CN112630333B - Two-dimensional GC-GC/MS system and application thereof in volatile substance detection - Google Patents
Two-dimensional GC-GC/MS system and application thereof in volatile substance detection Download PDFInfo
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- CN112630333B CN112630333B CN202011477597.0A CN202011477597A CN112630333B CN 112630333 B CN112630333 B CN 112630333B CN 202011477597 A CN202011477597 A CN 202011477597A CN 112630333 B CN112630333 B CN 112630333B
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Abstract
The invention discloses a two-dimensional GC-GC/MS system and application thereof in detecting volatile substances. The two-dimensional GC-GC/MS system is set as follows: the first gas chromatography column is connected with valves which are respectively connected with the flow paths of the following 1) and 2), or respectively connected with the flow paths of the following 3) or 4): 1) the back flushing component is arranged between the first damping column and the second damping column so as to ensure that only helium communicated by the back flushing component enters the mass spectrometer when the chromatographic column is replaced; 2) a second gas chromatography column and detector; 3) the back flushing component is arranged between the second gas chromatographic column and the second damping column so as to ensure that only helium communicated with the back flushing component enters the mass spectrometer when the chromatographic column is replaced; 4) a third damping column and a detector. The method can perform targeted separation aiming at the co-outflow peak on the first gas chromatographic column, and can also retain information such as retention time of other chromatographic peaks on the first gas chromatographic column.
Description
Technical Field
The invention relates to a two-dimensional GC-GC/MS system and application thereof in detecting volatile substances, belonging to the technical field of gas chromatography-tandem mass spectrometry.
Background
Volatile substances are important quality indexes of aromatic plants and also important factors influencing the quality of flowers, fruits and wines. The accurate analysis of the aromatic substances is beneficial to the breeding of new varieties of resource plants and the improvement of the quality of related products. For the test of plant volatile components, because the plant volatile components contain a plurality of substances with quite similar structures or the same boiling points, all the components are difficult to be well separated at one time by using one chromatographic column, for example, the corresponding isomers can be separated only by using a chiral chromatographic column.
The main chemical substance qualitative means of the original instrument GC-QqQ/MS is a quadrupole, the mass analyzer only has unit mass resolution, and the mass resolution is insufficient and is easily interfered by m/z similar ions, so that the front-stage chromatographic separation has great influence on the efficiency. At present, the interference of a substrate with a target cannot be further distinguished by obtaining the exact mass number of the target. The original instrument gas chromatography is only provided with one chromatographic column, the replacement of chromatographic columns with different properties can interfere the separation of other substances, the complete sample information is influenced, the repeatability among samples is influenced, and the maintenance cost and time of the instrument are increased by repeatedly replacing the chromatographic columns. Therefore, there is a need to provide a new two-dimensional GC-GC/MS system, which is expected to replace the chromatographic column without vacuum unloading, so as to switch the substances that are not easily separated on the one-dimensional chromatographic column to the chromatographic column with other properties, so that the substances that do not reach the baseline separation are better separated, thereby improving the accuracy of qualitative and quantitative analysis.
Disclosure of Invention
The invention aims to provide a two-dimensional GC-GC/MS system, which can well separate substances with all components which are difficult to be well separated at one time by adopting one chromatographic column, find new substances in a common flow peak, improve the accuracy of qualitative and quantitative determination and provide an accurate and efficient detection method in the detection of volatile substances of aromatic plants, fruits and processed products thereof.
