CN107991421A - The liquid phase chromatography analytical method of diazepam content in a kind of detection blood - Google Patents
The liquid phase chromatography analytical method of diazepam content in a kind of detection blood Download PDFInfo
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- CN107991421A CN107991421A CN201810075226.6A CN201810075226A CN107991421A CN 107991421 A CN107991421 A CN 107991421A CN 201810075226 A CN201810075226 A CN 201810075226A CN 107991421 A CN107991421 A CN 107991421A
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Abstract
The method of diazepam drug concentration is that standard solution is demarcated using liquid chromatography analysis equipment and UV detector in present invention detection blood, it is y=a*x+b that fitting, which obtains calibration curve equation, take blood sample to be measured, in the blood to be detected after treatment, equally sample to be measured is detected using liquid chromatography analysis equipment and UV detector, obtain blood y values to be measured, blood y to be measured is substituted into calibration curve equation, object relative concentration x in blood sample to be measured is obtained by calculation, internal standard compound working solution concentration is known, thus the diazepam drug concentration in blood to be detected is calculated in the sample, internal standard method is combined by the present invention with high performance liquid chromatography, improve the accuracy of quantitative result, eliminate systematic error；And analysis time is effectively shortened, make detection process easy to be quick, the diazepam blood concentration of patient's body is monitored more conducively in clinical treatment.
Description
Technical field
The present invention relates in the clinical therapeutic drug monitoring technical field of antipsychotics, more particularly to a kind of detection blood
The method of diazepam drug concentration.
Background technology
Diazepam (diazepam), it is also known as stable, be common benzodiazepine, clinically frequently as anesthesia before
The part of medication, adjuvant drug and General anesthesia.In addition, be also commonly used to control muscle spasmus and twitch, particularly with
Other antiepileptics share, and treat epileptic attack.Diazepam molecular formula is C16H13ClN2O, concrete structure formula are as shown in Figure 1.
As the common sedative of clinic, diazepam mainly carries out oxidative metabolism through liver cytochrome P 450 enzyme effect in vivo, has
Genetic polymorphism.Since its individual difference is big, therapeutic window is narrow, and toxic side effect is closely related with blood concentration, thus into promoting circulation of blood medicine
Concentration monitor is of great significance for instructing clinical rational drug use, reduction adverse reaction, improving curative effect.
At present, the assay method for the diazepam reported both at home and abroad has HPLC, HPLC-MS/MS, GC-MS, GC-ECD, consolidates
Phase extraction-UV Derivative Spectrophotometry detection, TLC-UV, enzyme-linked immunization etc..Wherein, import reagent used in immunization is expensive,
And used kit specificity is slightly worse；TLC-UV methods take time and effort, it is difficult to which great amount of samples is analyzed；Liquid chromatography mass joins
Usage analysis cost is higher, high to personnel and site requirements, is unsuitable for extensively carrying out in small-middle hospital；High performance liquid chromatography
Have the advantages that it is accurate it is sensitive, specificity is strong, favorable reproducibility, cost are low etc., it has also become the main side of measure diazepam blood concentration
Method.But used high performance liquid chromatography pre-treatment operation is more complicated at present, and analysis time is longer, and consumption organic solvent is more
The problems such as.
The content of the invention
For above-mentioned technical problem, it is an object of the present invention to provide a kind of liquid phase color for detecting diazepam drug concentration in blood
Spectral analysis method, makes the measure of diazepam blood concentration quick and precisely, and shortens sample analysis time, and the solvent in analysis
Flow velocity is very low, reduces the use of organic solvent, beneficial to environmental protection.
