心型脂肪酸结合蛋白纯品含量的柱前衍生超高效液相色谱方法研究

doi:10.3969/j.issn.1000-1158.2023.07.21

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摘要心型脂肪酸结合蛋白(H-FABP)被酸水解后,以氨基酸混合溶液国家标准物质为标准,γ-氨基丁酸为内标,经柱前衍生剂处理,再采用超高效液相色谱法可测定其中天冬氨酸、谷氨酸、组氨酸、精氨酸4种衍生物的量,进而推算出H-FABP纯品的含量。将此方法的测定结果与同位素稀释质谱法测定H-FABP含量的结果相比较,可验证所建方法的准确性。实验显示,在最优条件下,所建立的柱前衍生超高效液相色谱(UPLC)法中H-FABP含量测定结果为0.1113mg/mL,相对标准偏差为2.03%,扩展不确定度为0.0027mg/mL(k=2),检出限和定量限分别为1.16×10^-3mg/mL和3.86×10^-3mg/mL。此外,该方法成本较低且溯源链清晰,可作为一种补充验证方法用于纯品H-FABP标准物质的定值。

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	崔宏恩
	冯速
	叶人元
	刘阳
	方帅
	朱茂林
	樊晓琳

关键词 ：计量学, 心型脂肪酸结合蛋白, 定量分析, 柱前衍生超高效液相色谱法, 同位素稀释质谱法

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.

Key words： metrology heart-type fatty acid-binding protein mass fraction pre-column derivatization ultra performance liquid chromatography high performance liquid chromatography-isotope dilution mass spectrometry

收稿日期: 2023-01-09 发布日期: 2023-07-17

PACS:

TB99

基金资助:江苏省市场监督管理局科技计划(KJ185602)

作者简介: 崔宏恩(1987-),男,安徽亳州人,江苏省计量科学研究院高级工程师,主要从事生物计量和标准物质等方面的研究。Emial:cuihongen@163.com

引用本文:

崔宏恩,冯速,叶人元,刘阳,方帅,朱茂林,樊晓琳. 心型脂肪酸结合蛋白纯品含量的柱前衍生超高效液相色谱方法研究[J]. 计量学报, 2023, 44(7): 1141-1146.
CUI Hong-en,FENG Su,YE Ren-yuan,LIU Yang,FANG Shuai,ZHU Mao-lin,FAN Xiao-lin. Determination of the Mass Fraction of Pure Heart-type Fatty Acid-binding Protein by Pre-column Derivatization UPLC. Acta Metrologica Sinica, 2023, 44(7): 1141-1146.

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