WO2010090257A1

WO2010090257A1 - Solvent supply apparatus for liquid chromatography mass spectrometer, reagent bottle, solvent and method for supplying solvent

Info

Publication number: WO2010090257A1
Application number: PCT/JP2010/051609
Authority: WO
Inventors: 徹夏目; 洋中山; 輝康新井
Original assignee: 独立行政法人産業技術総合研究所; 独立行政法人理化学研究所; 日京テクノス株式会社
Priority date: 2009-02-05
Filing date: 2010-02-04
Publication date: 2010-08-12
Also published as: JP2010181271A; JP5526346B2

Abstract

Disclosed is a technique whereby, in analyzing proteins and the like by using a liquid chromatography mass spectrometer (LC-MS), a lowering in analysis sensitivity, which is caused by a chemical noise or a lowering in ionization efficiency due to a trace substance contained in a solvent or a trace substance present in the laboratory environment, can be prevented. A solvent supply apparatus (1) for an LC-MS which comprises, for a reagent bottle (2) in which a solvent to be used as a mobile phase in the LC-MS is stored and the opening of which is sealed with a metal foil, an aspiration nozzle (13) which is penetrated through the metal foil and inserted into the reagent bottle so as to suck up the solvent; a vacuum cap (14) which is in contact with the opening of the reagent bottle (2) and cover the same and in which a through hole for penetrating the aspiration nozzle (13) to be inserted into the reagent bottle (2) is formed; and a vacuum pump which is connected to the vacuum cap (14) and sucks up air in the space formed between the vacuum cap (14) covering the opening of the reagent bottle (2) and the metal foil.

Description

Solvent supply device, reagent bottle and solvent for liquid chromatograph mass spectrometer, and solvent supply method

The present invention relates to a solvent supply device, a reagent bottle and a solvent, and a solvent supply method for a liquid chromatograph mass spectrometer. More specifically, the present invention relates to a solvent supply device for a liquid chromatograph mass spectrometer that can prevent a decrease in analysis sensitivity due to chemical noise.

In the fields of biotechnology, medicine, drug discovery, food, etc., proteome analysis, peptome analysis, glycome analysis, metabolome analysis that comprehensively analyzes proteins, peptides, sugar chains, various metabolites, etc. are now being performed . For these analyses, a liquid chromatograph mass spectrometer (referred to as “LC-MS”) that combines a liquid chromatograph (hereinafter referred to as “LC”) and a mass spectrometer (referred to as “MS”) is widely used. As the sample volume becomes smaller and the analysis throughput becomes higher, LC-MS with higher sensitivity has been demanded.

In LC-MS, first, detection target substances such as proteins, peptides, sugar chains, and various metabolites contained in a sample solution are separated by LC. The separated detection target substance is ionized and then introduced into the MS, and the ion mass, dissociated ion mass, and ion intensity are measured. Based on the obtained mass spectrum (mass spectrum), the detection target substance is identified and the structure is determined.

At this time, if a substance other than the detection target substance (hereinafter referred to as “foreign substance”) is introduced in the process until the detection target substance is introduced into the MS, S / It is conceivable that the analysis sensitivity of the LC-MS decreases due to a decrease in N (signal noise ratio) or a decrease in the ionization efficiency of the detection target substance. Such a decrease in analysis sensitivity due to contamination by foreign substances can be a problem particularly when analyzing an extremely small amount of a detection target substance.

In relation to the present invention, Patent Document 1 discloses, as a technique for reducing chemical noise due to contamination by foreign substances, “mixed metal ions present as a background in the eluent in the analysis of metal ions by ion chromatography. An ion chromatograph eluent characterized by having a small amount of contaminating ions to enable analysis of trace ions that are difficult to measure due to background interference by removing the " .

Japanese Patent Laid-Open No. 7-5159

As a foreign substance that causes a decrease in the analytical sensitivity of LC-MS, for example, a trace substance mixed in a solvent used as a mobile phase of LC-MS, a trace substance in a laboratory environment, and the like can be considered.

