CN117491542B - Kit and method for detecting psychotropic drugs in dried blood slices - Google Patents
Kit and method for detecting psychotropic drugs in dried blood slices Download PDFInfo
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- CN117491542B CN117491542B CN202311842071.1A CN202311842071A CN117491542B CN 117491542 B CN117491542 B CN 117491542B CN 202311842071 A CN202311842071 A CN 202311842071A CN 117491542 B CN117491542 B CN 117491542B
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- 238000000034 method Methods 0.000 title claims abstract description 65
- 238000001514 detection method Methods 0.000 claims abstract description 45
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- 229960004038 fluvoxamine Drugs 0.000 claims description 40
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
- G01N30/7233—Mass spectrometers interfaced to liquid or supercritical fluid chromatograph
- G01N30/724—Nebulising, aerosol formation or ionisation
- G01N30/7266—Nebulising, aerosol formation or ionisation by electric field, e.g. electrospray
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8679—Target compound analysis, i.e. whereby a limited number of peaks is analysed
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N2030/042—Standards
- G01N2030/045—Standards internal
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
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- Chemical & Material Sciences (AREA)
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- Life Sciences & Earth Sciences (AREA)
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Abstract
The invention discloses a kit and a method for detecting psychotropic drugs in dried blood slices. In order to improve the release rate of the psychotropic drugs in the dried blood slices, the invention firstly provides a pretreatment kit and a pretreatment method for the dried blood slice samples. On the basis, the invention also provides a kit and a method for detecting the psychotropic drugs in the dried blood slices based on liquid chromatography-tandem mass spectrometry. By using the pretreatment kit and the pretreatment method, the psychotropic drugs contained in the dried blood sample can be fully released, the sensitivity of subsequent detection is greatly improved, and the quantitative lower limit of the fluorovoxamine and the like which are sensitive to chromatographic conditions can be obviously reduced. In addition, when the pretreatment kit and the pretreatment method are used for pretreating the sample, the extraction efficiency of the substrates in different batches is higher, and when the differences among the clinical patient sample individuals are faced, the detection results can be consistent, and the accuracy of the clinical detection results is ensured.
Description
Technical Field
The invention belongs to the technical field of blood concentration detection. More particularly, relates to a kit and a method for detecting psychotropic drugs in dried blood slices.
Background
Although the psychotropic drugs have important roles in treating or relieving mental diseases and syndromes, the clinical application of the psychotropic drugs still has a plurality of problems. For example: (1) Mental diseases are chronic diseases, long-term administration is needed, and adverse reactions of mental medicines are more; (2) The individual difference of psychology medicine pharmacokinetics is obvious, and the blood concentration of the same dosage can be different by several times to tens times; (3) The curative effect index of the mental medicine is not concise enough and is influenced by subjective factors of doctors; (4) Toxic reactions of mental drugs can appear hidden and are not easily distinguished from exacerbation of symptoms of the disease; (5) The blood concentration of the psychotropic drugs is related to the curative effect and toxic and side effects, and the curative effect is poor due to too high or too low. Therefore, how to select and use the psychotropic drugs and ensure the effectiveness and safety of the psychotropic drug treatment is an important problem facing the psychotropic treatment.
Therapeutic drug monitoring (therapeutic drug monitoring, TDM) is a technique that achieves optimal efficacy and minimal adverse effects and personalized treatment by analyzing drugs, biomarkers, etc. in biological samples. The national institute of clinical application expert consensus for therapeutic drug monitoring in the psychiatric department and the national institute of pharmaceutical detection consensus for AGNP neuropsychiatric pharmacology indicate that therapeutic drug monitoring is an important means for realizing individualized and accurate drug treatment of mental diseases. In view of the long administration time, great adverse reaction, and blood concentration and related curative effect and toxic and side effects of the psychotropic drugs, regular and reasonable therapeutic drug monitoring is necessary to be implemented on the psychotropic drugs.
Currently, TDM of psychotropic drugs is mostly based on conventional detection matrices, such as plasma, serum, whole blood, etc. However, the detection matrix needs to be collected and separated by professionals and needs to be refrigerated for preservation, so that the collection is inconvenient, the detection frequency is high, the blood collection amount is large due to the long TDM period, the psychological and physical burden of a patient is easy to increase, and the timely monitoring of the blood concentration is difficult to realize. The dry blood sampling technology is a method for obtaining a sample to be tested after dripping blood on a filter paper sheet and naturally drying. Compared with a whole blood matrix sample, the dry blood sheet sampling has the advantages of small required sample size, convenient collection, minimally invasive, low cost, easier preservation and transportation, and the like, has good biological safety, and is a sampling technology with great application prospect. However, there are a number of challenges associated with dry blood tablet-based TDM of psychotropic drugs. For example: because the blood sample is adsorbed on the filter paper, when the drug content in the dried blood sample is detected, the problem of insufficient release of the sample exists. Meanwhile, the sample quantity collected in the dry blood sheet is small, and compared with a serum sample with the same concentration, the response intensity of the dry blood sheet sample detection is low, and the requirement on the detection sensitivity is higher. In addition, the dry blood sample has a large relative contact area with air during storage and transportation, is susceptible to environmental temperature and humidity, and may cause degradation of certain chemically unstable drugs.
In the prior art, there is a liquid chromatography tandem mass spectrometry detection method for detecting bile acid, fat-soluble vitamin or 25-hydroxy vitamin D and the like contained in a dry blood sample. However, the methods for extracting and releasing different substances in the dried blood sample are different, and the liquid chromatography-tandem mass spectrometry detection method of the dried blood sample disclosed in the prior art is not suitable for detecting psychotropic drugs in the dried blood sample. In summary, a method and a kit for detecting the psychotropic drugs in the dried blood slices, which have high sample release rate and good detection sensitivity, are developed, and have important significance on TDM of the psychotropic drugs.
Disclosure of Invention
The invention provides a pretreatment kit and a pretreatment method for a dried blood sample, and provides a kit and a method for detecting psychotropic drugs in dried blood based on liquid chromatography-tandem mass spectrometry based on the pretreatment kit and the pretreatment method.
