CN108132315A - A kind of liquid phase chromatography analytical method for detecting blood citalopram content - Google Patents
A kind of liquid phase chromatography analytical method for detecting blood citalopram content Download PDFInfo
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- CN108132315A CN108132315A CN201810075227.0A CN201810075227A CN108132315A CN 108132315 A CN108132315 A CN 108132315A CN 201810075227 A CN201810075227 A CN 201810075227A CN 108132315 A CN108132315 A CN 108132315A
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The method of present invention detection blood citalopram drug concentration is that standard solution is demarcated using liquid chromatography analysis equipment and fluorescence detector, fitting obtains calibration curve equation as y=a*x+b, 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 fluorescence 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 Citalopram drug concentration in blood to be detected in the sample is calculated, internal standard method is combined by the present invention with high performance liquid chromatography, improve the accuracy of quantitative result, eliminate systematic error；Analysis time is shortened, makes detection process easy to be quick, the Citalopram 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 antidepressant Citalopram concentration.
Background technology
Citalopram (citalopram), molecular formula C20H21FN2O, particular chemical are illustrated in fig. 1 shown below, and are belonged to
Selective serotonin reuptake inhibitor, since to receptor-selective height, the less serious Cardiovascular Toxicity of generation is not easy to produce
Raw drug interaction, patient compliance are preferable, it has also become antidepressant first-line drug, in addition, being clinically also used to ease pain, guard against
The treatment of wine and senile dementia and multiple infarct dementia.Influenced by gene pleiomorphism, individual vivo medicine concentration difference compared with
Greatly, blood concentration and clinical efficacy and adverse reaction are closely related, in accurate Medical Era, need to monitor patient's blood concentration simultaneously
Drug dose is adjusted according to blood concentration, to realize personalized medicine.Therefore, blood citalopram concentration is measured to ensureing
Its curative effect of medication plays an important roll.
The assay method of common Citalopram blood concentration mainly include Capillary Zone Electrophoresis with Electrochemical luminescence method, efficiently
Liquid chromatogram-fluorescence detection, HPLC- UV detection method, high performance liquid chromatography Mass Spectrometry etc..Wherein, it is high
Effect liquid phase chromatogram Mass Spectrometry cost is higher, high to personnel and site requirements, is unsuitable for extensively carrying out in small-middle hospital etc.,
Therefore the method for detection human plasma citalopram concentration is mainly high performance liquid chromatography, but Citalopram exists both at home and abroad at present
Blood concentration in human body is relatively low, and the operation of method pre-treatment at present is more complicated, and it is not accurate enough that quantified by external standard method easily leads to result,
In addition, it is also longer there are analysis time, consume the problems such as organic solvent is more.
Invention content
For above-mentioned technical problem, it is an object of the present invention to provide a kind of sides for detecting blood citalopram drug concentration
Internal standard method is combined by method with high performance liquid chromatography, makes the measure of Citalopram blood concentration quick and precisely, and largely
Upper shortening sample analysis time, and solvent flow rate is relatively low in analysis, reduces the use of organic solvent, conducive to environmental protection.