The invention provides a two-dimensional GC-GC/MS system which comprises a first gas chromatographic column, a second gas chromatographic column, a first damping column, a second damping column, a third damping column, a valve, a mass spectrometer, a detector and a back flushing component, wherein the first gas chromatographic column is connected with the second damping column;
the first gas chromatography column is connected with the valves which are respectively connected with the flow paths of the following 1) and 2), or respectively connected with the flow paths of the following 3) or 4):
1) the back flushing assembly is arranged between the first damping column and the second damping column, so that helium introduced by the back flushing assembly enters the mass spectrometer through the second damping column without allowing air to enter the mass spectrometer when the first damping column is replaced by the second gas chromatographic column;
the flow path is connected with the first gas chromatograph for one-dimensional chromatographic analysis;
2) the second gas chromatography column and the detector;
3) the back flushing assembly is arranged between the second gas chromatographic column and the second damping column so as to ensure that helium introduced by the back flushing assembly enters the mass spectrometer through the second damping column without allowing air to enter the mass spectrometer when the second gas chromatographic column is better taken as the first damping column;
the flow path is connected with the first gas chromatograph for performing two-dimensional chromatographic analysis;
4) the third damping column and the detector.
In the two-dimensional GC-GC/MS system, the valve can be a deans switch valve; the detector may be a FID detector;
the first gas chromatographic column is an HP-5MS gas chromatographic column, and the second gas chromatographic column is a DB-WAX gas chromatographic column;
the length of the first damping column is 0.7 m; the length of the second damping column is 2 m; the length of the third damping column is 0.85 m;
the specific lengths of the damping columns at the three different positions can be determined according to the adopted gas chromatographic column.
When the two-dimensional GC-MS system is used for detecting volatile substances in resource plants, a normally open blowback assembly is needed for keeping the pressure of the system.
During detection, the DeansSwitch valve is set to be closed, a sample to be detected enters the mass spectrometer from the sample inlet through the first gas chromatographic column, the first damping column and the second damping column, and chromatographic and mass spectrum data of the sample on the HP-5MS are obtained; selecting a section without baseline separation, and waiting for two-dimensional GC-GC construction;
and under the non-vacuum-unloading state, the second gas chromatographic column is placed at the original position of the first damping column, the third damping column is placed at the original position of the second gas chromatographic column, and the construction of the two-dimensional GC-GC system is completed, wherein the process is about 10-30 minutes due to different proficiency degrees of the instrument. The starting point of the selected area time is set to be closed, and the end point of the selected area time is set to be opened, so that the selected area enters the mass spectrometer through the first gas chromatographic column, the second gas chromatographic column and the second damping column, and the substances in other periods enter the detector through the third damping column after coming out of the first gas chromatographic column.
After a two-dimensional system is constructed, because the pressure of the system changes, the outflow of each component changes compared with that of a one-dimensional system, and the change time is finely adjusted according to an adopted chromatographic column and a damping column and generally changes by 0.1-0.2 min.
The two-dimensional GC-GC/MS system is combined with a back flushing system, the chromatographic column can be replaced under the state of not unloading vacuum, and the operation that vacuum pumping is required for more than 4 hours when vacuum is unloaded once is avoided.
The invention is suitable for the following volatile substances which can be separated and detected by gas chromatography: fragrant plant, flower, essential oil, hydrolat, perfume, cosmetics, fruit juice, wine, etc.
The two-dimensional GC-GC/MS system adopts a center cutting mode, can realize maximum chromatographic separation aiming at the target object, and greatly increases the quantitative reliability of the target object. During a chromatographic run, the effluent from a first gas chromatography column is transferred over a second gas chromatography column having a different stationary phase for a specified period of time, such that the target and co-effluent impurities in the transferred chromatographic peak are completely separated on the second column. By adopting the scheme, the co-flow peak on the first gas chromatographic column can be separated in a targeted manner, and meanwhile, information such as retention time of other chromatographic peaks on the first gas chromatographic column can be retained.
Drawings
FIG. 1 is a schematic flow diagram of a prior art GC/MS system.
FIG. 2 is a schematic flow diagram (one-dimensional) of a GC-GC/MS system constructed in accordance with the present invention.
FIG. 3 is a schematic flow diagram (two-dimensional) of a GC-GC/MS system constructed in accordance with the present invention.
