Citation: SUN Ze-Peng, WANG Hong-Bin, CHEN Jin-Chao, SONG De-Wei, LI Hong-Mei, XIAO Peng. Quantitative Analysis of Recombinant N-terminal Pro-brain Natriuretic Peptide by Liquid Chromatography Coupled with Isotope Dilution Mass Spectrometry[J]. Chinese Journal of Analytical Chemistry, ;2023, 51(1): 120-129. doi: 10.19756/j.issn.0253-3820.221162 shu

Quantitative Analysis of Recombinant N-terminal Pro-brain Natriuretic Peptide by Liquid Chromatography Coupled with Isotope Dilution Mass Spectrometry

SUN Ze-Peng ^1,2 ,
WANG Hong-Bin ^1,, ,
CHEN Jin-Chao ² ,
SONG De-Wei ^2,3 ,
LI Hong-Mei ^2,3 ,
XIAO Peng ^2,3,,

1.
College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China;
2.
National Institute of Metrology, Beijing 100029, China;
3.
Key Laboratory of Chemical Metrology and Applications on Nutrition and Health for State Market Regulation, Beijing 100029, China

Corresponding author: WANG Hong-Bin, XIAO Peng,
Received Date: 2 April 2022
Revised Date: 13 October 2022

Fund Project: Supported by the National Key Research and Development of China(No.2019YFF0216505) and the Basic Research Funds of National Institute of Metrology, China (Nos.AKYZD2115-2, AKYGH2002).

