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. 2021 Nov 16;10(11):2814.
doi: 10.3390/foods10112814.

Simultaneous Determination of Purines and Uric Acid in Chinese Chicken Broth Using TFA/FA Hydrolysis Coupled with HPLC-VWD

Manli Wu  1   2   3   4 , Wangang Zhang  1   2   3   4 , Xixi Shen  1   2   3   4 , Wei Wang  1   2   3   4
Affiliations

Simultaneous Determination of Purines and Uric Acid in Chinese Chicken Broth Using TFA/FA Hydrolysis Coupled with HPLC-VWD

Manli Wu et al. Foods. .

Abstract

Chinese chicken broth is well known for its outstanding nutritional value and flavor, widely consumed in China. This study was designed to develop a sensitive and accurate high-performance liquid chromatography-variable wavelength detector (HPLC-VWD) method to simultaneously determine purines and uric acid in Chinese chicken broth for gout and hyperuricemia dietary management. Chromatographic separation was performed on an Agilent TC-C18 (2) column (4.6 mm × 250 mm, 5.0 µm), using 0.02 M KH2PO4 (pH 4.0) as a mobile phase. Sample pretreatment was optimized to enable the extraction of all analytes from Chinese chicken broth. The optimal pretreatment conditions were chicken broth-60% trifluoroacetic acid (TFA)/20% formic acid (FA) (1:1, v/v) in a volume ratio of 1:3 and hydrolysis for 40 min at 85 °C in a water bath. The limits of detection (LODs) and limits of quantification (LOQs) of the purines and uric acid were 0.58-1.71 µg/L and 1.92-5.70 µg/L, respectively. The recoveries were 91-101%, with the relative standard deviations (RSDs) lower than 3%. The complete method has been successfully applied to determine purines and uric acid in various Chinese chicken soups obtained from different provinces in China.

Keywords: Chinese chicken broth; HPLC; acid hydrolysis; gout; hyperuricemia; purine; uric acid.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Exogenous purines metabolic pathways.
Figure 2
Figure 2
UV full wavelength scan of purine bases and uric acid.
Figure 3
Figure 3
High-performance liquid chromatography (HPLC) profiles of four purine bases and uric acid at 254 nm. (a) Mixed standard working solution (5 mg/L); (b) Chinese chicken soup sample. Column: Agilent TC-C18 (2) (4.6 mm × 250 mm, 5.0 µm). Mobile phase: 0.2 M KH2PO4 buffer solution, pH = 4. Flow rate: 1.0 mL/min. Column temperature: 30 °C, injection column: 10 µL. Peaks: 1, Uric acid; 2, Guanine; 3, Hypoxanthine; 4, Xanthine; 5, Adenine.
Figure 4
Figure 4
Effect of different factors on extraction rate using the trifluoroacetic acid/formic acid (TFA/FA) method. (a) Hydrolysis temperature; (b) Hydrolysis time; (c) Chicken broth-TFA/FA (1:1, v/v) volume ratio; (d) TFA concentration; (e) FA concentration. The left vertical axis corresponds to the extraction rate of each analyte of adenine, guanine, hypoxanthine, xanthine and uric acid, and the right vertical axis corresponds to the total extraction rate of the five analytes.
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