FIG. 4 is a chromatogram for detecting rose aroma using the GC-GC/MS system of the invention.
FIGS. 5 and 6 are chromatograms of detection of peony aroma using the GC-GC/MS system of the present invention.
Detailed Description
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
As shown in fig. 1, which is a schematic view of a flow path system of a conventional GC/MS system, a gas chromatograph is directly connected to a mass spectrometer.
The present invention is improved on the basis of the GC/MS system shown in FIG. 1, and as shown in FIG. 2, an HP-5MS gas chromatographic column is connected with a deansSwitch valve which is respectively connected with the following flow paths 1) and 2):
1) the device comprises a damping column of 0.7m, a damping column of 2m and a Mass Spectrometer (MS), wherein a back flushing component is arranged between the damping column of 0.7m and the damping column of 2m so as to ensure that only helium communicated by the back flushing component enters the mass spectrometer instead of air when the chromatographic column is replaced, and the flow path is connected with an HP-5MS gas chromatographic column for one-dimensional chromatographic analysis;
2) DB-WAX gas chromatography column and FID detector.
To maintain system pressure, a normally open blowback assembly is required.
During detection, a DeansSwitch valve is set to be closed, and a sample enters MS from a sample inlet through a first chromatographic column HP-5MS and damping columns of 0.7m and 2m to obtain chromatographic and mass spectrum data of the sample on the HP-5 MS. The sections without baseline separation were selected and awaited for two-dimensional GC-GC construction.
As shown in FIG. 3, which is a schematic flow diagram of a GC-GC/MS system constructed according to the present invention, the construction process is as follows: under the non-vacuum-unloading state, the DB-WAX gas chromatographic column is placed at the original position of a damping column of 0.7m, the damping column of 0.85m is placed at the original position of the DB-WAX gas chromatographic column, and the construction of the two-dimensional GC-GC system is completed, wherein the process is about 10-30 minutes due to different proficiency levels of instruments. The following flow path connections 3) and 4) are formed:
3) the back flushing component is arranged between the DB-WAX gas chromatographic column and the 2m damping column so as to ensure that only helium communicated with the back flushing component enters the mass spectrometer instead of air when the chromatographic column is replaced, and two-dimensional chromatographic analysis is carried out;
4) a 0.85m damping column and FID detector.
The starting point of the time of the selected region during one-dimensional detection is set as closed, the time end point of the selected region is set as open, so that the selected region enters a mass spectrum through the first chromatographic column HP-5MS, the second chromatographic column DB-WAX and the damping columns of 2m, and substances in other periods enter the damping columns of 0.85m after coming out of the HP-5MS and enter the FID detector.
By adopting the GC-GC/MS system, the co-flow peak on the first gas chromatographic column can be separated in a targeted manner, and information such as retention time of other chromatographic peaks on the first gas chromatographic column can be retained.
Volatile substances in resource plants are detected by using the GC-GC/MS system constructed by the invention.
Detection of rose fragrance substance (liquid sample injection)
5uL of rose essential oil, adding 1mL of n-hexane, and sampling 1uL of the sample.
1. One-dimensional gas phase detection in the system shown in FIG. 2
To maintain the vacuum state of the MS, a normally open blowback assembly is required.
The one-dimensional gas phase detection method comprises the following steps:
3: 1 split-flow sample injection. Keeping the column temperature at 40 ℃ for 0.5 min; then heating to 60 ℃ at a speed of 25 ℃/min, and keeping for 2 min; then the temperature is raised to 250 ℃ at the speed of 6 ℃/min and kept for 10 min. DeansSwitch is set to: and off.
The sample was passed from the inlet through a first chromatography column, HP-5MS, and a 0.7m and 2m damping column into the MS, and chromatographic and mass spectral data on the HP-5MS was obtained for the sample, as shown in the upper panel of FIG. 4. Select the segments without baseline separation: 8.1-8.4 min, 18.9-19.2 min and 24.2-24.4 min, and waiting for the construction of the two-dimensional GC-GC.
