CN105869984B - High-resolution quadrupole rod time of-flight mass spectrometer quality axle correcting fluid and its application method - Google Patents
High-resolution quadrupole rod time of-flight mass spectrometer quality axle correcting fluid and its application method Download PDFInfo
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- CN105869984B CN105869984B CN201610238237.2A CN201610238237A CN105869984B CN 105869984 B CN105869984 B CN 105869984B CN 201610238237 A CN201610238237 A CN 201610238237A CN 105869984 B CN105869984 B CN 105869984B
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- correcting fluid
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- acetonitrile
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- formic acid
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- H01J49/00—Particle spectrometers or separator tubes
- H01J49/0009—Calibration of the apparatus
Abstract
The invention discloses a kind of high-resolution quadrupole rod time of-flight mass spectrometer quality axle correcting fluid, by hydroxybenzoic acid, ethyl-para-hydroxybenzoate, galuteolin, chlorogenic acid, enoxolone, Paeoniflorin, benzoylpaeoniflorin and Panax Notoginseng saponin R1Dissolving is prepared in a solvent；Solvent is selected from least one in water, methyl alcohol, formic acid, acetonitrile；In correcting fluid, the mass fraction of each solute is as follows:1~3 part of hydroxybenzoic acid；1~3 part of ethyl-para-hydroxybenzoate；1~3 part of galuteolin；1~3 part of chlorogenic acid；1~3 part of enoxolone；1~3 part of Paeoniflorin；1~3 part of benzoylpaeoniflorin；Panax Notoginseng saponin R11~3 part；The mass-volume concentration of each solute is 0.1~10 μ g/ml in correcting fluid.The formula of correcting fluid disclosed by the invention is simple, low cost, and solution state stability inferior is high, and the mass spectrographic correction error of firsts and seconds is respectively less than 5ppm.
Description
Technical field
The present invention relates to the technical field of mass spectral analysis, more particularly to a kind of high-resolution quadrupole rod time of-flight mass spectrometer matter
Amount axle correcting fluid and its application method.
Background technology
High-resolution quadrupole rod time of-flight mass spectrometer (QTOF) is the mass spectrograph that a class can obtain accurate molecular weight, can be inferred
Go out chemical structural formula, the qualitative ability than Low Resolution Mass Spectra is more strengthened, be widely used to organic molecule structural analysis, natural
The scientific domains such as drug ingedient identification, drug metabolite identification and polypeptide protein quality analysis.
Mass spectrograph tof tube quality axle (hereinafter referred to as " quality axle ") degree of accuracy is most important to the reliability of experimental result,
It is corrected and its degree of accuracy can shift over time, thus before requirement experiment and in experiment, influence quality axle offset
Factor have a lot, mainly including power supply, environment temperature, humidity, vacuum and instrumental pollution etc..
In order to ensure the accuracy of quality axle, prior art (such as SCIEX companies quadrupole rod time-of-flight mass spectrometry instrument
Equipment) using the strategy do not calibrated in correction, experiment before experiment, the method can avoid the ion depression effect of correcting fluid, and it lacks
Point is that, with the extension at correction interval, the degree of accuracy is just difficult to ensure that.Therefore, prior art advises every 2 hours or every 5 samples
Once, to ensure the accuracy of quality axle, but frequently correction can bring the larger problem of correcting fluid consumption for correction.
According to previous experiences, once experiment need to consume 10~20ml of correcting fluid, the commercially available price of correcting fluid be about 1700 yuan/
100ml, therefore the single operating cost of high-resolution mass spectrometer is very high.In serial experiments such as metabolism group, process control
In, correcting fluid consumption is huger, and operating cost accounting is higher to have become common recognition.
And existing commercially available calibration solution has two defects:
First, mass-to-charge ratio is poor for the compound stability in the solution of m/z144 and m/z403；
Second, the secondary correction Shortcomings with m/z403 as parent ion, low quality section (100~600) second order mses from
The quality error of son meets the requirements (within 5ppm), and the quality error of high-quality section (more than 600) is bigger than normal, and with molecule quantitative change
Greatly, error becomes larger.