It is a first object of the present invention to provide a pretreatment kit for a dry blood sample.
A second object of the present invention is to provide a method for pretreatment of a dried blood sheet sample.
The third object of the invention is to provide the application of the pretreatment kit or the pretreatment method in liquid chromatography tandem mass spectrometry detection of psychotropic drugs in dried blood slices.
The fourth object of the invention is to provide a kit for detecting psychotropic drugs in dried blood slices based on liquid chromatography-tandem mass spectrometry.
The fifth object of the invention is to provide a method for detecting psychotropic drugs in dried blood slices based on liquid chromatography-tandem mass spectrometry.
The above object of the present invention is achieved by the following technical scheme:
The pretreatment kit for the dry blood sample is constructed by continuous optimization. The pretreatment kit can further improve the release efficiency of the psychotropic drugs in the dry blood sample, and improve the sensitivity of liquid chromatography-tandem mass spectrometry detection of the psychotropic drugs in the dry blood sample. On the basis, the invention also provides a kit and a method for detecting the psychotropic drugs in the dried blood slices based on liquid chromatography-tandem mass spectrometry. By using the kit and the method, quantitative detection of sulpiride, perphenazine, fluvoxamine, trazodone, blonanserin and Fu Liu sitagliptin in a dry blood sample is realized, the detection limit is low, and the monitoring of the psychotropic drugs is facilitated.
The invention provides a pretreatment kit for a dried blood sample, which comprises an extracting solution and a sample releasing agent.
Specifically, the extracting solution consists of 5-10% by volume of dimethyl sulfoxide, 5-10% by volume of chloroform, 1-10% by volume of formic acid and the balance of water.
Preferably, the volume percentage of the dimethyl sulfoxide is 5-8%, the volume percentage of the chloroform is 5-6%, the volume percentage of the formic acid is 1-5%, and the balance is water.
More preferably, the volume percentage of dimethyl sulfoxide is 8%, the volume percentage of chloroform is 6%, the volume percentage of formic acid is 5%, and the balance is water.
Specifically, the sample releasing agent is methanol or consists of methanol and acetonitrile, and the volume ratio of the methanol to the acetonitrile is 1:0-1.
More specifically, the volume ratio of the methanol to the acetonitrile is 1:0.1-1.
The invention also provides a pretreatment method of the dried blood sample, which comprises the following steps: firstly, carrying out ultrasonic extraction on a dried blood sample by using the extracting solution, then adding the sample releasing agent into the solution after ultrasonic extraction for extraction, and reserving supernatant.
Specifically, the dried blood sheet samples are dried blood sheet sample pore sheets with the pore diameter of 3-6 mm, and the number of the dried blood sheet sample pore sheets is 1-3; the volume of the extracting solution is 30-60 mu L; the volume of the sample releasing agent is 100-150 mu L.
Specifically, the treatment time of the ultrasonic extraction is 1 min; the ultrasound conditions were 30 kHz. And adding a sample releasing agent after the ultrasonic treatment is finished, and carrying out oscillation extraction for 3-5 min.
Specifically, the dry blood sheet of the invention is human whole blood filter paper dry blood sheet or human serum filter paper dry blood sheet.
The extracting solution can perform primary extraction on the psychotropic drug components with different dissolution properties in the dried blood tablet sample, and the drugs in the sample can be fully released by adding the extracting solution before adding the sample releasing agent. In addition, the formic acid with a certain content is added into the extracting solution, so that the medicine can be primarily acidified, the extraction efficiency of the medicine can be increased, the mass spectrum response intensity of the medicine can be further enhanced in a positive ion scanning mode in the subsequent mass spectrum analysis, and the detection sensitivity can be improved. Therefore, the invention claims the application of the pretreatment kit or the pretreatment method in liquid chromatography tandem mass spectrometry detection of psychotropic drugs in dried blood slices.
The invention also provides a kit for detecting the psychotropic drugs in the dried blood slices based on liquid chromatography-tandem mass spectrometry, which comprises the pretreatment kit, a stabilizer, an internal standard substance of the detected psychotropic drugs, a dried blood slice standard substance of the detected psychotropic drugs and a dried blood slice quality control substance.
Specifically, the psychotropic drugs include sulpiride, perphenazine, fluvoxamine, trazodone, blonanserin and Fu Liu sitagliptin.
Specifically, the stabilizer consists of a preservative proclin300, vitamin C and polyethylene glycol; the volume ratio of proclin300 to vitamin C to polyethylene glycol is 1: 10-20: 1 to 1.2.
Preferably, the volume ratio of proclin300, vitamin C and polyethylene glycol is 1:10:1.
Specifically, the concentration of vitamin C is 25 mg/mL.
Specifically, the internal standard substance is deuterated substance of the detected psychotropic drug.
More specifically, the internal standard is sulpiride-D3, perphenazine-D4, fluvoxamine-D3, trazodone-D6, blonanserin-D5 and vosulbactam-D8.
In addition to the above reagents, the kit contains a mobile phase additive 1 and a mobile phase additive 2.
Specifically, the mobile phase additive 1 is ammonium acetate; the mobile phase additive 2 is formic acid.
Optionally, the kit also comprises a 96-well plate (such as a V-shaped or U-shaped bottom 96-well plate), an aluminum foil sealing film and an operation instruction.
Related applications of the kit of the invention are also within the scope of the invention. The kit is applied to the preparation of products for detecting one or more mental medicines of sulpiride, perphenazine, fluvoxamine, trazodone, blonanserin and Fu Liu sitagliptin in human whole blood filter paper dried blood slices.
As an alternative embodiment, in the kit of the present invention, the standard sample of the dry blood tablet and the quality control product of the dry blood tablet of the detected psychotropic drug may be replaced with the standard sample of the detected psychotropic drug and filter paper. The standard sample and the filter paper can be used for preparing the dry blood tablet standard sample and the dry blood tablet quality control product.
