19-Hydroxyprostaglandin B2 as an internal standard for on-line extraction-high-performance liquid chromatography analysis of lipoxygenase products.
Variable recovery of polar compounds is a potential pitfall in the on-line extraction-reversed-phase HPLC analysis of lipoxygenase products. Therefore, the addition of a polar internal standard to biological samples, which permits the assessment of the recovery of the polar components in individual samples, appears to be essential. 19-Hydroxyprostaglandin (PG) B2 was found to be a suitable compound for this application. It was easily prepared from deproteinized human semen by base-catalyzed dehydration of 19-hydroxy-PGE2, and purified in a single step by reversed-phase HPLC. Similarly to PGB2, already used as internal in HPLC, 19-hydroxy-PGB2 has excellent chemical stability and carries a strong uv chromophore that enables detection at 280 nm. Most importantly, it is eluted just prior to the polar arachidonic acid metabolites 20-hydroxy- and 20-carboxy-leukotriene B4, 12-oxo-dodecatrienoic acid and the lipoxines A and B. The measurement of the ratio of PGB2 and 19-hydroxy-PGB2 used in combination as internal standards in HPLC analysis of lipoxygenase products, provides a simple and reliable way to assess sample to sample recovery of the polar components when using on-line extraction procedures.
Borgeat
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P
Inflammation and Immunology-Rheumatology Research Unit, Le Centre Hospitalier de l'Université Laval, Sainte-Foy, Quebec, Canada.
Picard
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S
D000001:Calcimycin / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D006893:Hydroxyeicosatetraenoic Acids / Q000032:analysis; D007962:Leukocytes / Q000201:enzymology; D007975:Leukotriene B4 / Q000032:analysis; D044045:Lipoxins; D008084:Lipoxygenase / Q000378:metabolism*; D011453:Prostaglandins / Q000032:analysis*; D011456:Prostaglandins B / Q000032:analysis*
D006893:Hydroxyeicosatetraenoic Acids; D044045:Lipoxins; D011453:Prostaglandins; D011456:Prostaglandins B; C040527:lipoxin A4; D007975:Leukotriene B4; D000001:Calcimycin; C056827:19-hydroxyprostaglandin B2; C042651:lipoxin B4; D008084:Lipoxygenase