2. Two-dimensional gas phase detection in the system shown in FIG. 3
The two-dimensional gas phase detection method comprises the following steps:
3: 1 split-flow sample injection. Keeping the column temperature at 40 ℃ for 0.5 min; then heating to 60 ℃ at a speed of 25 ℃/min, and keeping for 2 min; then the temperature is raised to 250 ℃ at the speed of 6 ℃/min and kept for 15 min. DeansSwitch is set to: 0min is opened; 8.3min off, 8.6min on; 19.1min off, 19.4min on; and 24.4min off and 24.6min on (after a two-dimensional system is constructed, compound outflow is 0.2 second slower than that in one dimension because of the change of system pressure, so that 0.2 second is needed to be added in the two-dimensional switching, and the time is finely adjusted according to the changed chromatographic column and the damping column).
According to the set program, the time starting point of the selected area is set to be closed, and the time end point of the selected area is set to be opened, so that the selected area enters the mass spectrum through the first chromatographic column HP-5MS, the second chromatographic column DB-WAX and the damping column of 2m, and the chromatographic and mass spectrum data of the substance in the selected area on the DB-WAX are obtained, as shown in the middle graph in FIG. 4. Other time periods the material was discharged from the HP-5MS and then passed into a 0.85m damped column into the FID detector, and the resulting chromatographic data is shown in the lower graph of FIG. 4.
As can be seen from FIG. 4, the components that did not reach the baseline separation on the HP-5MS column were transferred to the DB-WAX column to obtain a complete separation while retaining information such as retention times of other chromatographic peaks on the HP-5MS column.
Detection of peony aromatic substance (solid phase micro-extraction sample injection)
Taking 1g of peony petals in a 20mL headspace bottle, balancing for 20min at 40 ℃, inserting an SPME extraction head, extracting for 30min at 40 ℃, and taking out the extraction head for sample injection.
1. One-dimensional gas phase detection in the system shown in FIG. 2
To maintain the vacuum state of the MS, a normally open blowback assembly is required.
The one-dimensional gas phase detection method comprises the following steps:
the sample inlet temperature is 250 ℃, the extraction head is resolved for 3min at the sample inlet, and the sample is injected without shunting. Keeping the column temperature at 55 ℃ for 3 min; then, the temperature is raised to 210 ℃ at a rate of 3 ℃/min and kept for 1 min. Deanswitch settings are: and off.
The sample was passed from the sample inlet through a first chromatography column, HP-5MS, and a 0.7m and 2m damping column into the MS, and chromatographic and mass spectral data of the sample on the HP-5MS was obtained, as shown in the upper panel of FIG. 5. Select the segments without baseline separation: 10.7-11.3 min, 11.3-11.8 min, 22-22.5 min and 47-48 min, and waiting for the construction of the two-dimensional GC-GC.
2. Two-dimensional gas phase detection in the system shown in FIG. 3
1) Two-dimensional gas phase detection method (MDHX01)
The sample inlet temperature is 250 ℃, the extraction head is resolved for 3min at the sample inlet, and the sample is injected without shunting. Keeping the column temperature at 55 ℃ for 3 min; then the temperature is raised to 210 ℃ at the speed of 3 ℃/min and kept for 15 min. DeansSwitch is set to: 0min is opened; closing for 10.9min and opening for 11.5 min; switch off for 22.2min, switch on for 22.7 min; 47.2min off, 48.2min on.
According to the above-mentioned set procedure, the time start point of the selected region is set as closed, and the time end point of the selected region is set as open, so that the selected region will enter into mass spectrum through the first chromatographic column HP-5MS, the second chromatographic column DB-WAX and the damping column of 2m, and obtain the chromatogram and mass spectrum data of the selected region material on DB-WAX, as shown in the middle diagram of FIG. 5. Other time periods the material was discharged from the HP-5MS and then passed into a 0.85m damped column into the FID detector, and the resulting chromatographic data is shown in the lower graph of FIG. 5.
