Pañak K C, Giorgieri S A, Díaz L E, Ruiz O A
Cátedra de Química Analítica Instrumental, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Argentina.
Electrophoresis. 1997 Oct;18(11):2047-9. doi: 10.1002/elps.1150181128.
A reproducible, rapid procedure for the simultaneous quantitative separation of S-adenosylmethionine and S-adenosylhomocysteine by capillary zone electrophoresis has been developed. Separations were performed by using an uncoated capillary of 60 cm effective length and 50 microm ID, 40 mM sodium phosphate buffer, pH 2.50, as background electrolyte solution, and 30 kV. On-line detection was carried out at 254 nm. Under the conditions selected we resolved a standard solution containing S-adenosylmethionine and S-adenosylhomocysteine in a run time shorter than 8 min. A mass detection limit in the range of 10 fmol was achieved. Adenosyl-L-methionine, S[methyl-3H] has also been assayed under the same experimental conditions. Other related compounds did not show interference, including those derived from the hydrolysis of S-adenosylmethionine. The present method allows simultaneous determination of these compounds, which play an important role in many microbiological and enzymatic research studies.
已开发出一种通过毛细管区带电泳同时定量分离S-腺苷甲硫氨酸和S-腺苷高半胱氨酸的可重现、快速方法。分离使用有效长度为60 cm、内径为50 µm的未涂层毛细管,40 mM磷酸钠缓冲液(pH 2.50)作为背景电解质溶液,电压为30 kV。在254 nm处进行在线检测。在所选条件下,我们在不到8分钟的运行时间内分离了含有S-腺苷甲硫氨酸和S-腺苷高半胱氨酸的标准溶液。实现了10 fmol范围内的质量检测限。在相同实验条件下还对腺苷-L-甲硫氨酸(S-[甲基-³H])进行了测定。其他相关化合物未显示干扰,包括那些由S-腺苷甲硫氨酸水解产生的化合物。本方法允许同时测定这些在许多微生物学和酶学研究中起重要作用的化合物。