A method for determining blood drug concentration of risperidone and paliperidone by two-dimensional liquid phase method

Abstract

The present application relates to risperidone and paliperidone blood concentration determination technical field, specifically refers to a kind of two-dimensional liquid phase method for determining the method for determining risperidone and paliperidone blood concentration, specifically comprising the following steps: selecting two-dimensional liquid phase instrument: Hunan Demet instrument Co., Ltd. FLC2701SPD-M30A full-automatic two-dimensional liquid phase system;Select calibration and quality control product source: Hunan Demet instrument Co., Ltd. Calibration product batch number: 20221219, quality control product batch number: 20230221;Chromatographic conditions are as follows: one-dimensional column: SXI 3.5*25mm*5um;Intermediate column: SCB 3.0*10mm*2.8um;Two-dimensional column: MSCB 3.0*50mm*3.3um.The present application has the advantages that: step design is reasonable, simple operation, low cost, while it can guarantee the accuracy and reliability of the determination of risperidone paliperidone blood concentration analysis result, for blood concentration detection work, promote two-dimensional liquid phase method for clinical detection, from saving cost, accelerate inspection TAT have higher value, have good popularization prospect and economic value.

Description

Technical Field

[0001] This invention relates to the field of risperidone and paliperidone blood drug concentration determination technology, specifically to a two-dimensional liquid chromatography method for determining risperidone and paliperidone blood drug concentrations. Background Technology

[0002] Risperidone, an organic compound with the chemical formula C23H27FN4O2, is a psychiatric medication used to treat acute and chronic schizophrenia. It is particularly effective for positive and negative symptoms and their associated affective symptoms (such as anxiety and depression). It can also alleviate affective symptoms associated with schizophrenia. For patients who respond to acute treatment, this medication can continue to exert its clinical efficacy during maintenance therapy.

[0003] Paliperidone, with the chemical formula 3-[2-[4-(6-fluoro-1,2-benzoxazol-3-yl)piperidin-1-yl]ethyl]-9-hydroxy-2-methyl-6,7,8,9-tetrahydropyrido[1,2-a]pyrimidin-4-one and molecular formula C23H27FN4O3, has a density of 1.45 g / cm3 and is a grayish-white to pale orange solid. Clinically, it is used to treat schizophrenia and schizoaffective disorder. Paliperidone is the major metabolite of risperidone. Its mechanism of action is not fully understood, but it is currently believed to be mediated through the combined antagonism of central dopamine 2 (D2) receptors and serotonin 2 (5HT2A) receptors. Paliperidone is also an antagonist of α1 and α2 adrenergic receptors and H1 histamine receptors, which may explain some of its other effects. Paliperidone has no affinity for cholinergic muscarinic receptors or β1- and β2-adrenergic receptors. In vitro, the pharmacological effects of (+)- and (-)-paliperidone enantiomers are similar.

[0004] Two-dimensional liquid chromatography (2D liquid chromatography) is less expensive than mass spectrometry (MS) with self-calibrated internal standards. However, due to the difference in detection sensitivity compared to mass spectrometry, it has difficulty detecting low concentrations of drugs. The sample injection volume is relatively large, the retention time is generally 5-10 minutes, the detection speed is slow, and the efficiency is not high. Currently, the range of drugs that can be detected is limited. However, for newly established laboratories with small sample volumes and limited detection needs, since the detection instruments and detection costs are lower than those of mass spectrometry isotope methods and self-calibrated internal standards, 2D liquid chromatography can be promoted first, and then upgraded to a new mass spectrometry detection system after three to five years, depending on the needs. Summary of the Invention

[0005] The purpose of this invention is to provide a two-dimensional liquid chromatography method for determining the blood concentrations of risperidone and paliperidone, in order to solve the problems mentioned in the background art.

