CN210690497U - Chromatographic system with detector correction - Google Patents

Abstract

The utility model provides a chromatographic system with detector calibration function, include: the device comprises a sample injection flow channel (L1), an analysis flow channel (L2), a waste liquid flow channel (L4), a multi-way valve and a quantitative ring, wherein the quantitative ring is connected to any two ports of the multi-way valve to form a connecting flow channel (L3); the sample injection flow channel (L1), the analysis flow channel (L2) and the waste liquid flow channel (L4) are respectively connected to any one port of the multi-way valve; and the automatic comparison liquid flow channel (L5) is also included, the automatic comparison liquid flow channel (L5) comprises an automatic comparison liquid storage bottle (B) and an automatic comparison pump (P1) connected to the automatic comparison liquid flow channel, and the automatic comparison liquid flow channel is also connected to any one of the rest ports of the multi-way valve. The chromatographic system can well correct the instability of the detection system by introducing the substance which is the same as the target substance as the self-comparison internal standard, and has the advantages of simple operation method, rapid determination, high sensitivity, good accuracy, strong stability and low cost.

Description

Chromatographic system with detector correction

Technical Field

The utility model relates to a chromatogram quantitative analysis specifically belongs to the chromatography instrument field.

Background

Liquid chromatography-mass spectrometry (LS-MS/MS) is also called as liquid chromatography-mass spectrometry technology, and takes liquid chromatography as a separation system and mass spectrometry as a detection system. After a sample is ionized in a mass spectrum ion source, ion fragments are separated according to the mass number through a mass analyzer of a mass spectrum, and a mass spectrogram is obtained through ion induction equipment. The liquid chromatography-mass spectrometry reflects the complementation of the advantages of the chromatogram and the mass spectrum, combines the advantages of high separation capability of the chromatogram on a complex sample and high selectivity, high sensitivity and capability of providing relative molecular weight of MS, and is widely applied to a plurality of fields such as pharmaceutical analysis, food analysis, environmental analysis and the like.

LS-MS/MS suffers from problems of cracking in the source of the compound, inhibition or enhancement of ionization efficiency, formation of ion adducts, etc., which cause the mass spectrometry to be extremely unstable, and in addition, mass number drift and vacuum degree change affect the stability of the mass spectrometry. Common correction means include a conventional internal standard method and a stable isotope internal standard method, but the above methods have the following problems:

1. the conventional internal standard method is difficult to find a compound which is consistent with a target object in structural properties and the like.

2. In the stable isotope internal standard method, the synthesis process of most isotope internal standards is complex, and large-scale industrial synthesis is difficult, so that the purchase price of the stable isotope internal standard method is very expensive, and the stable isotope and a target object have certain difference in physicochemical properties, so that when the state of a mass spectrum changes too much, the peak area change of the stable isotope is inconsistent with the peak area change of the target object, and therefore, the stable isotope is difficult to perform a good correction effect under many conditions.

Therefore, it is urgent and important to develop a technical means or method for maximally correcting the instability of the mass spectrum.

SUMMERY OF THE UTILITY MODEL

In view of the deficiencies of the prior art, it is an object of the present invention to provide a chromatography system with calibration function for correcting instability of a detection system. According to the chromatographic system, a substance which is the same as a target substance is introduced as a self-comparison internal standard, and the substance is introduced into the measurement system by using time difference, so that the instability of the detection system is corrected, the time for searching for a conventional internal standard substance is greatly reduced, the research and development speed and efficiency are improved, the problems of high cost and the like caused by using a stable isotope as the internal standard can be avoided, and the chromatographic system is more accurate and stable. The system is not only suitable for mass spectrometry detectors, but also for the correction of other unstable detectors, such as fluorescence detectors, electrochemical detectors, etc.