As the trace substances mixed in the solvent, first, foreign substances mixed in the solvent manufacturing process are assumed. This includes aerosol particles in the environment where the solvent production apparatus is placed, and dust generated by wear of the production apparatus. In addition, as the trace substance mixed in the solvent, a substance that comes from the reagent bottle and is mixed into the solvent after the solvent is accommodated in the reagent bottle can be considered. For example, when a plastic container is used for the reagent bottle, a plasticizer or unpolymerized monomer contained in the plastic may be eluted and mixed in the solvent. Further, even when a glass container is used for the reagent bottle, since plastic containers are often used for the lid of the reagent bottle, elution of the plasticizer and unpolymerized monomer may occur.

In addition, contamination of trace substances in the laboratory environment may be caused by aerosol particles in the air or dust adhering to the reagent bottle inside the reagent bottle when the reagent bottle is replaced when the solvent is supplied to the LC-MS. This may be caused by mixing or adhering to the LC-MS nozzle. In order to prevent the entry of trace substances in the laboratory environment, it is conceivable to perform the analysis with the LC-MS placed in a clean environment such as a clean room. It is necessary to provide a proper facility.

Therefore, the present invention prevents a decrease in analysis sensitivity caused by a chemical noise or a decrease in ionization efficiency due to a trace substance mixed in a solvent or a trace substance in a laboratory environment in analysis of a protein or the like using LC-MS. The main purpose is to provide technology that can be used.

In order to solve the above problems, the present invention contains a solvent used as a mobile phase of a liquid chromatograph mass spectrometer, and is inserted through the metal foil into a reagent bottle whose mouth is sealed with the metal foil. A suction nozzle that sucks the solvent, a vacuum cap that contacts and covers the mouth of the reagent bottle, and has a hole through which the suction nozzle inserted into the reagent bottle is inserted, and is connected to this vacuum cap And a vacuum pump for sucking air in a space formed between the vacuum cap covering the mouth of the reagent bottle and the metal foil, and a solvent supply device for a liquid chromatograph mass spectrometer. According to this solvent supply device, the substance that generates dust from the metal foil when the suction nozzle penetrates the metal foil is sucked by the vacuum pump to prevent the substance from being mixed into the solvent. Can do.
In this solvent supply apparatus, it is preferable that the suction nozzle is made of a corrosion-resistant metal. Thereby, it can prevent that a metal ion elutes in a solvent from a suction nozzle.
In this solvent supply device, the suction nozzle and the vacuum cap are supplied with an air supply port for supplying outside air, and a filter unit provided in a flow path of outside air supplied from the air supply port. It is desirable to dispose in an enclosure provided with an exhaust port for exhausting air and an openable / closable door. Thereby, it is possible to prevent aerosol particles and dust from being mixed inside the reagent bottle or adhering to the suction nozzle during the reagent bottle replacement operation when the solvent is replenished.
The solvent supply device moves the suction nozzle and the vacuum cap with respect to the reagent bottle arranged at a predetermined position in the casing so that the vacuum cap comes into contact with the mouth of the reagent bottle, and the suction nozzle is A driving means is provided that is inserted into a reagent bottle through a metal foil that seals the mouth.
The solvent supply apparatus may further include a first detection unit that detects a reagent bottle disposed at a predetermined position in the housing. In this case, the vacuum pump can be configured to start or stop suction based on a detection signal from the first detection means. Furthermore, the solvent supply apparatus may include a second detection unit that detects opening and closing of the door body. In this case, the drive means is configured to control the movement of the suction nozzle and the vacuum cap based on the detection signal from the second detection means and the detection signal from the first detection means. Can do.
In addition, the present invention provides a liquid chromatograph mass spectrometer equipped with this solvent supply device.

Next, the present invention relates to a reagent bottle for containing a solvent used as a mobile phase of a liquid chromatograph mass spectrometer, which comprises a metal foil that seals the mouth portion, and the mouth portion that is covered from the outside of the metal foil. There is provided a glass reagent bottle having a lid, and a lid having a hole inserted through a suction nozzle that is inserted through a metal foil and sucks a solvent. This reagent bottle is preferably used together with the solvent supply apparatus according to the present invention.
In this reagent bottle, the metal foil is preferably formed of a corrosion-resistant metal. Thereby, it can prevent that a metal ion elutes in a solvent from metal foil.

Furthermore, the present invention also provides a solvent used as a mobile phase of a liquid chromatograph mass spectrometer, which is prepared in a clean environment capable of preventing contamination of substances that cause chemical noise. This solvent is housed in the reagent bottle according to the present invention and is preferably used together with the solvent supply apparatus according to the present invention.