N-terminal pro-brain natriuretic peptide (NT-proBNP) is an essential biomarker for diagnosis of heart failure. Currently, since reference materials of NT-proBNP are unavailable globally, value transfer activities are hard to implement from the higher level to the end-users, which makes terminal clinical testing results inconsistent and incomparable. To achieve standard detection of NT-proBNP, in this work, recombinant NT-proBNP was chosen as a target during traceable approach development. Firstly, the immune capacity and relative molecular weight were tested and varied through Western Blot assay and mass spectrometric methods. Next, two kinds of isotope dilution mass spectrometry were adopted for the absolute quantification of NT-proBNP. The results suggested that the protein hydrolysis reaction reached equilibrium in 48 h. NT-proBNP was calculated to be 79.62 μg/g with an relative standard deviation (RSD) of 0.89% by detection of valine, isoleucine and arginine. On the other hand, the isotope dilution mass spectrometry method was used to analyze signature peptides produced by enzymatic digestion. NT-proBNP was calculated to be 76.04 μg/g with an RSD of 1.85% by detection of the two peptides under the conditions of enzymatic digestion for 24h and at mass ratio of 25∶1 of NT-proBNP to protease. The quantitative analysis results from the two methods were consistent with each other. As the amino acids applied in quantifications were SI-traceable primary reference materials, all the results were also traceable. Such approaches were candidates for NT-proBNP reference material development.
- Biomarker of heart failure,
- Nterminal pro-brain natriuretic peptide,
- Quantitative analysis,
- Isotope dilution mass spectrometry,
- Traceability
1. [1]
  AMBROSY A P, FONAROW G C, BUTLER J, CHIONCEL O, GREENE S J, VADUGANATHAN M, NODARI S, LAM C S P, SATO N, SHAH A N, GHEORGHIADE M. J. Am. Coll. Cardiol., 2014, 63(12):1123-1133.
2. [2]
  METRA M, LUCIOLI P. Eur. J. Heart. Fail, 2020, 22(4):759.
3. [3]
  EMDIN M, PASSINO C, PRONTERA C, FONTANA M, POLETTI R, GABUTTI A, MAMMINI C, GIANNONI A, ZYW L, ZUCCHELLI G, CLERICO A. Clin. Chem., 2007, 53(7):1289-1297.[4] GOETZE J P. Clin. Chem., 2004, 50(9):1503-1510.[5] VASILE V C, JAFFE A S. Clin. Chem., 2017, 63(1):50-58.
4. [4]
  CANTINOTTI M, STORTI S, PARRI M S, MURZI M, CLERICO A. Clin. Chem., 2009, 55(7):1438-1440.
5. [5]
6. [6]
7. [7]
8. [8]
9. [9]
10. [10]
  MUNOZ A, KRAL R, SCHIMMEL H. Anal. Biochem., 2011, 408(1):124-131.
11. [11]
12. [12]
  JEONG J S, LIM H M, KIM S K, KU H K, OH K H, PARK S R. J. Chromatogr. A, 2011, 1218(38):6596-6602.
13. [13]
14. [14]
1. [1]
  Wei Peng , Baoying Wen , Huamin Li , Yiru Wang , Jianfeng Li . Exploration and Practice on Raman Scattering Spectroscopy Experimental Teaching. University Chemistry, 2024, 39(8): 230-240. doi: 10.3866/PKU.DXHX202312062
2. [2]
  Yaling Chen . Basic Theory and Competitive Exam Analysis of Dynamic Isotope Effect. University Chemistry, 2024, 39(8): 403-410. doi: 10.3866/PKU.DXHX202311093
3. [3]
  Ling Bai , Limin Lu , Xiaoqiang Wang , Dongping Wu , Yansha Gao . Exploration and Practice of Teaching Reforms in “Quantitative Analytical Chemistry” under the Perspective of New Agricultural Science. University Chemistry, 2024, 39(3): 158-166. doi: 10.3866/PKU.DXHX202308101
4. [4]
  Mi Wen , Baoshuo Jia , Yongqi Chai , Tong Wang , Jianbo Liu , Hailong Wu . Improvement of Fluorescence Quantitative Analysis Experiment: Simultaneous Determination of Rhodamine 6G and Rhodamine 123 in Food Using Chemometrics-Assisted Three-Dimensional Fluorescence Method. University Chemistry, 2025, 40(4): 390-398. doi: 10.12461/PKU.DXHX202405147
5. [5]
  Zian Lin , Yingxue Jin . Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI-MS) for Disease Marker Screening and Identification: A Comprehensive Experiment Teaching Reform in Instrumental Analysis. University Chemistry, 2024, 39(11): 327-334. doi: 10.12461/PKU.DXHX202403066
6. [6]
  Min Gu , Huiwen Xiong , Liling Liu , Jilie Kong , Xueen Fang . Rapid Quantitative Detection of Procalcitonin by Microfluidics: An Instrumental Analytical Chemistry Experiment. University Chemistry, 2024, 39(4): 87-93. doi: 10.3866/PKU.DXHX202310120
