Takasu A, Ohya K, Sano M
J Chromatogr. 1983 Feb 11;272(2):243-50.
A selected ion monitoring (SIM) method has been devised for the determination of metabolites of dydrogesterone, 20 alpha-hydroxy-9 beta,10 alpha-pregna-4,6-dien-3-one (DHD) and DHD glucuronide, in plasma. Using testosterone as an internal standard (IS), DHD and IS were extracted with n-hexane and were purified by means of magnesium oxide column chromatography. The purified DHD and IS were converted to their diheptafluorobutyryl derivatives (DHD diHFB and testosterone diHFB) with heptafluorobutyric anhydride in acetone for analysis by SIM. SIM was carried out with a 2% OV-17 column (1 m) at 230 degrees C by monitoring the molecular ions of the derivatives (m/z 706 for DHD diHFB, m/z 680 for testosterone diHFB). DHD was determined from a calibration curve using a peak area method. The determination limit of the devised method was about 5 ng DHD per ml of plasma and the reproducibility was within +/- 6% of the coefficient of variation for 30 ng of DHD per ml of plasma or above.
已设计出一种选择离子监测（SIM）方法，用于测定血浆中炔诺孕酮的代谢物、20α-羟基-9β,10α-孕甾-4,6-二烯-3-酮（DHD）和DHD葡糖醛酸。以睾酮作为内标（IS），用正己烷萃取DHD和IS，并通过氧化镁柱色谱法进行纯化。纯化后的DHD和IS在丙酮中与七氟丁酸酐反应转化为其二七氟丁酰衍生物（DHD二HFB和睾酮二HFB），用于SIM分析。SIM在230℃下使用2% OV-17柱（1 m）进行，通过监测衍生物的分子离子（DHD二HFB为m/z 706，睾酮二HFB为m/z 680）。采用峰面积法根据校准曲线测定DHD。所设计方法的测定限约为每毫升血浆5 ng DHD，对于每毫升血浆30 ng DHD及以上，重现性在变异系数的±6%以内。