Specifically, the preparation method of the dried blood tablet standard sample and the dried blood tablet quality control product comprises the following steps: uniformly mixing a standard product containing the detected psychotropic drugs with healthy human whole blood, preparing a human whole blood sample, diluting the human whole blood sample to a required concentration by using healthy human whole blood, and then dripping the diluted human whole blood sample onto filter paper for drying; the whole blood of the healthy person can be replaced by serum of the healthy person.
Specifically, the drop volume of the human whole blood sample is 50-100 mu L.
Specifically, the human whole blood sample contains the stabilizer of the invention; the mass of the stabilizer is 4-6% of the volume of the whole blood of the healthy person.
More specifically, the mass of the stabilizer is 5% of the volume of whole blood of the healthy person used.
The stabilizer disclosed by the invention is added in the preparation process of the dried blood sample standard and the quality control product, so that the prepared dried blood sample standard and quality control product can be kept stable in the long-term storage process. The combined action of multiple components in the stabilizer can enhance the adsorptivity of a blood or serum sample and a filter paper sheet, keep the stability of a dry blood sample matrix, prevent the oxidation of a medicine with unstable chemical properties in the sample, and greatly reduce the degradation speed.
Taking the kit for detecting the psychotropic drugs sulpiride, perphenazine, fluvoxamine, trazodone, blonanserin and Fu Liu sitagliptin as an example. In the kit, dry blood tablet standards of the psychotropic drugs sulpiride, perphenazine, fluvoxamine, trazodone, blonanserin and Fu Liu sitagliptin comprise blank samples and dry blood tablet standards C1-C7; wherein, the concentration of the six kinds of mental drugs contained on the dried blood tablet standard sample C1 is respectively as follows: sulpiride 15 ng/mL, perphenazine 0.2 ng/mL, fluvoxamine 10 ng/mL, trazodone 20 ng/mL, blonanserin 2 ng/mL, and voathixetine 1 ng/mL;
The concentrations of the six psychotropic drugs contained on the dried blood tablet standard C2 were respectively: sulpiride 30 ng/mL, perphenazine 0.4 ng/mL, fluvoxamine 20 ng/mL, trazodone 40 ng/mL, blonanserin 4 ng/mL, and voathixetine 2 ng/mL;
The concentrations of the six psychotropic drugs contained on the dried blood tablet standard C3 were respectively: sulpiride 300 ng/mL, perphenazine 4 ng/mL, fluvoxamine 200 ng/mL, trazodone 400 ng/mL, blonanserin 40 ng/mL, and vothiacetin 20 ng/mL;
the concentrations of the six psychotropic drugs contained on the dried blood tablet standard C4 were: sulpiride 600 ng/mL, perphenazine 8 ng/mL, fluvoxamine 400 ng/mL, triadimefon 800 ng/mL, blonanserin 80 ng/mL, and voathixetine 40 ng/mL;
The concentrations of the six psychotropic drugs contained on the dried blood tablet standard C5 were: sulpiride 900 ng/mL, perphenazine 12 ng/mL, fluvoxamine 600 ng/mL, trazodone 1200 ng/mL, blonanserin 120 ng/mL, and vothiacetin 60 ng/mL;
The concentrations of the six psychotropic drugs contained on the dried blood tablet standard C6 were: sulpiride 1800 ng/mL, perphenazine 24 ng/mL, fluvoxamine 1200 ng/mL, trazodone 2400 ng/mL, blonanserin 240 ng/mL, and vothiacetin 120 ng/mL;
The concentrations of the six psychotropic drugs contained on the dried blood tablet standard C7 were: sulpiride 2250 ng/mL, perphenazine 30 ng/mL, fluvoxamine 1500 ng/mL, triadimefon 3000 ng/mL, blonanserin 300 ng/mL, and voathiacin 150 ng/mL.
The dry blood tablet quality control product comprises low, medium and high concentration dry blood tablet quality control products; wherein, the concentration of the six kinds of mental drugs contained on the low concentration dry blood tablet quality control product is respectively as follows: sulpiride 45 ng/mL, perphenazine 0.6 ng/mL, fluvoxamine 30 ng/mL, triadimefon 60 ng/mL, blonanserin 6 ng/mL, and voathixetine 3 ng/mL;
The concentration of the six kinds of mental drugs contained on the medium concentration dry blood tablet quality control product is respectively as follows: sulpiride 450 ng/mL, perphenazine 6 ng/mL, fluvoxamine 300 ng/mL, trazodone 600 ng/mL, blonanserin 60 ng/mL, and vothiacetin 30 ng/mL;
The concentration of the six kinds of mental medicines contained on the high-concentration dry blood tablet quality control product is respectively as follows: shupiride 1500 ng/mL, perphenazine 20 ng/mL, fluvoxamine 1000 ng/mL, trazodone 2000 ng/mL, blonanserin 200 ng/mL, and vothiacetin 100 ng/mL.
The invention also provides a method for detecting the psychotropic drugs in the dried blood slices based on liquid chromatography-tandem mass spectrometry, wherein the psychotropic drugs comprise sulpiride, perphenazine, fluvoxamine, trazodone, blonanserin and Fu Liu sitagliptin;
specifically, the method comprises the steps of:
S1, pretreatment of a dried blood sheet sample: the sample release agent in the pretreatment kit is used for respectively extracting a dry blood sample to be detected, a dry blood sample standard sample and a dry blood sample quality control product, and supernatant fluid is reserved; or the pretreatment method is used for respectively treating the dry blood sample to be tested, the dry blood sample standard and the dry blood sample quality control product; adding deuterated substances of the detected psychotropic drugs to the sample release agent as internal standards before adding the sample release agent;
S2, quantitative detection of a dried blood sample: and (3) quantitatively detecting and analyzing the supernatant obtained by the step (S1) by using a liquid chromatography-tandem mass spectrometry method.
Specifically, in step S2, the internal standard dry powder and the sample releasing agent are mixed in proportion before the sample releasing agent is used for treatment, so that the concentration of the sulpiride-D3 internal standard is 1000 ng/mL, the concentration of the perphenazine-D4 internal standard is 5 ng/mL, the concentration of the fluvoxamine-D3 internal standard is 300 ng/mL, the concentration of the trazodone-D6 internal standard is 500 ng/mL, the concentration of the blonanserin-D5 internal standard is 100 ng/mL, and the concentration of the vosulbactam-D8 internal standard is 100 ng/mL.