The content of the invention
The invention discloses a kind of high-resolution quadrupole rod time of-flight mass spectrometer quality axle correcting fluid, the correcting fluid is matched somebody with somebody
Side is simple, low cost, and solution state stability inferior is high, and the error that first mass spectrometric quality axle is corrected in the negative ion mode is less than
5ppm, during correction second order mses quality axle, correcting range is substantially improved, and meets now high-resolution quadrupole rod flight time mass spectrum
Instrument carries out the demand of quality axle correction.
A kind of high-resolution quadrupole rod time of-flight mass spectrometer quality axle correcting fluid, by hydroxybenzoic acid, para hydroxybenzene first
Acetoacetic ester, galuteolin, chlorogenic acid, enoxolone, Paeoniflorin, benzoylpaeoniflorin and Panax Notoginseng saponin R1Dissolving is made in a solvent
It is standby to obtain；
Described solvent is selected from least one in water, methyl alcohol, formic acid, acetonitrile；
In the correcting fluid, the mass fraction of each solute is as follows:
The mass-volume concentration of each solute is 0.1~10 μ g/ml in the correcting fluid.
The present invention by the suitable correcting fluid of compound concentration be used for first mass spectrometric quality axle correction, and using mass-to-charge ratio compared with
Big ion is parent ion, and second order mses quality axle is corrected with its daughter ion.The correcting fluid can not only well correct one-level
Mass spectrum quality axle, the correcting range of second order mses quality axle is also expanded be twice the above, and this correction formula of liquid is simple, matter
Amount stabilization, it is easy to implement, it is adaptable to produce in batches.
Preferably, described solvent includes acetonitrile-water mixed solvent, formic acid-Methanol+Water, formic acid-second
Nitrile-water mixed solvent.
Preferably, in the acetonitrile-water mixed solvent, acetonitrile is 4 with the volume ratio of water:1；
In the formic acid-Methanol+Water, the volume ratio of formic acid, methyl alcohol and water is 0.002:1:1；
In the formic acid-acetonitrile-water mixed solvent, the volume ratio of formic acid, acetonitrile and water is 0.005:4:1.
Preferably, by etc. the hydroxybenzoic acid of quality, ethyl-para-hydroxybenzoate, galuteolin, chlorogenic acid, Radix Glycyrrhizae time
Acid, Paeoniflorin, benzoylpaeoniflorin and Panax Notoginseng saponin R1Acetonitrile-water mixed solvent is dissolved in, acetonitrile is 4 with the volume ratio of water:
1；
The gross mass volumetric concentration of the correcting fluid is 80 μ g/ml.
It is hydroxybenzoic acid, ethyl-para-hydroxybenzoate, galuteolin, chlorogenic acid, sweet preferably, in the correcting fluid
Careless hypo acid, Paeoniflorin, benzoylpaeoniflorin and Panax Notoginseng saponin R1Mass ratio be 1:3:1:3:1:3:1:3；
Solvent is formic acid-Methanol+Water, and the volume ratio of formic acid, methyl alcohol and water is 0.002:1:1；
In the correcting fluid, the mass-volume concentration of hydroxybenzoic acid, galuteolin, enoxolone and benzoylpaeoniflorin
It is 0.833 μ g/ml.
Ethyl-para-hydroxybenzoate, chlorogenic acid, Paeoniflorin and Panax Notoginseng saponin R1Mass-volume concentration be 2.5 μ g/ml.
Preferably, by etc. the hydroxybenzoic acid of quality, ethyl-para-hydroxybenzoate, galuteolin, chlorogenic acid, Radix Glycyrrhizae time
Acid, Paeoniflorin, benzoylpaeoniflorin and Panax Notoginseng saponin R1It is dissolved in formic acid-acetonitrile-water mixed solvent, formic acid, acetonitrile and water
Volume ratio be 0.005:4:1；
The gross mass volumetric concentration of the correcting fluid is 8 μ g/ml.
