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- E-ISSN 2093-8950
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Simultaneous Determination of Five Porphyrins in Human Urine and Plasma Using High Performance Liquid Chromatography-Tandem Mass Spectrometry
Mass Spectrometry Letters / Mass Spectrometry Letters, (P)2233-4203; (E)2093-8950
2014, v.5 no.2, pp.42-48
https://doi.org/10.5478/MSL.2014.5.2.42
(International Scientific Standards Co. Ltd.)
(International Scientific Standards Co. Ltd.)
(The Korea Institute for children's Social Development and Rudolph Child Research Center)
(Institute for EONE Laboratories)
(Institute for EONE Laboratories)
(International Scientific Standards Co. Ltd.)
(International Scientific Standards Co. Ltd.)
(Kangwon National University)
(International Scientific Standards Co. Ltd.)
(International Scientific Standards Co. Ltd.)
(Chung-Ang University)
(International Scientific Standards Co. Ltd.)
(The Korea Institute for children's Social Development and Rudolph Child Research Center)
(Institute for EONE Laboratories)
(Institute for EONE Laboratories)
(International Scientific Standards Co. Ltd.)
(International Scientific Standards Co. Ltd.)
(Kangwon National University)
(International Scientific Standards Co. Ltd.)
(International Scientific Standards Co. Ltd.)
(Chung-Ang University)
Hur,
Y., Tae,
S., Koh,
Y., Hong,
S., Yoon,
Y. H., Jang,
H., Kim,
S., Kim,
K. H., Kang,
S. W., Lee,
Y., &
Han,
S.
B.
(2014). Simultaneous Determination of Five Porphyrins in Human Urine and Plasma Using High Performance Liquid Chromatography-Tandem Mass Spectrometry. Mass Spectrometry Letters, 5(2), 42-48, https://doi.org/10.5478/MSL.2014.5.2.42
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Abstract
A specific and sensitive liquid chromatography-electrospray ionization tandem mass spectrometry method (LC-ESIMS/MS) was developed and validated for the simultaneous quantification of porphyrins (coproporphyrin, pentacarboxylporphyrin,hexacarboxylporphyrin, heptacarboxylporphyrin, and uroporphyrin) in human plasma and urine. Acidified plasma samplesand urine samples were prepared by using liquid-liquid extraction using ethyl acetate and protein precipitation with acetonitrile,respectively. The separation was achieved onto a Synergi Fusion RP column (150 mm × 2.0 mm, 4 μm) with a gradient elutionof mobile phase A (0.1% formic acid in 2 mmol/L ammonium acetate, v/v) and mobile phase B (20% methanol in acetonitrile, v/v) at a flow rate of 450 μL/min. Porphyrins and the internal standard (IS), coproporphyrin I-15N4, were detected by a tandemmass spectrometer equipped with an electrospray ion source operating in positive ion mode. Multiple reaction monitoring(MRM) transitions of the protonated precursor ions and the related product ions were optimized to increase selectivity and sensitivity. The proposed method was validated by assessing selectivity, linearity, limit of quantification (LOQ), precision, accuracy,recovery, and stability. The calibration curves were obtained in the range of 0.1-100 nmol/L and the LOQs were estimated as0.1 nmol/L for all porphyrins. Results obtained from the validation study of porphyrins showed good accuracy, precision, recovery,and stability. Finally, the proposed method was successfully applied to clinical studies on the autism spectrum disorder(ASD) diagnosis of 203 Korean children.
- keywords
- Porphyrins, tandem mass spectrometry, autism, diagnosis, plasma, urine
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- Submission Date
- 2014-05-01
- Revised Date
- 2014-05-01
- Accepted Date
- 2014-06-01
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