Specifically, the chromatographic conditions used in step S2 are: chromatographic column: agela Durashell C8 chromatographic columns; mobile phase: comprises a mobile phase A and a mobile phase B; mobile phase A is an aqueous solution containing 5 mmol/L ammonium acetate and 0.1% formic acid; mobile phase B is methanol; column temperature: 40. the temperature is lower than the temperature; sample cell temperature: 8. the temperature is lower than the temperature; sample injection volume: 10. mu L; the elution mode is linear gradient elution, and the linear gradient elution program is shown in the following table:
The mass spectrum conditions are as follows: detecting by adopting a multi-reaction monitoring (MRM) mode; in the negative ion mode, an ESI ion source is adopted, the atomization air flow is 3L/min, the heating air flow is 10L/min, the drying air flow is 10L/min, the interface temperature is 350 ℃, and the DL temperature is 200 ℃; the parameters for the multiple reaction monitoring are shown in the following table, To quantify ion pairs:
The invention adopts a gradient elution mode, and ammonium acetate and formic acid are added as mobile phase additives, so that the method can ensure that the medicine with weak retention on a chromatographic column, such as sulpiride, has normal peak shape and smaller half peak width, and ensures that the quantitative determination has better repeatability and accuracy. For components such as blonanserin, fu Liuxi and the like which elute on the chromatographic column, the efficiency of protonation is improved and the response intensity of the lower limit point of quantification is enhanced by optimizing the elution gradient.
On the basis, the invention greatly enhances the response of the medicine with low quantitative lower limit and low ionization efficiency by optimizing the ion source parameter and the mass spectrum MRM scanning parameter, so that the medicine can meet the requirement of quantitative determination.
The method also comprises the establishment of a standard curve; the method comprises the following steps: simultaneously processing the dried blood sample standard sample and the quality control sample along with the dried blood sample to be detected, and analyzing the processed dried blood sample by using a liquid phase mass spectrometer; the peak areas of sulpiride, perphenazine, fluvoxamine, trazodone, blonanserin, fu Liu sittin and corresponding internal standard substances of each concentration are recorded respectively, the ratio of the peak areas of the medicines to the corresponding internal standard substances is taken as an ordinate, the corresponding concentration of the medicines is taken as an abscissa, and a standard curve equation of the medicines is obtained through linear regression; substituting the peak area ratio of each drug and the corresponding internal standard substance measured by the sample to be measured into a standard curve equation, and calculating the content of each psychotropic drug in the dried blood slices to be measured.
The invention has the following beneficial effects:
In order to improve the release rate of the psychotropic drugs in the dried blood slices, the invention firstly provides a pretreatment kit and a pretreatment method for the dried blood slice samples. On the basis, the invention also provides a kit and a method for detecting the psychotropic drugs in the dried blood slices based on liquid chromatography-tandem mass spectrometry. By using the pretreatment kit and the pretreatment method, the psychotropic drugs contained in the dried blood sample can be fully released, the sensitivity of subsequent detection is greatly improved, and the quantitative lower limit of the fluvoxamine, the voxamine and the blonanserin which are sensitive to chromatographic conditions can be obviously reduced.
In addition, when the pretreatment kit and the pretreatment method are used for pretreatment of the quality control samples, the extraction efficiency of the substrates of different batches is higher, and when the differences among clinical patient samples are faced, the detection results can be consistent, and the accuracy of the clinical detection results is ensured.
Drawings
FIG. 1 is an ion flow chromatogram of sulpiride and its internal standard in dried blood C3 (whole blood) as described in example 4.
FIG. 2 is an ion flow chromatogram of trazodone and its internal standard in dried blood C3 (whole blood) as described in example 4.
FIG. 3 is an ion flow chromatogram of fluvoxamine and its internal standard in dried blood piece standard C3 (whole blood) as described in example 4.
FIG. 4 is an ion-flow chromatogram of the detection of perphenazine and its internal standard in dried blood C3 (whole blood) as described in example 4.
FIG. 5 is an ion flow chromatogram of the detection of the vomeropherin and its internal standard in dried blood sheet standard C3 (whole blood) as described in example 4.
FIG. 6 is an ion flow chromatogram of bloodpiece standard C3 (whole blood) for bloodpiece and its internal standard as described in example 4.
FIG. 7 is an ion flow chromatogram of sulpiride and its internal standard in dried blood piece standard C3 (serum) as described in example 4.
FIG. 8 is an ion flow chromatogram of trazodone and its internal standard in dried blood piece standard C3 (serum) as described in example 4.
FIG. 9 is an ion flow chromatogram of fluvoxamine and its internal standard in dried blood piece standard C3 (serum) as described in example 4.
FIG. 10 is an ion-flow chromatogram of the detection of perphenazine and its internal standard in dried blood sample C3 (serum) as described in example 4.
FIG. 11 is an ion flow chromatogram of the detection of the presence of vomeropherin and its internal standard in dried blood piece standard C3 (serum) as described in example 4.
FIG. 12 is an ion flow chromatogram of bloodpiece standard C3 (serum) for bloodpiece and its internal standard as described in example 4.
Detailed Description
The invention is further illustrated in the following drawings and specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
Reagents and materials used in the following examples are commercially available unless otherwise specified.
Example 1A kit for detecting psychotropic drugs in dried blood slices based on liquid chromatography tandem mass spectrometry
The embodiment provides a kit for detecting psychotropic drugs in dried blood slices based on liquid chromatography-tandem mass spectrometry, which comprises the pretreatment kit (comprising an extracting solution and a sample releasing agent) disclosed by the invention, an internal standard substance, a dried blood slice quality control substance and the like; the psychotropic drugs are sulpiride, perphenazine, fluvoxamine, trazodone, blonanserin and Fu Liu sitagliptin. Specifically, the kit of the invention contains the components shown in Table 1.
Table 1 composition of the kit
Note that: the kit needs to be stored at 2-8 ℃ in a dark place.
