Add time:07/31/2019         Source:sciencedirect.com

A method for the quantitative determination of 4-hydroxy-3-methoxyphenyl glycol and its conjugates in human cerebrospinal fluid has been developed. Unconjugated 4-hydroxy-3-methoxyphenyl glycol and its conjugates after hydrolysis (81% efficiency) were reacted with trifluoroacetic anhydride. The derivative containing three trifluoroacetyl groups was analyzed by mass fragmentography. 4-Hydroxy-3-methoxyphenyl glycol labelled with two deuterium atoms was synthesized and used as internal standard. The molecular ions m/e 472 and 474 in the mass spectra were used for monitoring 4-hydroxy-3-methoxyphenyl glycol and labelled 4-hydroxy-3-methoxyphenyl glycol, respectively. This sensitive and specific method allows the quantitative determination of unconjugated and conjugated 4-hydroxy-3-methoxyphenyl glycol in 2 ml of the same cerebrospinal fluid specimen in concentrations down to 1 ng/ml. The standard deviation for the determination of unconjugated 4-hydroxy-3-methoxyphenyl glycol was ± 7% in the concentration range 2–36 ng/ml. Synthetic 4-hydroxy-3-methoxyphenyl glycol sulphate added to cerebrospinal fluid (15 ng/ml) was recovered in a yield of 99.3 ± 8.0%.

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