PSEUDO INTERNAL STANDARD METHOD, DEVICE AND APPLICATION FOR MASS SPECTROMETRY QUANTITATIVE ANALYSIS

• US 20180269047A1
• Filed: 08/30/2016
• Published: 09/20/2018
• Est. Priority Date: 09/21/2015
• Status: Active Grant

• Current Assignee: Guangdong Lianjie Biotechnology Co., Ltd.
• Original Assignee: Guangdong Lianjie Biotechnology Co., Ltd.
• Inventors: ZHU, Janshon
• Granted Patent: US 10,643,829 B2
• Time in Patent Office: Days
• CPC Class Codes: G01N 2030/042 Standards
• CPC Class Codes: G01N 2030/8822 involving blood
• CPC Class Codes: G01N 30/06 Preparation
• CPC Class Codes: G01N 30/20 using a sampling valve
• CPC Class Codes: G01N 30/36 in high pressure liquid systems
• CPC Class Codes: G01N 30/72 Mass spectrometers mass spectrometers per se H01J49/00
• CPC Class Codes: G01N 30/7233 interfaced to liquid or supercritical fluid chromatograph interfaces in general for introducing or extracting samples to be analysed with specially adapted mass spectrometer, see H01J49/04
• CPC Class Codes: G01N 30/86 Signal analysis
• CPC Class Codes: G01N 30/8665 for calibrating the measuring apparatus
• CPC Class Codes: G01N 30/88 Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
• CPC Class Codes: H01J 49/0009 Calibration of the apparatus

Abstract

A pseudo internal standard method, device and application for mass spectrometry quantitative analysis is disclosed.

Claims

1-12. -12. (canceled)

13. A method of pseudo internal standard for mass spectrometry quantitative analysis, the method comprising:

weighing a standard substance to prepare standard solutions of different concentrations using a blank substrate of a sample to be tested;

then weighing the same standard substance to prepare internal standard solutions of one to a plurality of concentrations using the blank substrate of the sample to be tested;

wherein an internal standard series have the same concentration range and span as said standard solutions;

processing said standard solutions and said internal standard solutions of the different concentrations to standard samples and internal standard samples via sample processing steps;

injecting said standard samples and said internal standard samples into a chromatography-mass spectrometry system to perform analysis and detection;

plotting a ratio of detected peak areas between said standard sample and said internal standard sample versus concentrations of said standard samples to establish a calibration curve quantified by method of pseudo internal standard; and

processing the unknown sample by the same sample processing steps then following the same steps as standard sample analysis to inject the sample to be detected and the internal standard into the chromatography-mass spectrometry system to perform analysis and detection, the peak area ratio of the sample over the internal standard is then compared with the calibration curve to determine the concentration of the unknown sample.

14. The method of claim 13, further comprising:

pausing an injector for a while after each completion of injection of said standard sample; and

waiting for a time required for a chromatographic peak to be complete;

wherein said internal standard sample goes through a chromatographic column of the chromatography-mass spectrometry system;

wherein said standard sample and said internal standard sample are separated and analyzed in the same detection cycle.

15. The method of claim 13, wherein:

said internal standard sample is injected after each completion of injection of said standard sample; and

said internal standard sample directly enters the mass spectrometer in a manner of flow injection to perform detection bypassing said chromatographic column of said chromatography-mass spectrometry system.

16. The method of claim 15, wherein when said internal standard sample is injected using the flow injection, a flow rate of the flow injection is 20% lower than a flow rate of the mobile phase in the sample analysis, or the flow of spray gas is increased from 60 L/min to 70 to 100 L/min.

17. The method of claim 13, wherein after each completion of injection of said standard sample, an internal standard sample with a concentration closest to a concentration of said standard sample is injected after a chromatographic peak of the standard sample is detected, and the internal standard sample directly enters the mass spectrometer in a manner of flow injection to perform determination bypassing said chromatographic column of said chromatography-mass spectrometry system.

18. A device of pseudo internal standard for mass spectrometry quantitative analysis, the device comprising:

a chromatographic mobile phase and a mobile phase conveying device;

a sample injector connected to the mobile phase conveying device;

a chromatographic column connected to the sample injector;

a mobile phase switching device matched with the chromatographic column;

a mass spectrometer connected to the chromatographic column; and

a software system configured to control the aforementioned hardware to implement a pseudo internal standard mass spectrometry quantitative process;

wherein said sample injector includes an injection needle and an injection valve connected to said injection needle configured to switch between a sample-loading state and a sample-injection state, said injection needle and said injection valve are connected through a sample loop.

19. The device of claim 18, wherein said mobile phase switching device includes two three-way switching valves with one connected to the front and the other connected to the back of said chromatographic column.

20. The device of claim 18, wherein said mobile phase switching device includes a four-way switching valve connected to said chromatographic column.

21. The device of claim 20, further comprising:

an internal standard sample injection device connected to said four-way switching valve;

wherein said internal standard sample injection device includes;

an internal standard sample injector connected to said four-way switching valve;

a second mobile phase conveying device connected to said internal standard sample injector; and

a second mobile phase storage device connected to said second mobile phase conveying device.

22. The device of claim 18, wherein said mobile phase switching device includes a six-way switching valve connected to the chromatographic column.

23. The device of claim 22, wherein said six-way switching valve includes a six-way injection valve in an internal standard sample injector configured to inject said internal standard sample.

24. A method of using the device of claim 18, wherein the device is applied to blood sample analyses.