The present invention uses following technical scheme：
A kind of method of detection blood citalopram drug concentration of the present invention, wherein：It includes the following steps：
(1) calibration of standard solution
First by 20 μ L of the standard working solution of at least three kinds various concentrations respectively with 10 μ L internal standards working solutions and by 1: 1 first
170 μ L of alcohol and water composition redissolve liquid and are mixed and made at least three kinds of standard solution, and above-mentioned standard solution is respectively 1200- in rotating speed
Under 2000rpm after vortex mixing 30s-2min, using liquid chromatography analysis equipment and fluorescence detector to above-mentioned standard solution into
Row detection obtains the Citalopram and internal standard chromatogram of above-mentioned at least three kinds standard solution, in above-mentioned Citalopram and internal standard color
Standard target object peak area and internal standard compound peak area are respectively obtained in spectrogram, with above-mentioned at least three standard targets object peak area with
Ordinate y of the ratio of internal standard compound peak area as canonical plotting is dense with above-mentioned standard working solution concentration and internal standard working solution
Ratio, that is, relative concentration of degree is the abscissa x as canonical plotting, will detect gained at least three groups of data above into line
Property return, fitting obtains calibration curve equation as y=a*x+b, and obtains weight coefficient a and b；The standard working solution is west
Phthalein Pulan solution, the internal standard working solution are cyheptamide solution；
(a) preparation of standard working solution：
Accurately weighing Citalopram standard items 3.63mg is placed in 10ml volumetric flasks, and the methanol for being 0%-25% with water content is molten
Liquid is dissolved, and constant volume obtains standard reserving solution A, i.e., the Citalopram solution of a concentration of 284 μ g/mL, by standard in 10ml
The dilution for the methanol solution that storing solution A is 25%-60% with water content is diluted, and is containing 0.1-1.5 μ g/mL west respectively
Each standard working solution is configured in the range of phthalein Pulan, and is preserved under the conditions of -80 DEG C；
(b) preparation of internal standard working solution：
Accurately weighing cyheptamide standard items 10mg is placed in 10mL volumetric flasks, is that 0%-25% methanol dissolves with water content,
And constant volume obtains standard reserving solution B, by the dilution of the standard reserving solution B methanol solutions for being 25%-60% with water content in 10mL
Liquid is diluted, and obtains the internal standard working solution of a concentration of 20 μ g/mL, 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, supernatant is taken to obtain serum or blood
It is spare to before analyzing that slurry, 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) it pipettes internal standard working solution described in 10 μ L steps (b) with liquid-transfering gun and in 1.5ml centrifuge tubes, then adds in 200
Serum or blood plasma described in μ L step (2), be vortexed concussion mixing 30s-2min under the rotating speed of 1200-2000rpm；
(d) it pipettes 20 μ L acetonitriles with liquid-transfering gun to add in the centrifuge tube of step (c), the whirlpool under the rotating speed of 1200-2000rpm
Then rotation mixing 1-1.5min adds in the 10 μ L of NaOH solution of 10mol/L, under the rotating speed of 1200-2000rpm in centrifuge tube
After vortex mixed 1-1.5min, the methyl tertiary butyl ether(MTBE) extractant 1mL containing 10%-30% n-hexanes is added, in 1200-
Vortex mixing 2-5min under the rotating speed of 2000rpm, to achieve the purpose that fully to extract, then in the rotating speed of 10000-15000rpm
Lower high speed centrifugation 4-6min, obtains supernatant；
(e) 900 μ L of step (d) supernatant is taken to be put into another 1.5ml centrifuge tube, use N at normal temperatures2Slowly drying；
(f) the 200 μ L of redissolution liquid being made of 1: 1 first alcohol and water are added in into the centrifuge tube of the above-mentioned drying of step (e),
Then under 1200-2000rpm rotating speeds after vortex mixing 2-5min, then the high speed centrifugation 4- under the rotating speed of 10000-15000rpm
6min, it is sample to be tested to obtain supernatant；
(4) detection of sample to be tested
It is detected, obtained using liquid chromatography analysis equipment and the fluorescence detector sample to be measured to above-mentioned steps (f)
The Citalopram of above-mentioned sample to be measured and internal standard chromatogram obtain object to be measured in above-mentioned Citalopram and internal standard chromatogram
The ratio y of object to be measured object peak area and internal standard compound peak area is substituted into above-mentioned steps by object peak area and internal standard compound peak area
(1) in calibration curve equation, object relative concentration x in detected sample, internal standard compound working solution concentration is obtained by calculation
It is known, the Citalopram drug concentration in blood to be detected in the sample is thus calculated；
A kind of method of detection blood citalopram drug concentration of the present invention, wherein：The above-mentioned standard working solution
The concentration of contained Citalopram in the quasi- working solution of concentration index, internal standard working solution concentration refer to contained cycloheptyl in internal standard working solution
The concentration of meter Te；
A kind of method of detection blood citalopram drug concentration of the present invention, wherein：Seven are used in step (1)
The standard working solution of kind various concentration；
A kind of method of detection blood citalopram drug concentration of the present invention, wherein：Seven kinds of various concentrations
Standard working solution is respectively the Citalopram solution containing 0.1,0.2,0.4,0.8,1.0,1.2,1.5 μ g/mL concentration；
A kind of method of detection blood citalopram drug concentration of the present invention, wherein：In step (a) and step (b)
It is dissolved to obtain standard reserving solution A and standard reserving solution B with methanol respectively, with the dilution for the methanol solution that water content is 50%
Liquid is diluted respectively, configures each standard working solution and internal standard working solution, and the extractant described in step (d) is by 15: 85
N-hexane and methyl tertiary butyl ether(MTBE) composition extractant；
A kind of method of detection blood citalopram drug concentration of the present invention, wherein：The high performance liquid chromatograph
Used pot strainer is 1/16 0.5M of SSI COL PRE-FILTER WATER；
A kind of method of detection blood citalopram drug concentration of the present invention, wherein：The high performance liquid chromatograph
Used chromatographic column is the AccucoreC18 of Thermo companies；
A kind of method of detection blood citalopram drug concentration of the present invention, wherein：The high performance liquid chromatograph
The column temperature of setting is 35 DEG C；
A kind of method of detection blood citalopram drug concentration of the present invention, wherein：The high performance liquid chromatograph
Used mobile phase is the acetonitrile-water containing 0.1% formic acid and 10mM sodium dihydrogen phosphates, and using gradient elution, and sample size is
30μL；
A kind of method of detection blood citalopram drug concentration of the present invention, wherein：The water content is volume ratio
Water content.