7. [7]
  Xiaowei TANG , Shiquan XIAO , Jingwen SUN , Yu ZHU , Xiaoting CHEN , Haiyan ZHANG . A zinc complex for the detection of anthrax biomarker. Chinese Journal of Inorganic Chemistry, 2024, 40(10): 1850-1860. doi: 10.11862/CJIC.20240173
8. [8]
  Zongyuan Chen , ChunSheng Shi , Yiwen Li , Ganlin Zu , Qiang Jin , Haishan Wang , Fujun Wang , Dekun Yan , Zhijun Guo , Wangsuo Wu . Measurement of Uranium Isotopes in Environmental Water Samples by Alpha-Spectroscopy: Design of an Undergraduate Radiochemistry Experiment. University Chemistry, 2025, 40(4): 353-358. doi: 10.12461/PKU.DXHX202406103
9. [9]
  Cheng Zheng , Shiying Zheng , Yanping Zhang , Shoutian Zheng , Qiaohua Wei . Synthesis, Copper Content Analysis, and Luminescent Performance Study of Binuclear Copper (I) Complexes with Isomeric Luminescence Shift: A Comprehensive Chemical Experiment Recommendation. University Chemistry, 2024, 39(7): 322-329. doi: 10.3866/PKU.DXHX202310131
10. [10]
  Junjie Zhang , Yue Wang , Qiuhan Wu , Ruquan Shen , Han Liu , Xinhua Duan . Preparation and Selective Separation of Lightweight Magnetic Molecularly Imprinted Polymers for Trace Tetracycline Detection in Milk. University Chemistry, 2024, 39(5): 251-257. doi: 10.3866/PKU.DXHX202311084
11. [11]
  Yanhui Zhong , Ran Wang , Zian Lin . Analysis of Halogenated Quinone Compounds in Environmental Water by Dispersive Solid-Phase Extraction with Liquid Chromatography-Triple Quadrupole Mass Spectrometry. University Chemistry, 2024, 39(11): 296-303. doi: 10.12461/PKU.DXHX202402017
12. [12]
  Xilin Zhao , Xingyu Tu , Zongxuan Li , Rui Dong , Bo Jiang , Zhiwei Miao . Research Progress in Enantioselective Synthesis of Axial Chiral Compounds. University Chemistry, 2024, 39(11): 158-173. doi: 10.12461/PKU.DXHX202403106
13. [13]
  Yonghui Wang , Weilin Chen , Yangguang Li . Knowledge Construction of “Solubility of Inorganic Substances” in Elemental Chemistry Teaching. University Chemistry, 2024, 39(4): 261-267. doi: 10.3866/PKU.DXHX202312102
14. [14]
  Shijie Li , Ke Rong , Xiaoqin Wang , Chuqi Shen , Fang Yang , Qinghong Zhang . Design of Carbon Quantum Dots/CdS/Ta₃N₅ S-Scheme Heterojunction Nanofibers for Efficient Photocatalytic Antibiotic Removal. Acta Physico-Chimica Sinica, 2024, 40(12): 2403005-. doi: 10.3866/PKU.WHXB202403005
15. [15]
  Ke QIAO , Yanlin LI , Shengli HUANG , Guoyu YANG . Advancements in asymmetric catalysis employing chiral iridium (ruthenium) complexes. Chinese Journal of Inorganic Chemistry, 2024, 40(11): 2091-2104. doi: 10.11862/CJIC.20240265
16. [16]
  Ruiying WANG , Hui WANG , Fenglan CHAI , Zhinan ZUO , Benlai WU . Three-dimensional homochiral Eu(Ⅲ) coordination polymer and its amino acid configuration recognition. Chinese Journal of Inorganic Chemistry, 2025, 41(5): 877-884. doi: 10.11862/CJIC.20250052
17. [17]
  Conghao Shi , Ranran Wang , Juli Jiang , Leyong Wang . The Illustration on Stereoisomers of Macrocycles Containing Multiple Chiral Centers via Tröger Base-based Macrocycles. University Chemistry, 2024, 39(7): 394-397. doi: 10.3866/PKU.DXHX202311034
18. [18]
  Rong Tian , Yadi Yang , Naihao Lu . Comprehensive Experimental Design of Undergraduate Students Based on Interdisciplinarity: Study on the Effect of Quercetin on Chlorination Activity of Myeloperoxidase. University Chemistry, 2024, 39(8): 247-254. doi: 10.3866/PKU.DXHX202312064
19. [19]
  Xiaoning TANG , Junnan LIU , Xingfu YANG , Jie LEI , Qiuyang LUO , Shu XIA , An XUE . Effect of sodium alginate-sodium carboxymethylcellulose gel layer on the stability of Zn anodes. Chinese Journal of Inorganic Chemistry, 2024, 40(8): 1452-1460. doi: 10.11862/CJIC.20240191
20. [20]
  Qin Hou , Jiayi Hou , Aiju Shi , Xingliang Xu , Yuanhong Zhang , Yijing Li , Juying Hou , Yanfang Wang . Preparation of Cuprous Iodide Coordination Polymer and Fluorescent Detection of Nitrite: A Comprehensive Chemical Design Experiment. University Chemistry, 2024, 39(8): 221-229. doi: 10.3866/PKU.DXHX202312056

Get Citation

PDF

XML

Metrics

PDF Downloads(14)
Abstract views(841)
HTML views(71)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142