DEVELOPMENT AND VALIDATION OF HPLC-MS/MS METHOD OF DETERMINATION OF A NEW DERIVATIVE OF VALPROLEIC ACID AND 1,3,4-THIADIAZOLE IN RABBIT BLOOD PLASMAFOR PHARMACOKINETIC STUDY

DOI: https://doi.org/10.29296/25877313-2020-08-04
Issue: 
8
Year: 
2020

A.S. Malygin Post-graduate Student, Tver State Medical University (Tver) ORCID ID 0000-0003-1955-5105 E-mail: tgmu-nauka@mail.ru N.S. Popov Ph.D. (Pharm.), Assistant, Tver State Medical University (Tver) ORCID ID 0000-0002-1792-7414 M.A. Demidova Dr.Sc. (Med.), Professor, Tver State Medical University (Tver) ORCID ID 0000-0002-0270-8809 N.A. Shatokhina Ph.D. (Med.), Associate Professor, Tver State Medical University (Tver) ORCID ID 0000-0001-6970-1313

Objective. To develop and validate the HPLC-mass spectrometric method for determining a new derivative of valproic acid and 1,3,4-thiadiazole in rabbit plasma. Material and methods. The object of the study was N-(5-ethyl-1,3,4-thiadiazol-2-yl) -2-propylpentanamide (valprazolamide) – a new antiepileptic drug from the group of thiadiazole derivatives of valproic acid. Valprazolamide was determined in rabbit plasma using an Agilent 1260 Infinity II high perfor-mance liquid chromatograph (Agilent Technologies, Germany). An AB Sciex QTrap 3200 MD triple quadrupole mass spectrometer (AB Sciex, Singapore) with an electrospray ion source (Turbo V with a TurboIonSpray probe) was used as a detector. Selectivity, carry over, linearity, accuracy, precision, ma-trix effectand recovery were evaluated for the developed method. Results. A method of HPLC-mass spectrometric determination of valprazolamide in rabbit plasma was developed (Phenomenex Synergi C18 analytical column 4 μm 2.0 × 50 mm, gradient elution: 0–1 minute 10% aqueous acetonitrile solution; 1–3 minutes linear increase in concentration acetonitrile in a mixture up to 90%; 3–4 minutes isocratic section with an acetonitrile concentration of 90%; 4–5 minutes conditioning the column with a 10% solu-tion; flow rate of the mobile phase – 0.6 ml / min; volume of sample injected – 20 μl; total time gradient elution – 5 min t; mass spectrometric detec-tion). Mass detection conditions: positive polarization, voltage of the electrospray 5500.0 V, declustering potential 41.0; the curtain gas pressure is 20.0 psi, the spray gas is 40.0 psi, the input potential is 3.5 V. The MRM transitions for valprazolamide were m/z 256.1 → m/z 130.1 and m/z 81.0. Acetazo-lamide (m/z 223.1 → m/z 73.0) was used as an internal standard. Conclusion. The analytical range of the method for determining valprazolamide is 1–1000 ng/ml. The developed method is selective, accurate, preci-sion and linear, it meets the requirements for the validation of bioanalytical methodsin all respects.

Keywords: 
HPLC-MS/MS
valproic acid
1
3
4-thiadiazole
antiepileptic drugs
mass spectrometry

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