Advantageous effect of the present invention：
The method of detection blood citalopram drug concentration of the present invention, by internal standard method and high performance liquid chromatography
It is combined, improves the accuracy of quantitative result, eliminate systematic error；Analysis time is shortened, makes detection process easy to be quick,
The Citalopram blood concentration of patient's body is monitored more conducively in clinical treatment, the personalization for Citalopram is given
Medicine, the generation offer experiment basis for reducing toxicity.
Description of the drawings
Fig. 1 Citalopram chemical structural formulas；
Fig. 2 is embodiment Plays solution citalopram and its internal standard chromatogram；Wherein, 1- Citaloprams；2- cycloheptyls
Meter Te；
Fig. 3 is mark-on serum or plasma sample citalopram and its internal standard chromatogram in embodiment；Wherein, the western phthaleins of 1- are general
It is blue；2-
Cyheptamide；
Below in conjunction with specific embodiments and the drawings, the invention will be further described.
Specific embodiment
The method of the detection blood citalopram drug concentration of the present invention, it includes the following steps：
(1) calibration of standard solution
First by 20 μ L of the standard working solution of seven kinds of various concentrations respectively with 10 μ L internal standards working solutions and by 1: 1 methanol and
170 μ L of water composition redissolve liquid and are mixed and made into seven kinds of standard solution, in rotating speed are respectively whirlpool under 2000rpm by above-mentioned standard solution
After revolving mixing 1min, above-mentioned standard solution is detected using liquid chromatography analysis equipment and fluorescence detector, is obtained above-mentioned
The Citalopram and internal standard chromatogram of seven kinds of standard solution, standard mesh is respectively obtained in above-mentioned Citalopram and internal standard chromatogram
Object peak area and internal standard compound peak area are marked, is made respectively with the ratio of above-mentioned seven kinds of standard target object peak areas and internal standard compound peak area
For the ordinate y of canonical plotting, divided with ratio, that is, relative concentration of above-mentioned standard working solution concentration and internal standard working solution concentration
Not as the abscissa x of canonical plotting, seven groups of data of gained will be detected above and carry out linear regression, fitting obtains standard curve
Equation is y=a*x+b, and obtains weight coefficient a and b；The standard working solution is Citalopram solution, the internal standard work
Liquid is cyheptamide solution；
(a) preparation of standard working solution：
Accurately weighing Citalopram standard items 3.63mg is placed in 10ml volumetric flasks, is dissolved with methanol solution, and constant volume
In 10ml, standard reserving solution A is obtained, i.e., standard reserving solution A is by the Citalopram solution of a concentration of 284 μ g/mL with water content
The dilution of 50% methanol solution is diluted, and is containing 0.1,0.2,0.4,0.8,1.0,1.2,1.5 μ g/mL concentration respectively
Citalopram solution, and preserved under the conditions of -80 DEG C；
(b) preparation of internal standard working solution：
It is accurate to weigh cyheptamide standard items 10mg and be placed in 10mL volumetric flasks, dissolved with methanol solution, and constant volume in
10mL obtains standard reserving solution B, and standard reserving solution B with the dilution for the methanol solution that water content is 50% is diluted, is obtained
To the internal standard working solution of a concentration of 20 μ g/mL, 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, supernatant is taken to obtain serum or blood
It is spare to before analyzing that slurry, 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) it pipettes internal standard working solution described in 10 μ L steps (b) with liquid-transfering gun and in 1.5ml centrifuge tubes, then adds in 200
