Haginaka J, Yasuda H, Uno T, Nakagawa T
J Chromatogr. 1986 Apr 25;377:269-77. doi: 10.1016/s0378-4347(00)80782-3.
A high-performance liquid chromatographic method with fluorimetric detection has been developed for the determination of clavulanate in human urine and plasma. Clavulanate in plasma samples was ultrafiltered using YMT membrane and reacted with benzaldehyde in phosphate buffer solution (pH 3.8) at 100 degrees C for 20 min. Clavulanate in urine samples was filtered with a polyacrylate membrane after ten-fold dilution, and reacted under the same conditions as those for plasma samples. The fluorescent product thus formed from clavulanate was separated from ordinary components of plasma and urine on a reversed-phase C18 column followed by fluorimetric detection (lambda ex = 386 nm, lambda em = 460 nm). The within- and between-run precisions were of the order of 4.02% (n = 10) and 6.23% (n = 5) for plasma samples at a level of 0.67 microgram/ml. The detection limit was as low as 10 ng/ml in plasma samples with a 50-microliter injection. Coexisting ticarcillin, amoxicillin or 1-amino-4-hydroxybutan-2-one (which is a metabolite of clavulanate in rat and dog) did not interfere in the clavulanate assay.
已开发出一种带荧光检测的高效液相色谱法，用于测定人尿液和血浆中的克拉维酸。血浆样品中的克拉维酸使用YMT膜进行超滤，并在100℃下于pH 3.8的磷酸盐缓冲溶液中与苯甲醛反应20分钟。尿液样品中的克拉维酸在十倍稀释后用聚丙烯酸酯膜过滤，并在与血浆样品相同的条件下反应。由克拉维酸形成的荧光产物在反相C18柱上与血浆和尿液的普通成分分离，随后进行荧光检测（激发波长λex = 386 nm，发射波长λem = 460 nm）。对于浓度为0.67微克/毫升的血浆样品，批内和批间精密度分别为4.02%（n = 10）和6.23%（n = 5）。进样量为50微升时，血浆样品中的检测限低至10纳克/毫升。共存的替卡西林、阿莫西林或1-氨基-4-羟基丁-2-酮（这是克拉维酸在大鼠和狗体内的代谢产物）不干扰克拉维酸的测定。