The present invention uses following technical scheme：
A kind of method for detecting diazepam drug concentration in blood of the present invention, wherein：It comprises the following steps：
(1) calibration of standard solution
First by the 10 μ L of standard working solution of at least three kinds various concentrations first with 10 μ L internal standards working solutions and 80 μ L respectively
Alcohol is mixed and made at least three kinds of standard solution, and above-mentioned standard solution is vortexed in the case where rotating speed is 1200-2000rpm mixes 30s- respectively
After 1min, above-mentioned standard solution is detected using liquid chromatography analysis equipment and UV detector, draws above-mentioned at least three
The diazepam and internal standard chromatogram of kind standard solution, respectively obtain standard target thing peak in above-mentioned diazepam and internal standard chromatogram
Area and internal standard compound peak area, standard is used as using the ratio of above-mentioned at least three standard targets thing peak area and internal standard compound peak area
The ordinate y of curve map, is marked using ratio, that is, relative concentration of above-mentioned standard working solution concentration and internal standard working solution concentration to be used as
The abscissa x of directrix curve figure, detection gained at least three groups of data carry out linear regression by more than, and fitting obtains calibration curve equation
For y=a*x+b, and draw weight coefficient a and b；The standard working solution is diazepam solution, and the internal standard working solution is chlorine
Nitrazepam solution；
(a) preparation of standard working solution：
Accurately weighing diazepam standard items 5mg is placed in 5ml volumetric flasks, and the methanol solution for being 0%-25% with water content carries out
Dissolving, and constant volume obtains standard reserving solution A, the methanol solution for being 0%-25% with water content by standard reserving solution A in 5ml
Dilution is diluted, and configures each standard working solution in the range of containing 0.5ug/mL-50ug/mL diazepams respectively, and
Preserved under the conditions of -20 DEG C；
(b) preparation of internal standard working solution：
Accurately weighing Clonazepam standard items 5mg is placed in 5mL volumetric flasks, is that 0%-25% methanol dissolves with water content, and
Constant volume obtains standard reserving solution B in 5mL, by the dilution of the standard reserving solution B methanol solutions for being 0%-25% with water content into
Row dilution, obtains concentration and is the internal standard working solution of 10ug/mL, and is preserved under the conditions of -20 DEG C；
(2) centrifugation of blood is detected
Blood to be detected at least 5ml is taken, 10min is centrifuged in the case where centrifugal speed is 3500rpm, takes supernatant to obtain serum or blood
Slurry, it is spare to before analyzing that above-mentioned serum or blood plasma are placed in the lower preservation of -20 DEG C of freezings；
(3) sample to be tested is handled
(c) pipette internal standard working solution described in 10 μ L steps (b) with liquid-transfering gun and in 1.5ml centrifuge tubes, then add 100
Serum or blood plasma described in μ L step (2), be vortexed concussion mixing 20s-1min under the rotating speed of 1200-2000rpm；
(d) ethyl acetate that 800uL is pipetted with liquid-transfering gun is added in the centrifuge tube of step (c), 1200-2000rpm's
Be vortexed concussion mixing 4-6min under rotating speed, then the high speed centrifugation 4-6min under the rotating speed of 10000-15000rpm, obtains supernatant；
(e) take step (d) supernatant 750uL to be put into another 1.5ml centrifuge tube, use N at normal temperatures2Slowly drying；
(f) methanol of 100 μ L is added into the centrifuge tube of the above-mentioned drying of step (e), is then turned in 1200-2000rpm
Speed is lower to be vortexed after mixing 30s-1min, then the high speed centrifugation 4-6min under the rotating speed of 10000-15000rpm, obtains supernatant i.e.
For sample to be tested；
(4) detection of sample to be tested
It is detected, is drawn using liquid chromatography analysis equipment and the UV detector sample to be measured to above-mentioned steps (f)
The diazepam and internal standard chromatogram of above-mentioned sample to be measured, obtain object peak to be measured in above-mentioned diazepam and internal standard chromatogram
Area and internal standard compound peak area, above-mentioned steps (one) are substituted into by the ratio y of object peak area to be measured and internal standard compound peak area
In calibration curve equation, object relative concentration x in detected sample is obtained by calculation, internal standard compound working solution concentration is known
, the diazepam drug concentration in blood to be detected is thus calculated in the sample；
A kind of method for detecting diazepam drug concentration in blood of the present invention, wherein：The above-mentioned standard working solution concentration
Refer to the concentration of contained diazepam in standard working solution, internal standard working solution concentration refers to contained Clonazepam in internal standard working solution
Concentration；
A kind of method for detecting diazepam drug concentration in blood of the present invention, wherein：In step (1) using seven kinds not
With the standard working solution of concentration；
A kind of method for detecting diazepam drug concentration in blood of the present invention, wherein：The standard of seven kinds of various concentrations
Working solution be respectively containing 0.625,1.25,2.50,5.00,10.00,20.00, the diazepam solution of 50.00ug/mL concentration；
A kind of method for detecting diazepam drug concentration in blood of the present invention, wherein：In step (a) and (b), standard storage
Standby liquid A and standard internal standard storing solution B is dissolved with methanol to be prepared, and the dilution for preparing standard working solution and internal standard working solution is first
Alcohol；
A kind of method for detecting diazepam drug concentration in blood of the present invention, wherein：The UV detector is VANQISH
Detector, its Detection wavelength are 230nm, and pot strainer is 1290 Infinity inline filter of Agilent；
A kind of method for detecting diazepam drug concentration in blood of the present invention, wherein：The high performance liquid chromatograph is made
Chromatographic column is the Zorbax Eclipse PlusC18 of Agilent companies；
A kind of method for detecting diazepam drug concentration in blood of the present invention, wherein：The high performance liquid chromatograph is set
Column temperature be 30 DEG C；
A kind of method for detecting diazepam drug concentration in blood of the present invention, wherein：The high performance liquid chromatograph is made
It is methanol-water with mobile phase, the sample size of high performance liquid chromatograph is 5 μ L, uses gradient elution program, flow velocity 0.7ml/
min；
A kind of method for detecting diazepam drug concentration in blood of the present invention, wherein：The water content is volumetric(al) moisture content.
