Derivatization to improve thermospray HPLC/MS sensitivity for the determination of prostaglandins and thromboxane B2

Derivatization of prostaglandins and thromboxane B2 using diethyl amino ethyl chloride and diethyl amino ethyl bromide improved thermospray HPLC/MS sensitivity. The derivatization was evaluated for prostaglandin A1, A2, D2, E2, F1 alpha, F2 alpha, and thromboxane B2. The derivatization reaction, thermospray operating conditions, and mode of detection were optimized to produce the most intense [M + H]+ or [M - H]- ions for the derivative. Derivatization was better than 99% complete in 1 hour at 75 degrees C. No thermal degradation of the prostaglandins was observed. Positive thermospray ionization proved the mode most sensitive, enabling detection from 10 to 300 pg of each prostaglandin under multiple ion detection. The thermospray spectra exhibited intense [M + H]+ ions for the derivative with a few fragment ions from sequential losses of water from the [M + H]+ ion. Detection of a prostaglandin metabolite in plasma over the concentration range from 3 ppm to 30 ppb was possible using this derivatization.