The technical scheme of the utility model is that:

chromatographic system with detector correction comprising:

the sample injection flow passage is used for injecting samples;

an analysis flow channel for sample analysis;

a waste liquid channel for discharging waste liquid;

the multi-way valve is used for connecting and switching each flow passage;

the quantitative ring is connected to any two ports of the multi-way valve to form a connecting flow channel;

the sample injection flow channel, the analysis flow channel and the waste liquid flow channel are respectively connected to any one port of the multi-way valve;

the automatic comparison device also comprises a self comparison liquid channel, wherein the self comparison liquid channel comprises a self comparison liquid storage bottle and a self comparison pump connected to the self comparison liquid channel, and the self comparison liquid channel is also connected to any one of the rest ports of the multi-way valve; the self-comparing pump is a pump with a cavity volume less than or equal to 200 microliters.

Preferably, the multi-way valve is a six-way valve, the sample injection channel is communicated with a starting port of the six-way valve, the analysis channel is communicated with a first port of the six-way valve, the waste liquid channel is communicated with a third port of the six-way valve, the connection channel is communicated with a second port and a fifth port of the six-way valve, and the automatic liquid comparison channel is communicated with a fourth port of the six-way valve.

According to the preferable scheme, the refrigerating device is arranged outside the liquid comparing storage bottle, so that the liquid comparing storage bottle can be cooled and treated more stably.

Preferably, the liquid comparing flow passage is further provided with a filter, and the filter is positioned on the downstream flow passage of the liquid comparing pump and can filter impurities in the liquid comparing flow passage.

Preferably, the self-comparing pump comprises an injection pump, a diaphragm pump, a peristaltic pump, a plunger pump and the like, and the volume of the cavity is preferably less than or equal to 200 microliters.

Preferably, the sample injection flow channel is provided with at least one mobile phase storage bottle, a chromatographic pump and a sample injector.

In a preferred scheme, an analysis column is arranged on the analysis flow channel, and the rear end of the analysis flow channel is connected with a detector.

The technical principle is as follows: the principle of the mass spectrum system is that liquid is used as a mobile phase, a high-pressure infusion system is adopted, a mixture carried by the mobile phase such as a single solvent with different polarities or a mixed solvent with different proportions, a buffer solution and the like is pumped into a chromatographic column filled with a stationary phase, and due to the difference of the properties and the structures of components in the mixture and the difference of the acting force generated between the components and the stationary phase, the mixture is repeatedly distributed and balanced between the components and the stationary phase along with the movement of the mobile phase, so that the components are retained by the stationary phase for different time, and then flow out of the stationary phase in sequence according to a certain sequence and enter a detector for detection, and the analysis of a sample is realized. After an automatic mass spectrum correction structure is introduced, the self-comparison liquid is stored in the six-way valve in advance by controlling the starting of the self-comparison pump. After sample injection, the self-comparing liquid is added into the system through switching of the internal structure of the six-way valve after a certain time (generally within 1 min), and finally the self-comparing liquid and a sample to be detected are separated through the same chromatographic conditions and chromatographic columns, and peaks are sequentially generated, so that the effect of correcting and calibrating the target is achieved.

The self-comparison liquid takes a target substance as an internal standard substance, the dissolving solution of the self-comparison liquid is the same as the mobile phase output by the chromatographic pump, and the concentration of the self-comparison liquid is in the linear range of the target substance to be detected. The introduction of the self-calibration solution increases the selectivity of the internal standard, and particularly for some targets without isotopes, isomers and similar structural properties, the self-calibration solution can be well used as the internal standard to play a correction role.

Compared with the prior art, the utility model has the advantages that:

1. according to the chromatographic system, a substance which is the same as a target substance is introduced as a self-comparison internal standard, and the substance is introduced into the measurement system by using time difference, so that the instability of the detection system is corrected, the time for searching for a conventional internal standard substance is greatly reduced, the research and development speed and efficiency are improved, the problems of high cost and the like caused by using a stable isotope as the internal standard can be avoided, and the chromatographic system is more accurate and stable.

2. The automatic correction system avoids a series of problems of inconsistent internal standard addition amount and the like caused by factors such as personnel, measuring instruments and the like in the traditional internal standard addition mode, simplifies the development process of the chromatographic method, is not limited by the types of chromatographic columns and mobile phases any more, ensures the consistency of the internal standard addition, and greatly ensures the correction effect of the internal standard.