2) Two-dimensional gas phase detection method (MDHXQ2, since two peaks of 10.7-11.3 min and 11.3-11.8 min are too close to each other (as two upper half of figure 6), a second two-dimensional method is selected to be established to cut the peak of 11.3-11.8 min)
The sample inlet temperature is 250 ℃, the extraction head is resolved for 3min at the sample inlet, and the sample is injected without shunting. Keeping the column temperature at 55 ℃ for 3 min; then, the temperature is raised to 210 ℃ at a rate of 3 ℃/min and kept for 6 min. DeansSwitch is set to: 0min is opened; switch off for 11.5min and switch on for 12.0 min. The chromatographic data obtained is shown in the lower part of fig. 6, wherein the upper part is a one-dimensional chromatogram.
As can be seen from FIGS. 5 and 6, the components that did not reach the baseline separation on the HP-5MS column were transferred to the DB-WAX column to obtain a complete separation while retaining information such as retention time of other chromatographic peaks on the HP-5MS column.
Claims (4)
1. A two-dimensional GC-GC/MS system comprises a first gas chromatographic column, a second gas chromatographic column, a first damping column, a second damping column, a third damping column, a valve, a mass spectrometer, a FID detector and a back flushing component;
the valve is a DeansSwitch valve;
the first gas chromatographic column is an HP-5MS gas chromatographic column;
the second gas chromatographic column is a DB-WAX gas chromatographic column;
the length of the first damping column is 0.7 m;
the length of the second damping column is 2 m;
the length of the third damping column is 0.85 m;
the first gas chromatography column is connected with the valves which are respectively connected with the flow paths of the following 1) and 2), or respectively connected with the flow paths of the following 3) or 4):
1) the back flushing assembly is arranged between the first damping column and the second damping column, so that helium introduced by the back flushing assembly enters the mass spectrometer through the second damping column without allowing air to enter the mass spectrometer when the first damping column is replaced by the second gas chromatographic column;
2) the second gas chromatography column and the FID detector;
3) the back flushing component is arranged between the second gas chromatographic column and the second damping column, so that helium introduced by the back flushing component enters the mass spectrometer through the second damping column without allowing air to enter the mass spectrometer when the second gas chromatographic column is replaced by the first damping column;
4) the third damping column and the FID detector.
2. Use of the two-dimensional GC-GC/MS system of claim 1 for volatile substance detection of aromatic plants, fruits and their processed products.
3. A method of detecting a volatile substance in a sample, comprising the steps of:
s1, closing the valve in the two-dimensional GC-MS system of claim 1 to enable the blowback assembly to be in a normally open state; a sample to be detected enters from a sample inlet, sequentially passes through the first gas chromatographic column, the first damping column and the second damping column, then enters the mass spectrometer, so that chromatographic and mass spectrum data of the sample to be detected are obtained, and a section which does not reach baseline separation is determined;
s2, replacing the second gas chromatographic column with the third damping column and replacing the first damping column with the second gas chromatographic column in a non-vacuum unloading state; the sample to be detected enters from the sample inlet, the valve is controlled, and the substance in the section determined in the step S1 sequentially passes through the first gas chromatographic column, the second damping column and the mass spectrometer to obtain the chromatogram and mass spectrum data of the section; materials in other sections sequentially pass through the first gas chromatography column and the third damping column and then enter the FID detector;
wherein the DeansSwitch valve is closed at the time start of the zone where the baseline split is not reached as determined at step S1, and the DeansSwitch valve is opened at the time end of the zone.
4. The method of claim 3, wherein: the sample to be detected is volatile substance which can be separated and detected by gas chromatography, and comprises aromatic plant, flower, essential oil, hydrolat, perfume, cosmetics, fruit juice and wine.
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