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Determination of the Mass Fraction of Pure Heart-type Fatty Acid-binding Protein by Pre-column Derivatization UPLC |
CUI Hong-en1,FENG Su2,YE Ren-yuan2,LIU Yang2,FANG Shuai1,ZHU Mao-lin1,FAN Xiao-lin1 |
1. Jiangsu Institute of Metrology, Nanjing, Jiangsu 210023, China
2. Nanjing Vazyme Biotech Co., Ltd., Nanjing, Jiangsu 210000, China |
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Abstract After the heart type fatty acid binding protein (H-FABP) is hydrolyzed by acid hydrolysis, the amount of four derivatives of aspartic acid, glutamic acid, histidine and arginine can be determined, and then the content of pure H-FABP can be calculated. During the determination process, the national standard substance of amino acid mixed solution was used as the standard, γ-aminobutyric acid was used as the internal standard, and treated with pre column derivatizer. Comparing the results of this method with those of isotope dilution mass spectrometry for the determination of H-FABP content can verify the accuracy of the established method. The experiment showed that under the optimal conditions, the H-FABP content in the established pre column derivatization ultra high performance liquid chromatography (UPLC) method was determined to be 0.1113mg/mL. The relative standard deviation was 2.03%, the expanded uncertainty was 0.0027mg/mL (k=2), and the detection limit and quantification limit were 1.16×10-3mg/mL and 3.86×10-3mg/mL, respectively. In addition, this method has a low cost and clear traceability chain, which may serve as a supplementary validation method for the calibration of pure H-FABP reference materials.
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Received: 09 January 2023
Published: 17 July 2023
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