[0006] To solve the above-mentioned technical problems, the present invention provides the following technical solution: a method for determining the blood concentrations of risperidone and paliperidone using a two-dimensional liquid chromatography method, specifically including the following steps:

[0007] 1) Selected two-dimensional liquid chromatography instrument: Hunan Demeter Instrument Co., Ltd. FLC2701SPD-M30A fully automatic two-dimensional liquid chromatography system;

[0008] 2) Source of calibrators and quality control products: Hunan Demeter Instrument Co., Ltd., calibrator batch number: 20221219, quality control product batch number: 20230221;

[0009] 3) The chromatographic conditions are as follows:

[0010] One-dimensional chromatographic column: SXI 3.5*25mm*5um;

[0011] Intermediate column: SCB 3.0*10mm*2.8um;

[0012] Two-dimensional column: MSCB 3.0*50mm*3.3um;

[0013] One-dimensional reagent: CAA-1D mobile phase model CAA-1B;

[0014] Two-dimensional reagent: BPI3-2A: API3-2A: MPI-2A: OPI-2A = 20:60:0:20;

[0015] Auxiliary reagent: pure water;

[0016] Processing method ORG-1 processing temperature: 50℃;

[0017] Detection wavelength CH1: 276nm; Sample volume: 150uL;

[0018] CH2:286nm Method Name: Risperidone Paliperidone - Secondary Simplex Method - 150uL.lcm;

[0019] Flow rates: Pump A: 0.7 ml / min; Pump B: 0.01 ml / min; Pump C: 0.6 ml / min;

[0020] Time program: The secondary simplex constant pressure pump stops automatically.

[0021] The advantages of this invention are: reasonable step design, simple operation, and low cost. At the same time, it can ensure the accuracy and reliability of the blood drug concentration analysis results of risperidone and paliperidone. For blood drug concentration detection, promoting the use of two-dimensional liquid chromatography for clinical testing has high value in terms of cost saving and speeding up the testing of TAT, and has good prospects for promotion and economic value. Attached Figure Description

[0022] Figure 1 This is a time program diagram for measuring risperidone and paliperidone blood drug concentrations.

[0023] Figure 2 This is a confirmation LC2 chromatogram of risperidone and paliperidone blood drug concentration measurement. Paliperidone peak area: 431142; risperidone peak area: 557298.

[0024] Figure 3 This is a confirmation graph of the online blood concentration measurement of risperidone and paliperidone. Paliperidone peak area: 389011; risperidone peak area: 499325.

[0025] Figure 4 This is a confirmation graph of the standard curve for measuring risperidone blood concentration.

[0026] Figure 5 This is a confirmation graph of the standard curve for measuring paliperidone blood concentration.

[0027] Figure 6 The QCL chart confirms the blood drug concentration quality control results for risperidone and paliperidone: Deviation: paliperidone 1.0%, risperidone 6.9%.

[0028] Figure 7 The QCM chart confirms the blood drug concentration quality control results for risperidone and paliperidone: Deviation: paliperidone -1.19%, risperidone 3.83%.

[0029] Figure 8 The QCH chart confirms the blood drug concentration quality control results for risperidone and paliperidone: Deviation: paliperidone 0.0001%, risperidone 3.8%. Detailed Implementation

[0030] For the purposes of the detailed description below, it should be understood that the invention may take various alternative variations and sequences of steps unless expressly stated otherwise. Furthermore, except in any operational instance, or otherwise indicated, all figures representing the amounts of ingredients used, for example, in the specification and claims, should be understood to be modified in all cases by the term “about.” Therefore, unless indicated to the contrary, the numerical parameters set forth in the following specification and appended claims are approximations varying with the desired performance to be obtained according to the invention. It is not at all an attempt to limit the application of the doctrine of equivalents to the scope of the claims; each numerical parameter should be interpreted at least according to the number of significant figures reported and by applying ordinary rounding techniques.

[0031] Although the numerical ranges and parameters illustrating the broad scope of the invention are approximate, the values ​​listed in the specific examples are reported as precisely as possible. However, any numerical value inherently contains some error that is necessarily caused by the standard deviation found in their respective test measurements.

[0032] Furthermore, it should be understood that any numerical range described herein is intended to include all subranges falling within it. For example, the range “1 to 10” is intended to include all subranges between the minimum value 1 and the maximum value 10, i.e., a minimum value equal to or greater than 1 and a maximum value equal to or less than 10.

