Development of a sensitive fluorescent derivatization reagent 1,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate (BCEOC) and its application for determination of amino acids from seeds and bryophyte plants using high-performance liquid chromatography with fluorescence detection and identification with electrospray ionization mass spectrometry
文献类型：期刊论文
| 作者 | You, JM; Ming, YF; Shi, YW; Zhao, XN; Suo, YR; Wang, HL; Li, YL; Sun, J |
| 刊名 | talanta
 |
| 出版日期 | 2005-12-15 |
| 卷号 | 68期号:2页码:448-458 |
| 关键词 | derivatization HPLC amino acids 1,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate (BCEOC) |
| 合作状况 | 其它 |
| 中文摘要 | a pre-column derivatization method for the sensitive determination of amino acids and peptides using the tagging reagent 1,2-benzo-3,4dihydrocarbazole-9-ethyl chloroformate (bceoc) followed by high-performance liquid chromatography with fluorescence detection has been developed. identification of derivatives was carried out by liquid chromatography/electrospray ionization mass spectrometry (lc/esi-ms/ms). the chromophore of 2-(9-carbazole)-ethyl chloroformate (ceoc) reagent was replaced by 1,2-benzo-3,4-dihydrocarbazole functional group, which resulted in a sensitive fluorescence derivatizing reagent bceoc. bceoc can easily and quickly label peptides and amino acids. derivatives are stable enough to be efficiently analyzed by high-performance liquid chromatography. the derivatives showed an intense protonated molecular ion corresponding m/z (m + h)(+) under electrospray ionization (esi) positive-ion mode with an exception being tyr detected at negative mode. the collision-induced dissociation of protonated molecular ion formed a product at m/z 246.2 corresponding to the cleavage of c-o bond of bceoc molecule. studies on derivatization demonstrate excellent derivative yields over the ph 9.0-10.0. maximal yields close to 100% are observed with a 3-4-fold molar reagent excess. derivatives exhibit strong fluorescence and extracted detzvatization solution with n-hexane/ethyl acetate (10:1, v/v) allows for the direct injection with no significant interference from the major fluorescent reagent degradation by-products, such as 1,2-benzo-3,4-dihydrocarbazole-9-ethanol (bdc-oh) (a major by-product), mono- 1,2-benzo-3,4-dihydrocarbazole-9-ethyl carbonate (bceoc-oh) and bis-(1,2-benzo-3,4-dihydrocarbazole-9-ethyl) carbonate (bceoc)(2). in addition, the detection responses for bceoc derivatives are compared to those obtained with previously synthesized 2-(9-carbazole)-ethyl chloroformate (ceoc) in our laboratory. the ratios ac(bceoc)/ac(ceoc) = 2.05-6.51 for fluorescence responses are observed (here, ac is relative fluorescence response). separation of the derivatized peptides and amino acids had been optimized on hypersil bds c-18 column. detection limits were calculated from 1.0 pmol injection at a signal-to-noise ratio of 3, and were 6.3 (lys)-177.6 (his) fmol. the mean interday accuracy ranged from 92 to 106% for fluorescence detection with mean %cv < 7.5. the mean interday precision for all standards was < 10% of the expected concentration. excellent linear responses were observed with coefficients of > 0.9999. good compositional data could be obtained from the analysis of derivatized protein hydrolysates containing as little as 50.5 ng of sample. therefore, the facile bceoc derivatization coupled with mass spectrometry allowed the development of a highly sensitive and specific method for the quantitative analysis of trace levels of amino acids and peptides from biological and natural environmental samples. (c) 2005 elsevier b.v. all rights reserved. |
| 英文摘要 | a pre-column derivatization method for the sensitive determination of amino acids and peptides using the tagging reagent 1,2-benzo-3,4dihydrocarbazole-9-ethyl chloroformate (bceoc) followed by high-performance liquid chromatography with fluorescence detection has been developed. identification of derivatives was carried out by liquid chromatography/electrospray ionization mass spectrometry (lc/esi-ms/ms). the chromophore of 2-(9-carbazole)-ethyl chloroformate (ceoc) reagent was replaced by 1,2-benzo-3,4-dihydrocarbazole functional group, which resulted in a sensitive fluorescence derivatizing reagent bceoc. bceoc can easily and quickly label peptides and amino