High-resolution quadrupole rod time of-flight mass spectrometer quality axle is carried out the invention also discloses using above-mentioned correcting fluid
First mass spectrometric correction is carried out under the method for correction, negative ion mode, with m/z137.02442 (hydroxybenzoic acid), m/
Z165.05572 (ethyl-para-hydroxybenzoate), m/z285.04046 (cyanidenon), m/z353.08782 (chlorogenic acid), m/
Z469.33233 (enoxolone), m/z525.16136 (Paeoniflorin+formic acid), m/z629.18758 (benzoylpaeoniflorins+first
Acid), m/z931.52719 (Panax Notoginseng saponin Rs1) as reference ion, correct first mass spectrometric tof tube quality axle.
Under negative ion mode, second order mses correction is carried out, with m/z931.52710 as parent ion, add the fragmentation energy of 60V
Amount, using m/z161.04555, m/z475.37928, m/z637.43211 and m/z931.52719 as reference ion, corrects two
Level mass spectrum tof tube quality axle.
Compared with prior art, the invention has the advantages that:
1st, formula components are easy to get, and configuration is simple, low cost；
2nd, under negative ion mode, first mass spectrometric quality axle is corrected using the calibration solution, error is less than 5ppm, with commercially available product
The effect of product is consistent；During correction second order mses quality axle, the error in the range of m/z (100~1200) is less than 5ppm, commercially available prod
Error only in the range of m/z (100~600) is less than 5ppm, and correcting range is substantially improved.
Brief description of the drawings
Fig. 1 is one-level anion quality axle correction mass spectrogram in embodiment 3；
Fig. 2 is two grades of anion quality axle correction mass spectrograms in embodiment 3.
Specific embodiment
With reference to embodiment, the present invention is further described.
Embodiment 1
Weigh P-hydroxybenzoic acid, ethyl-para-hydroxybenzoate, cyanidenon, chlorogenic acid, enoxolone, Paeoniflorin, benzene
Formyl Paeoniflorin and Panax Notoginseng saponin R1Each 30mg, (acetonitrile, water volume ratio are 4 to be dissolved in 3L acetonitrile-aqueous solutions:1) in, it is configured to
Total concentration is the correcting fluid of 80 μ g/ml, and will correct liquid pump using CDS enters ion gun (flow velocity 0.05ml/min), after atomization ionization,
Into mass spectrum.Under negative ion mode, one-level timing is done, respectively with m/z137.02442, m/z165.05572, m/
Z285.04046, m/z353.08782, m/z469.33233, m/z525.16136, m/z629.18758, m/z931.52719 make
It is reference ion, carries out first quality axle correction.M/z931.52710 is parent ion, the fragmentation energies of 60V is added, with m/
Z161.04555, m/z475.37928, m/z637.43211, m/z931.52719 are that reference ion corrects second mass axle.
The condition of gathered data is as follows after correction:Chromatographic column is Agilent Eclipse Plus-C18(100mm×4.6mm
i.d.,1.8μm)；It is mobile phase A with 0.1% formic acid solution, is Mobile phase B, linear gradient elution, stream with 0.1% formic acid acetonitrile
Speed is 0.7ml/min；Column temperature is 30 DEG C；Detection wavelength is 210nm；The μ l of 10mg/ml ginseng extracts sample introduction 5, Mass Spectrometry Conditions
Triple TOF 5600+ flight time LC-MS instrument:Anion scan pattern；Sweep limits:m/z 100-2000；Atomization
Gas (GS1):50psi；Atomization gas (GS2):50psi；Gas curtain gas (CUR):30psi；Ion source temperature (TEM):550 DEG C (negative)；
Ion source voltage (IS):- 4500V (negative)；One-level is scanned:Remove cluster voltage:100V；Focus voltage:10V；Two grades of scannings:Using
IDA patterns, CID energy is 30,50 and 70V.
Data measured is shown in Table 1, as shown in Table 1, one through peakview software analysis, the quality error of several typical compounds
The quality error of level and second order mses is respectively less than 5ppm, meets the quality requirement of high resolution mass spectrum data.