3. The utility model discloses a chromatographic system makes the detector obtain maximum correction, has improved the stability of mass spectrum quantitative detection, can let the fine detection project of being applied to high risk height of mass spectrum system go, for example fields such as clinical medication monitoring.

Drawings

FIG. 1 is a schematic diagram of a chromatographic system with detector calibration according to the present invention;

FIG. 2 is a diagram of the initial operating state of the self-comparing module during the operation of the system;

FIG. 3 is a diagram of an operational state of the self-comparing module during operation of the system;

FIG. 4 is another operating state diagram of the self-comparing module during system operation;

FIG. 5 is a chromatogram of cyclosporine and an internal standard, the retention time of the cyclosporine being 2.464min, the retention time of the internal standard being 2.984 (from the internal standard) and 2.919 (cyclosporine A-d4), respectively.

FIG. 6 is a graph of a cyclosporin standard curve.

FIG. 7 is a graph showing the results of analysis of blood cyclosporin concentrations in blood samples;

FIG. 8 is a graph showing the results of another analysis of the plasma concentration of cyclosporin in a blood sample;

wherein: s1, mobile phases I, S2, S3, mobile phases III and DGU, a degasser, a P1 auto-comparison pump, SIL, an auto-sampler, a P2 chromatographic pump, a V1 six-way valve, a C filter, a C2 chromatographic column, a D detector, a W waste liquid bottle, a Loop ring, a quantitative ring, a refrigerating device, a B auto-comparison liquid, an L1 sample injection flow channel, an L2 analysis flow channel, an L3 connecting flow channel, an L4 waste liquid flow channel, an L5 auto-comparison liquid flow channel, a 0 starting port, a 1 first port and a 2; a second port, 3; a third port, 4; a fourth port, 5; a fifth port.

Detailed Description

The detailed structure of the present invention will be further described with reference to the accompanying drawings and the detailed description.

Example 1

As shown in fig. 1: chromatographic system with detector correction comprising: the device comprises a sample injection flow channel L1, an analysis flow channel L2, a waste liquid flow channel L4, a multi-way valve and a quantitative ring, wherein the quantitative ring is connected to any two ports of the multi-way valve to form a connecting flow channel L3; from the comparison flow path L5.

The multi-way valve is a six-way valve, the sample injection flow channel L1 is communicated with a starting port 0 of the six-way valve, the analysis flow channel L2 is communicated with a first port 1 of the six-way valve, the waste liquid flow channel L4 is communicated with a third port 3 of the six-way valve, the connecting flow channel L3 is communicated with a second port 2 and a fifth port 5 of the six-way valve, and the comparing flow channel L5 is communicated with a fourth port 4 of the six-way valve.

The self-comparing liquid flow channel L5 comprises a self-comparing liquid storage bottle B and a self-comparing pump P1 connected to the self-comparing liquid flow channel, wherein the self-comparing pump P1 is a small-volume diaphragm pump, and the cavity volume is 25 microliter.

And a refrigerating device A is arranged outside the self-comparing liquid storage bottle B.

The self-comparing liquid flow passage L5 is also provided with a filter C which is positioned on the downstream flow passage of the self-comparing pump P1.

And three mobile phase storage bottles, a chromatographic pump P2 and a sample injector SIL are arranged on the sample injection flow channel L1.

An analytical column C2 is arranged on the analytical flow channel L2, and a detector D is connected to the rear end of the analytical flow channel L2.

Description of the working process:

step 1) is shown in fig. 2: in an initial state, the chromatographic pump P2 is started, the sample injection channel LI is communicated with the analysis channel L2 through the six-way valve, and the analysis channel is communicated with the detector D.

Step 2) is shown in fig. 3; when the self-comparing module is in a working state, after a certain time (generally within 1min, for example, 0.5min), the self-comparing pump P1 is started, and the self-comparing flow channel L5 is communicated with the fourth port 4 of the six-way valve, is communicated with the connecting flow channel L3 through the fifth port 5, and is communicated with the waste flow channel L4 through the second port 2. At this time, the self-comparative liquid is stored in the quantitative ring connecting the flow passage L3.

