A Comparative Study Of Modern Analysis Methods For The Determination Of Quetiapine Fumarate In Biological Samples

Schizophrenia is one of the most common mental disorders.It’s pathogenesis is not clearly yet.The course of this disease is very long,and always recurrent attacks.This disease has brought a lot of trouble to people.Atypical antipsychotics quetiapine fumarate（QF）is the preferred drug for the treatment of various types of schizophrenia,because of its excellent curative effect,less side effects and various drug combination.With the development of pharmaceutics and other related technologies,QF extended release formulation began to widely used in clinical.The object of this thesis is QF.We have used three methods to research the preclinical pharmacokinetic in Beagle dogs after oral administration 200 mg of QXR（proprietary）,QXR（generics）and QIR（proprietary）,respectively.Three methods are HPLC-QqQ-MS/MS,HPLC-TOF-MS and FESI-CZE-PDA,respectively.In addition,we have studied the bioequivalence of generic QF extended release formulation,which is manufaxtured by Hefei medical professional pharmaceutical companies.What’s more,the differences of the three methods in determination of biological samples have compared and evaluated,to provide any reference for method selection to determination of the biological samples in the future.This thesis can be divided into the following three parts: 1.Development and validation of HPLC-QqQ-MS/MS method for determination of quetiapine fumarate and it’s bioequivalence study of generic extended release formulation in Beagle dogsIn this section,we have established a HPLC-QqQ-MS/MS method,which is one of the most commonly used drug analysis method,to determination quetiapine in Beagle dog plasma.The drug and internal standard were separated using a Dikma Diamonsil C18（5 μm,100 mm x 4.6 mm）column.The flow rate was 1 mL/min with 3:7 aplit ratio.The gradient elution procedure was used with methanol and water containing 0.1% formic acid.The peak time of drug and IS were between 3.0 3.5 min,and the peak shape is good too.Per sample analysis time was 5.5 min.Beagle dog plasma samples were prepared by methanol precipitation method.There is no interference of plasma endogenous substances.The methodology validation is qualified.This method has been successfully applied to the bioequivalence study of generic extended release formulation in Beagle dogs.To compare with the single doses and multidoses results: the main pharmacokinetic characteristics of t_max,C_max,AUC_0²4 and F_0²4（%）in six Beagle dogs after oral administration 200 mg of QXR（proprietary）and QXR（generics）were no difference;while,there were significant differences between QXR（generics）and QIR（proprietary）.The concentration of quetiapine in Beagle dog plasma began to stabilize at the five day during the one week multidoses test.There was no obvious accumulation in Beagle dogs.2.Development and validation of HPLC-TOF-MS method for determination of quetiapine fumarate and it’s preclinical pharmacokinetic study in Beagle dogsTime of flight mass spectrometry has the characteristics of high resolution,high measurement precision,broad linear range and fast analysis speed and so on.In this chapter,we have developed a HPLC-TOF-MS method to quantitative quetiapine in Beagle dog plasma.Using Agilent C18（3.5 μm,100 mm x 3.0 mm）column to separate the drug and IS within 3 min.Methanol and water containing 0.1% formic acid（60:40,v/v）were adopted as the mobile phase at flow rate of 0.3 mL/min.The pretreatment method was as the same as chapter one,there is no plasma endogenous substance and no residual phenomenon.Methodological evaluation results of this method are in conformity with the relevant requirements of the FDA about biological sample analysis in vivo.This method of HPLC-TOF-MS were successfully applied to the pharmacokinetic study of QXR（proprietary）and QIR（proprietary）in Beagle dogs.3.Development and validation of FESI-CZE-PDA method for determination of quetiapine fumarate and it’s preclinical pharmacokinetic study in Beagle dogsIn this chapter,we have established a FESI-CZE-PDA method with high sensitivity to measurement the concentration of quetiapine.The background electrolyte was composed of 50 mM NaH₂PO₄-H₃PO₄（pH 2.5）.The separation voltage was 13 kV.The sample was introduced by electrokinetic（5 k V × 10 s）modes.And the detection wavelength was set at 210 nm.We used organic solvent online stacking amplification technology by choosing 65% acetonitrile solution as the sample solvent enhanced the sensitivity of quetiapine about 40–50 folds in total,and overcomed the drawback of PDA detector.At the same time,we have adopted a liquid-liquid extraction pretreatment to optimum the quantitative limit of QTP to 1 ng/m L.A rapid and high-throughput sample preparation was applied in 96-well deep format plate in order to save processing time.The methodology validation is qualified.This FESI-CZE-PDA method has been applied to the pharmacokinetic study of QXR（proprietary）and QIR（proprietary）in Beagle dogs.This thesis have established three methods for determination of QTP in biological samples,such as HPLC-QqQ-MS/MS,HPLC-TOF-MS and FESI-CZE-PDA.All of the methods are stable and reliable,and can be used in the pharmacokinetic study of QTP.