Beneficial effect of the present invention：
The method of diazepam drug concentration in detection blood of the present invention, by internal standard method and high performance liquid chromatography phase
With reference to improving the accuracy of quantitative result, eliminate systematic error；And analysis time is effectively shortened, make detection process easy
Quickly, the diazepam blood concentration of patient's body is monitored more conducively in clinical treatment, is given for the personalization of diazepam
Medicine, the generation offer experiment basis for reducing toxicity.
Brief description of the drawings
Fig. 1 diazepam chemical structural formulas；
Fig. 2 is diazepam and its internal standard chromatogram in embodiment Plays solution；
Fig. 3 is diazepam and its internal standard chromatogram in mark-on serum in embodiment or plasma sample；
Below in conjunction with specific embodiments and the drawings, the invention will be further described.
Embodiment
The method of diazepam drug concentration comprises the following steps in the detection blood of the present invention：
(1) calibration of standard solution
Methanol of the 10 μ L of standard working solution of seven kinds of various concentrations respectively with 10 μ L internal standards working solutions and 80 μ L is mixed first
Seven kinds of standard solution are made in conjunction, and above-mentioned standard solution after the mixing 1min that is vortexed under rotating speed is 1400rpm, uses liquid phase color respectively
Spectrum analysis instrument and UV detector are detected above-mentioned standard solution, draw the diazepam of above-mentioned seven kinds of standard solution and interior
Chromatogram is marked, standard target thing peak area and internal standard compound peak area are respectively obtained in above-mentioned diazepam and internal standard chromatogram, with
Ordinate y of the ratio of above-mentioned at least three standard targets thing peak area and internal standard compound peak area as canonical plotting, the above
The ratio i.e. relative concentration for stating standard working solution concentration and internal standard working solution concentration is the abscissa x as canonical plotting, will
Detection gained at least three groups of data carry out linear regressions above, and it is y=a*x+b that fitting, which obtains calibration curve equation, and is drawn
Weight coefficient a and b；The standard working solution is diazepam solution, in the quasi- working solution of standard working solution concentration index containedly
The concentration that west is dissolved；The internal standard working solution is Clonazepam solution, and internal standard working solution concentration refers to contained chlorine in internal standard working solution
The concentration of nitrazepam；
(a) preparation of standard working solution：
Accurately weighing diazepam standard items 5mg is placed in 5ml volumetric flasks, is dissolved with methanol, and constant volume is obtained in 5ml
Standard reserving solution A, standard reserving solution A is diluted with methanol, be respectively configured out the concentration containing diazepam for 0.625,
1.25th, 2.50,5.00,10.00, the 20.00, standard working solution of 50.00ug/mL, and preserved under the conditions of -20 DEG C；
(b) preparation of internal standard working solution：
Accurately weighing Clonazepam standard items 5mg is placed in 5mL volumetric flasks, is dissolved with methanol, and constant volume is obtained in 5mL
To standard reserving solution B, standard reserving solution B is diluted with methanol, obtain concentration be 10ug/mL internal standard working solution, and-
Preserved under the conditions of 20 DEG C；
(2) centrifugation of blood is detected
Blood to be detected at least 5ml is taken, 10min is centrifuged in the case where centrifugal speed is 3500rpm, takes supernatant to obtain serum or blood
Slurry, it is spare to before analyzing that above-mentioned serum or blood plasma are placed in the lower preservation of -20 DEG C of freezings；
(3) sample to be tested is handled
(c) pipette internal standard working solution described in 10 μ L steps (b) with liquid-transfering gun and in 1.5ml centrifuge tubes, then add 100
Serum or blood plasma described in μ L step (2), be vortexed concussion mixing 30s under the rotating speed of 2000rpm；
(d) ethyl acetate that 800uL is pipetted with liquid-transfering gun is added in the centrifuge tube of step (c), under the rotating speed of 2000rpm
