Established and Optimized the Measurement of Serum Troponin I Using Liquid Chromatography Tandem Mass Spectrometry

doi:10.6023/A19010032

Acta Chim. Sinica ›› 2019, Vol. 77 ›› Issue (6): 539-544.DOI: 10.6023/A19010032 Previous Articles Next Articles

Article

血清肌钙蛋白I液相色谱串联质谱法建立及优化

李亚男, 季伙燕, 王天一, 沈蕾, 施秀英, 王建新

南通大学附属医院医学检验科南通 226001

投稿日期:2019-01-18 发布日期:2019-05-08
通讯作者: 王建新 E-mail:wwbft@126.com
基金资助:
项目受国家重点研发计划项目（No.2017YFF0205401）资助.

Established and Optimized the Measurement of Serum Troponin I Using Liquid Chromatography Tandem Mass Spectrometry

Li Yanan, Ji Huoyan, Wang Tianyi, Shen Lei, Shi Xiuying, Wang Jianxin

Department of Laboratory Medicine, Affiliated Hospital of Nantong University, Nantong 226001

Received:2019-01-18 Published:2019-05-08
Supported by:
Project supported by the National Key Research and Development Program of China (No. 2017YFF0205401).

1. Established and Optimized the Measurement of Serum Troponin I Using Liquid Chromatography Tandem Mass Spectrometry.PDF(181KB)

Abstract

Cardiac troponin I (cTnI) is one of the most popular biomarkers for the diagnosis of acute myocardial injury (AMI) in patients. In this study, a novel method was developed and optimized for quantification of cTnI in human serum by immunoaffinity enrichment combining isotope dilution liquid chromatography tandem mass spectrometry. cTnI was first captured from human serum with immunomagnetic beads conjugated with the monoclonal antibody, and then enzymatically hydrolyzed into peptides after a series of operation, including denaturation, reduction, acetylation, digestion and purification. Subsequently, enzymatic peptides were separated by passing through Symmetry Shield C18 column at a speed of 0.2 mL/min with 0.1% acid acetonitrile solution and 0.1% acid aqueous solution as mobile phases under gradient elution. A specific peptide, NITEIADLTQK, was selected and quantified. The qualitative analysis was achieved by three ion transitions under multiple reaction monitoring (MRM) when the isotopically labeled peptide, NITEIAD[(¹³C₆,¹⁵N)L]TQK, was used as a reference. After the optimal conditional experiment, results showed that the surrogate peptide was separated out well at about 4.95 min with little interference. Ranging from 10 ng/mL to 600 ng/mL, a good linearity was shown with correlation coefficients all above 0.99. The limit of detection (LOD) and the limit of quantification (LOQ) were estimated to be 2.5 ng/mL, 8.32 ng/mL, respectively. This method also provided good accuracy (relative bias of three concentrations diluted from SRM2921 were between -7.94% and -6.49%) and repeatability (total relative standard deviations of three concentration were 6.43%, 3.18% and 2.75%, respectively). Carry-over rates were estimated between -0.47% and 0.04%. The novel assay successfully determined five specimens from AMI patients with concentrations from 16.38 ng/mL to 557.53 ng/mL. Our results demonstrate that this method can be applied for determination of serum cTnI in AMI patients with high selectivity, low carry-over rates, good repeatability and good accuracy, which helps to establish candidate reference measurement procedure of serum cTnI.

Key words: liquid chromatography tandem mass spectrometry, troponin I, surrogate peptides, quantification, immunomagnetic beads

Cite this article

Li Yanan, Ji Huoyan, Wang Tianyi, Shen Lei, Shi Xiuying, Wang Jianxin. Established and Optimized the Measurement of Serum Troponin I Using Liquid Chromatography Tandem Mass Spectrometry[J]. Acta Chim. Sinica, 2019, 77(6): 539-544.