Serum or blood plasma described in μ L step (2), be vortexed concussion mixing 1min under the rotating speed of 2000rpm；
(d) 20 μ L acetonitriles are pipetted with liquid-transfering gun to add in the centrifuge tube of step (c), is vortexed under the rotating speed of 2000rpm mixed
1min is closed, the NaOH solution 10 μ L, the vortex mixed 1min under the rotating speed of 2000rpm of 10mol/L are then added in centrifuge tube
Afterwards, the methyl tertiary butyl ether(MTBE) extractant 1mL containing 15% n-hexane, the vortex mixing 3min under the rotating speed of 2000rpm are added,
To achieve the purpose that fully to extract, then the high speed centrifugation 5min under the rotating speed of 12000rpm, obtains supernatant；
(e) 900 μ L of step (d) supernatant is taken to be put into another 1.5ml centrifuge tube, use N at normal temperatures2Slowly drying；
(f) the 200 μ L of redissolution liquid being made of 1: 1 first alcohol and water are added in into the centrifuge tube of the above-mentioned drying of step (e),
Then under 2000rpm rotating speeds after vortex mixing 3min, then the high speed centrifugation 5min under the rotating speed of 12000rpm, obtain supernatant
As sample to be tested；
(4) detection of sample to be tested
It is detected, obtained using liquid chromatography analysis equipment and the fluorescence detector sample to be measured to above-mentioned steps (f)
The Citalopram of above-mentioned sample to be measured and internal standard chromatogram obtain object to be measured in above-mentioned Citalopram and internal standard chromatogram
The ratio y of object to be measured object peak area and internal standard compound peak area is substituted into above-mentioned steps by object peak area and internal standard compound peak area
(1) in calibration curve equation, object relative concentration x in detected sample, internal standard compound working solution concentration is obtained by calculation
It is known, the Citalopram drug concentration in blood to be detected in the sample is thus calculated.
Pot strainer used in high performance liquid chromatograph is 1/16 0.5M of SSI COL PRE-FILTER WATER.
Chromatographic column used in high performance liquid chromatograph is the AccucoreC18 of Thermo companies, and column temperature is 35 DEG C, high performance liquid chromatography
Instrument uses mobile phase as the acetonitrile-water containing 0.1% formic acid and 10mM sodium dihydrogen phosphates, and using gradient elution, sample size
For 30 μ L.
If without specified otherwise, the water content in the application is the water content of volume ratio.
1 liquid chromatogram gradient elution program of table
| Time/min | Flow velocity (mL/min) | Water (0.1%FA, 10mM NaH2PO4)/% | Acetonitrile/% |
| 0.0 | 0.7 | 70 | 30 |
| 4.0 | 0.7 | 70 | 30 |
| 4.1 | 0.8 | 70 | 30 |
| 4.2 | 0.9 | 70 | 30 |
| 6.0 | 0.9 | 70 | 30 |
| 6.8 | 0.9 | 10 | 90 |
| 7.0 | 0.7 | 10 | 90 |
| 10.0 | 0.7 | 10 | 90 |
2 fluorescence detector of table is set
| Channel | Excitation wavelength (nm) | Launch wavelength (nm) |
| 1 | 236 | 306 |
| 2 | 249 | 302 |
| 3 | 235 | 290 |
| 4 | 240 | 300 |
Technical method demonstration is as follows in the present embodiment：
First, the linear relationship and quantitative limit of this method
By the Citalopram mark song working solution of each concentration of 20 μ L of above-mentioned preparation, 10 μ L internal standard working solutions are added in, and
It adds in and redissolves liquid (methanol: water=1: 1) 170 μ L mixing sample introductions by the present embodiment determination condition, are surveyed from low to high by concentration
It is fixed, it is mapped with quantitative chromatographic peak area-concentration, obtains standard curve, the results showed that the range of linearity and quantitative limit of Citalopram
It is as follows：
(1) detection limit (LOD)：0.70ng/mL.
