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19-Hydroxyprostaglandin B2 as an Internal Standard for On-Line Extraction-High-Performance Liquid Chromatography Analysis of Lipoxygenase Products

19-Hydroxyprostaglandin B2 as an Internal Standard for On-Line Extraction-High-Performance Liquid Chromatography Analysis of Lipoxygenase Products

P Borgeat, S Picard

Anal Biochem. 1988 Jun;171(2):283-9.

PMID: 2841889

Abstract:

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP80434828	20-Carboxy-leukotriene B4		80434-82-8	Price

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