Quantitative analysis method for sulfur hexafluoride in animal biological matrix

Abstract

The invention discloses a quantitative analysis method for sulfur hexafluoride in an animal biological matrix, which comprises the following steps: S1, sample pretreatment: taking a sample and putting the sample into a pre-vacuumized headspace bottle, and adding internal standard gas; s2, headspace balance: after the headspace temperature is 40 DEG C and the balance time is 5 minutes, sucking a sample through an automatic sample injector, and accurately transferring 1mL of sulfur hexafluoride gas to a GC-MS / MS system through quantitative exchange; s3, chromatographic separation: a GS-GASPRO chromatographic column is adopted, the helium flow rate is 1.2 mL / min, and a gradient heating program is adopted; s4, mass spectrometric detection: adopting an SRM mode, and performing SF6 monitoring: ion pair 127-89, CE24; and C3F8 monitoring: ion pairs from 169 to 69, CE8. According to the method, a whole blood and exhaled air synchronous analysis method is created for the first time, and matrix difference is eliminated through an internal standard correction and pressure balance technology.

Description

Technical Field

[0001] The present invention relates to the technical field of analytical chemistry, and specifically to a method for quantitative analysis of sulfur hexafluoride in animal biological matrices. Background Art

[0002] As an inert gas contrast agent, sulfur hexafluoride has important applications in medical imaging. In animal experiments, it is necessary to accurately monitor its metabolic kinetics in whole blood and exhaled breath. However, existing detection methods have problems such as complex pretreatment, insufficient sensitivity, or inability to adapt to different biological matrices (such as whole blood and gas) simultaneously.

[0003] Traditional detection methods (such as single quadrupole GC-MS) have low sensitivity and are easily interfered by the matrix. The extraction efficiency of SF6 in whole blood samples is low, and for exhaled breath samples, due to low concentration and large volume, complex enrichment steps are required. Existing methods have a long analysis time (>15 minutes), a narrow linear range (0.1 - 50 μg / mL), and lack a standardized process for animal experiments. Summary of the Invention

[0004] The purpose of the present invention is to provide a method for quantitative analysis of sulfur hexafluoride in animal biological matrices to solve the problems in the prior art.

[0005] To achieve the above purpose, the present invention provides the following technical solution: A method for quantitative analysis of sulfur hexafluoride in animal biological matrices, comprising the following steps:

[0006] S1. Sample pretreatment: Take the sample in a pre-evacuated headspace vial and add an internal standard gas;

[0007] S2. Headspace equilibration: The headspace temperature is 40°C. After 5 minutes of equilibration, aspirate the sample through an autosampler and accurately transfer 1 mL of sulfur hexafluoride gas to the GC-MS / MS system through a quantitative transfer;

[0008] S3. Chromatographic separation: Use a GS-GASPRO chromatographic column, with a helium flow rate of 1.2 mL / min and a gradient temperature program;

[0009] S4. Mass spectrometry detection: Use the SRM mode. For SF6 monitoring: ion pair 127→89, CE24; for C3F8 monitoring: ion pair 169→69, CE8.

[0010] Preferably, the sample in S1 is a whole blood sample. Take 0.5 mL of whole blood in a pre-evacuated headspace vial and add 100 μL of perfluoropropane internal standard gas.

[0011] Preferably, the sample in S1 is an exhaled breath sample. Take 4.0 mL of exhaled breath in a pre-evacuated headspace vial and add 100 μL of perfluoropropane internal standard gas.

[0012] Preferably, in step S2, the temperature of the quantitative loop during headspace equilibration is 50°C. Combining with the pressure equilibration module of the Tri plus 500 headspace sampler, the repeatability of the 1 mL injection volume is ensured.

[0013] Preferably, in step S3, the split ratio during chromatographic separation is 20:1, and the gradient temperature program is: hold at 45°C for 6 min → increase to 100°C at 50°C / min → hold for 3 min.

[0014] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0015] 1. The first synchronous analysis method for whole blood and exhaled breath, eliminating matrix differences through internal standard calibration and pressure equilibration techniques.

[0016] 2. The single-sample analysis is completed in 10.1 minutes, with the efficiency increased by 30%.

[0017] 3. The SRM mode of the Trace 1300 / TSQ9000 system is adopted to reduce the cross-interference to <0.1%. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The drawings are used to provide a further understanding of the present invention, and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention, but do not constitute a limitation to the present invention. In the drawings:

[0019] Figure 1 is the flow chart of the present invention;

[0020] Figure 2 is the SRM chromatogram of SF6 and C3F8 of the present invention;

[0021] Figure 3 is the standard curve graph of the whole blood sample of the present invention;

[0022] Figure 4 is the standard curve graph of the exhaled breath sample of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0023] To make the objectives, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Apparently, the described embodiments are some, but not all, of the embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art without creative efforts based on the embodiments of the present invention belong to the scope of protection of the present invention. Therefore, the following detailed description of the embodiments of the present invention provided in the drawings is not intended to limit the scope of the claimed invention, but merely represents selected embodiments of the present invention.

