9-anthryldiazomethane (ADAM) を用いた高速液体クロマトグラフィー (HPLC) による妊産婦尿中プロスタグランジンおよびその代謝産物測定の問題点
1994 年 46 巻 5 号 p. 569-578
詳細
1994 年 46 巻 5 号 p. 569-578
Prostaglandins (PGs) play an important role in pregnancy, labor and delivery, especially in the onset and progress of labor. We measured and identified PGs and their metabolites in the urine during pregnancy, labor, and the puerperium. PGs and their metabolites were measured using derivatization with anthryldiazomethane (ADAM) and high performance liquid chromatography (HPLC).
Urine samples were collected from pregnant women at various stage of gestation, postpartum women (1, 3 and 5 days after delivery), three nonpregnant women and two men.
PGs and their metabolites were extracted from urine with octadecylsilyl silica by the method of Powell and re-extracted to remove substances that interfered with derivatization using ADAM. The ADAM derivatives of PGs and their metabolites were applied to reverse phase HPLC under condition A (solvent system, methanol : water 75 : 25 (v/v); flow rate, 1ml/min) and condition B (solvent system, acetonitrile : water : phosphoric acid 60 : 39.9 : 0.1 (v/v/v); flow rate, 1ml/min) and detected using a fluorescence spectrophotometer (EX 365 nm, EM 412 nm).
Specific chromatographic peaks were observed in urine samples obtained from the pregnant women after more than five weeks of gestation, but were not observed in the samples obtained from nonpregnant women and men. In addition, the specific urinary peaks in the pregnant women disappeared within five days after delivery. The retention times of the specific chromatographic peaks were 54 min, 58 min, 62 min, 67 min, 71 min under condition A, while the times were 37 min, 39 min, 43 min, 57 min, and 61 min under condition B.
PGE2, PGF2α, 15-keto-PGE2, 15-keto-PGF2α, 13, 14-dihydro-15-keto-PGE2, 13, 14-dihydro-15-keto-PGF2α, metabolites of estrogen and progesterone, malonic acid, oxalic acid, and hippuric acid were also studied using delivertization with ADAM and HPLC. However, none of the peaks of these substances were consistent with the specific peaks in the urine of pregnant women. These findings suggest that the urinary peaks obtained in our pregnant women were pregnancy-specific. However, identification of the substances that produce these pregnancy-specific peaks has not yet been achieved and further studies are needed for this purpose. [Adv Obstet Gynecol 46 (5); 569-578, 1994 (H6. 9)]