Finally, according to the present invention, a solvent used as a mobile phase contained in a reagent bottle whose mouth is sealed with a metal foil is sucked with a suction nozzle inserted into the reagent bottle through the metal foil, and subjected to liquid chromatography. Provided is a solvent supply method for a liquid chromatograph mass spectrometer, which includes a step of feeding the liquid to a graph mass spectrometer.
The solvent supply method further includes a step of sucking a substance that generates dust from the metal foil with a vacuum pump when the suction nozzle penetrates the metal foil, and preventing the substance from being mixed into the solvent; And a step of preparing the solvent in a clean environment capable of preventing mixing of substances causing chemical noise and storing the solvent in the reagent bottle made of glass.

According to the present invention, in analysis of proteins and the like using LC-MS, it is possible to prevent a decrease in analysis sensitivity due to a chemical substance noise or a decrease in ionization efficiency due to a trace substance mixed in a solvent or a trace substance in a laboratory environment. Possible technology is provided.

It is a schematic diagram explaining operation | movement of the solvent supply apparatus 1 which concerns on this invention, and the structure of the reagent bottle 2 which concerns on this invention. It is a schematic diagram explaining the structure of the solvent supply apparatus 1 which concerns on this invention. (A) shows a side view and (B) shows a front view. It is a schematic diagram explaining operation | movement of the solvent supply apparatus 1 which concerns on this invention.

Hereinafter, preferred embodiments for carrying out the present invention will be described with reference to the drawings. In addition, embodiment described below shows an example of typical embodiment of this invention, and, thereby, the range of this invention is not interpreted narrowly.

1. Solvent First, the solvent according to the present invention will be described. This solvent is a solvent used as a mobile phase for LC-MS (generally also called “eluent”), and is characterized by being prepared in a clean environment that can prevent contamination of substances that cause chemical noise. It is what.

As described above, one of the factors that lower the analytical sensitivity of LC-MS is aerosol particles and dust mixed in the solvent manufacturing process. Thus, by preparing a solvent in a clean environment from which aerosol particles and dust have been removed, a solvent free from chemical substances and contamination of trace substances that cause a reduction in ionization efficiency is provided.

This solvent can be prepared in a clean environment created by a so-called “clean room” or “clean box”. In the industrial field, clean rooms and clean boxes are used to create a clean environment in which manufacturing equipment is placed in the manufacture of precision machines and semiconductors. In general, in these clean rooms, etc., outside air that has passed through a high-performance air filter called “HEPA filter” or “ULPA filter” with a very fine filter is sent to the inside, and the supplied air is circulated or By discharging to the outside, aerosol particles and dust in the internal environment are removed, creating a clean environment. Furthermore, in order to prevent the inflow of aerosol particles and dust from the outside to the internal environment, the internal pressure is set to be positive with respect to the external pressure.

The solvent according to the present invention is also prepared in a clean environment using a clean room or the like or equipment equivalent thereto. HEPA filters are listed in the Japanese Industrial Standards (JIS standard) as “Air with a particle collection rate of 99.97% or more for particles with a rated air volume of 0.3 μm and an initial pressure loss of 245 Pa or less. “Filter”. In addition, the ULPA filter is the same standard as “an air filter with a particle collection rate of 99.9995% or more for particles with a rated air volume of 0.15 μm and an initial pressure loss of 245 Pa or less”. It is prescribed. By preparing the solvent in a clean environment such as these HEPA filter or ULPA filter, it is possible to obtain a solvent free from contamination by trace substances that cause a reduction in chemical noise and ionization efficiency in LC-MS analysis.

As the solvent according to the present invention, a wide variety of solvents used as mobile phases for LC-MS can be mentioned. Specifically, water and organic solvents such as acetonitrile, methanol and isopropyl alcohol are widely used. In addition, the solvent according to the present invention may include formic acid, acetic acid, trifluoroacetic acid, triethylamine, hexafluoroisopropanol (HEIP), ammonium acetate, etc. used by adding to water or an organic solvent.

As described above, the solvent according to the present invention is a solvent that does not contain trace substances in the solvent production process. However, when supplying this solvent, it prevents the trace substances from the reagent bottles from entering the solvent. Therefore, it is desirable that the reagent bottle is supplied in a reagent bottle according to the present invention described below.