[0024] Example 1

[0025] Please refer to Figures 1-3 , in the embodiment of the present invention, a method for quantitative analysis of sulfur hexafluoride in animal biological matrix includes the following steps:

[0026] S1. Sample pretreatment: Using whole blood samples, take 0.5 mL of whole blood into a pre-evacuated headspace vial, and add 100 μL of perfluoropropane (C3F8) internal standard gas;

[0027] S2. Headspace equilibration: The headspace temperature is 40 °C. After 5 minutes of equilibration time, aspirate the sample through an autosampler, and accurately transfer 1 mL of sulfur hexafluoride gas to the GC-MS / MS system through a quantitative loop;

[0028] S3. Chromatographic separation: Using a GS-GASPRO chromatographic column, the helium flow rate is 1.2 mL / min, with a gradient temperature program;

[0029] S4. Mass spectrometry detection: Using the SRM mode, for SF6 monitoring: ion pair 127→89, CE24; for C3F8 monitoring: ion pair 169→69, CE8, to achieve precise separation of characteristic ion pairs.

[0030] In the headspace equilibration in S2, the temperature of the quantitative loop is 50 °C. Combining with the pressure equilibration module of the Tri plus 500 headspace sampler, ensure the repeatability of the 1 mL injection volume.

[0031] In the chromatographic separation in S3, the split ratio is 20:1, and the gradient temperature program is: hold at 45 °C for 6 minutes → rise to 100 °C at 50 °C / min → hold for 3 minutes.

[0032] Analyze according to the above chromatographic-mass spectrometry conditions, and calculate the SF6 concentration by the internal standard method using Chromeleon 7 software.

[0033] Method validation data

[0034] Linear range: 0.121 - 121.400 ng / mL;

[0035] Limit of detection (LOD): 0.121 ng / mL;

[0036] Precision: Intra-day / inter-day RSD are both < 10%; Recovery rate: 92.7 - 111.5%, and the results are shown in the following table.

[0037]

[0038]

[0039]

[0040] Example 2

[0041] Please refer to Figure 1 、 2 Figure 4. In an embodiment of the present invention, a method for quantitative analysis of sulfur hexafluoride in an animal biological matrix includes the following steps:

[0042] S1. Sample pretreatment: Use exhaled breath samples. Take 4.0 mL of exhaled breath into a pre-evacuated headspace vial, and add 100 μL of perfluoropropane (C3F8) internal standard gas to avoid dilution error;

[0043] S2. Headspace equilibration: The headspace temperature is 40 °C. After 5 min of equilibration time, aspirate the sample through an autosampler, and accurately transfer 1 mL of sulfur hexafluoride gas to the GC-MS / MS system through a quantitative loop;

[0044] S3. Chromatographic separation: Use a GS-GASPRO chromatographic column, helium flow rate of 1.2 mL / min, and a gradient temperature program;

[0045] S4. Mass spectrometry detection: Use the SRM mode. For SF6 monitoring: ion pair 127→89, CE 24; for C3F8 monitoring: ion pair 169→69, CE 8, to achieve precise separation of characteristic ion pairs.

[0046] In the headspace equilibration in S2, the temperature of the quantitative loop is 50 °C. Combining with the pressure equilibration module of the Tri plus 500 headspace sampler ensures the repeatability of the 1 mL injection volume.

[0047] In the chromatographic separation in S3, the split ratio is 20:1, and the gradient temperature program is: hold at 45 °C for 6 min → rise to 100 °C at 50 °C / min → hold for 3 min.

[0048] Analyze according to the above chromatographic-mass spectrometry conditions, and calculate the SF6 concentration by the internal standard method using Chromeleon 7 software.

[0049] Method validation data

[0050] Linear range: 0.015 - 30.350 ng / mL;

[0051] Limit of detection (LOD): 0.015 ng / mL;

[0052] Precision: Intra-day / inter-day RSD are both < 10%; Recovery rate: 90.3 - 105.6%, and the results are shown in the following table.

[0053]

[0054]

[0055] Finally, it should be noted that the above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or perform equivalent replacements for some of the technical features. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A method for quantitative analysis of sulfur hexafluoride in animal biological matrices, characterized in that: The following steps are involved: S1. Sample pretreatment: Take the sample into a pre-vacuumed headspace bottle and add internal standard gas; S2. Headspace balance: The headspace temperature is 40°C. After a 5-min balance time, the sample is drawn through an automatic sampler, and 1 mL of sulfur hexafluoride gas is accurately transferred to the GC-MS / MS system through quantitative exchange. S3, chromatographic separation: using GS-GASPRO column, helium flow rate 1.2mL / min, gradient temperature program; S4. Mass spectrometry detection: Using SRM mode, SF6 monitoring: ion pair 127→89, CE24; C3F8 monitoring: ion pair 169→69, CE8.

2. The method for quantitative analysis of sulfur hexafluoride in an animal biological matrix according to claim 1, characterized in that: The sample in S1 is a whole blood sample. 0.5 mL of whole blood is taken into a pre-vacuumed headspace bottle, and 100 μL of perfluoropropane internal standard gas is added.

3. The method for quantitative analysis of sulfur hexafluoride in an animal biological matrix according to claim 1, characterized in that: The sample in S1 is an exhaled breath sample, 4.0 mL of exhaled breath is taken into a pre-vacuumed headspace bottle, and 100 μL of perfluoropropane internal standard gas is added.

4. The method for quantitative analysis of sulfur hexafluoride in an animal biological matrix according to claim 1, characterized in that: The temperature of the quantitative loop in the headspace balance in S2 is 50° C., combined with the pressure balance module of the Tri plus 500 headspace injector, to ensure the repeatability of the 1 mL injection volume.

5. The method for quantitative analysis of sulfur hexafluoride in an animal biological matrix according to claim 1, characterized in that: The split ratio during chromatographic separation in S3 is 20:1, and the gradient temperature program is: 45°C for 6 min → 50°C / min to 100°C → for 3 min.

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