CN117890519A - Method for determining purity of fluorine-18F marked PSMA radioactive drug by high performance liquid chromatography - Google Patents
Method for determining purity of fluorine-18F marked PSMA radioactive drug by high performance liquid chromatography Download PDFInfo
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- WROMPOXWARCANT-UHFFFAOYSA-N tfa trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F.OC(=O)C(F)(F)F WROMPOXWARCANT-UHFFFAOYSA-N 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
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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
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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
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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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/30—Control of physical parameters of the fluid carrier of temperature
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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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/32—Control of physical parameters of the fluid carrier of pressure or speed
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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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
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- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
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- 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
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- G01N30/8624—Detection of slopes or peaks; baseline correction
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- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
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- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/30—Control of physical parameters of the fluid carrier of temperature
- G01N2030/3007—Control of physical parameters of the fluid carrier of temperature same temperature for whole column
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- 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
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
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Abstract
The invention relates to the technical field of analysis and detection, in particular to a method for measuring the radiochemical purity of fluorine-18F marked PSMA radioactive drugs by using a high performance liquid chromatography. The mobile phase of the method comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is an aqueous solution of trifluoroacetic acid, the mobile phase B is an acetonitrile solution of the trifluoroacetic acid, and gradient elution is adopted. The detection method provided by the invention can effectively determine the radiochemical purity of the fluorine-18F marked PSMA radioactive drug, the radioactive main component and the radioactive chemical impurity can be effectively separated, the separation degree is more than 1.5, meanwhile, the method has excellent specificity, accuracy and sensitivity, realizes the accurate detection of the radioactive purity of the fluorine-18F marked PSMA radioactive drug, and provides technical reference for establishing reasonable radioactive drug quality standard.
Description
Technical Field
The invention relates to the technical field of analysis and detection, in particular to a method for measuring the radiochemical purity of fluorine-18F marked PSMA radioactive drugs by using a high performance liquid chromatography.
Background
The fluorine-18F labeled PSMA radiopharmaceutical is a fluorine [ sic ] 18 F]A radioactive diagnostic medicine of a nuclide labeled Prostate Specific Membrane Antigen (PSMA) ligand is used for PET imaging of prostate cancer patients of PSMA positive focus, and radiochemical purity is a key quality control item of the radioactive medicine, and the radioactive quantity of a radionuclide in a specific chemical form accounts for the proportion of the total radioactive quantity of the radionuclide, so the radioactive chemical purity is one of effective control indexes of fluorine-18F labeled PSMA radioactive medicine, and the level determines clinical diagnostic effects. The main factor affecting the purity of the radiochemical is the content of the radiochemical impurities, and the sources of the radiochemical impurities are as follows: unreacted complete fluorine-18F ion, intermediate fluorine-18F labeled precursor in synthesis process, incomplete purification of fluorine-18F labeled intermediate, fluorine-18F in synthesis processLabeling other structural impurities, fluorine-18F labeling impurities caused by radiolysis, and the like. The radioactive chemical impurities may affect the distribution of the medicine in the body, interfere with the diagnostic or therapeutic effect, or cause unnecessary radiation damage to other organs of the human body, so that the radiochemical purity of the medicine must be strictly controlled, and the quality, safety and effectiveness of the medicine are ensured.
The existing method for measuring the radiochemical purity is more, the most commonly used method is paper chromatography, thin layer chromatography and high performance liquid chromatography, the paper chromatography and the thin layer chromatography are simple in measurement and are mainly used for measuring few impurity types with larger polarity difference between the impurities and main components, and the defects of low separation efficiency and discrimination capability exist for samples with complex components. The high performance liquid chromatography measures the radiochemical purity by utilizing an ultraviolet-visible light split detector connected in series with a radioactive detector, on one hand, the method can effectively separate impurities with larger or smaller polarity difference from various impurities, has the advantages of high sensitivity, good specificity, high repeatability and the like, and on the other hand, the method connects the ultraviolet detector and the radioactive detector in series, so that the qualitative identification of the radiochemical impurities in the radiochemical purity analysis process of the radiopharmaceuticals is realized.