2. Reagent Bottle A reagent bottle according to the present invention will be described with reference to FIG. FIG. 1 is a schematic diagram for explaining the operation of the solvent supply apparatus according to the present invention to be described later. As the operation of the solvent supply device will be described in detail later, the configuration of the reagent bottle indicated by reference numeral 2 in FIG. 1 will be described here.

In the figure, the reagent bottle denoted by reference numeral 2 is a reagent bottle for containing a solvent (reference numeral S in the figure) used as a mobile phase of LC-MS, and is made of glass and has a metal mouth portion. It is characterized by being sealed with a foil (reference numeral 21).

As described above, one of the factors that lowers the analytical sensitivity of LC-MS is a plasticizer or an unpolymerized monomer mixed from a reagent bottle. Therefore, first, the reagent bottle is made of a glass container to eliminate elution of the plasticizer and unpolymerized monomer into the solvent, which is a problem when a plastic container is used. Further, by sealing the mouth portion of the reagent bottle 2 with a metal foil 21 having a diameter substantially the same as the opening diameter of the mouth portion, elution of a plasticizer or the like which becomes a problem when a plastic lid is used is also possible. Exclude. Therefore, by accommodating the solvent according to the present invention described above in the reagent bottle 2 and supplying it, it is possible to provide a solvent free from the mixing of plasticizers and unpolymerized monomers that cause chemical noise and ionization efficiency reduction.

The metal foil 21 is formed of a corrosion-resistant metal, that is, a metal or an alloy that is hardly corroded. The metal foil 21 seals the mouth of the reagent bottle 2 to prevent foreign substances from entering the inside. However, when the metal foil 21 itself is corroded by a solvent, formic acid or the like added to the solvent, the metal foil 21 was corroded. Metal ions are eluted from the metal foil 21 into the solvent and cause chemical noise. In order to prevent this, the metal foil 21 is desirably formed of a corrosion-resistant metal, specifically, titanium, chromium, zirconia, stainless steel, gold, or the like.

Titanium, chromium, zirconia, and stainless steel oxides are very stable and hardly corroded, and form an oxide film (so-called “passive film”) on the metal surface in the air. This passive film formed by the combination of metal with oxygen is a thin and dense film, which protects the inside of the metal from acid corrosion and oxidation, and exhibits strong corrosion resistance. Titanium, etc. in the pool bear diagram (potential-pH diagram) is a “passive zone” where the progress of corrosion stops due to the formation of a passive film compared to the “corrosion zone” where corrosion progresses, and very stable and corrosive. There is a wide "dead zone (stable zone)" that is difficult to do. For this reason, by forming the metal foil 21 with titanium or the like, the elution of metal ions into the solvent due to the corrosion of the metal foil 21 can be prevented. Titanium, chromium, and zirconia can be alloys with aluminum, copper, iron, manganese, molybdenum, and the like, respectively, and the ratio of chromium, nickel, and the like contained in stainless steel can be set as appropriate.

The metal foil 21 can be formed of gold other than titanium, chromium, zirconia, and stainless steel. Gold creates an immunity that is the dead zone of the pool bear diagram, and is very stable and resistant to corrosion. For this reason, by forming the metal foil 21 with gold, elution of metal ions into the solvent due to corrosion of the metal foil 21 can be prevented. In addition, as the material of the metal foil 21, a corrosion-resistant metal can be widely used. For example, Hastelloy (registered trademark) containing nickel as a main component and molybdenum or chromium can be used.

The metal foil 21 has an opening diameter at the mouth of the reagent bottle 2 and a diameter substantially the same as the diameter of the metal foil 21, and a lid 22 that covers the mouth of the reagent bottle 2 from the outside of the metal foil 21; The mouth portion is sealed so as to be pressed against the mouth portion of the reagent bottle 2 by an annular packing 23 interposed between the lid 22 and the metal foil 21. That is, when the reagent bottle 2 containing the solvent is sealed, the metal foil 21 is put on the mouth portion, and the lid 22 is screwed into the mouth portion with the annular packing 23 placed on the mouth portion. The reagent bottle 2 is sealed by bringing 21 into close contact with the mouth. At this time, the annular packing 23 may be integrally provided inside the lid 22.