Be vortexed concussion mixing 5min, then the high speed centrifugation 5min under the rotating speed of 12000rpm, obtains supernatant；
(e) take step (d) supernatant 750uL to be put into another 1.5ml centrifuge tube, use N at normal temperatures2Slowly drying；
(f) methanol of 100 μ L is added into the centrifuge tube of the above-mentioned drying of step (e), then the whirlpool under 2000rpm rotating speeds
After rotation mixes 1min, then the high speed centrifugation 5min under the rotating speed of 12000rpm, it is sample to be tested to obtain supernatant；
(4) detection of sample to be tested
It is detected, is drawn using liquid chromatography analysis equipment and the UV detector sample to be measured to above-mentioned steps (f)
The diazepam and internal standard chromatogram of above-mentioned sample to be measured, obtain object peak to be measured in above-mentioned diazepam and internal standard chromatogram
Area and internal standard compound peak area, above-mentioned steps (one) are substituted into by the ratio y of object peak area to be measured and internal standard compound peak area
In calibration curve equation, object relative concentration x in detected sample is obtained by calculation, internal standard compound working solution concentration is known
, the diazepam drug concentration in blood to be detected is thus calculated in the sample.
UV detector is VANQISH detectors, its Detection wavelength is 230nm, and pot strainer is Agilent 1290
Infinity inline filter；Chromatographic column used in high performance liquid chromatograph is the Zorbax of Agilent companies
Eclipse PlusC18；Column temperature is 30 DEG C, and it is methanol-water that high performance liquid chromatograph, which uses mobile phase, high performance liquid chromatograph
Sample size be 5 μ L, use gradient elution program, flow velocity 0.7ml/min.
If without specified otherwise, the water content is volumetric(al) moisture content in this application.
| Time/min | Methanol/% | Water/% |
| 0.00 | 65 | 35 |
| 2.50 | 65 | 35 |
| 2.51 | 70 | 30 |
| 4.50 | 70 | 30 |
| 6.00 | 80 | 20 |
1 liquid chromatogram elution program of table
Technical method demonstration is as follows in the present embodiment：
First, the linear relationship and quantitative limit of this method
By the diazepam standard working solution of each concentration of 10 μ L of above-mentioned preparation, 10 μ L internal standard working solutions are added, and add
Enter 80 μ L of methanol and mix sample introduction, by the present embodiment determination condition, be measured from low to high by concentration, with quantitative chromatographic peak face
Product-concentration is mapped, and obtains standard curve, the results showed that the range of linearity and quantitative limit of diazepam are as follows：
(1) test limit (LOD)：0.0079μg/mL.
(2) quantitative limit (LOQ)：0.0263μg/mL.
(3) range of linearity：
Diazepam is linear good in the range of 0.0625 μ g/mL to 5.000 μ g/mL, coefficient R2＞ 0.99.
2nd, the rate of recovery and precision of this method
Taking diazepam standard working solution to be configured to, high, medium and low 3 kinds of concentration carries out sample recovery rate experiment and precision is real
Test, be measured by the present embodiment method, replicate analysis measure 3 batches, its rate of recovery and precision are respectively such as table 2.Its it is low,
Average recovery rate in the range of 3 middle and high pitch-based spheres is 99.9%~102.7%, relative standard deviation for 1.91%~
2.63%, it the results are shown in Table 2.
| Mark-on amount | 0.125μg/mL | 0.500μg/mL | 2.000μg/mL |
| Average recovery rate | 107.0% | 100.6% | 100.8% |
| Precision RSD | 1.13% | 1.85% | 2.38% |
2 diazepam recovery of standard addition of table and precision
Summary checking test, the test limit of the present embodiment, all technical such as the rate of recovery and precision meet
It is required that diazepam drug concentration in method detection blood, reappearance is good, and sample recovery rate is high, improves the standard of testing result
Exactness.
Diazepam and its interior target chromatogram are shown in Fig. 3 in serum or plasma sample, diazepam and its internal standard in standard solution
Chromatogram see Fig. 2, the retention time of diazepam and diazepam is respectively 5.33min and 3.03min, this is understood by Fig. 2 and Fig. 3
Using diazepam as internal standard, systematic error is eliminated embodiment method, and the identification of target compound is more accurate, and analysis time
Short, small, the high specificity of interference.
