Is pre‐heat necessary for the measurement of 8‐oxo‐7,8‐dihydroguanosine and 8‐oxo‐7,8‐dihydro‐2′‐deoxyguanosine in urine samples

Abstract

Background

It is currently unclear for the necessary of pre‐heating urine samples for the accurate determination of 8‐oxo‐7,8‐dihydroguanosine (8‐oxoG) and 8‐oxo‐7,8‐dihydro‐2′‐deoxyguanosine (8‐oxodG). Thus, we conducted this study to evaluate the effect of pre‐heat (i.e., to 37°C) on the accurate measurement of 8‐oxoG and 8‐oxodG in frozen urine samples.

Methods

Random urine samples from six healthy volunteers, six patients with renal dysfunction, and six patients with systematic diseases such as diabetes were collected, split, and stored at −80°C for up to 1 month. The frozen samples were thawed at room temperature (RT) or 37°C for different time, 10‐fold diluted with ddH2O containing 1% formic acid, and determined by self‐established LC–MS/MS method coupled with an ACQUITY™ Primer HSS T3 column.

Results

Thawing the samples at RT for 30 or 120 min, or at 37°C for 15 or 90 min did not affect the determination of 8‐oxoG and 8‐oxodG in urine samples. Moreover, no significant difference between thawing the urine samples at RT and 37°C was found after storing at −80°C for 1–3 months.

Conclusion

It is not always necessary to pre‐heat the frozen urine samples to release 8‐oxoG and 8‐oxodG from precipitates, which is associated with different pre‐treatment and determination methods.

Abstract

For both apparently healthy volunteers and patients with kidney dysfunction or other systematic diseases, no significant difference between thawing the frozen urine samples at RT and 37°C was found for the accurate determinations of urinary 8‐oxoG and 8‐oxodG with this pre‐treatment and determination method.