When the metal foil 21 is adhered to the mouth by the lid 22 and the annular packing 23 and the reagent bottle 2 is sealed, there is a problem when the metal foil 21 is adhered and sealed to the mouth using an adhesive or the like. It is possible to eliminate the dissolution of the adhesive component in the solvent.

Further, the lid 22 has a function of preventing the metal foil 21 from being damaged due to accidental contact with the surface of the metal foil 21 or the solvent leakage due to the damage during the transportation and storage of the reagent bottle 2 after containing the solvent. 21 also functions to prevent dust from adhering to the surface. In addition, the material of the lid | cover 22 and the annular packing 23 is not specifically limited, The plastic resin used normally may be sufficient.

A hole 221 through which the suction nozzle 13 provided in the solvent supply device described below is inserted is formed in the upper surface of the lid 22. The diameter of the hole 221 is substantially the same as or slightly larger than the diameter of the suction nozzle 13 so that the suction nozzle 13 can be inserted. Hereinafter, the lid 22 is referred to as a “hole cap 22”. In order to prevent damage to the metal foil 21 and adhesion of dust to the surface of the metal foil 21 during transportation and storage, the hole 221 of the perforated cap 22 is temporarily closed. A protective seal may be provided.

3. Configuration of Solvent Supply Device (3-1) Subsequently, the configuration of the solvent supply device for LC-MS according to the present invention will be described with reference to FIG. 1 and FIG. 2A is a side view of the solvent supply apparatus, and FIG. 2B is a front view.

As mentioned above, one of the factors that lowers the analytical sensitivity of LC-MS is that it can be mixed into the reagent bottle during the replacement of the reagent bottle when the solvent is supplied to the LC-MS, or the nozzle of the LC-MS Aerosol particles and dust adhered to the surface. The solvent supply device according to the present invention has a function of preventing chemical noise and ionization efficiency from being reduced due to the prevention of mixing of aerosol particles or the like into the reagent bottle and adhesion to the nozzle. Hereafter, each structure of a solvent supply apparatus is demonstrated in order.

In FIG. 2, the solvent supply device denoted by reference numeral 1 is a device that is connected to the LC-MS and supplies mobile phase solvent thereto, and is in contact with the suction nozzle 13 and the mouth of the reagent bottle 2. And a vacuum cap 14 (not shown) connected to the vacuum cap 14.

The suction nozzle 13 is inserted into the mouth of the reagent bottle 2 (see FIG. 1) according to the present invention containing a solvent, and functions to suck the internal solvent and send it to the LC-MS. In the figure, reference numeral 131 denotes a tube (tube) for sending the solvent sucked by the suction nozzle 13 to the LC-MS.

The suction nozzle 13 is formed of a corrosion-resistant metal like the metal foil 21 of the reagent bottle 2 described above. This is because if the suction nozzle 13 is corroded by the solvent, formic acid or the like added to the solvent, metal ions are eluted from the corroded suction nozzle 13 into the solvent, causing chemical noise.

As shown in FIG. 1B, the vacuum cap 14 is in contact with and covers the mouth of the reagent bottle 2 (here, the perforated cap 22), and a space K is formed between the vacuum cap 14 and the metal foil 21. Form. A braking piece 144 that contacts the mouth of the reagent bottle 2 is provided around the inner peripheral surface of the vacuum cap 14, and the vacuum cap 14 is brought into contact with the mouth of the reagent bottle 2 by the braking piece 144. A space K is formed between the metal foil 21 and the metal foil 21.

In the drawing, reference numeral 143 denotes a tube connected to the vacuum pump at one end and connected to the vacuum cap 14 at the other end. The tube 143 has a suction port 142 that communicates with the space K at the connection to the vacuum cap 14, and the vacuum pump sucks air in the space K from the suction port 142.

The upper surface of the vacuum cap 14 is provided with a hole 141 through which the suction nozzle 13 is inserted, like the hole cap 22 of the reagent bottle 2 described above. The diameter of the hole 141 is substantially the same as or slightly larger than the diameter of the suction nozzle 13 so that the suction nozzle 13 can be inserted. The material of the vacuum cap 14 may be a commonly used plastic resin.