Disclosure of Invention
The invention provides a method for measuring the purity of the radioactive drug of fluorine-18F marked PSMA by high performance liquid chromatography, which has better specificity, accuracy, stability and reliability and can be used for measuring the purity of the radioactive drug of fluorine-18F marked PSMA and controlling the quality.
Specifically, the invention provides the following technical scheme:
a method for measuring the radiochemical purity of fluorine-18F marked PSMA by high performance liquid chromatography includes that mobile phase A is aqueous solution of trifluoroacetic acid and mobile phase B is acetonitrile solution of trifluoroacetic acid.
Preferably, the concentration of trifluoroacetic acid in the mobile phase A is 0.05-0.20%.
Preferably, the concentration of trifluoroacetic acid in the mobile phase B is 0.05-0.20%.
Preferably, gradient elution is employed, and in a preferred embodiment, based on 100% total volume of the mobile phase,
at 0-6 min, the volume of the mobile phase A is reduced from 88% to 85%, and the volume of the mobile phase B is increased from 12% to 15%;
at the time of 6-13 min, the volume of the mobile phase A is 85%, and the volume of the mobile phase B is 15%;
at 13-16 min, the volume of the mobile phase A is reduced from 85% to 55%, and the volume of the mobile phase B is increased from 15% to 45%;
at 16-21 min, the volume of the mobile phase A is gradually decreased from 55% to 0%, and the volume of the mobile phase B is gradually increased from 45% to 100%;
at 21-24 min, the volume of the mobile phase A is 0%, and the volume of the mobile phase B is 100%;
at 24-26 min, the volume of the mobile phase A is increased from 0% to 88%, and the volume of the mobile phase B is decreased from 100% to 12%;
at 26-31 min, the volume of the mobile phase A is 88%, and the volume of the mobile phase B is 12%.
In another preferred embodiment, the total volume of the mobile phase is 100%,
at 0-15 min, the volume of the mobile phase A is 85%, and the volume of the mobile phase B is 15%;
at 15-20 min, the volume of the mobile phase A is reduced from 85% to 80%, and the volume of the mobile phase B is increased from 15% to 20%;
at 20-30 min, the volume of the mobile phase A is reduced from 80% to 30%, and the volume of the mobile phase B is increased from 20% to 70%;
at 30-35 min, the volume of the mobile phase A is 30%, and the volume of the mobile phase B is 70%;
at 35-50 min, the volume of the mobile phase A is gradually decreased from 30% to 0%, and the volume of the mobile phase B is gradually increased from 70% to 100%;
at 50-52 min, the volume of the mobile phase A is 0%, and the volume of the mobile phase B is 100%;
at 52-55 min, the volume of the mobile phase A is increased from 0% to 85%, and the volume of the mobile phase B is decreased from 100% to 15%;
at 55-65 min, the volume of the mobile phase A is 85%, and the volume of the mobile phase B is 15%.
The high performance liquid chromatography is a common analysis and detection method in the field, has better accuracy and stability, and therefore, the invention provides a method for detecting the radioactive purity of the fluorine-18F marked PSMA radioactive drug by adopting the high performance liquid chromatography.
When high performance liquid chromatography is adopted for detection, a common chromatographic column is a C18 reversed phase chromatographic column, a common mobile phase is an organic solvent such as methanol, acetonitrile, tetrahydrofuran, trifluoroacetic acid (TFA) and the like, and a common elution mode is isocratic elution. However, since many radioactive impurities with similar polarities may exist in the fluorine-18F-labeled PSMA radiopharmaceuticals, it is difficult to separate the radioactive main component from the radioactive impurities with similar polarities under the conventional analysis conditions of the above-mentioned high performance liquid chromatography.
Through a large number of experiments, the invention discovers that the fluorine-18F marked PSMA radioactive drug has better solubility in acetonitrile, so that the solvent adopted in the analysis of a final preparation is acetonitrile, and further discovers that the peak type can be obviously improved by adding a proper amount of trifluoroacetic acid into a mobile phase. Meanwhile, in order to separate components with similar polarities, a specific gradient elution mode is adopted.