Step 3) is shown in fig. 4; and (3) switching the six-way valve, wherein the starting port 2 is communicated with the fifth port 5, the first port 1 is communicated with the second port 2, so that the sample injection channel LI and the connecting channel L3 are communicated with the analysis channel L2, the self-comparison liquid stored in the quantitative ring is brought into the chromatographic column, and the self-comparison liquid and the sample to be detected are separated by the same chromatographic conditions and the chromatographic column to sequentially generate peaks, thereby performing the function of correcting and calibrating the target.

Example 2 concrete application case

According to the experiment, cyclosporine is selected as a test medicine, an internal standard method, an isotope internal standard method and an external standard method are respectively adopted for investigating indexes such as linearity, sensitivity, specificity, accuracy and precision, and the feasibility, reliability and stability of a liquid chromatography/mass spectrometry system with a detector correction function are verified through result comparison of three quantitative modes.

1. Instrument for measuring the position of a moving object

The chromatographic system with detector calibration as described in example 1 was used and the mass spectrometric detection system used Shimadzu LCMS-8050 CL clinical mass spectrometry.

A chromatographic pump: shimadzu LC-30AD CL

Sample injector: shimadzu SIL-30AC MP CL

A controller: shimadzu CBM-20A CL

A detector: shimadzu LCMS-8050 CL

A workstation: shimadzu Lab-Solution

2. Conditions of analysis

A chromatographic column: germany MN Nucleodur CN-RP chromatography column (4.6X 100mm, 5 μm);

mobile phase: the mobile phase was water (containing 2mmol/L formic acid): methanol 35:65 (V/V);

flow rate: the flow rate of the chromatographic pump is 1.0 mL/min-1

Column temperature: 55 deg.C

Sample introduction amount: 1 μ L

Characteristic ion: 1224.90>1224.75 (quantitative)

1224.90>1189.20 (nature)

Internal standard: cyclosporin A-d4(100 ng/ml); cyclosporine (self-comparing liquid 10ng/ml)

The correction function of the system detector is controlled by a time program of a chromatographic workstation, manual intervention is not needed, and high-automation rapid analysis is realized.

The main part conditions in each process are shown in table 1.

TABLE 1 time program

3. Preparation of the solution

3.1 preparation of Standard solution

Accurately weighing 0.95mg of cyclosporine reference substance, dissolving with methanol, and fixing the volume to a proper volume to obtain a cyclosporine mother solution with the concentration of 945.0ug mL < -1 >, and storing in a refrigerator at the temperature of-76 ℃. When in use, the working solution is diluted to the required concentration. Precisely sucking appropriate amount of above mother liquor, and diluting with human serum to obtain low concentration (LQC), medium concentration (MQC) and high concentration (HQC) quality control samples with concentration of 25, 100, and 400 ng.mL < -1 >.

3.2 preparation of internal standard solution

Taking a certain amount of prepared cyclosporine standard solution, taking a mobile phase as a matrix, preparing 50ml of self-comparison solution with the concentration of 10ng/ml, and storing in a refrigerator at the temperature of 24 ℃. Can be directly used when needed.

Accurately weighing 1.0mg of cyclosporine A-d4 reference substance, dissolving with methanol, and metering to a proper volume to obtain a concentration of 40.0 ug/mL^-1The mother liquor of the cyclosporine A-d4 is stored in a refrigerator at the temperature of-76 ℃. When in use, the solution is diluted to 4.0 ug/mL^-1The working fluid of (1).

4. Sample pretreatment method

4.1 external Standard method or self-comparison internal Standard method

Accurately sucking 90% acetonitrile 1000ul to 1.5ml EP tube, accurately adding 400ul serum, vortex and shake for 1min, high speed centrifuging (14500r min-1) for 8 min, and collecting 1000ul supernatant, and testing on computer.

4.2 pretreatment in isotope internal Standard method

400ul of serum is taken to be put into an EP tube of 1.5mL, 10ul of cyclosporine A-d4 internal standard working solution with the concentration of 4.0ug mL < -1 > is added, and the mixture is mixed evenly. Accurately sucking 1000ul of 90% acetonitrile into serum, vortexing and shaking for 1min, centrifuging at high speed (14500r min-1) for 8 min, and collecting 1000ul of supernatant for computer test.