Table 1
Embodiment 2
Weigh P-hydroxybenzoic acid 10mg, ethyl-para-hydroxybenzoate 30mg, cyanidenon 10mg, chlorogenic acid 30mg, sweet
Careless hypo acid 10mg, Paeoniflorin 30mg, benzoylpaeoniflorin 10mg and Panax Notoginseng saponin R130mg, is dissolved in 12L formic acid-methanol-water
(formic acid, methyl alcohol, the volume ratio of water are 0.002:1:1) solution is configured to correcting fluid, and solution is pumped into ion gun (stream using CDS
Fast 0.5ml/min), after atomization ionization, into mass spectrum.Under negative ion mode, one-level timing is done, m/ is used respectively
z137.02442、m/z165.05572、m/z285.04046、m/z353.08782、m/z469.33233、m/z525.16136、
M/z629.18758, m/z931.52719 carry out first quality axle correction as reference ion.M/z931.52710 for it is female from
Son, adds the fragmentation energies of 60V, is with m/z161.04555, m/z475.37928, m/z637.43211, m/z931.52719
Reference ion corrects second mass axle.
, with embodiment 1, data measured is through peakview software analysis, the quality error of several typical compounds for analysis method
2 are shown in Table, as shown in Table 2, the mass spectrographic quality error of firsts and seconds is respectively less than 5ppm, the quality for meeting high resolution mass spectrum data will
Ask.
Table 2
Embodiment 3
Weigh P-hydroxybenzoic acid 10mg, ethyl-para-hydroxybenzoate 10mg, cyanidenon 10mg, chlorogenic acid 10mg, sweet
Careless hypo acid 10mg, Paeoniflorin 10mg, benzoylpaeoniflorin 10mg and Panax Notoginseng saponin R110mg, is dissolved in 10L formic acid-acetonitrile-water
(formic acid, acetonitrile, the volume ratio of water are 0.005:4:1) solution is configured to correcting fluid, and solution is pumped into ion gun (stream using CDS
Fast 0.5ml/min), after atomization ionization, into mass spectrum.Under negative ion mode, one-level timing is done, m/ is used respectively
z137.02442、m/z165.05572、m/z285.04046、m/z353.08782、m/z469.33233、m/z525.16136、
M/z629.18758, m/z931.52719 carry out first quality axle correction as reference ion.M/z931.52710 for it is female from
Son, adds the fragmentation energies of 60V, is with m/z161.04555, m/z475.37928, m/z637.43211, m/z931.52719
Reference ion corrects second mass axle.
, with embodiment 1, data measured is through peakview software analysis, the quality error of several typical compounds for analysis method
3 are shown in Table, as shown in Table 3, the mass spectrographic quality error of firsts and seconds is respectively less than 5ppm, the quality for meeting high resolution mass spectrum data will
Ask.
Table three
Claims (8)
1. a kind of high-resolution quadrupole rod time of-flight mass spectrometer quality axle correcting fluid, it is characterised in that
By hydroxybenzoic acid, ethyl-para-hydroxybenzoate, galuteolin, chlorogenic acid, enoxolone, Paeoniflorin, benzoyl Chinese herbaceous peony
Glycosides and Panax Notoginseng saponin R1Dissolving is prepared in a solvent；
Described solvent is selected from least one in water, methyl alcohol, formic acid, acetonitrile；
In the correcting fluid, the mass fraction of each solute is as follows:
The mass-volume concentration of each solute is 0.1~10 μ g/ml in the correcting fluid.
2. high-resolution quadrupole rod time of-flight mass spectrometer quality axle correcting fluid according to claim 1, it is characterised in that
Described solvent includes acetonitrile-water mixed solvent, formic acid-Methanol+Water or formic acid-acetonitrile-water mixed solvent.
3. high-resolution quadrupole rod time of-flight mass spectrometer quality axle correcting fluid according to claim 2, it is characterised in that
In the acetonitrile-water mixed solvent, acetonitrile is 4 with the volume ratio of water:1；
In the formic acid-Methanol+Water, the volume ratio of formic acid, methyl alcohol and water is 0.002:1:1；
In the formic acid-acetonitrile-water mixed solvent, the volume ratio of formic acid, acetonitrile and water is 0.005:4:1.