The mixed internal standard dry powders described in Table 1 contained sulpiride-D3 dry powder 500 ng, perphenazine-D4 dry powder 2.5 ng, fluvoxamine-D3 dry powder 150 ng, trazodone-D6 dry powder 250 ng, blonanserin-D5 dry powder 50 ng and vomitoxin-D8 dry powder 50 ng.
The standard sample of the dried blood slices in the table 1 can be replaced by dry powder or solution containing six kinds of psychotropic drug standard substances, and filter paper or a blood sample collection card (consisting of blood sample collection filter paper and information card), and the standard sample of the dried blood slices is prepared in actual use. The preparation method of the dried blood piece standard sample comprises the following steps: firstly, uniformly mixing a solution containing six psychotropic drug standard substances with healthy human whole blood, preparing a human whole blood sample, diluting the human whole blood sample to a required concentration by using the healthy human whole blood, and then dripping the diluted human whole blood sample onto filter paper for drying; the whole blood of the healthy person can be replaced by serum of the healthy person. The preparation method of the quality control product of the dried blood slices is the same as that of the standard sample of the dried blood slices.
Taking a blood sample collection card (Kaipu Biochemical Co., ltd.) as an example, 50-100. Mu.L of human whole blood sample or diluted sample was collected per blood sample. The volume of the dry human whole blood sample in the same batch of the kit is about the same, and is generally 75 mu L.
Specifically, the dried blood piece standard sample comprises blank samples (blank, the rest operations are the same except that the blank does not contain the psychotropic drugs) and dried blood piece standard samples C1-C7; wherein, the concentration of the six kinds of mental drugs contained on the dried blood tablet standard sample C1 is respectively as follows: sulpiride 15 ng/mL, perphenazine 0.2 ng/mL, fluvoxamine 10 ng/mL, trazodone 20 ng/mL, blonanserin 2 ng/mL, and voathixetine 1 ng/mL;
The concentrations of the six psychotropic drugs contained on the dried blood tablet standard C2 were respectively: sulpiride 30 ng/mL, perphenazine 0.4 ng/mL, fluvoxamine 20 ng/mL, trazodone 40 ng/mL, blonanserin 4 ng/mL, and voathixetine 2 ng/mL;
The concentrations of the six psychotropic drugs contained on the dried blood tablet standard C3 were respectively: sulpiride 300 ng/mL, perphenazine 4 ng/mL, fluvoxamine 200 ng/mL, trazodone 400 ng/mL, blonanserin 40 ng/mL, and vothiacetin 20 ng/mL;
the concentrations of the six psychotropic drugs contained on the dried blood tablet standard C4 were: sulpiride 600 ng/mL, perphenazine 8 ng/mL, fluvoxamine 400 ng/mL, triadimefon 800 ng/mL, blonanserin 80 ng/mL, and voathixetine 40 ng/mL;
The concentrations of the six psychotropic drugs contained on the dried blood tablet standard C5 were: sulpiride 900 ng/mL, perphenazine 12 ng/mL, fluvoxamine 600 ng/mL, trazodone 1200 ng/mL, blonanserin 120 ng/mL, and vothiacetin 60 ng/mL;
The concentrations of the six psychotropic drugs contained on the dried blood tablet standard C6 were: sulpiride 1800 ng/mL, perphenazine 24 ng/mL, fluvoxamine 1200 ng/mL, trazodone 2400 ng/mL, blonanserin 240 ng/mL, and vothiacetin 120 ng/mL;
The concentrations of the six psychotropic drugs contained on the dried blood tablet standard C7 were: sulpiride 2250 ng/mL, perphenazine 30 ng/mL, fluvoxamine 1500 ng/mL, triadimefon 3000 ng/mL, blonanserin 300 ng/mL, and voathiacin 150 ng/mL.
The dry blood tablet quality control materials in table 1 comprise low, medium and high concentration dry blood tablet quality control materials; wherein, the concentration of the six kinds of mental drugs contained on the low concentration dry blood tablet quality control product is respectively as follows: sulpiride 45 ng/mL, perphenazine 0.6 ng/mL, fluvoxamine 30 ng/mL, triadimefon 60 ng/mL, blonanserin 6 ng/mL, and voathixetine 3 ng/mL;
The concentration of the six kinds of mental drugs contained on the medium concentration dry blood tablet quality control product is respectively as follows: sulpiride 450 ng/mL, perphenazine 6 ng/mL, fluvoxamine 300 ng/mL, trazodone 600 ng/mL, blonanserin 60 ng/mL, and vothiacetin 30 ng/mL;
The concentration of the six kinds of mental medicines contained on the high-concentration dry blood tablet quality control product is respectively as follows: shupiride 1500 ng/mL, perphenazine 20 ng/mL, fluvoxamine 1000 ng/mL, trazodone 2000 ng/mL, blonanserin 200 ng/mL, and vothiacetin 100 ng/mL.
In order to improve the stability of the dried blood sample and the dried blood sample quality control product, a stabilizer is also added in the preparation of the human whole blood sample; the mass of the stabilizer is 0.5 percent of the whole blood of the healthy person. The stabilizer consists of the following components: preservative proclin300: vitamin C: polyethylene glycol = 1:10:1.
The concentration of the vitamin C is 25 mg/mL; preservative Proclin300 was purchased from Sigma under the accession number 48912-U; polyethylene glycol was purchased from Sigma under the accession number 202398-5G.
Example 2A kit for detecting psychotropic drugs in dried blood slices based on liquid chromatography tandem mass spectrometry
The kit of this example differs from example 1 in that the stabilizing agent consists of the following components: preservative proclin300: vitamin C: polyethylene glycol = 1:15:1.
Example 3A kit for detecting psychotropic drugs in dried blood slices based on liquid chromatography tandem mass spectrometry
The kit of this example differs from example 1 in that the stabilizing agent consists of the following components: preservative proclin300: vitamin C: polyethylene glycol = 1:20:1.2.