(2) quantitative limit (LOQ)：2.33ng/mL.
(3) range of linearity：
Citalopram is linear good in the range of 10ng/mL to 150ng/mL, coefficient R2＞ 0.99.
2nd, the rate of recovery and precision of this method
Citalopram standard working solution is taken to be configured to, and high, medium and low 3 kinds of concentration carries out sample recovery rate experiment and precision is real
It tests, is measured by the present embodiment method, replicate analysis measures 3 batches, and the rate of recovery and precision are respectively such as the following table 3.Its
Average recovery rate in the range of 3 basic, normal, high pitch-based spheres is 92.40%~97.89%, and relative standard deviation is
1.20%~1.48%, detailed results see the table below 3.
3 Citalopram recovery of standard addition of table and precision
| Mark-on amount | 20ng/mL | 80ng/mL | 120ng/mL |
| Average recovery rate | 97.89% | 97.10% | 92.40% |
| Precision RSD | 1.48% | 1.37% | 1.20% |
Summary verification test, the detection limit of the present embodiment, all technicals such as the rate of recovery and precision meet
It is required that method detects blood citalopram drug concentration, reproducibility is good, and sample recovery rate is high, improves testing result
Accuracy.Serum or plasma sample citalopram and its interior target chromatogram are shown in Fig. 2, standard solution citalopram and its interior
Target chromatogram is shown in Fig. 3, and the retention time of Citalopram and cyheptamide is respectively 3.953min and 7.393min, by Fig. 2 and
Fig. 3 understands the present embodiment method using cyheptamide as internal standard, and systematic error is eliminated, subject to the identification more of target compound
Really, and analysis time is short, interferes small, high specificity.
Embodiment described above is only that the preferred embodiment of the present invention is described, not to the model of the present invention
It encloses and is defined, under the premise of design spirit of the present invention is not departed from, those of ordinary skill in the art are to the technical side of the present invention
The various modifications and improvement that case is made should all be fallen into the protection domain that claims of the present invention determines.
Claims (10)
- A kind of 1. method for detecting blood citalopram drug concentration, it is characterised in that：It includes the following steps：(1) calibration of standard solutionFirst by 20 μ L of the standard working solution of at least three kinds various concentrations respectively with 10 μ L internal standards working solutions and by 1: 1 methanol and 170 μ L of water composition redissolve liquid and are mixed and made at least three kinds of standard solution, and above-mentioned standard solution is respectively 1200- in rotating speed Under 2000rpm after vortex mixing 30s-2min, using liquid chromatography analysis equipment and fluorescence detector to above-mentioned standard solution into Row detection obtains the Citalopram and internal standard chromatogram of above-mentioned at least three kinds standard solution, in above-mentioned Citalopram and internal standard color Standard target object peak area and internal standard compound peak area are respectively obtained in spectrogram, with above-mentioned at least three standard targets object peak area with Ordinate y of the ratio of internal standard compound peak area as canonical plotting is dense with above-mentioned standard working solution concentration and internal standard working solution Ratio, that is, relative concentration of degree is the abscissa x as canonical plotting, will detect gained at least three groups of data above into line Property return, fitting obtains calibration curve equation as y=a*x+b, and obtains weight coefficient a and b；The standard working solution is west Phthalein Pulan solution, the internal standard working solution are cyheptamide solution；(a) preparation of standard working solutionIt is accurate to weigh Citalopram standard items 3.63mg and be placed in 10ml volumetric flasks, with the methanol solution that water content is 0%-25% into Row dissolving, and constant volume obtains standard reserving solution A, i.e., the Citalopram solution of a concentration of 284 μ g/mL, by standard inventory in 10ml The dilution for the methanol solution that liquid A is 25%-60% with water content is diluted, respectively general containing the western phthaleins of 0.1-1.5 μ g/mL Each standard working solution is configured, and preserved under the conditions of -80 DEG C in blue range；(b) preparation of internal standard working solution：Accurately weighing cyheptamide standard items 10mg is placed in 10mL volumetric flasks, is that 0%-25% methanol dissolves, and fixed with water content Be dissolved in 10mL, obtain standard reserving solution B, by standard reserving solution B with the dilution of