Furthermore, the solvent supply device 1 has a drive unit (not shown) for moving the suction nozzle 13 and the vacuum cap 14 with respect to the reagent bottle 2. This driving means makes the vacuum cap 14 contact the mouth of the reagent bottle 2 as shown in FIG. 1 (B), and the metal foil that seals the mouth of the suction nozzle as shown in FIG. 1 (C). It functions to penetrate through 21 and to be inserted into the reagent bottle 2.

The liquid feeding pump, vacuum pump, and driving means for sucking and feeding the solvent by the suction nozzle 13 are provided in the electrical space 112 of the housing 11 constituting the main body of the solvent supply device 1. On the other hand, the suction nozzle 13 and the vacuum cap 14 are disposed in the internal space 111 of the housing 11. The housing 11 is the clean box described above, and the solvent supply device 1 includes a cleaning mechanism for maintaining the internal space 111 in a clean environment.

The cleaning mechanism can have the same configuration as a normally used clean box. The cleaning mechanism is disposed in the electrical space 112 of the main body 11 and is provided, for example, in an air supply port that supplies the outside air of the housing 11 to the internal space 111 and a flow path of the outside air supplied from the air supply port. And an exhaust port for exhausting the supplied air to the outside of the housing 11. The air discharged from the exhaust port may be re-introduced into the internal space 111 from the intake port and circulated. In FIG. 2, illustration of the air supply port, the filter unit, and the exhaust port is omitted.

The filter unit is configured by combining the above-described HEPA filter or the like and one or more air filters having a coarser grain and lower performance as necessary. By arranging a HEPA filter or the like in the flow path of the outside air supplied from the air supply port, it is possible to remove the aerosol particles and dust in the outside air and maintain the internal space 111 of the housing 11 in a clean environment. .

The air supply port is provided above the housing 11, preferably on the top surface of the internal space 111. Further, it is preferable that the exhaust port is provided below the back surface portion (electrical space 112 side) of the housing 11. By providing the intake port and the exhaust port at these positions, it is possible to form an airflow flowing from the upper side to the downstream side in the internal space 111 of the housing 11. This airflow functions to prevent outside air including aerosol particles and dust from flowing into the internal space 111 when the door 12 described below is opened and closed (details will be described later).

An openable / closable door 12 for taking in and out the reagent bottle 2 in and out of the internal space 111 is provided on the front surface of the housing 11. The casing 11 has an airtight structure in which air does not enter and exit other than the above-described intake and exhaust ports when the door body 12 is closed. Here, a case where the door body 12 is configured as a slide cover that opens and closes by sliding in the vertical direction on the front portion of the housing 11 will be described as an example. However, the door body 12 is not limited to a slide cover, and may be, for example, a right-opening or left-opening door that is opened and closed by a hinge (hinge), or a double door. It is preferable to provide a packing for maintaining airtightness at a portion where the door body 12 and the housing 11 are in contact with each other. Hereinafter, the “door body 12” is referred to as a “slide cover 12”.

Open / close of the slide cover 12 is detected by an open / close sensor (not shown). In addition, the solvent supply device 1 includes a reagent bottle sensor 15 for detecting the reagent bottle 2 arranged at a predetermined position in the internal space 111 and a liquid level sensor for detecting the remaining amount of the solvent in the reagent bottle 2. 16 is provided. In the present invention, the reagent bottle sensor 15 is also referred to as “first detection means”, and the open / close sensor is also referred to as “second detection means”.

The above-described vacuum pump starts or stops suction based on the detection signal of the reagent bottle 2 from the reagent bottle sensor 15. The above-described driving means controls the movement of the suction nozzle 13 and the vacuum cap 14 based on detection signals from the reagent bottle sensor 15 and the open / close sensor.

(3-2) Apparatus Operation Next, the operation of the solvent supply apparatus 1 will be described with reference to FIGS. FIG. 3 shows a side view of the solvent supply apparatus.

First, as shown in FIG. 2, in order to set the reagent bottle 2 containing the solvent in the solvent supply device 1, the slide cover 12 is slid upward to open and the reagent bottle 2 is inserted into the opened internal space 111. , Arranged at a predetermined position.

At this time, the air flow flowing from the upper side to the lower side is formed in the internal space 111 by the above-described cleaning mechanism, so that the outside air containing aerosol particles and dust flows back into the internal space 111 that has been opened. Can be prevented. Further, the cleaning mechanism can more reliably prevent the atmospheric pressure of the internal space 111 from being set to a positive pressure with respect to the outside and the inflow of aerosol particles and the like.

