Gentili D, Zucchetti M, Conter V, Masera G, D'Incalci M
Mario Negri Institute, Milan, Italy.
J Chromatogr B Biomed Appl. 1994 Jul 1;657(1):47-52. doi: 10.1016/0378-4347(94)80068-5.
The antileukaemic efficacy of L-asparaginase is related to the ability of the enzyme to induce the complete disappearance from plasma of L-asparagine, an amino acid essential to lymphoblastic leukaemia cells. It is not feasible to monitor L-asparagine plasma levels in patients under L-asparaginase treatment using the usual analytical procedures as the enzyme continues the hydrolysis of L-asparagine after blood sampling and during plasma extraction. A method was therefore developed for the determination of L-asparagine in patients receiving L-asparaginase. Sulphosalicylic acid is added to blood samples to deproteinize and inactivate L-asparaginase rapidly. The samples are then analysed by HPLC using a Novapack C18 column and fluorescence detection. With the same method L-asparagine is determined in blood cells and, by difference, plasma levels are calculated. This method is highly specific and sufficiently simple and sensitive for clinical use.
L-天冬酰胺酶的抗白血病疗效与该酶使血浆中L-天冬酰胺完全消失的能力有关,L-天冬酰胺是淋巴细胞白血病细胞所必需的一种氨基酸。使用常规分析方法监测接受L-天冬酰胺酶治疗患者的血浆L-天冬酰胺水平是不可行的,因为在采血后和血浆提取过程中,该酶会持续水解L-天冬酰胺。因此,开发了一种用于测定接受L-天冬酰胺酶治疗患者体内L-天冬酰胺的方法。向血样中加入磺基水杨酸以快速使蛋白质变性并使L-天冬酰胺酶失活。然后使用Novapack C18柱和荧光检测通过高效液相色谱法对样品进行分析。用相同方法测定血细胞中的L-天冬酰胺,并通过差值计算血浆水平。该方法特异性高,对于临床应用而言足够简单且灵敏。