methanol solution that water content is 25%-60% into Row dilution, obtains the internal standard working solution of a concentration of 20 μ g/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, supernatant is taken 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 in 200 μ L steps Suddenly serum or blood plasma described in (two), be vortexed concussion mixing 30s-2min under the rotating speed of 1200-2000rpm；(d) 20 μ L acetonitriles are pipetted with liquid-transfering gun to add in the centrifuge tube of step (c), is vortexed under the rotating speed of 1200-2000rpm mixed 1-1.5min is closed, the 10 μ L of NaOH solution of 10mol/L are then added in centrifuge tube, are vortexed under the rotating speed of 1200-2000rpm After mixing 1-1.5min, the methyl tertiary butyl ether(MTBE) extractant 1mL containing 10%-30% n-hexanes is added, in 1200- Vortex mixing 2-5min under the rotating speed of 2000rpm, to achieve the purpose that fully to extract, then in the rotating speed of 10000-15000rpm Lower high speed centrifugation 4-6min, obtains supernatant；(e) 900 μ L of step (d) supernatant is taken to be put into another 1.5ml centrifuge tube, use N at normal temperatures2Slowly drying；(f) the 200 μ L of redissolution liquid being made of 1: 1 first alcohol and water are added in into the centrifuge tube of the above-mentioned drying of step (e), then Under 1200-2000rpm rotating speeds after vortex mixing 2-5min, then the high speed centrifugation 4- under the rotating speed of 10000-15000rpm 6min, it is sample to be tested to obtain supernatant；(4) detection of sample to be testedIt is detected, obtained above-mentioned using liquid chromatography analysis equipment and the fluorescence detector sample to be measured to above-mentioned steps (f) The Citalopram of sample to be measured and internal standard chromatogram obtain object to be measured object peak in above-mentioned Citalopram and internal standard chromatogram The ratio y of object to be measured object peak area and internal standard compound peak area is substituted into above-mentioned steps (one) by area 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 Citalopram drug concentration in blood to be detected in the sample is thus calculated.
- 2. the method for detection blood citalopram drug concentration as described in claim 1, it is characterised in that：The above-mentioned mark The concentration of contained Citalopram, internal standard working solution concentration refer to institute in internal standard working solution in the quasi- quasi- working solution of working solution concentration index Concentration containing cyheptamide.
- 3. the method for detection blood citalopram drug concentration as claimed in claim 2, it is characterised in that：In step (1) The middle standard working solution using seven kinds of various concentrations.
- 4. the method for detection blood citalopram drug concentration as claimed in claim 3, it is characterised in that：Described seven kinds are not Standard working solution with concentration is respectively that the Citalopram containing 0.1,0.2,0.4,0.8,1.0,1.2,1.5 μ g/mL concentration is molten Liquid.
- 5. the method for detection blood citalopram drug concentration as claimed in claim 4, it is characterised in that：In step (a) It is dissolved to obtain standard reserving solution A and standard reserving solution B with methanol respectively with step (b), with the methanol that water content is 50% The dilution of solution is diluted respectively, configures each standard working solution and internal standard working solution, the extractant described in step (d) It is by 15：The extractant of 85 n-hexane and methyl tertiary butyl ether(MTBE) composition.
- 6. the method for detection blood citalopram drug concentration as claimed in claim 5, it is characterised in that：The efficient liquid Pot strainer used in chromatography is 1/16 0.5M of SSI COL PRE-FILTER WATER.
- 7. the method for detection blood citalopram drug concentration as claimed in claim 5, it is characterised in that：The efficient liquid Chromatographic column used in chromatography is the AccucoreC18 of Thermo companies.
- 8. the method for detection blood citalopram drug concentration as claimed in claim 5, it is characterised in that：The efficient liquid The column temperature of chromatography setting is 35 DEG C.
- 9. the method for detection blood citalopram drug concentration as claimed in claim 5, it is characterised in that：The efficient liquid Chromatography uses mobile phase as the acetonitrile-water containing 0.1% formic acid and 10mM sodium dihydrogen phosphates, and uses gradient elution, Sample size is 30 μ L.
- 10. the method for the detection blood citalopram drug concentration as described in one of claim 6 to 9, it is characterised in that：Institute State the water content that water content is volume ratio.
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