[0033] Example

[0034] A two-dimensional liquid chromatography method for determining the blood concentrations of risperidone and paliperidone specifically includes the following steps:

[0035] 1) Selected two-dimensional liquid chromatography instrument: Hunan Demeter Instrument Co., Ltd. FLC2701SPD-M30A fully automatic two-dimensional liquid chromatography system;

[0036] 2) Source of calibrators and quality control products: Hunan Demeter Instrument Co., Ltd., calibrator batch number: 20221219, quality control product batch number: 20230221;

[0037] 3) The chromatographic conditions are as follows:

[0038] One-dimensional chromatographic column: SXI 3.5*25mm*5um;

[0039] Intermediate column: SCB 3.0*10mm*2.8um;

[0040] Two-dimensional column: MSCB 3.0*50mm*3.3um;

[0041] One-dimensional reagent: CAA-1D mobile phase model CAA-1B;

[0042] Two-dimensional reagent: BPI3-2A: API3-2A: MPI-2A: OPI-2A = 20:60:0:20;

[0043] Auxiliary reagent: pure water;

[0044] Processing method ORG-1 processing temperature: 50℃;

[0045] Detection wavelength CH1: 276nm; Sample volume: 150uL;

[0046] CH2:286nm Method Name: Risperidone Paliperidone - Secondary Simplex Method - 150uL.lcm;

[0047] Flow rates: Pump A: 0.7 ml / min; Pump B: 0.01 ml / min; Pump C: 0.6 ml / min;

[0048] Time program: The secondary simplex constant pressure pump stops automatically.

[0049] in conclusion

[0050] ● The transfer recovery rate of risperidone was 90.7%, and the transfer recovery rate of paliperidone was 90.2%, which meets the transfer recovery rate requirement of <90%-110%>.

[0051] ● Risperidone standard curve R 2 =0.9999761, Paliperidone standard curve R 2 =0.9999176 meets the methodological validation requirements;

[0052] ● The quality control results meet the method validation requirements, and the chromatograms are pure and free of interference.

[0053] The comparison between two-dimensional liquid chromatography (2D-LC) and mass spectrometry (MS / ISO) for determining risperidone and paliperidone blood concentrations concluded that MS / ISO is the reference method for blood drug concentration detection. Comparison of the research data showed a high degree of consistency between the two-dimensional liquid chromatography and IOSO results. With a CV of 10%, only 5 samples with lower concentrations showed 2D-LC results outside the acceptable range compared to the IOSO results out of range. This indicates that 2D-LC can be used for routine clinical detection of risperidone and paliperidone blood concentrations.

[0054] In summary, for blood drug concentration detection, mass spectrometry isotope method is used for research, while promoting mass spectrometry self-calibration internal standard method and two-dimensional liquid chromatography method for clinical testing is of great value in terms of cost saving and speeding up TAT testing.

[0055] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A method for determining the blood concentrations of risperidone and paliperidone using two-dimensional liquid chromatography, characterized in that, Specifically, the following steps are included: 1) Selected two-dimensional liquid chromatography instrument: Hunan Demeter Instrument Co., Ltd. FLC2701 SPD-M30A fully automatic two-dimensional liquid chromatography system; 2) Source of calibrators and quality control products: Hunan Demeter Instrument Co., Ltd., calibrator batch number: 20221219, quality control product batch number: 20230221; 3) The chromatographic conditions are as follows: One-dimensional chromatographic column: SXI 3.5*25mm*5um; Intermediate column: SCB 3.0*10mm*2.8um; Two-dimensional column: MSCB 3.0*50mm*3.3um; One-dimensional reagent: CAA-1D mobile phase model CAA-1B; Two-dimensional reagent: BPI3-2A: API3-2A: MPI-2A: OPI-2A = 20:60:0:20; Auxiliary reagent: pure water; Processing method ORG-1 processing temperature: 50℃; Detection wavelength CH1: 276nm; Sample volume: 150uL; CH2:286nm Method Name: Risperidone Paliperidone - Secondary Simplex Method - 150uL.lcm; Flow rates: Pump A: 0.7 ml / min; Pump B: 0.01 ml / min; Pump C: 0.6 ml / min; Time program: The secondary simplex constant pressure pump stops automatically.

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