When an aqueous solution of trifluoroacetic acid is selected, and an acetonitrile solution of trifluoroacetic acid is used as the mobile phase a and the mobile phase B, respectively, the radioactive main component and other radioactive impurities can be separated according to the gradient elution procedure, and the separation degree is more than 1.5. Compared with other existing detection methods, the method provided by the invention can accurately determine the radiochemical purity of the fluorine-18F marked PSMA radioactive drug.
In order to further improve the accuracy and stability of the detection result, other conditions of the high performance liquid chromatography are optimized, and the following optimization scheme is obtained:
the chromatographic column adopts octadecyl bonded silica gel as filler; further, the size of the column is preferably 4.6 mm. Times.250 mm, 5. Mu.m, more preferably LiChroCART C18,4.6 mm. Times.250 mm, 5. Mu.m. The chromatographic column can separate neutral, acidic and weak basic compounds and is well suited for use in the separation system of the present invention.
The flow rate of the mobile phase is 0.8-1.2 mL/min. Studies have shown that controlling the flow rate within this range facilitates retention of the radioactive main component of the fluorine-18F labeled PSMA radiopharmaceutical within a suitable range.
The column temperature is 20-30 ℃.
The detection wavelength is 220-254 nm; studies have shown that setting the detection wavelength in the above range achieves the best absorption intensity for the control solution of fluorine-18F labeled PSMA radiopharmaceutical.
As one of the specific embodiments of the present invention, the method for determining the radiochemical purity of fluorine-18F labeled PSMA radiopharmaceuticals comprises the steps of:
a) Preparing blank solution
Taking sodium chloride injection and ethanol according to the volume ratio of 9:1 to prepare a blank solution; or taking the water solution as a blank solution;
b) Preparing reference substance solution
Taking a proper amount of a reference substance of the fluorine-18F marked PSMA radioactive drug, and adding water to dissolve the reference substance to prepare a reference substance solution with a certain concentration;
c) Preparing test solution
Taking fluorine-18F labeled PSMA radioactive drug, and diluting to a proper concentration by using a blank solution;
d) Measurement method
Injecting the blank solution, the reference solution and the sample solution into a chromatograph respectively, and recording a chromatogram; and calculating the radiochemical purity of the sample by an internal standard method according to the peak area ratio in the chromatogram of the sample.
The beneficial effects of the invention are as follows:
the detection method provided by the invention can effectively separate the main component and the radioactive impurity in the fluorine-18F marked PSMA radioactive drug at the same time, and has high analysis speed and separation degree of more than 1.5. The method is verified in terms of systematic applicability, specificity, linearity, range and durability according to the method verification guidelines of the analysis method 9101 of the edition 2020 of Chinese pharmacopoeia, and the method is proved to have good accuracy, stability and reliability and can be used for the determination and control of the amplification purity of the fluorine-18F marked PSMA radiopharmaceuticals. The establishment of the analysis method realizes accurate detection of the radioactive purity of the fluorine-18F marked PSMA radioactive drug, and plays an important guiding role in the development and optimization of prescription technology.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following description will briefly explain the drawings needed in the embodiments or the prior art, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings can be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a system applicability solution chromatogram (ultraviolet signal) of test example 1.
FIG. 2 is a blank solution chromatogram (radioactive signal) of test example 2.
FIG. 3 is a blank solution chromatogram (UV signal) of test example 2.
FIG. 4 is a chromatogram (UV signal) of the control solution of test example 2.
FIG. 5 shows a chromatogram (radioactive signal) of a sample solution of test example 2.
FIG. 6 shows the structural formula of the R, S-isomer of test example 2.
FIG. 7 shows the structural formula of the S, R-isomer of test example 2.
FIG. 8 is a deprotection solution (radioactive signal) of the fluorine-18F labeled precursor of test example 2.
FIG. 9 is a linear and range chromatogram (radioactive signal) of the test sample solution of test example 3.
Detailed Description
The present invention is described in further detail below with reference to specific examples, but is not intended to limit the scope of the present invention.
