S.S. Babkina a*, N.A. Ulakhovich b**, E.P. Medyantseva b***, A.R. Gataulina b****
aMIREA – Russian Technological University, Moscow, 119454 Russia
bKazan Federal University, Kazan, 420008 Russia
E-mail: *email@example.com, **Nikolay.Ulakhovich@kpfu.ru, ***firstname.lastname@example.org, ****email@example.com
Received October 23, 2019
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For citation: Babkina S.S., Ulakhovich N.А., Medyantseva E.P., Gataulina A.R. Bioaffinity-based method for catecholamines determination with amperometric DNA sensor. Uchenye Zapiski Kazanskogo Universiteta. Seriya Estestvennye Nauki, 2020, vol. 162, no. 1, pp. 5–16. doi: 10.26907/2542-064X.2020.1.5-16. (In Russian)
A bioaffinity-based method for the determination of adrenaline and dopamine involving immobilized renatured DNA incorporated into the amperometric biosensor was developed.
With the help of spectrophometry and voltammetry data, it was found that the composition of complexes formed with catecholamines and their stability constants were changing with concentration from [КАL2] to [КА2L], where КА – catecholamine, L – base pare, and lg βeff = 10.1–13.5. The high affinity of adrenaline and dopamine to the immobilized renatured DNA molecule was revealed, thereby making it possible to run effective concentration of KA from the analyzed solutions of low concentration including multicomponent fluids.
The minimum reporting level was found to be 5.5·10–9 (adrenaline) and 2.0·10–8 (dopamin) mol/L, respectively. The proposed method is characterized by high sensitivity, reproducibility (relative accuracy does not exceed 5%), and single determination time shorter than 30 min.
Keywords: adrenaline, amperometric sensor, bioaffinity-based method, biosensor, DNA, dopamine, catecholamines
Fig. 1. Changes in the mole fraction of adrenaline – r-IDNA complexes: 1 – changes in the mole fraction of the [AD2L] complex; 2 – changes in the mole fraction of the [ADL2] complex; cDNA = 0.01 mg/mL, acetate buffer, pH 4.9.
Fig. 2. Changes in the mole fraction of dopamine – r-IDNA complexes: 1 – changes in the mole fraction of the [DA2L] complex; 2 – changes in the mole fraction of the [DАL2] complex; cDNA = 0.01 mg/mL, acetate buffer, pH 4.9.
Fig. 3. Dependence of the oxidation current of AD and DA on the preconcentration time of catecholamines on the biosensor containing r-IDNA: 1 – cAD = 1·10–7 mol/L; 2 – cDA = 5·10–6 mol/L.
Fig. 4. Dependence of the oxidation current of AD and DA on the reactivation time of the biosensor containing r-IDNK: 1 – cAD = 1·10–7 mol/L; 2 – cDA = 5·10–6 mol/L.
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