4. high-resolution quadrupole rod time of-flight mass spectrometer quality axle correcting fluid according to claim 3, it is characterised in that
By etc. quality hydroxybenzoic acid, ethyl-para-hydroxybenzoate, galuteolin, chlorogenic acid, enoxolone, Paeoniflorin, benzoyl
Paeoniflorin and Panax Notoginseng saponin R1Acetonitrile-water mixed solvent is dissolved in, acetonitrile is 4 with the volume ratio of water:1；
The gross mass volumetric concentration of the correcting fluid is 80 μ g/ml.
5. high-resolution quadrupole rod time of-flight mass spectrometer quality axle correcting fluid according to claim 3, it is characterised in that
In the correcting fluid, hydroxybenzoic acid, ethyl-para-hydroxybenzoate, galuteolin, chlorogenic acid, enoxolone, Paeoniflorin, benzene first
Acyl Paeoniflorin and Panax Notoginseng saponin R1Mass ratio be 1:3:1:3:1:3:1:3；
Solvent is formic acid-Methanol+Water, and the volume ratio of formic acid, methyl alcohol and water is 0.002:1:1；
In the correcting fluid, the mass-volume concentration of hydroxybenzoic acid, galuteolin, enoxolone and benzoylpaeoniflorin is
0.833μg/ml。
Ethyl-para-hydroxybenzoate, chlorogenic acid, Paeoniflorin and Panax Notoginseng saponin R1Mass-volume concentration be 2.5 μ g/ml.
6. high-resolution quadrupole rod time of-flight mass spectrometer quality axle correcting fluid according to claim 3, it is characterised in that
By etc. quality hydroxybenzoic acid, ethyl-para-hydroxybenzoate, galuteolin, chlorogenic acid, enoxolone, Paeoniflorin, benzoyl
Paeoniflorin and Panax Notoginseng saponin R1It is dissolved in formic acid-acetonitrile-water mixed solvent, the volume ratio of formic acid, acetonitrile and water is 0.005:
4:1；
The gross mass volumetric concentration of the correcting fluid is 8 μ g/ml.
7. the correcting fluid described in a kind of utilization claim 1~6 any claim is to high-resolution quadrupole rod flight time mass spectrum
The method that instrument quality axle is corrected, it is characterised in that first mass spectrometric correction is carried out under negative ion mode, with m/
z137.02442、m/z165.05572、m/z285.04046、m/z353.08782、m/z469.33233、m/z525.16136、
M/z629.18758 and m/z931.52719 corrects first mass spectrometric tof tube quality axle as reference ion.
8. the method being corrected to high-resolution quadrupole rod time of-flight mass spectrometer quality axle according to claim 7, its
It is characterised by, under negative ion mode, carries out second order mses correction, with m/z931.52710 as parent ion, adds the fragmentation energy of 60V
Amount, using m/z161.04555, m/z475.37928, m/z637.43211 and m/z931.52719 as reference ion, corrects two
Level mass spectrum tof tube quality axle.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US5872357A (en) * | 1997-05-30 | 1999-02-16 | Hewlett-Packard Company | Mass spectrometry calibration using homogeneously substituted fluorinated triazatriphosphorines |
| CN1898674A (en) * | 2003-10-20 | 2007-01-17 | 瑟诺生物科学有限责任公司 | Methods for calibrating mass spectrometry (ms) and other instrument systems and for processing ms and other data |
| CN104597114A (en) * | 2015-01-21 | 2015-05-06 | 华中师范大学 | Mass correction kit and mass correction method for low-mass area of high-resolution mass spectrometer in negative ion mode |
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| US9257267B2 (en) * | 2011-03-31 | 2016-02-09 | Dh Technologies Development Pte. Ltd. | Composition, method, and kit for calibrating a mass spectrometer |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5872357A (en) * | 1997-05-30 | 1999-02-16 | Hewlett-Packard Company | Mass spectrometry calibration using homogeneously substituted fluorinated triazatriphosphorines |
| CN1898674A (en) * | 2003-10-20 | 2007-01-17 | 瑟诺生物科学有限责任公司 | Methods for calibrating mass spectrometry (ms) and other instrument systems and for processing ms and other data |
| CN104597114A (en) * | 2015-01-21 | 2015-05-06 | 华中师范大学 | Mass correction kit and mass correction method for low-mass area of high-resolution mass spectrometer in negative ion mode |
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