Export EndNote|Reference Manager|ProCite|BibTeX|RefWorks

share this article

References

[1] Stillman, A. E.; Oudkerk, M.; Bluemke, D.; Bremerich, J.; Esteves, F. P.; Garcia, E. V.; Gutberlet, M.; Hundley, W. G.; Jerosch-Herold, M.; Kuijpers, D.; Kwong, R. K.; Nagel, E.; Lerakis, S.; Oshinski, J.; Paul, J. F.; Underwood, R.; Witersperger, B. J.; Rees, M. R. Int. J. Cardiovasc. Imaging 2011, 27, 7.
[2] Soetkamp, D.; Raedschelders, K.; Mastali, M.; Sobhani, K.; Bairey Merz, C. N.; Van Eyk, J. Expert Rev. Proteomics 2017, 14, 973.
[3] Boriani, G.; Biffi, M.; Cervi, V.; Bronzetti, G.; Magagnoli, G.; Zannoli, R.; Branzi, A. Chest. 2000, 118, 342.
[4] Panteghini, M.; Bunk, D. M.; Christenson, R. H.; Katrukha, A.; Porter, R. A.; Schimmel, H.; Wang, L.; Tate, J. R. Clin. Chem. Lab. Med. 2008, 46, 1501.
[5] Tate, J. R.; Bunk, D. M.; Christenson, R. H.; Katrukha, A.; Noble, J. E.; Porter, R. A.; Schimmel, H.; Wang, L.; Panteghini, M. Pathology 2010, 42, 402.
[6] Hoofnagle, A. N.; Wener, M. H. J. Immunol. Methods 2009, 347, 3.
[7] Noble, J. E.; Bunk, D. M.; Christenson, R. H.; Cole, K. D.; He, H. J.; Katrukha, A. G.; Panteghini, M.; Porter, R. A.; Schimmel, H.; Tate, J. R.; Wang, L. Clin. Chem. Lab. Med. 2010, 48, 1603.
[8] Guo, Q.-Z.; Du, Z.-X. Chin. J. Chem. 2019, 29, 1922.
[9] Chen, L.-N.; Song, F.-R.; Zheng, Z.; Xing, J.-P.; Liu, Z.-Q.; Liu, S.-Y. Acta Chim. Sinica 2012, 70, 843(in Chinese). (陈丽娜, 宋凤瑞, 郑重, 邢俊鹏, 刘志强, 刘淑莹, 化学学报, 2012, 70, 843.)
[10] Agger, S. A.; Marney, L. C.; Hoofnagle, A. N. Clin. Chem. 2010, 56, 1804.
[11] Kuhn, E.; Wu, J.; Karl, J.; Liao, H.; Zolg, W.; Guild, B. Proteomics 2004, 4, 1175.
[12] Keshishian, H.; Addona, T.; Burgess, M.; Mani, D. R.; Shi, X.; Kuhn, E.; Sabatine, M. S.; Gerszten, R. E.; Carr, S. A. Mol. Cell. Proteomics 2009, 8, 2339.
[13] Kuhn, E.; Addona, T.; Keshishian, H.; Burgess, M.; Mani, D. R.; Lee, R. T.; Sabatine, M. S.; Gerszten, R. E.; Carr, S. A. Clin. Chem. 2009, 55, 1108.
[14] Li, X.-Q.; Yang, Z.; Zhang, Q.-H. J. Chin. Mass Spectrom. Soc. 2013, 34, 338(in Chinese). (李秀琴, 杨总, 张庆合, 质谱学报, 2013, 34, 338.)
[15] Anderson, N. L. Clin. Chem. 2010, 56, 177.
[16] Anderson, N. L.; Anderson, N. G. Mol. Cell. Proteomics 2002, 1, 845.
[17] Williams, D. K.; Muddiman, D. C. J. Proteome Res. 2009, 8, 1085.
[18] Ji, H.; Wang, J.; Ju, S.; Cong, H.; Wang, X.; Su, J.; Wang, H. J. Chromatogr. B:Anal. Technol. Biomed. Life Sci. 2017, 1059, 49.
[19] Mann, M.; Hendrickson, R. C.; Pandey, A. Annu. Rev. Biochem. 2001, 70, 437.
[20] Zhao, M.; Wu, F. Protein Expression Purif. 2015, 116, 120.
[21] Rice, R. H.; Means, G. E. Biochim. Biophys. Acta 1977, 492, 316.
[22] Nash, D. C.; Chase, H. A. J. Chromatogr. A 1998, 807, 185.

血清肌钙蛋白I液相色谱串联质谱法建立及优化

Established and Optimized the Measurement of Serum Troponin I Using Liquid Chromatography Tandem Mass Spectrometry

PDF

Supporting Info.

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 4

Recommended Articles

Metrics

Comments

[1]	Wang Dabin, Zhao Lixia, Guo Lianghong, Zhang Hui, Wan Bin, Yang Yu. Online Quantification of O₂^·- and H₂O₂ and Their Formation Kinetics in Ultraviolet (UV)-Irradiated Nano-TiO₂ Suspensions by Continuous Flow Chemiluminescence [J]. Acta Chim. Sinica, 2015, 73(5): 388-394.
[2]	Yin Xuefei, Liu Xiaohui, Shen Huali, Jin Hong, Yang Pengyuan. Whole Proteome of Fusobacterium nucleatum and Quantitative Analysis of Proteomic Change in Cancer Environment [J]. Acta Chim. Sinica, 2015, 73(4): 337-342.
[3]	Zhu Fengjun, Zhang Jing, Zheng Saijing, Liu Baizhan, Guo Yinlong. Determination of Small Molecular Aldehydes in Cigarette Smoke by Extraction and Derivatization in Single Drop [J]. Acta Chimica Sinica, 2012, 70(12): 1332-1336.
[4]	Chen Lina, Song Fengrui, Zheng Zhonga, Xing Junpeng, Liu Zhiqiang, Liu Shuying. Studies on the Determination Method of Pesticide Multi-residues in Ginseng by Ultra Performance Liquid Chromatography Tandem Mass Spectrometry [J]. Acta Chimica Sinica, 2012, 70(07): 843-851 .