5. Method verification

5.1 chromatographic behavior and specificity Studies

Under the chromatographic conditions, the specificity of the method is examined by measuring the blank blood sample, the standard solution and the blood sample after the drug administration of the cyclosporine and the blood sample after the drug administration of the subject.

5.2 examination of Linear Range and quantitative Limit (LOQ)

The cyclosporine working solution with a series of concentrations is prepared by referring to the relevant guidelines of the national institute of Clinical Laboratory Standardization (CLSI), the cyclosporine working solution is processed and measured according to the method, when the external standard method is used for quantification, the linear regression equation is obtained by drawing a working curve by taking the area of the cyclosporine peak in the sample serum as the ordinate and the concentration as the abscissa, and the linear range of the cyclosporine is inspected. When the internal standard method is used for quantification, a working curve is drawn by taking the ratio of the area of the cyclosporine peak in the sample-added serum to the area of the internal standard peak as a vertical coordinate and the concentration as a horizontal coordinate, an obtained linear regression equation is used for inspecting the linear range of cyclosporine.

5.3 degree of daily precision and daytime precision

And (3) processing and measuring quality control samples with high, medium and low cyclosporine horizontal concentrations, setting 5 groups of parallels for each concentration in each quantitative mode, substituting the measured result into a linear regression equation to calculate the concentration, and comparing the concentration with the concentration theoretically added to obtain the addition recovery rate and the variation coefficient of the method, the inspection accuracy and the precision in days. Samples of the above concentrations were prepared and measured for 3 consecutive days, and the method was examined for daytime precision. And synchronously inspecting the change conditions of the peak areas of the target substance, the internal standard substance and the peak area ratio of the two peak areas of the medium-concentration level quality control sample under three quantitative methods.

5.4 investigation of sample stability

Taking quality control samples with high, medium and low cyclosporine horizontal concentrations, performing repeated freeze thawing for 0, 1, 2, 3 times (-76 ℃), and then determining to investigate the freeze thawing stability of the plasma sample; measuring after the plasma sample is placed for 8 hours at room temperature, and investigating the placing stability of the plasma sample; placing the cyclosporine working curve and the treatment fluid of the quality control samples with three concentrations in an automatic sample injector (10 ℃), repeatedly injecting and analyzing within 12h, and inspecting the storage stability of the sample treatment fluid; the quality control samples with three concentrations are stored under the freezing condition of minus 76 ℃, and are respectively taken out before being placed (0 days) and after being stored for 30 days, 60 days and 90 days in a freezing way, and then are treated and analyzed after being unfrozen, and the freezing storage stability of the plasma sample is examined.

6 results and discussion

6.1 chromatographic behavior and specificity

The experimental result shows that the endogenous substances and other impurities in the blood plasma do not interfere with the separation and determination of the sample. The retention time of cyclosporine was 2.464min, the retention time of the internal standard was 2.984 (from internal standard) and 2.919 (cyclosporine A-d4), respectively, and the chromatogram is shown in FIG. 5.

6.2 Linear Range and quantitative limits

The prepared 9.63, 24.08, 48.16, 96.32, 240.8 and 481.6ng/mL-1 series gradient cyclosporine working solutions are respectively processed and measured according to the method, an external standard method uses the concentration of cyclosporine in the sample serum as a horizontal coordinate and the peak area as a vertical coordinate to draw a working curve, an internal standard method uses the ratio of the peak area of cyclosporine in the sample serum to the peak area of an internal standard as a vertical coordinate and the concentration as a horizontal coordinate to draw a working curve, and the obtained linear regression equation is shown in FIG. 2. The results show that: when the cyclosporine is 9.63-481.6ngmL^-1Within the range, the concentration and peak area (peak area ratio) are in a good linear relationship, as shown in FIG. 6.