acids. derivatives are stable enough to be efficiently analyzed by high-performance liquid chromatography. the derivatives showed an intense protonated molecular ion corresponding m/z (m + h)(+) under electrospray ionization (esi) positive-ion mode with an exception being tyr detected at negative mode. the collision-induced dissociation of protonated molecular ion formed a product at m/z 246.2 corresponding to the cleavage of c-o bond of bceoc molecule. studies on derivatization demonstrate excellent derivative yields over the ph 9.0-10.0. maximal yields close to 100% are observed with a 3-4-fold molar reagent excess. derivatives exhibit strong fluorescence and extracted detzvatization solution with n-hexane/ethyl acetate (10:1, v/v) allows for the direct injection with no significant interference from the major fluorescent reagent degradation by-products, such as 1,2-benzo-3,4-dihydrocarbazole-9-ethanol (bdc-oh) (a major by-product), mono- 1,2-benzo-3,4-dihydrocarbazole-9-ethyl carbonate (bceoc-oh) and bis-(1,2-benzo-3,4-dihydrocarbazole-9-ethyl) carbonate (bceoc)(2). in addition, the detection responses for bceoc derivatives are compared to those obtained with previously synthesized 2-(9-carbazole)-ethyl chloroformate (ceoc) in our laboratory. the ratios ac(bceoc)/ac(ceoc) = 2.05-6.51 for fluorescence responses are observed (here, ac is relative fluorescence response). separation of the derivatized peptides and amino acids had been optimized on hypersil bds c-18 column. detection limits were calculated from 1.0 pmol injection at a signal-to-noise ratio of 3, and were 6.3 (lys)-177.6 (his) fmol. the mean interday accuracy ranged from 92 to 106% for fluorescence detection with mean %cv < 7.5. the mean interday precision for all standards was < 10% of the expected concentration. excellent linear responses were observed with coefficients of > 0.9999. good compositional data could be obtained from the analysis of derivatized protein hydrolysates containing as little as 50.5 ng of sample. therefore, the facile bceoc derivatization coupled with mass spectrometry allowed the development of a highly sensitive and specific method for the quantitative analysis of trace levels of amino acids and peptides from biological and natural environmental samples. (c) 2005 elsevier b.v. all rights reserved. |
| 学科主题 | 生物科学 |
| WOS标题词 | science & technology ; physical sciences |
| 类目[WOS] | chemistry, analytical |
| 研究领域[WOS] | chemistry |
| 关键词[WOS] | precolumn derivatization ; 9-fluorenylmethyl chloroformate ; capillary-electrophoresis ; biogenic-amines ; chloride ; hydroxyproline ; derivatives |
| 收录类别 | ISTP ; SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000233491100046 |
| 公开日期 | 2009-12-09 |
| 源URL | [http://ir.nwipb.ac.cn/handle/363003/1353]  |
| 专题 | 西北高原生物研究所_中国科学院西北高原生物研究所 |
| 作者单位 | 1.Qufu Normal Univ, Dept Chem, Qufu 273165, Peoples R China 2.Chinese Acad Sci, NW Plateau Inst Biol, Xining, Peoples R China |
| 推荐引用方式 GB/T 7714 | You, JM,Ming, YF,Shi, YW,et al. Development of a sensitive fluorescent derivatization reagent 1,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate (BCEOC) and its application for determination of amino acids from seeds and bryophyte plants using high-performance liquid chromatography with fluorescence detection and identification with electrospray ionization mass spectrometry[J]. talanta,2005,68(2):448-458. |
| APA | You, JM.,Ming, YF.,Shi, YW.,Zhao, XN.,Suo, YR.,...&Sun, J.(2005).Development of a sensitive fluorescent derivatization reagent 1,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate (BCEOC) and its application for determination of amino acids from seeds and bryophyte plants using high-performance liquid chromatography with fluorescence detection and identification with electrospray ionization mass spectrometry.talanta,68(2),448-458. |
| MLA | You, JM,et al."Development of a sensitive fluorescent derivatization reagent 1,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate (BCEOC) and its application for determination of amino acids from seeds and bryophyte plants using high-performance liquid chromatography with fluorescence detection and identification with electrospray ionization mass spectrometry".talanta 68.2(2005):448-458. |
入库方式： OAI收割
来源：西北高原生物研究所
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