The reagent bottle 2 arranged at a predetermined position in the internal space 111 is detected by the reagent bottle sensor 15, and a detection signal is output to the vacuum pump. In response to this output, the vacuum pump starts suction.

After the reagent bottle 2 is set, as shown in FIG. 3, the slide cover 12 is slid down and closed to make the internal space 111 airtight. When the slide cover 12 is closed, a detection signal from the open / close sensor is output to the driving means. The driving means receives the output from the open / close sensor and the output of the detection signal of the reagent bottle 2 from the reagent bottle sensor 15, and lowers the suction nozzle 13 and the vacuum cap 14 with respect to the arranged reagent bottle 2 (FIG. Middle, see arrow F).

As a result, first, as shown in FIG. 1 (B), the brake piece 144 of the lowered vacuum cap 14 comes into contact with the mouth of the reagent bottle 2 (here, the perforated cap 22), and the vacuum cap 14 A space K is formed between the metal foil 21 and the metal foil 21. The air in the space K is sucked by the vacuum pump that has started sucking, and is discharged out of the space K from the suction port 142 of the pipe 143 connected to the vacuum cap 14.

Subsequently, as shown in FIG. 1C, the lowered suction nozzle 13 is inserted through the

holes

141 and 221 formed in the upper surfaces of the vacuum cap 14 and the perforated cap 22, respectively. The metal foil 21 that seals the mouth portion passes through and is inserted into the reagent bottle 2. At this time, in order to make it easy for the suction nozzle 13 to penetrate the metal foil 21, the thickness of the metal foil 21 is desirably about 1 to 100 μm, preferably about 1 to 20 μm.

When dust is attached to the metal foil 21 when the suction nozzle 13 penetrates, the dust attached to the metal foil 21 in the space K is caused by vibration or impact generated when the suction nozzle 23 penetrates the metal foil 21. In the reagent bottle 2 and may adhere to the suction nozzle 13. In FIG. 1C, symbol D indicates a substance (dust generation substance) that generates dust from the metal foil 21 when the suction nozzle 23 penetrates the metal foil 21.

The dusting substance D mixed into the reagent bottle 2 or adhering to the suction nozzle 13 may cause chemical noise or a decrease in ionization efficiency. In the solvent supply device 1, even when the dust generation material D is generated when the suction nozzle 23 penetrates the metal foil 21, the dust generation material D is removed together with the air in the space K by the vacuum pump that has started the suction. Since it can be sucked out and quickly discharged out of the space K and removed, it is possible to prevent the dusting substance D from mixing into the reagent bottle 2 or adhering to the suction nozzle 13.

Here, if the pressure in the space K is too low due to suction by the vacuum pump, the solvent S in the reagent bottle 2 may be sucked into the space K and the tube 143. In order to avoid this, the diameter of the hole 141 of the vacuum cap 14 is formed to be slightly larger than the diameter of the suction nozzle 13, and a gap is generated between the vacuum cap 14 and the suction nozzle 13 when the suction nozzle 13 is inserted. It is desirable to prevent the pressure in the space K from dropping too much.

When the tip of the suction nozzle 13 is inserted to the vicinity of the bottom surface of the reagent bottle 2, the suction of the solvent is started, and the solution is sent to the LC-MS through the tube 131. For example, the solvent is fed in response to a signal from the LC-MS, and at any timing until the remaining amount of the solvent in the reagent bottle 2 detected by the liquid level sensor 16 becomes less than a predetermined value. Can be done.

As described above, in the solvent supply device 1, the reagent bottle 2 and the suction nozzle 13 are disposed in the internal space 111 from which aerosol particles and dust are removed, and are attached to the metal foil 21 of the reagent bottle 2. Chemical dust and ionization efficiency due to aerosol particles and dust mixed into the reagent bottle 2 and attached to the suction nozzle 13 during the reagent bottle replacement operation by sucking and removing even a small amount of dust with a vacuum pump Decline can be prevented. Therefore, by using the solvent and reagent bottle 2 according to the present invention described above in combination with the solvent supply device 1, a solvent free from chemical noise and contamination of substances that cause a reduction in ionization efficiency is supplied to the LC-MS. It becomes possible to perform highly sensitive analysis.