6.3 accuracy and precision of the method

And (3) observing the change conditions of the peak areas of the target substance, the internal standard substance and the peak area ratio of the two peak areas of the medium-concentration-level quality control sample under three quantitative methods respectively. The results of examining the recovery rate, the intra-day precision and the inter-day precision of the three concentration level quality control samples in three quantitative manners are shown in tables 2 to 10. The variation coefficients of the peak areas of the target substance, the peak area of the internal standard substance and the peak area ratio of the target substance to the internal standard substance in the test result of the medium-concentration quality control substance under the isotope internal calibration method are respectively 13.59%, 8.43% and 14.90%. The accuracy of the quality control sample of each concentration level is 93.0-102.6%, and the variation coefficient in day and day is less than 16.9%. The variation coefficients of the peak areas of the target substance, the peak areas of the internal standard substance and the peak area ratio of the target substance to the internal standard substance in the test result of the medium-concentration quality control substance under the self-internal standard quantitative method are respectively 16%, 17% and 3%. The accuracy of the quality control sample of each concentration level is 99.2-103.7%, and the variation coefficient in day and day is less than 3.2%. The variation coefficient of the peak area of the target object in the test result of the medium-concentration quality control product under the external standard quantitative method is 25.10%. The accuracy of the quality control sample of each concentration level is 92.6-102.1%, and the variation coefficient in day and day is less than 34.4%.

Through comparison of the three quantitative methods, the coefficient of variation of the peak areas of the target substance and the internal standard substance under the three quantitative methods is larger, but the coefficient of variation of the ratio of the peak area of the target substance to the peak area of the internal standard substance in the quantitative method compared with the internal standard substance is the lowest, the coefficient of variation in the day and the day is the lowest, the correction capability of the analysis method is the strongest, the stability is the highest, and the method meets the methodological requirements of bioequivalence research.

TABLE 2 variation of peak areas and ratios of target and internal standard in isotope internal standard method

Table 3 isotope internal standard quantitation recovery and intraday precision (n ═ 5)

Table 4 isotope internal standard quantitation method daytime precision (n ═ 5)

TABLE 5 variation of peak areas and ratios of target and internal standard in self-comparison internal standard method

Table 6 self-calibration standard quantitation recovery and intraday precision (n ═ 5)

TABLE 7 self-calibration standard quantitation method precision in day (n ═ 5)

TABLE 8 Peak area Change of target in external Standard method

Table 9 external standard quantitative method recovery and in-day precision (n ═ 5)

Table 10 external standard quantitative method daytime precision (n ═ 5)

6.4 sample stability

The experimental results show that: cyclosporine in plasma does not undergo significant changes during 3 freeze-thaw cycles; the difference between the measured value of the quality control sample after 8 hours at room temperature and the measured value of the quality control sample after 0 hour is within +/-6 percent, and the variation coefficients are less than 7 percent, which shows that the cyclosporine in the blood plasma has stability within 8 hours at room temperature; the cyclosporine working curve and the quality control sample in the analysis batch repeatedly analyzed within 12 hours after the treatment solution is placed in the automatic sample injector (10 ℃) meet the requirement of accurate quantification, which indicates that the sample treatment solution has stability within 12 hours of storage of the automatic sample injector; cyclosporine plasma samples were stable within 90 days of storage at-76 ℃ freezing.

6.5 actual sample analysis

The drug concentration of cyclosporine in the plasma of a patient is determined by using a self-comparison internal standard quantification method which is verified by comparison, the drug concentration in the plasma of 30 patients collected after a certain time by using the cyclosporine is determined, and typical chromatograms are shown in figures 7 and 8, wherein the blood concentration of the cyclosporine in a blood sample of figure 7 is 87.5 ng/mL < -1 >, and the concentration of the cyclosporine in a blood sample of figure 8 is 98.5 ng/mL < -1 >. The blood concentration monitoring of the cyclosporine can guide the clinical implementation of individualized administration in time.

