[Determination of 3-nitropropionic acid in plasma by high-performance liquid chromatography-tandem mass spectrometry method].

Objective

To establish a rapid method for detecting 3-nitropropionic acid (3-NPA) in plasma using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) .

Methods

In May 2024, the plasma samples were extracted by precipitating proteins with methanol∶ acetonitrile (volume ratio 1∶1). Then, they were analyzed using a C18 chromatographic column with a gradient elution using a 0.1% formic acid aqueous solution and acetonitrile as the mobile phase, in the electrospray negative ion mode (ESI-) and the multiple reaction monitoring mode (MRM). The method was quantified using external standards. The linearity range, sensitivity, accuracy, and precision of the method were verified.

Results

The HPLC-MS/MS method for measuring 3-NPA in plasma exhibited a good linear relationship within the concentration range of 1-50 μg/L, with a correlation coefficient of 0.9996. The detection limit of the method was 0.1 μg/L, and the quantitative lower limit was 0.3 μg/L. The recovery rate of the spiked samples ranged from 92.0% to 103.4%, and the intra-and inter-day precisions were 2.4%-9.1% and 2.8%-7.3%, respectively.

Conclusion

The HPLC-MS/MS method is simple, rapid, sensitive and accurate, and is suitable for the qualitative and quantitative analysis of 3-NPA in the plasma of patients with sugarcane poisoning or occupational exposed population.