages4
    JournalAnnals of Clinical Biochemistry
    Volume49
    Issue number3
    DOIs
    Publication statusPublished - May 2012

    Fingerprint

    High pressure liquid chromatography
    Prolactin
    Chromatography
    High Pressure Liquid Chromatography
    Gel Chromatography
    Gels
    Tromethamine
    Serum
    Area Under Curve
    Assays
    Flow rate
    polymeric prolactin
    Personnel
    Plasmas
    Injections

    ASJC Scopus subject areas

    • Clinical Biochemistry

    Cite this

    Bell, D. A., Hoad, K., Leong, L., Bakar, J. A., Sheehan, P., & Vasikaran, S. D. (2012). A high pressure liquid chromatography method for separation of prolactin forms. Annals of Clinical Biochemistry, 49(3), 285-288. https://doi.org/10.1258/acb.2011.011209

    A high pressure liquid chromatography method for separation of prolactin forms. / Bell, Damon A.; Hoad, Kirsten; Leong, Lillian; Bakar, Juwaini Abu; Sheehan, Paul; Vasikaran, Samuel D.

    In: Annals of Clinical Biochemistry, Vol. 49, No. 3, 05.2012, p. 285-288.

    Research output: Contribution to journalArticle

    Bell, DA, Hoad, K, Leong, L, Bakar, JA, Sheehan, P & Vasikaran, SD 2012, 'A high pressure liquid chromatography method for separation of prolactin forms', Annals of Clinical Biochemistry, vol. 49, no. 3, pp. 285-288. https://doi.org/10.1258/acb.2011.011209
    Bell, Damon A. ; Hoad, Kirsten ; Leong, Lillian ; Bakar, Juwaini Abu ; Sheehan, Paul ; Vasikaran, Samuel D. / A high pressure liquid chromatography method for separation of prolactin forms. In: Annals of Clinical Biochemistry. 2012 ; Vol. 49, No. 3. pp. 285-288.
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