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| Title: | Simultaneous determination of 3-notrotyrosine, o-, m-, and p-tyrosine in urine samples by HPLC-UV with precolumn cloud point extraction |
| Author (s): | Du, Ming Wu, Wei Ercal, Nuran Ma, Yinfa |
| Department/Lab Affiliations: | Center for Environmental Science and Technology (CEST) Chemistry Environmental Research Center |
| Keywords: | 3-Nitrotyrosine Tyrosine |
| Issue Date: | 2004 |
| Publisher: | Elsevier |
| Citation: | Du, Ming Wei Wu, Yinfa Ma Simultaneous determination of 3-notrotyrosine, o-, m-, and p-tyrosine in urine samples by HPLC-UV with precolumn cloud point extraction, J. of Chromatography B, Vol. 803, 2004, pp. 321-329, 2004. |
| Abstract: | Stable 3-nitro tyrosine (3-NO2-Tyr), o-, m-, and p-tyrosine isomers induced by oxidation of tyrosine residues in protein were considered important biomarkers for the existence of toxic oxidizing agents peroxynitrite (ONOO−) and OH√, which could lead to such diseases as acute lung injury, neurodegenerative disorders, atherosclerosis, cancers and many other diseases. Therefore, development of an accurate, simple and sensitive method to simultaneously detect o-, m-, and p-tyrosine and 3-NO2-Tyr is necessary. Fluorescence detection is highly sensitive to o-, m-, and p-tyrosine, but it cannot be used to detect 3-NO2-Tyr, due to the strong fluorescence-quenching characteristic of the NO2 group. In this study, we developed a highly sensitive reversed HPLC–UV method, combined with pre-column cloud point extraction (CPE), to simultaneously determine o-, m-, and p-tyrosine and 3-NO2-Tyr. The procedure included derivatization of a sample with 6-aminoquinolyl-N-hydroxy-succinimidyl carbomate (AccQ) at 0.20 mol/l borate buffer (pH 8.80) for 30 min at 70 °C, and pre-concentration with surfactant cloud point extraction. The surfactant-rich phase was then diluted with deionized water and injected directly into the to HPLC column for analysis. A C18 column (3.9 mm i.d. × 300 mm) was used for gradient elution separation at 25 °C and the detection wavelength was at 254 nm. Nineteen general amino acids showed no interference. The detection limits of p-, o-, m-Tyr and 3-NO2-Tyr were between 5 and 15 nmol/l. The linear range was from 0.05 to not, vert, similar100 μmol/l. |
| Type: | Article - Journal text |
| In Title: | Journal of Chromatography B |
| Copyright Notice: | Pre-print: author can archive with restrictions;Restriction: This does not include Cell Press; Post-print: author can archive; This material is presented to ensure timely dissemination of scholarly and technical work. Copyright and all rights therein are retained by authors or by other copyright holders. All persons copying this information are expected to adhere to the terms and constraints invoked by each author's copyright. In most cases, these works may not be reposted without the explicit permission of the copyright holder. FULL COPYRIGHT INFORMATION: |
| Publisher URL: | |
| Link to this page: |
| title | Simultaneous determination of 3-notrotyrosine, o-, m-, and p-tyrosine in urine samples by HPLC-UV with precolumn cloud point extraction |
| contributor.author | Du, Ming |
| contributor.author | Wu, Wei |
| contributor.author | Ercal, Nuran |
| contributor.author | Ma, Yinfa |
| contributor.deptlab | Center for Environmental Science and Technology (CEST) |
| contributor.deptlab | Chemistry |
| contributor.deptlab | Environmental Research Center |
| subject | 3-Nitrotyrosine |
| subject | Tyrosine |
| date.issued | 2004 |
| publisher | Elsevier |
| identifier.citation | Du, Ming Wei Wu, Yinfa Ma Simultaneous determination of 3-notrotyrosine, o-, m-, and p-tyrosine in urine samples by HPLC-UV with precolumn cloud point extraction, J. of Chromatography B, Vol. 803, 2004, pp. 321-329, 2004. |
| identifier.pub.URI | |
| description.abstract | Stable 3-nitro tyrosine (3-NO2-Tyr), o-, m-, and p-tyrosine isomers induced by oxidation of tyrosine residues in protein were considered important biomarkers for the existence of toxic oxidizing agents peroxynitrite (ONOO−) and OH√, which could lead to such diseases as acute lung injury, neurodegenerative disorders, atherosclerosis, cancers and many other diseases. Therefore, development of an accurate, simple and sensitive method to simultaneously detect o-, m-, and p-tyrosine and 3-NO2-Tyr is necessary. Fluorescence detection is highly sensitive to o-, m-, and p-tyrosine, but it cannot be used to detect 3-NO2-Tyr, due to the strong fluorescence-quenching characteristic of the NO2 group. In this study, we developed a highly sensitive reversed HPLC–UV method, combined with pre-column cloud point extraction (CPE), to simultaneously determine o-, m-, and p-tyrosine and 3-NO2-Tyr. The procedure included derivatization of a sample with 6-aminoquinolyl-N-hydroxy-succinimidyl carbomate (AccQ) at 0.20 mol/l borate buffer (pH 8.80) for 30 min at 70 °C, and pre-concentration with surfactant cloud point extraction. The surfactant-rich phase was then diluted with deionized water and injected directly into the to HPLC column for analysis. A C18 column (3.9 mm i.d. × 300 mm) was used for gradient elution separation at 25 °C and the detection wavelength was at 254 nm. Nineteen general amino acids showed no interference. The detection limits of p-, o-, m-Tyr and 3-NO2-Tyr were between 5 and 15 nmol/l. The linear range was from 0.05 to not, vert, similar100 μmol/l. |
| type | Article - Journal |
| type.DCMIType | text |
| type.status | Postprint |
| rights | Pre-print: author can archive with restrictions;Restriction: This does not include Cell Press; Post-print: author can archive; |
| rights | This material is presented to ensure timely dissemination of scholarly and technical work. Copyright and all rights therein are retained by authors or by other copyright holders. All persons copying this information are expected to adhere to the terms and constraints invoked by each author's copyright. In most cases, these works may not be reposted without the explicit permission of the copyright holder. |
| rights.URI | |
| relation.isPartOf | Journal of Chromatography B |
| date.available | 2008-06-12T21:27:11Z |
| identifier.persist.URI |