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  • Determination of the oxidative stress biomarker urinary 8-hydroxy-2'-deoxyguanosine by automated on-line in-tube solid-phase microextraction coupled with liquid chromatography-tandem mass spectrometry.

Determination of the oxidative stress biomarker urinary 8-hydroxy-2'-deoxyguanosine by automated on-line in-tube solid-phase microextraction coupled with liquid chromatography-tandem mass spectrometry.

Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (2015-09-10)
Hiroyuki Kataoka, Keisuke Mizuno, Eri Oda, Akihiro Saito
ABSTRACT

A simple and sensitive method for the determination of 8-hydroxy-2'-deoxyguanosine (8-OHdG), a marker of oxidative DNA damage in human urine, was developed using automated on-line in-tube solid-phase microextraction (SPME) coupled with stable isotope-dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS). Creatinine was also analyzed simultaneously to normalize urine volume by the in-tube SPME LC-MS/MS method, and 8-OHdG and creatinine were separated within 3 min using a Zorbax Eclipse XDB-C8 column. Electrospray MS/MS for these compounds was performed on an API 4000 triple quadruple mass spectrometer in the positive ion mode by multiple reaction monitoring. The optimum in-tube SPME conditions were 20 draw/eject cycles of 40 μL of sample at a flow rate of 200 μL/min using a Carboxen 1006 PLOT capillary column as an extraction device. The extracted compounds were easily desorbed from the capillary by passage of the mobile phase, and no carryover was observed. The calibration curve for 8-OHdG using its stable isotope-labeled internal standard was linear in the range of 0.05-10 ng/mL, and the detection limit was 8.3 pg/mL. The intra-day and inter-day precision (relative standard deviations) were below 3.1% and 9.6% (n=5), respectively. This method was applied successfully to the analysis of urine samples without any other pretreatment and interference peaks, with good recovery rates above 91% in spiked urine samples. The limits of quantification of 8-OHdG and creatinine in 0.1 mL urine samples were about 0.32 and 0.69 ng/mL (S/N=10), respectively. This method was utilized to assess the effects of smoking, green tea drinking and alcohol drinking on the urinary excretion of 8-OHdG.

MATERIALS
Product Number
Brand
Product Description

Supelco
Carboxen® 572 for Tobacco Smoke, Adsorbent Bed Wt: 300 mg, cartridge size 3 mL, pkg of 50 ea
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Carboxen® Adsorbent, matrix Carboxen® 572, 20-45 mesh, bottle of 10 g
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Carboxen® Adsorbent, matrix Carboxen® 569, 20-45 mesh, bottle of 500 g
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SPME fiber assembly Polydimethylsiloxane (PDMS), metal alloy fiber, df 100 μm(PDMS, needle size 23 ga, for use with autosampler
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Carboxen® 1000 Reversible SPE Tube, 200 mg/mL, pk 30
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ORBO-93 Sorbent Tube, ORBO tube I.D. × L 6 mm × 95 mm, Bed A 180 mg, Bed B 90 mg, 60/80 mesh, pkg of 25 ea
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SPME-OC Fiber Assembly, Polydimethylsiloxane/Divinylbenzene (PDMS/DVB), df 75 μm(Coating thickness includes 65 μm coating + 10 μm OC (overcoating), needle size 23 ga, pkg of 3 ea, fiber L 1 cm
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Carboxen® Adsorbent, matrix Carboxen® 564, 20-45 mesh, bottle of 10 g
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Carboxen® Adsorbent, matrix Carboxen® 569, 20-45 mesh, bottle of 10 g
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SPME Portable Field Sampler, coating PDMS
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Carboxen® Adsorbent, matrix Carboxen® 1000, 60-80 mesh, bottle of 10 g
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Carboxen® Adsorbent, matrix Carboxen® 1003, 40-60 mesh, bottle of 10 g
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Carboxen® Adsorbent, matrix Carboxen® 564, 20-45 mesh, pack of 144 × 290 mg
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Carboxen®-1006 PLOT Capillary GC Column, L × I.D. 30 m × 0.32 mm, average thickness 15 μm
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SPME fiber assembly Divinylbenzene/Carboxen/Polydimethylsiloxane (DVB/CAR/PDMS), needle size 23 ga, StableFlex, for use with manual holder or autosampler, fiber L 2 cm
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SPME fiber assembly Carboxen/Polydimethylsiloxane (CAR/PDMS), df 85 μm(CAR/PDMS, needle size 23 ga, StableFlex, for use with autosampler
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SPME fiber assembly Polydimethylsiloxane (PDMS), df 7 μm(PDMS, needle size 23 ga, for use with autosampler
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SPME fiber assembly polyacrylate (PA), df 85 μm(PA, for use with autosampler, needle size 23 ga
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SPME fiber assembly Divinylbenzene/Carboxen/Polydimethylsiloxane (DVB/CAR/PDMS), needle size 24 ga, StableFlex
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SPME fiber assembly Polydimethylsiloxane (PDMS), df 100 μm(PDMS, for use with autosampler, needle size 24 ga
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SPME Portable Field Sampler, coating CAR/PDMS
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SPME fiber assembly Polydimethylsiloxane (PDMS), df 7 μm(PDMS, needle size 24 ga, for use with manual holder
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SPME fiber assembly polyacrylate (PA), df 85 μm(PA, for use with manual holder, needle size 24 ga
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Carboxen® Adsorbent, matrix Carboxen® 563, 20-45 mesh, bottle of 10 g
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SPME fiber assembly polyacrylate (PA), df 85 μm(PA, for use with autosampler, needle size 24 ga
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SPME Sampling Stand, for use with 4 mL vials
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SPME fiber assembly Polydimethylsiloxane (PDMS), df 100 μm(PDMS, for use with manual holder, needle size 23 ga
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SPME fiber assembly Carboxen/Polydimethylsiloxane (CAR/PDMS), df 85 μm(CAR/PDMS, needle size 24 ga, StableFlex, for use with autosampler
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SPME fiber assembly Divinylbenzene/Carboxen/Polydimethylsiloxane (DVB/CAR/PDMS), needle size 23 ga, StableFlex, for use with autosampler
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SPME fiber assembly Divinylbenzene/Carboxen/Polydimethylsiloxane (DVB/CAR/PDMS), needle size 24 ga, for use with manual holder