In the present embodiment, the solvent supply device 1 is configured as a separate unit from the LC-MS, and is described as a device that supplies the solvent connected to the LC-MS. However, the solvent supply device 1 is integrated with the LC-MS. It may be a configured device.

Since the solvent supply device for a liquid chromatograph mass spectrometer according to the present invention can analyze proteins, peptides, sugar chains, various metabolites, etc. with high sensitivity, biotechnology, medicine, drug discovery, foods, etc. In the field, it can contribute to analysis in particular for trace substances.

DESCRIPTION OF SYMBOLS 1 Solvent supply apparatus 11 Housing | casing 12 Door body 13 Suction nozzle 14 Vacuum cap 142 Suction port 144 Brake piece 15 Reagent bottle sensor 16 Liquid level sensor 111 Internal space 112 Electrical space 2 Reagent bottle 21 Metal foil 22 Lid (hole opening cap)
D Dust generating material K Space S Solvent

Claims

A suction nozzle that contains a solvent used as a mobile phase of a liquid chromatograph mass spectrometer and has a mouth sealed with a metal foil, and is inserted through the metal foil and sucks the solvent.
A vacuum cap that is in contact with and covers the mouth of the reagent bottle and has a hole through which a suction nozzle inserted into the reagent bottle is inserted;
A solvent supply for a liquid chromatograph mass spectrometer, comprising: a vacuum pump connected to the vacuum cap and sucking air in a space formed between the vacuum cap covering the mouth of the reagent bottle and the metal foil apparatus.
The solvent supply device according to claim 1, wherein the suction nozzle is made of a corrosion-resistant metal.
An air supply port for supplying outside air, a filter unit provided in a flow path of outside air supplied from the air supply port, an exhaust port for exhausting the supplied air, and a door body that can be opened and closed The solvent supply device according to claim 2, wherein the suction nozzle and the vacuum cap are disposed in a housing provided with the above.
The suction nozzle and the vacuum cap are moved with respect to the reagent bottle arranged at a predetermined position in the housing, the vacuum cap is brought into contact with the mouth of the reagent bottle, and the mouth of the suction nozzle is sealed. The solvent supply apparatus according to claim 3, further comprising a driving unit that is inserted into the reagent bottle through the metal foil.
A first detection means for detecting a reagent bottle arranged at a predetermined position in the housing is provided, and the vacuum pump starts or stops aspiration based on a detection signal from the first detection means. 4. The solvent supply apparatus according to 4.
Comprising a second detection means for detecting opening and closing of the door body;
6. The solvent supply apparatus according to claim 5, wherein the driving unit controls movement of the suction nozzle and the vacuum cap based on a detection signal from the second detection unit and a detection signal from the first detection unit. .
A liquid chromatograph mass spectrometer equipped with the solvent supply device according to any one of claims 1 to 6.
A reagent bottle for containing a solvent used as a mobile phase of a liquid chromatograph mass spectrometer,
A metal foil that seals the mouth,
A glass reagent bottle having a lid covered with an opening from the outside of a metal foil and having a hole inserted through a metal foil and through which a suction nozzle for sucking a solvent is inserted.
The reagent bottle according to claim 8, wherein the metal foil is formed of a corrosion-resistant metal.
Solvent used as a mobile phase in a liquid chromatograph mass spectrometer, and prepared in a clean environment capable of preventing contamination of substances that cause chemical noise.
A metal foil that seals the mouth,
The mouth portion is covered from the outside of the metal foil, and is received in a glass reagent bottle having a lid that is inserted through the metal foil and has a hole through which a suction nozzle that sucks the solvent is inserted. The solvent according to claim 10.
A solvent used as a mobile phase contained in a reagent bottle whose mouth is sealed with a metal foil,
A solvent supply method for a liquid chromatograph mass spectrometer, comprising a step of sucking with a suction nozzle inserted through a metal bottle and penetrating the metal foil, and feeding the liquid to a liquid chromatograph mass spectrometer.
The solvent supply method according to claim 12, further comprising: sucking a substance that generates dust from the metal foil with a vacuum pump when the suction nozzle penetrates the metal foil to prevent the substance from being mixed into the solvent. .
14. The solvent supply method according to claim 13, further comprising a step of preparing the solvent in a clean environment capable of preventing mixing of substances causing chemical noise and storing the solvent in the reagent bottle made of glass.