Example 4 method for detecting psychotropic drugs in dried blood slices based on liquid chromatography-tandem mass spectrometry
The invention also provides a method for detecting the psychotropic drugs in the dried blood slices based on the liquid chromatography-tandem mass spectrometry by using the kit described in the embodiment 1, and the quantitative detection of the six psychotropic drugs (sulpiride, perphenazine, fluvoxamine, trazodone, blonanserin and Fu Liu sitagliptin) in the dried blood slice sample is realized.
The method comprises the following steps:
(1) Sampling a dry blood sample: punching a dried blood sheet standard sample, a quality control product and a dried blood sheet sample to be detected respectively to obtain hole sheets (with the aperture of 3-6 mm) with the same size, and transferring the hole sheets (1 sheet) obtained by punching into a clean EP tube;
(2) Sample treatment of dried blood slices: adding 10mL methanol into the mixed internal standard substance dry powder (internal standard) and uniformly mixing to prepare a sample release agent containing the internal standard; adding 100 mu L of sample release agent into an EP tube, respectively extracting a dried blood piece standard sample, a quality control product and a dried blood piece sample to be detected (oscillating extraction is 3-5 min), centrifuging 14000 rpm after extraction is finished for 8min, taking a supernatant and detecting the supernatant to be detected;
the internal standard is detected deuterated matters of six psychotropic drugs, and in the prepared sample release agent, the concentration of the sulpiride-D3 internal standard is 1000 ng/mL, the concentration of the perphenazine-D4 internal standard is 5 ng/mL, the concentration of the fluvoxamine-D3 internal standard is 300 ng/mL, the concentration of the trazodone-D6 internal standard is 500 ng/mL, the concentration of the blonanserin-D5 internal standard is 100 ng/mL, and the concentration of the vothixetine-D8 internal standard is 100 ng/mL;
(3) Dry blood sheet sample assay: analyzing the supernatant obtained by extraction in the step (2) by adopting a liquid chromatography tandem mass spectrometry; the chromatographic conditions and mass spectral conditions were as follows:
chromatographic conditions:
mobile phase, including mobile phase a and mobile phase B; wherein the mobile phase A consists of deionized water, ammonium acetate and formic acid; in the mobile phase A, the final concentration of ammonium acetate is 5 mmol/L, and the volume fraction of formic acid is 0.1%; mobile phase B is methanol;
Chromatographic column: agela Durashell C8 chromatographic column (5 μm, 3.0) 50 mm）；
Column temperature: 40. the temperature is lower than the temperature;
Sample cell temperature: 8. the temperature is lower than the temperature;
Sample injection volume: 10. mu L;
The elution mode was linear gradient elution, and the procedure of linear gradient elution was performed according to the following table 2:
TABLE 2 Linear gradient elution procedure
Mass spectrometry conditions:
detecting by adopting a multi-reaction monitoring (MRM) mode; the mass spectrum conditions are as follows: in the negative ion mode, an ESI ion source is adopted, the atomization air flow is 3L/min, the heating air flow is 10L/min, the drying air flow is 10L/min, the interface temperature is 350 ℃, and the DL temperature is 200 ℃;
The parameters of the MRM are shown in table 3 below, To quantify ion pairs:
TABLE 3 MRM parameters
In the experimental process, the dry blood sample standard sample and the quality control sample are processed simultaneously along with the dry blood sample to be tested, and the processed dry blood sample and quality control sample are analyzed by a liquid phase mass spectrometer; the peak areas of sulpiride, perphenazine, fluvoxamine, trazodone, blonanserin, fu Liu sittin and corresponding internal standard substances of each concentration are recorded respectively, the ratio of the peak areas of the medicines to the corresponding internal standard substances is taken as an ordinate, the corresponding concentration of the medicines is taken as an abscissa, and a standard curve equation of the medicines is obtained through linear regression; substituting the peak area ratio of each drug and the corresponding internal standard substance measured by the sample to be measured into a standard curve equation, and calculating the content of each psychotropic drug in the dried blood slices to be measured.
In this example, a dried blood piece standard sample C3 (whole blood) and a dried blood piece standard sample C3 (serum) (the preparation method is the same as that of the dried blood piece standard sample C3 (whole blood), and the whole blood of a healthy person is replaced with the serum of the healthy person) are used as detection samples, and the detection is performed by the method.
The ion flow chromatograms of the six psychotropic drugs and the internal standard thereof in the dried blood slice standard C3 (whole blood) and the dried blood slice standard C3 (serum) are respectively shown in figures 1 to 12; wherein, figures 1-6 are ion flow chromatograms of sulpiride, trazodone, fluvoxamine, perphenazine, voathiacin, blonanserin and internal standards thereof in a dried blood sample C3 (whole blood) in sequence; FIGS. 7-12 are ion flow chromatograms of sulpiride, trazodone, fluvoxamine, perphenazine, vomethidine, blonanserin and internal standards in sequence in dried blood sample C3 (serum). From figures 1-12, the detected six psychotropic drugs have good chromatographic peak shapes and no impurity peaks and tail, which shows that the kit and the method can be used for quantitative detection of dry blood tablet drugs.
Standard curve equations for six psychotropic drugs in dried blood slice standard C3 (whole blood) are shown in table 4. As shown in Table 4, the linear correlation coefficients r are all larger than 0.99, which indicates that the detection method of the invention can improve the reliability of quantitative detection results of dry blood sample.
TABLE 4 Linear concentration ranges, standard curve equations and linear correlation coefficients for six psychotropic drugs
Example 5 method for detecting psychotropic drugs in dried blood slices based on liquid chromatography-tandem mass spectrometry
The method of this example differs from the method of example 4 in that: the solvent used for preparing the sample releasing agent is not methanol, but is prepared from methanol and acetonitrile according to the volume ratio of 1:1, and the rest operations are the same.
Example 6 method for detecting psychotropic drugs in dried blood slices based on liquid chromatography-tandem mass spectrometry
The method of this example differs from the method of example 4 in that: before the sample release agent is used for treatment, an extracting solution consisting of dimethyl sulfoxide, chloroform, formic acid and water is prepared, wherein the content (v/v) of the dimethyl sulfoxide in the extracting solution is 5%, the content (v/v) of the chloroform is 5%, the content (v/v) of the formic acid is 1%, and the balance is water; adding 30 mu L of the extracting solution into a dried blood sample, and performing ultrasonic treatment (30 kHz) on the dried blood sample for 1 min; the dried blood sheet sample was then further extracted by adding a further 100 μl of sample release agent.
