| 摘要 |
PURPOSE:To perform the simultaneous analysis of creatine and creatinine with high accuracy, by applying the eluate, which was obtained by applying a specimen after protein removal treatment to reversed-phase partition chromatography, to reversed-phase partition chromatography of an acidic moving phase to measure the ultraviolet absorbancy of the eluate. CONSTITUTION:A specimen such as serum, to which a perchloric acid solution was added under mixing, is centrifugally separated to obtain a protein removed supernatant solution which is, in turn, flowed through a reversed-phase chromatographic column, wherein an octadecyl group or a trimethyl group was chemically bonded to a silica gel, by using a buffer solution with pH of 2.0-7.0 as a moving phase to obtain a creatin and creatinine-containing solution from which impurities in the specimen were removed. This eluate is applied to reversed-phase ion-pair chromatography by using a buffer solution, of which the pH is adjusted to 2.0-3.0 by phosphoric acid, as a moving phase to separate and elute creatin and creatinine. The ultraviolet absorbancy of this eluate is measured at a wavelength of 120nm. By this method, simultaneous analysis is enabled with high sensitivity and good reproducibility. |