Example 1
The present example provides a method for determining the radiochemical purity of fluorine-18F-labeled PSMA radiopharmaceuticals using high performance liquid chromatography, comprising the steps of:
a) Preparing blank solution
Taking sodium chloride injection and ethanol according to the volume ratio of 9:1 to prepare a blank solution
B) Preparing reference substance solution
Taking a proper amount of a reference substance of the fluorine-18F marked PSMA radioactive drug, and adding water to dissolve the reference substance to prepare a reference substance solution of 300 mug/ml;
c) Preparing test solution
Taking fluorine-18F labeled PSMA radiopharmaceuticals, and diluting to about 1.0mCi/ml by using a blank solution;
d) Measurement method
Injecting the blank solution, the reference solution and the sample solution into a chromatograph respectively, and recording a chromatogram; and calculating the radiochemical purity of the sample by an internal standard method according to the peak area ratio in the chromatogram of the sample.
Wherein:
chromatographic column: octadecyl bonded silica gel is used as filler; the column had a size of LiChroCART C18 (Merck), 4.6 mm. Times.250 mm, I.D,5 μm.
Mobile phase a:0.05% tfa in water;
mobile phase B: acetonitrile solution of 0.05% tfa;
column temperature: 20-30 ℃;
flow rate: 0.8-1.2 mL/min;
wavelength: 220-254 nm;
gradient elution procedure is as follows table 1:
TABLE 1
| Time/min | Mobile phase a/％ | Mobile phase B/% |
| 0 | 88 | 12 |
| 6 | 85 | 15 |
| 13 | 85 | 15 |
| 16 | 55 | 45 |
| 21 | 0 | 100 |
| 24 | 0 | 100 |
| 26 | 88 | 12 |
| 31 | 88 | 12 |
Instrument and reagent:
UltiMate3000 high performance liquid chromatograph, siemens; LB514 radioactive flow detector, berton; XPR205/A electronic balance, metrehler company; HPLC chromatographic column, liChroCART C18, 4.6mm.times.250 mm,5 μm.
A fluorine-18F labeled PSMA radiopharmaceutical control; acetonitrile (chromatographic purity, national drug, lot number: 20210804); trifluoroacetic acid (Allatin, lot I1902130); the ultrapure water is self-made by a laboratory ultrapure water machine.
Effect verification
The method of the embodiment is verified in a systematic applicability, linearity, range and durability method according to the method verification guidelines of the analysis method of 9101 of the edition 2020 of Chinese pharmacopoeia, which is specifically as follows:
the specific configuration process of each solution in example 1 and the experimental verification process of the method of the present invention are shown in experimental examples 1 to 4.
Experimental example 1 System applicability test
System applicability solution: and (3) taking a proper amount of a reference substance, and adding water to dissolve the reference substance to prepare a system applicability solution with the concentration of 300 mug/ml.
And (3) measuring: the chromatographic column uses LiChroCART C18,4.6mm ×250mm,5 μm, detection wavelength 220nm, flow rate 1.0mL/min, column temperature 25 ℃, flow mobile phase A:0.05% tfa in water; mobile phase B: acetonitrile solution of 0.05% tfa, gradient elution was used.
Gradient elution is as shown in table 2:
TABLE 2
| Time/min | Mobile phase a/% | Mobile phase B/% |
| 0 | 88 | 12 |
| 6 | 85 | 15 |
| 13 | 85 | 15 |
| 16 | 55 | 45 |
| 21 | 0 | 100 |
| 24 | 0 | 100 |
| 26 | 88 | 12 |
| 31 | 88 | 12 |
20 μl of the system applicability solution was injected into a high performance liquid chromatograph, and the chromatogram was recorded. The detection results are shown in Table 3, and the chromatograms are shown in FIG. 1.
Table 3 System applicability-control determination results
| Sequence number | Theoretical plate number (n) | Tailing factor |
| 1 | 16641 | 1.16 |
Conclusion: the PSMA peak n marked by fluorine-19F in the system applicability solution is 16641, and the requirement of more than or equal to 3500 is met; t is 1.16, and meets the requirement of not more than 2.0. The high performance liquid chromatography system provided by the invention has good applicability.