Example 7 method for detecting psychotropic drugs in dried blood slices based on liquid chromatography-tandem mass spectrometry
The method of this example differs from the method of example 6 in that: the extract contains dimethyl sulfoxide 8% (v/v), chloroform 6% (v/v), formic acid 5% (v/v), and water in balance.
Example 8 method for detecting psychotropic drugs in dried blood slices based on liquid chromatography-tandem mass spectrometry
The method of this example differs from the method of example 6 in that: the extract contains dimethyl sulfoxide 10% (v/v), chloroform 10% (v/v), formic acid 10% (v/v), and water in balance.
Comparative example 1
The method of this comparative example differs from the method of example 6 in that: the components of the extracting solutions are different, and the extracting solution in the comparative example is a mixed solution of formic acid and water; wherein the content of the formic acid is 5%.
Comparative example 2
The method of this comparative example differs from the method of example 6 in that: the components of the extraction solution used are different, and the extraction solution does not contain formic acid.
Comparative example 3
The method of this comparative example differs from the method of example 6 in that: and replacing chloroform in the extracting solution with acetone.
Comparative example 4
The method of this comparative example differs from the method of example 6 in that: the chloroform in the extracting solution is replaced by acetone, and dimethyl sulfoxide is not contained.
Comparative example 5
The method of this comparative example differs from the method of example 6 in that: the contents of all the components in the extracting solution are different, and the extracting solution in the comparative example consists of dimethyl sulfoxide, chloroform, formic acid and water; wherein the content (v/v) of dimethyl sulfoxide in the extracting solution is 15%, the content (v/v) of chloroform is 3%, the content (v/v) of formic acid is 5%, and the balance is water.
Comparative example 6
The method of this comparative example differs from the method of example 6 in that: different gradient elution modes are adopted. The manner of gradient elution in the method described in this comparative example is shown in Table 5.
TABLE 5 Linear gradient elution procedure
Comparative example 7
The kit of this comparative example differs from example 1 in that: the stabilizer is not added in the preparation process of the dry blood tablet quality control product, so that the influence of the stabilizer on the stability of the dry blood tablet sample is explored.
Comparative example 8
The kit of this comparative example differs from example 1 in that: the added stabilizing agent is different in the preparation process of the quality control product. The stabilizer used in this comparative example consisted of the following components: preservative proclin300: vitamin C: tertiary butanol=1: 10:1.
Comparative example 9
The kit of this comparative example differs from example 1 in that: the added stabilizing agent is different in the preparation process of the quality control product. The stabilizer used in this comparative example consisted of the following components: preservative proclin300: vitamin C: polyethylene glycol = 1:20:2.
Test example 1 test of test effects of the methods described in examples 4 to 8 and comparative examples 1 to 6
The high concentration quality control materials in the kits of example 1 were examined by the methods of examples 4 to 8 and comparative examples 1 to 6, respectively, to determine the detection effect by detecting the peak areas obtained (a larger peak area means a relatively good detection effect). The data of the detection effect are summarized in Table 6.
TABLE 6 detection effect data of the methods described in examples 4 to 8 and comparative examples 1 to 6
From the data in Table 6, it can be seen that the method of the present invention can detect six kinds of psychotropic drugs (sulpiride, perphenazine, fluvoxamine, trazodone, blonanserin and Fu Liu sittin) in the dried blood sample, and the method of the present invention can improve the release efficiency of the dried blood sample and the response intensity of the detection.
Test example 2 stability test of the kits of examples 1 to 3 and comparative examples 7 to 9
The high concentration quality control in the test kits of examples 1 to 3 and comparative examples 7 to 9 were used as test samples for stability test, the samples were placed at 40 ℃ and 75% humidity, two samples were arranged in parallel for each test condition, and after 14 days of standing, peak areas of psychotropic drug responses in the samples were measured by the method of example 6, each sample was measured in parallel for 2 times, and the average value of the peak areas was calculated to investigate the stability of the test kits, and the results are shown in table 7.
Table 7 results of stability test of the kits described in examples 1 to 3 and comparative examples 7 to 9
In a stability test experiment, compared with a dry blood tablet quality control product without adding a stabilizer, the stabilizer provided by the invention can effectively reduce the degradation rate of medicines with unstable chemical properties, such as fluvoxamine and perphenazine, and the components in the stabilizer and the proportion of the components have important influence on improving the stability of the medicines.
Test example 3 quantitative limit test
The present invention selects samples of different concentrations near the limit of quantitation (calibration curve C1 point concentration), and the limit of quantitation was tested as described in examples 4 and 7 and comparative examples 3 and 6, with 5 replicates of each sample requiring CV.ltoreq.20% and accuracy within.+ -. 15%, and the results are shown in Table 8 (units: ng/mL).
Table 8 limit of quantitation test data for the methods described in examples 4, 7 and comparative examples 3, 6
As can be seen from the data in Table 8, the detection sensitivity can be greatly improved by the pretreatment of the extracting solution provided by the invention in combination with the sample release agent, and the quantitative lower limit of perphenazine with lower limit of reference concentration range, fluvoxamine, voathiacin and blonanserin which are sensitive to chromatographic conditions can be obviously reduced by ionization efficiency and chromatographic retention. The selection of the chromatographic elution gradient provided by the invention plays an important role in obtaining the expected detection result.
Test example 4
In the invention, 6 batches of human whole blood from different sources are collected and used as matrixes, dry blood sheet low-concentration quality control products are prepared and quantitatively detected in the modes of examples 4 and 7 and comparative examples 3 and 6, each sample is repeatedly measured for 2 times to calculate average value, and the variation coefficient CV (%) of the detection results of the 6 batches of human whole blood matrixes from different sources is calculated, and the results are shown in the following table 9.