Embodiment described above is only that the preferred embodiment of the present invention is described, not to the model of the present invention
Enclose and be defined, on the premise of design spirit of the present invention is not departed from, technical side of the those of ordinary skill in the art to the present invention
The various modifications and improvement that case is made, should all fall into the protection domain that claims of the present invention determines.
Claims (10)
- A kind of 1. method for detecting diazepam drug concentration in blood, it is characterised in that：It comprises the following steps：(1) calibration of standard solutionMethanol of the 10 μ L of standard working solution of at least three kinds various concentrations respectively with 10 μ L internal standards working solutions and 80 μ L is mixed first At least three kinds of standard solution are made in conjunction, and above-mentioned standard solution is vortexed in the case where rotating speed is 1200-2000rpm mixes 30s-1min respectively Afterwards, above-mentioned standard solution is detected using liquid chromatography analysis equipment and UV detector, draws above-mentioned at least three kinds marks The diazepam and internal standard chromatogram of quasi- solution, respectively obtain standard target thing peak area in above-mentioned diazepam and internal standard chromatogram With internal standard compound peak area, standard curve is used as using the ratio of above-mentioned at least three standard targets thing peak area and internal standard compound peak area The ordinate y of figure, using ratio, that is, relative concentration of above-mentioned standard working solution concentration and internal standard working solution concentration to be bent as standard The abscissa x of line chart, detection gained at least three groups of data carry out linear regression by more than, and it is y that fitting, which obtains calibration curve equation, =a*x+b, and draw weight coefficient a and b；The standard working solution is diazepam solution, and the internal standard working solution is chlorine nitre Dissolve solution in west；(a) preparation of standard working solution：Accurately weighing diazepam standard items 5mg is placed in 5ml volumetric flasks, and the methanol solution for being 0%-25% with water content carries out molten Solution, and constant volume obtains standard reserving solution A, by the dilute of the standard reserving solution A methanol solutions for being 0%-25% with water content in 5ml Release liquid to be diluted, configure each standard working solution in the range of containing 0.5ug/mL-50ug/mL diazepams respectively, and- Preserved under the conditions of 20 DEG C；(b) preparation of internal standard working solution：Accurately weighing Clonazepam standard items 5mg is placed in 5mL volumetric flasks, is that 0%-25% methanol dissolves with water content, and constant volume In 5mL, standard reserving solution B is obtained, the dilution of the standard reserving solution B methanol solutions for being 0%-25% with water content is carried out dilute Release, obtain concentration and be the internal standard working solution of 10ug/mL, and preserved under the conditions of -20 DEG C；(2) centrifugation of blood is detectedBlood to be detected at least 5ml is taken, 10min is centrifuged in the case where centrifugal speed is 3500rpm, takes supernatant to obtain serum or blood plasma, It is spare to before analyzing that above-mentioned serum or blood plasma are placed in the lower preservation of -20 DEG C of freezings；(3) sample to be tested is handled(c) internal standard working solution described in 10 μ L steps (b) is pipetted in 1.5ml centrifuge tubes with liquid-transfering gun, then add 100 μ L steps Suddenly serum or blood plasma described in (two), be vortexed concussion mixing 20s-1min under the rotating speed of 1200-2000rpm；(d) ethyl acetate that 800uL is pipetted with liquid-transfering gun is added in the centrifuge tube of step (c), in the rotating speed of 1200-2000rpm The lower concussion mixing 4-6min that is vortexed, then the high speed centrifugation 4-6min under the rotating speed of 10000-15000rpm, obtain supernatant；(e) take step (d) supernatant 750uL to be put into another 1.5ml centrifuge tube, use N at normal temperatures2Slowly drying；(f) methanol of 100 μ L is added into the centrifuge tube of the above-mentioned drying of step (e), then under 1200-2000rpm rotating speeds It is vortexed after mixing 30s-1min, then the high speed centrifugation 4-6min under the rotating speed of 10000-15000rpm, it is to treat to obtain supernatant Sample；(4) detection of sample to be testedIt is detected, is drawn above-mentioned using liquid chromatography analysis equipment and the UV detector sample to be measured to above-mentioned steps (f) The diazepam and internal standard chromatogram of sample to be measured, obtain object peak area to be measured in above-mentioned diazepam and internal standard chromatogram With internal standard compound peak area, the ratio y of object peak area to be measured and internal standard compound peak area is substituted into the standards of above-mentioned steps (one) In curvilinear equation, be obtained by calculation object relative concentration x in detected sample, internal standard compound working solution concentration be it is known, Thus the diazepam drug concentration in blood to be detected is calculated in the sample.
