A method for determining quetiapine blood concentration by mass spectrum self-calibration internal standard method

Abstract

The present application relates to quetiapine blood concentration determination technical field, specifically refers to a kind of mass spectrum self-calibration internal standard method determination quetiapine blood concentration method, specifically includes the following steps:1) select mass spectrometer instrument: hunan demit instrument limited company LC / MS, Ms-Mate9500 high performance liquid chromatography tandem mass spectrometry detection system;2) select calibration and quality control product source: hunan demit instrument limited company calibration batch number: 20221219, quality control product batch number: 20230221;3) chromatographic condition: chromatographic column: MSCB-2A3.0*50mm*2.5um (double column alternation);Quetiapine mass / charge ratio (M / Z): 384;Pretreatment: 200ul sample+500ulMARG-1, atomizer flow: 1.5L / min, characteristic ion: 384+heating block temperature: 350℃, flow rate: 0.4mL / min, dry gas flow: 10L / min, temperature: 45℃QarrayRF: 0V, sample amount: 1uL, ion source mode: ESI+.The present application has the advantages that: step design is reasonable, simple operation, low cost, while it can guarantee the accuracy and reliability of determination quetiapine blood concentration analysis result, with good popularization prospect and economic value.

Description

Technical Field

[0001] This invention relates to the field of quetiapine blood concentration determination technology, specifically to a method for determining quetiapine blood concentration using a mass spectrometry self-calibration internal standard method. Background Technology

[0002] Quetiapine, ① is used for all types of schizophrenia, effective not only for positive symptoms but also for some negative symptoms. ② It can also alleviate affective symptoms associated with schizophrenia, such as depression, anxiety, and cognitive deficits. It belongs to the dibenzo-oxazine class of atypical antipsychotics. It has affinity for 5-HT, dopamine D2, histamine, and H1-, α1-, and α2- receptors.

[0003] The internal standard method in mass spectrometry is a quantitative method. Its principle involves adding a specific weight of a pure substance as an internal standard to a specific amount of the analyte mixture. Then, mass spectrometry analysis is performed on the sample containing the internal standard to determine the peak area (or peak height) and relative correction factor of both the internal standard and the analyte. The percentage content of the analyte in the sample can then be calculated using a formula. The advantages of the internal standard method include: 1. It can correct for instrument fluctuation errors caused by analysis at different times. 2. It can correct for errors in the injection volume during analysis and eliminates interference from other components in the sample. 3. The internal standard method has a wide range of applications, especially when a standard reference is unavailable, making it particularly advantageous.

[0004] The in-house calibration method is a method for quantitative analysis of specific analytes in unknown samples by calibrating an internal standard at a known concentration in a laboratory setting. In the determination of quetiapine and clozapine, the in-house calibration method can determine the concentration of the standard internal standard by preparing a standard curve, and then use this standard curve to calibrate the high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS / MS) isotope internal standard method, thereby achieving accurate determination of quetiapine and clozapine. Both HPLC-MS / MS and the in-house calibration method are common quantitative analysis methods widely used in drug development, biomedical research, and clinical testing.

[0005] The advantages of self-calibrated internal standards are their simplicity, low cost, and ability to ensure the accuracy and reliability of analytical results. However, their disadvantages include the need for specialized knowledge and skills in preparing the standard curve, and the fact that the prepared standard curve may only be applicable to specific analytes and may not be accurate enough for other analytes. Currently, the main approach to addressing this issue is to prepare mixed internal standards containing multiple drugs, allowing for simultaneous use by various detection methods to detect multiple drugs. Due to its low cost and rapid operation, it has high application value in clinical laboratories with large sample sizes. Summary of the Invention

[0006] The purpose of this invention is to provide a method for determining the blood concentration of quetiapine using a mass spectrometry self-calibration internal standard method, in order to solve the problems mentioned in the background art.

[0007] To solve the above-mentioned technical problems, the technical solution provided by the present invention is: a method for determining the blood concentration of quetiapine using a mass spectrometry self-calibration internal standard method, specifically including the following steps:

[0008] 1) Mass spectrometry instrument selected: Hunan Demeter Instrument Co., Ltd. LC / MS, Ms-Mate9500 high performance liquid chromatography-tandem mass spectrometry detection system;

[0009] 2) Source of calibrators and quality control products: Hunan Demeter Instrument Co., Ltd., calibrator batch number: 20221219, quality control product batch number: 20230221;

[0010] 3) The chromatographic conditions are as follows:

[0011] Chromatographic column: MSCB-2A 3.0*50mm*2.5um (dual column alternation), column A serial number: 2DC21C035, column B serial number: 2DC21C036;

[0012] Quetiapine / charge ratio (M / Z): 384

[0013]

[0014] 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 results of quetiapine blood concentration analysis. For blood drug concentration detection, promoting the use of mass spectrometry self-calibration internal standard method for clinical testing has high value in terms of cost saving and speeding up TAT testing, and has good prospects for promotion and economic value.

[0015] As a preferred option, the column interface temperature is 350℃.

[0016] As a preferred method, the ratio of quetiapine reagent is: MMA-2A: NMD-2A: LPA-2A = 30:55:15, and the DL temperature is 250℃.

[0017] As a preferred option, MMA-2A batch number: 20231012-1, LP-2A batch number: 20230424-1, NMD-2A batch number: 20231017-1. Attached Figure Description

[0018] Figure 1 It is a blank smear with self-calibrated internal standard TIC plot.

[0019] Figure 2 It is the TIC spectrum of the calibrator STD3+ self-calibration internal standard.

[0020] Figure 3 This is the quetiapine correction curve and infographic.