Table 96 coefficient of variation of human dried blood sheet-like quality control of different sources
From the data in table 9, it can be seen that the pretreatment method provided by the invention is used for pretreating the quality control sample, and has higher extraction efficiency for different batches of matrixes, which indicates that the pretreatment reagent and the pretreatment method provided by the invention can effectively remove the matrix effect of the dried blood sample. When the differences among the clinical patient samples are faced, the detection results can be consistent, and the accuracy of the clinical detection results is ensured.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (7)
1. The pretreatment kit is characterized in that the pretreatment kit is a pretreatment kit for liquid chromatography tandem mass spectrometry detection of psychotropic drugs in dried blood, and the psychotropic drugs comprise sulpiride, perphenazine, fluvoxamine, trazodone, blonanserin and Fu Liu sitagliptin; the pretreatment kit comprises an extracting solution and a sample releasing agent; the extracting solution consists of dimethyl sulfoxide, chloroform, formic acid and water, wherein the volume percentage of the dimethyl sulfoxide is 5-10%, the volume percentage of the chloroform is 5-10%, the volume percentage of the formic acid is 1-10%, and the balance is water; the sample releasing agent is methanol or consists of methanol and acetonitrile, and the volume ratio of the methanol to the acetonitrile is 1:0-1.
2. The use of the pretreatment kit of claim 1 in liquid chromatography tandem mass spectrometry detection of psychotropic drugs in dried blood slices.
3. A kit for detecting a psychotropic drug in dried blood slices based on liquid chromatography-tandem mass spectrometry, which is characterized by comprising the pretreatment kit, a stabilizer, an internal standard substance of the psychotropic drug detected, a dried blood slice standard substance of the psychotropic drug detected and a dried blood slice quality control substance of the psychotropic drug detected according to claim 1, and further comprising a mobile phase additive 1 and a mobile phase additive 2; the stabilizer consists of proclin300, vitamin C and polyethylene glycol, wherein the volume ratio of proclin300 to vitamin C to polyethylene glycol is 1: 10-20: 1 to 1.2; the internal standard substance is deuterated substance of the detected psychotropic drug; the mobile phase additive 1 is ammonium acetate; the mobile phase additive 2 is formic acid.
4. The kit of claim 3, wherein the standard and quality control of the dried blood slices of the detected psychotropic drug are replaced with standard and filter paper of the detected psychotropic drug.
5. The kit according to claim 4, wherein the preparation method of the dried blood piece standard sample and the dried blood piece quality control product is as follows: uniformly mixing a standard product containing the detected psychotropic drugs with healthy human whole blood, preparing a human whole blood sample, diluting the human whole blood sample to a required concentration by using healthy human whole blood, and then dripping the diluted human whole blood sample onto filter paper for drying; the whole blood of the healthy person can be replaced by serum of the healthy person.
6. The kit of claim 5, wherein the human whole blood sample contains the stabilizing agent.
7. The method for detecting the psychotropic drugs in the dried blood slices based on liquid chromatography-tandem mass spectrometry is characterized by comprising the following steps of:
(1) Sampling a dry blood sample: punching the dried blood sheet standard sample, the quality control product and the dried blood sheet sample to be detected respectively to obtain hole sheets with the same size, and transferring the hole sheets obtained by punching into a clean container;
(2) Sample treatment of dried blood slices: adding the sample release agent in the pretreatment kit of claim 1 into the mixed internal standard dry powder, and uniformly mixing to prepare a sample release agent containing an internal standard; adding the extracting solution in the pretreatment kit of claim 1 into a dried blood sample, adding the sample release agent after ultrasonic treatment to continuously extract the dried blood sample, centrifuging after extraction, taking the supernatant, and measuring;
the internal standard is detected deuterated matters of six psychotropic drugs, and in the prepared sample release agent, the concentration of the sulpiride-D3 internal standard is 1000 ng/mL, the concentration of the perphenazine-D4 internal standard is 5 ng/mL, the concentration of the fluvoxamine-D3 internal standard is 300 ng/mL, the concentration of the trazodone-D6 internal standard is 500 ng/mL, the concentration of the blonanserin-D5 internal standard is 100 ng/mL, and the concentration of the vothixetine-D8 internal standard is 100 ng/mL;
(3) Dry blood sheet sample assay: analyzing the supernatant obtained by extraction by adopting a liquid chromatography tandem mass spectrometry; the chromatographic conditions are as follows: chromatographic column: agela Durashell C8 chromatographic columns; mobile phase: comprises a mobile phase A and a mobile phase B; mobile phase A is an aqueous solution containing 5 mmol/L ammonium acetate and 0.1% formic acid; mobile phase B is methanol; column temperature: 40. the temperature is lower than the temperature; sample cell temperature: 8. the temperature is lower than the temperature; sample injection volume: 10. mu L; the elution mode is linear gradient elution, and the linear gradient elution program is shown in the following table:
；
the mass spectrum conditions are as follows: detecting by adopting a multi-reaction monitoring mode; in the negative ion mode, an ESI ion source is adopted, the atomization air flow is 3L/min, the heating air flow is 10L/min, the drying air flow is 10L/min, the interface temperature is 350 ℃, and the DL temperature is 200 ℃; the parameters for the multiple reaction monitoring are shown in the following table, To quantify ion pairs:
；
The psychotropic drugs include sulpiride, perphenazine, fluvoxamine, trazodone, blonanserin and Fu Liu sitagliptin.
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| CN101398414A (en) * | 2008-10-15 | 2009-04-01 | 上海市公安局刑事侦查总队 | Method for qualitatively screening 242 kinds of compounds by liquid phase chromatography-mass spectra at the same times |
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| CN101398414A (en) * | 2008-10-15 | 2009-04-01 | 上海市公安局刑事侦查总队 | Method for qualitatively screening 242 kinds of compounds by liquid phase chromatography-mass spectra at the same times |
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| Title |
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| 基于液相色谱-二级质谱联用技术精神科药物中毒筛查平台的构建与临床应用;倪晓佳等;中国医院药学杂志;20170930;第37卷(第17期);1671-1674 * |
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