- 2. the method for diazepam drug concentration in blood is detected as claimed in claim 1, it is characterised in that：The above-mentioned standard The concentration of contained diazepam in the quasi- working solution of working solution concentration index, internal standard working solution concentration refer to contained in internal standard working solution The concentration of Clonazepam.
- 3. the method for diazepam drug concentration in blood is detected as claimed in claim 2, it is characterised in that：In step (1) Use the standard working solution of seven kinds of various concentrations.
- 4. the method for diazepam drug concentration in blood is detected as claimed in claim 3, it is characterised in that：Seven kinds of differences The standard working solution of concentration be respectively containing 0.625,1.25,2.50,5.00,10.00,20.00, the ground of 50.00ug/mL concentration Dissolve solution in west.
- 5. the method for diazepam drug concentration in blood is detected as claimed in claim 4, it is characterised in that：In step (a) and (b) in, standard reserving solution A and standard internal standard storing solution B are dissolved with methanol and prepared, and prepare standard working solution and internal standard working solution Dilution be methanol.
- 6. the method for diazepam drug concentration in blood is detected as claimed in claim 5, it is characterised in that：The ultraviolet detection Device is VANQISH detectors, its Detection wavelength is 230nm, and pot strainer is Agilent 1290Infinity inline filter。
- 7. the method for diazepam drug concentration in blood is detected as claimed in claim 5, it is characterised in that：The efficient liquid phase Chromatographic column used in chromatograph is the Zorbax Eclipse PlusC18 of Agilent companies.
- 8. the method for diazepam drug concentration in blood is detected as claimed in claim 5, it is characterised in that：The efficient liquid phase The column temperature that chromatograph is set is 30 DEG C.
- 9. the method for diazepam drug concentration in blood is detected as claimed in claim 5, it is characterised in that：The efficient liquid phase It is methanol-water that chromatograph, which uses mobile phase, and the sample size of high performance liquid chromatograph is 5 μ L, uses gradient elution program, flow velocity For 0.7ml/min.
- 10. the method for diazepam drug concentration in the detection blood as described in one of claim 5 to 9, it is characterised in that：It is described Water content is volumetric(al) moisture content.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108445113A (en) * | 2018-05-30 | 2018-08-24 | 北京和合医学诊断技术股份有限公司 | Detect the on-line solid phase extraction liquid phase chromatography analytical method of Clonazepam content in blood |
| CN110715996A (en) * | 2018-07-12 | 2020-01-21 | 中国科学院生态环境研究中心 | Method for full-automatic online detection of free-state small molecular compound in serum |
| CN113267580A (en) * | 2021-05-18 | 2021-08-17 | 中南民族大学 | Method for monitoring diazepam blood concentration |
| CN116297389A (en) * | 2023-02-15 | 2023-06-23 | 中国人民解放军空军特色医学中心 | Drug detection method capable of causing driving operation disability in biological sample at accident site |
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| CN108445113A (en) * | 2018-05-30 | 2018-08-24 | 北京和合医学诊断技术股份有限公司 | Detect the on-line solid phase extraction liquid phase chromatography analytical method of Clonazepam content in blood |
| CN108445113B (en) * | 2018-05-30 | 2022-07-26 | 北京和合医学诊断技术股份有限公司 | On-line solid phase extraction liquid phase chromatographic analysis method for detecting clonazepam content in blood |
| CN110715996A (en) * | 2018-07-12 | 2020-01-21 | 中国科学院生态环境研究中心 | Method for full-automatic online detection of free-state small molecular compound in serum |
| CN110715996B (en) * | 2018-07-12 | 2022-04-12 | 中国科学院生态环境研究中心 | Method for full-automatic online detection of free-state small molecular compound in serum |
| CN113267580A (en) * | 2021-05-18 | 2021-08-17 | 中南民族大学 | Method for monitoring diazepam blood concentration |
| CN116297389A (en) * | 2023-02-15 | 2023-06-23 | 中国人民解放军空军特色医学中心 | Drug detection method capable of causing driving operation disability in biological sample at accident site |
| CN116297389B (en) * | 2023-02-15 | 2023-12-26 | 中国人民解放军空军特色医学中心 | Drug detection method capable of causing driving operation disability in biological sample at accident site |
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