Experimental example 2 specificity test
Blank solution: taking sodium chloride injection and ethanol according to the volume ratio of 9:1 to prepare a blank solution;
control solution: dissolving a proper amount of reference substance in water to obtain a reference substance solution with the concentration of 300 mug/ml;
r, S-isomer solution of fluorine-19F labeled PSMA: dissolving a proper amount of reference substance in water at concentration of 300 μg/ml, with specific structure shown in figure 6;
fluorine-19F labeled PSMA S, R-isomer solution: dissolving a proper amount of reference substance in water at concentration of 300 μg/ml, and specifically shown in figure 7;
fluorine-18F labeled precursor deprotection solution (refers to the sample solution before purification, containing other radioactive impurities): the fluorine-18F labeled precursor deprotection solution was collected during the process and tested.
Test solution: and (5) taking fluorine-18F marked PSMA radioactive drug to obtain the product.
And after the baseline of the high performance liquid chromatograph is balanced, taking a blank solution, an R, S-isomer solution, an S, R-isomer solution, a reference substance solution, a fluorine-18F marked precursor deprotection solution and a sample solution according to specified chromatographic conditions, injecting 1 needle for each sample, and preserving a chromatogram. The results are shown in Table 4, and the chromatograms are shown in FIGS. 2-5 and 8.
Table 4 specificity
| Sequence number | Sample name | Has interference to main peak of sample solution |
| 1 | Blank solution | Without any means for |
| 3 | Reference substance solution | - |
| 5 | R, S-isomer solutions | Without any means for |
| 6 | S, R-isomer solutions | Without any means for |
| 7 | Deprotection solution for fluorine-18F labeled precursor | Without any means for |
| 8 | Test solution | - |
Conclusion: the retention time of the main peak of the test solution should be consistent with that of the main peak of the control solution. At the peak retention time of the main component of the fluorine-18F marked PSMA radioactive drug, the blank solution, the R, S-isomer solution and the S, R-isomer solution have no interference, other radioactive chemical impurities generated in the process have no interference on the determination of the radioactive main peak, and the separation degree is more than 1.5.
Experimental example 3 Linear and Range test
Test solution: the lower linear limit solution concentration is the quantitative limit level, and the upper linear limit is the radioactivity concentration under the condition of the proper parameters of the radioactivity detector. And in the range of the interval, diluting the test sample by using a sodium chloride injection-ethanol (9:1) mixed solution to obtain at least 5 concentration points, and drawing a curve by taking the main peak radioactive concentration as an abscissa and the main peak area as an ordinate.
And (3) taking 20 mu l of each of the sample solutions with different concentrations, injecting the sample solutions into a high performance liquid chromatograph, recording a chromatogram, taking the radioactive concentration of the sample solution as an abscissa and the peak area as an ordinate, and performing a linear regression equation. The results are shown in Table 5, and the linear and range chromatograms of the test solutions are shown in FIG. 9.
TABLE 5 linearity and Range
| Composition of the components | Standard curve | Linearity and range | r |
| 18F-PSMA | y＝85.614x+210.96 | 0.32～172.86MBq/mL | 0.9996 |
Conclusion: the linear range of radiochemical purity measurement is 0.32 MBq/mL-172.86 MBq/mL, the linear regression equation in the range is y=85.61x+210.90, the correlation coefficient r is 0.9996, and the requirement that r is more than or equal to 0.990 is met.
Experimental example 4 durability
Durability: after the baseline of the high performance liquid chromatograph is balanced, taking a control solution for 1 needle according to different chromatographic conditions (changing flow rate, column temperature and chromatographic columns of different batches), taking a sample for 1 needle of the test solution, recording a chromatogram, recording retention time of ultraviolet main peaks of the control solution and radioactive main peaks of the test solution, wherein RSD% of the ratio of the radioactive main peaks to the retention time of the control solution peaks in the test solution is less than or equal to 2%, radiochemical purity RSD% is less than or equal to 2%, and recording the chromatogram. The results are shown in Table 6.
Table 6 durability results
Conclusion: when the column temperature, flow rate, and single variable in the chromatographic column are slightly varied in the method, the ratio of the retention time of the main radioactive peak fluorine-18F-labeled PSMA in the test solution to the retention time of the fluorine-19F-labeled PSMA in the control solution (t 2 /t 1 ) The RSD% of the method is 1.38%, the radiochemical purity RSD% is 0.12%, and the requirement of less than or equal to 2% is met, so that the method is good in durability.
