ISSN print edition: 0366-6352 ISSN electronic edition: 1336-9075 Registr. No.: MK SR 9/7 Published monthly

Highly sensitive assay of dopamine and tyrosinase using benzoyl peroxide facilitated in-situ fluorogenic reaction

Yu Ding, Li-Hong Yang, Ji-Wei Shen, Yin-Mao Wei, and Chao-Zhan Wang

Key Laboratory of Synthetic and Natural Functional Molecule of the Ministry of Education, College of Chemistry and Materials Science, Northwest University, Xi’an, China

E-mail: czwang@nwu.edu.cn

Received: 21 December 2021  Accepted: 10 March 2022

Abstract:

In this work, simple, cost efficient, highly selective and sensitive assay methods for dopamine (DA) concentration and tyrosinase (TYR) activity were established based on a benzoyl peroxide (BPO) facilitated in situ “turn-on” fluorogenic reaction between resorcinol (RS) and DA, in which BPO was introduced in the system to accelerate DA oxidation, thereby realizing a faster reaction with RS to form the fluorescent substance, azamonardine, which was verified by HPLC, ¹H NMR, ¹³C NMR and high resolution mass spectrometry. The introduction of BPO remarkably improved the sensitivity, linearity range and response speed for assay of DA and TYR. For the determination of DA concentration, the concentration has a good linear relationship in the range of 4.0–1.2 μM, and the detection limit is as low as 0.8 nM, and it was successfully applied to measuring DA in human urine and serum samples. In addition, a fluorescent method for determining the TYR activity was further established based on the enzymatic oxidation of tyramine to DA, the detection limit is 0.0013 U mL⁻¹. And the method was used to determine TYR in human serum and screening of TYR inhibitors.

Keywords: Benzoyl peroxide; Oxidation; Tyrosinase; Dopamine; Fluorescence

Full paper is available at www.springerlink.com.

DOI: 10.1007/s11696-022-02169-1

Chemical Papers 76 (7) 4235–4244 (2022)