[0021] Figure 4 This is the QCM spectrum of quetiapine. Detailed Implementation

[0022] 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.

[0023] 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.

[0024] 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.

[0025] Example

[0026] A method for determining quetiapine blood concentration using a mass spectrometry self-calibrated internal standard method includes the following steps:

[0027] 1) Mass spectrometry instrument selected: Hunan Demeter Instrument Co., Ltd. LC / MS, Ms-Mate9500 high performance liquid chromatography-tandem mass spectrometry detection system;

[0028] 2) Source of calibrators and quality control products: Hunan Demeter Instrument Co., Ltd., calibrator batch number: 20221219, quality control product batch number: 20230221;

[0029] 3) The chromatographic conditions are as follows:

[0030] Chromatographic column: MSCB-2A 3.0*50mm*2.5um (dual column alternation), column A serial number: 2DC21C035, column B serial number: 2DC21C036;

[0031] Quetiapine / charge ratio (M / Z): 384

[0032]

[0033] As a preferred embodiment, the column interface temperature is 350°C.

[0034] As a preferred embodiment, the ratio of quetiapine reagent is: MMA-2A: NMD-2A: LPA-2A = 30:55:15, and the DL temperature is 250℃.

[0035] As a preferred embodiment, MMA-2A batch number: 20231012-1, LP-2A batch number: 20230424-1, NMD-2A batch number: 20231017-1.

[0036] Time Procedure: See Table 1 Internal Standard: MPS-5 Self-calibration Internal Standard Table 1 Time Procedure Table

[0037]

[0038]

[0039]

[0040] National Health Commission Clinical Laboratory Center

[0041] Statistical results of the 2024 interlaboratory quality assessment for monitoring psychotropic drugs.

[0042] Laboratory Name: Department of Laboratory Medicine, Third People's Hospital of Qujing City, Yunnan Province

[0043] Laboratory code: 128146

[0044] Measurement date: 2024-03-01 First statistical analysis date: 2024-04-17

[0045] Item: Quetiapine ng / ml

[0046]

[0047] Method: Two-dimensional liquid chromatography-mass spectrometry (2D LC-MS) Instrument: Hunan Demeter HPLC

[0048] Reagents: Hunan Demeter Company; Calibrators: Hunan Demeter Company

[0049] Conclusion: The chromatographic and mass spectrometry parameters were complete and reasonable; the quality control results met the requirements for method validation; and the chromatograms were pure and free of interference.

[0050] The high, medium and low concentrations were controlled by internal quality control. The target values ​​were provided by Hunan Demeter Company. The mass spectrometry blood drug concentration was calculated based on the calibration curve obtained by using the ratio of the peak area of ​​the sample to the area of ​​the internal standard (isotope or self-calibrated internal standard) as the x-axis and the concentration value as the y-axis. The retention time of the method for determining the blood concentration of quetiapine by mass spectrometry self-calibrated internal standard method of this invention is 2.159.

[0051] The comparison between high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS / MS) and isotope- and non-isotope methods for determining quetiapine blood concentrations concluded that the mass spectrometry isotope method is a reference method for blood drug concentration detection. Comparison of the research data showed a high degree of consistency between the self-calibrated internal standard method and the isotope method. With a CV of 10%, only 6 samples with lower concentrations showed results outside the range of the self-calibrated internal standard method out of range compared to the isotope method out of range. This indicates that the mass spectrometry self-calibrated internal standard method can be used for routine clinical detection of quetiapine blood concentrations.

[0052] High-performance liquid chromatography-tandem mass spectrometry (HPLC-MS-MS) is a highly sensitive and high-throughput analytical technique that can simultaneously detect and quantify multiple compounds in a sample. In the content determination of various drugs such as quetiapine and clozapine, HPLC-MS-MS can be used as a quantitative analysis method. By adding an internal standard of known concentration, the determination results can be calibrated, thereby improving the accuracy of the determination.

[0053] In conclusion, for blood drug concentration detection, mass spectrometry isotope method is used for research, and promoting the use of mass spectrometry self-calibration internal standard method for clinical testing is of great value in terms of cost saving and speeding up TAT testing.

[0054] 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 quetiapine blood concentration using a mass spectrometry self-calibration internal standard method, characterized in that, Specifically, the following steps are included: 1) Mass spectrometry instrument selected: Hunan Demeter Instrument Co., Ltd. LC / MS, Ms-Mate9500 high performance liquid chromatography-tandem mass spectrometry detection 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: Chromatographic column: MSCB-2A 3.0*50mm*2.5um (dual column alternation), column A serial number: 2DC21C035, column B serial number: 2DC21C036; Quetiapine / charge ratio (M / Z): 384 Pretreatment: 200µl sample + 500µl MARG-1 nebulizer; flow rate: 1.5L / min Characteristic ion: 384+ Heating block temperature: 350℃ Flow rate: 0.4 mL / min; Drying gas flow rate: 10 L / min Temperature: 45℃ Qarray RF: 0V Injection volume: 1 μL; Ion source mode: ESI+.

2. The method for determining quetiapine blood concentration using a mass spectrometry self-calibration internal standard method as described in claim 1, characterized in that: The column interface temperature is 350℃.

3. The method for determining quetiapine blood concentration using a mass spectrometry self-calibration internal standard method as described in claim 1, characterized in that: Quetiapine reagent ratio: MMA-2A: NMD-2A: LPA-2A = 30:55:15, DL temperature: 250℃.

4. The method for determining quetiapine blood concentration by mass spectrometry self-calibration internal standard method as described in claim 3, characterized in that: MMA-2A batch number: 20231012-1, LP-2A batch number: 20230424-1, NMD-2A batch number: 20231017-1.

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