Comparative example 1
The test provides a detection method which is different from the embodiment in that the method is a thin layer chromatography, the stationary phase is a silica gel G plate, and the developing agent is acetonitrile solution with a certain proportion. The results show that the major radioactive component of the fluorine-18F labeled PSMA radiopharmaceutical is poorly separated from the nearly polar radioactive impurities.
Comparative example 2
Comparative example 2 differs from example 1 only in that: gradient elution is not adopted, and the whole process adopts the volume ratio of 88: the mixed solution of mobile phase a and mobile phase B of 12 was eluted as a mobile phase. As a result, the separation degree of the adjacent chromatographic peaks is less than 1.5, and the main component and the adjacent impurity peaks cannot be effectively separated.
While the invention has been described in detail in the foregoing general description and with reference to specific embodiments thereof, it will be apparent to one skilled in the art that modifications and improvements can be made thereto. Accordingly, such modifications or improvements may be made without departing from the spirit of the invention and are intended to be within the scope of the invention as claimed.
Claims (10)
1. A method for determining the radiochemical purity of fluorine-18F marked PSMA radiopharmaceuticals by high performance liquid chromatography is characterized in that a mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is an aqueous solution of trifluoroacetic acid, and the mobile phase B is an acetonitrile solution of trifluoroacetic acid.
2. The process according to claim 1, wherein the concentration of trifluoroacetic acid in mobile phase a is between 0.05% and 0.20%.
3. The process according to claim 1, wherein the concentration of trifluoroacetic acid in mobile phase B is between 0.05% and 0.20%.
4. A process according to claim 1 to 3, wherein a gradient elution is used, based on 100% of the total volume of the mobile phase,
at 0-6 min, the volume of the mobile phase A is reduced from 88% to 85%, and the volume of the mobile phase B is increased from 12% to 15%;
at the time of 6-13 min, the volume of the mobile phase A is 85%, and the volume of the mobile phase B is 15%;
at 13-16 min, the volume of the mobile phase A is reduced from 85% to 55%, and the volume of the mobile phase B is increased from 15% to 45%;
at 16-21 min, the volume of the mobile phase A is gradually decreased from 55% to 0%, and the volume of the mobile phase B is gradually increased from 45% to 100%;
at 21-24 min, the volume of the mobile phase A is 0%, and the volume of the mobile phase B is 100%;
at 24-26 min, the volume of the mobile phase A is increased from 0% to 88%, and the volume of the mobile phase B is decreased from 100% to 12%;
at 26-31 min, the volume of the mobile phase A is 88%, and the volume of the mobile phase B is 12%.
5. A process according to claim 1 to 3, wherein a gradient elution is used, based on 100% of the total volume of the mobile phase,
at 0-15 min, the volume of the mobile phase A is 85%, and the volume of the mobile phase B is 15%;
at 15-20 min, the volume of the mobile phase A is reduced from 85% to 80%, and the volume of the mobile phase B is increased from 15% to 20%;
at 20-30 min, the volume of the mobile phase A is reduced from 80% to 30%, and the volume of the mobile phase B is increased from 20% to 70%;
at 30-35 min, the volume of the mobile phase A is 30%, and the volume of the mobile phase B is 70%;
at 35-50 min, the volume of the mobile phase A is gradually decreased from 30% to 0%, and the volume of the mobile phase B is gradually increased from 70% to 100%;
at 50-52 min, the volume of the mobile phase A is 0%, and the volume of the mobile phase B is 100%;
at 52-55 min, the volume of the mobile phase A is increased from 0% to 85%, and the volume of the mobile phase B is decreased from 100% to 15%;
at 55-65 min, the volume of the mobile phase A is 85%, and the volume of the mobile phase B is 15%.
6. A method according to any one of claims 1-3, wherein the chromatographic column employs octadecylsilane-bonded silica as a packing agent.
7. The method of claim 6, wherein the chromatographic column has a specification of 4.6mm x 250mm,5 μm.
8. A method according to any one of claims 1-3, characterized in that the flow rate of the mobile phase is 0.8-1.2 mL/min.
9. A process according to any one of claims 1 to 3, wherein the column temperature is 20 ℃ to 30 ℃.
10. A method according to any one of claims 1-3, wherein the detection wavelength is 220-254 nm.
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