Mci gel ca08f

The urinary 8-OHdG levels were determined according to our previous study.^{(23 (link))} Briefly, each urine sample was defrosted to room temperature and centrifuged at 8,500 × g for 5 min. A 50 µl portion of the urine supernatant was mixed with the same volume of a dilution solution, containing the ribonucleoside marker 8-hydroxyguanosine. Afterwards, a 20 µl portion of the prepared solution was fractionated and injected into the first HPLC column (MCI GEL CA08F, 1.5 × 150 mm; Mitsubishi Chemical, Tokyo, Japan). The chromatograph was recorded with a UV detector (Gilson UV/VIS-151, 235 nm). The fraction containing 8-OHdG was then automatically injected into the second HPLC column (Insertsil^TM ODS-3, 3 µm, 4.6 × 250 mm; GL Sciences Inc., Tokyo, Japan). The chromatograph was recorded with an electrochemical (EC) detector (Coulochem II; ESA, Chelmsford, MA). The 8-OHdG levels were expressed as the ratios to the urinary creatinine contents (UV detector at 235 nm).

We determined the urinary 8‐OHdG levels according to the method of Kasai et al.⁸ Briefly, urine samples were thawed and centrifuged, and then 50 μL of the supernatant was mixed with an equal volume of a dilution solution containing the ribonucleoside marker 8‐hydroxyguanosine. A 20‐µL portion of this mixture was injected into the high‐performance liquid chromatography (HPLC)‐1 column (MCI GEL CA08F, 1.5 × 150 mm; Mitsubishi Chemical). The chromatograph was recorded with a UV detector (Gilson UV/VIS‐151, 235 nm). The fraction containing 8‐OHdG was automatically injected into the HPLC‐2 column (Inertsil™ ODS‐3, 3 μm, 4.6 × 250 mm; GL Sciences, Inc). The 8‐OHdG was detected by an electrochemical detector (Coulochem II, ESA). The 8‐OHdG levels were expressed as the ratios to the urinary creatinine contents (UV detection at 235 nm).

The urinary 8-OHdG levels were determined based on the automated two-step separation method.11 (link) Briefly, each defrosted urine sample was centrifuged at 13,000 rpm for 5 minutes and 50 μL urine supernatant was mixed with the same volume of a dilution solution containing the ribonucleoside marker 8-hydroxyguanosine. A 20 μL aliquot of the diluted urine sample was injected to an automated high-performance liquid chromatography (HPLC) system. The anion exchange guard column (MCI GEL CA08F, 1.5 × 150 mm; Mitsubishi Chemical, Tokyo, Japan) was used in HPLC-1. The chromatograms were recorded by a Gilson UV detector (UV/VIS-151, 235 nm). The 8-OHdG fraction was collected, automatically injected into a reverse-phase column in HPLC-2 (Capcell Pak C18, 4.6 × 250 mm; GL Sciences Inc., Tokyo, Japan), and detected by an EC detector (Coulochem II; ESA, Chelmsford, MA, USA). 8-OHdG levels were normalized for urinary creatinine levels. Urinary creatinine was simultaneously analyzed with UV detector at 225 nm to adjust individual urine density.