7 conclusion

The feasibility, reliability and stability of the self-comparison internal standard method are verified through comparison of three comparison experiments of an external standard method, an isotope internal standard method and the self-comparison internal standard method. A method for measuring cyclosporine in blood plasma by a liquid chromatography/mass spectrometry system with a detector correction function is established. The method can complete the detection of cyclosporine in plasma within 3.5min, adopts (self-comparison) internal standard method for quantification, and has the method quantification limit of 9.63ng/mL, the linear range of 9.63-481.9ng/mL and the correlation coefficient of 0.9994. The result of the specificity investigation shows that the endogenous substances and other impurities in the blood plasma do not cause obvious interference to the analysis, and the continuous sample injection does not have the influence of the interference of subsequent chromatographic peaks. The method has the advantages that the daily precision is 1.1-2.0%, the daytime precision is 1.8-3.2%, the accuracy of quality control samples of each concentration level is 99.2-103.7%, and the requirement of accurate quantification of the drug concentration in blood plasma can be met; the stability experiment result shows that the sample is stable after being placed for 8 hours at room temperature, the concentration of cyclosporine does not have obvious change after 3 times of freeze-thaw cycle, and the cyclosporine does not have obvious change after the treated sample is placed in an automatic sample injector (10 ℃) for 12 hours. The method has the characteristics of simple pretreatment, rapid determination, high sensitivity, good accuracy, strong stability and the like, can meet the requirements of clinical report on timeliness, accuracy, determination range and the like, and can be used as an effective method for monitoring the concentration of a therapeutic drug and determining the cyclosporine sample in clinical pharmaceutical research. The system of the invention has strong drug determination capability, high automation technology and complete reliability technology, thus being simple and convenient to operate and strong in analysis compatibility.

The above description is a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto, and any person skilled in the art can substitute or change the technical solution of the present invention and its concept within the technical scope of the present invention.

Claims (7)

1. Chromatographic system with detector correction comprising:

a sample injection flow channel (L1) for injecting samples;

an analysis flow channel (L2) for sample analysis;

a waste liquid flow path (L4) for discharging waste liquid;

the multi-way valve is used for connecting and switching each flow passage;

a fixed-displacement ring connected to any two ports of the multi-way valve to form a connection flow path (L3);

the sample injection flow channel (L1), the analysis flow channel (L2) and the waste liquid flow channel (L4) are respectively connected to any one port of the multi-way valve;

the method is characterized in that: the automatic comparison liquid flow channel (L5) is further included, the automatic comparison liquid flow channel (L5) comprises an automatic comparison liquid storage bottle (B) and an automatic comparison pump (P1) connected to the automatic comparison liquid flow channel, and the automatic comparison liquid flow channel (L5) is also connected to any one of the rest ports of the multi-way valve; the self-comparing pump (P1) is a pump with the cavity volume less than or equal to 200 microliter.

2. The chromatography system with detector calibration function according to claim 1, wherein the multi-way valve is a six-way valve, the sample inlet channel (L1) is communicated with a start port (0) of the six-way valve, the analysis channel (L2) is communicated with a first port (1) of the six-way valve, the waste liquid channel (L4) is communicated with a third port (3) of the six-way valve, the connection channel (L3) is communicated with a second port (2) and a fifth port (5) of the six-way valve, and the self-comparison liquid channel (L5) is communicated with a fourth port (4) of the six-way valve.

3. The chromatography system with detector calibration function according to claim 1 or 2, wherein a refrigeration device (a) is provided outside the self-comparing liquid storage bottle (B).

4. The chromatography system with detector calibration function according to claim 1 or 2, wherein the self-comparing liquid flow path (L5) is further provided with a filter (C) located on the flow path downstream of the self-comparing pump (P1).

5. Chromatography system with detector calibration function according to claim 1 or 2, wherein the self-comparing pump (P1) comprises a syringe pump, a diaphragm pump, a peristaltic pump, a plunger pump.

6. The chromatography system with detector calibration function according to claim 1 or 2, wherein at least one of a mobile phase storage bottle, a chromatography pump (P2) and a Sample Injector (SIL) is provided in the sample injection channel (L1).

7. The chromatography system with detector calibration function according to claim 1 or 2, wherein the analysis flow channel (L2) is provided with an analysis column (C2), and the rear end of the analysis flow channel (L2) is connected with a detector (D).

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