Mass calibration kit and calibration method for low-mass region of high-resolution mass spectrometer in negative ion mode

Abstract

A mass calibration kit and a calibration method for a low-mass area of a high-resolution mass spectrometer in negative ion mode. The mass calibration kit comprises semiconductor nanometer material suspension, a free fatty acid standard solution and a MALDI sample target cleaning liquid. The mass calibration method comprises: adjusting a voltage difference between a sample target of the mass spectrometer and a slit to be 20 V; dripping the semiconductor nanometer material suspension on the surface of the sample target till a solvent is completely volatilized and dried; dripping the free fatty acid standard solution on the surface of a semiconductor nanometer material till the solvent is completely volatilized and dried; and putting the sample target in the mass spectrometer for mass calibration, wherein calibration coefficients obtained after the instrument calibration can be used for correcting a sample mass spectrometric detection result. The calibration kit can effectively correct a low-mass area of a MALDI mass spectrometer in negative ion mode; mass spectrum signals are free of background interference; accurate measurement of the mass of a small molecule compound can be realized; and a relative error is less than 6 ppm.

Claims

1. A mass calibration kit for a low-mass area of a high-resolution mass spectrometer in a negative ion mode, comprising: a semiconductor nanometer material suspension, a free fatty acid standard solution, and a MALDI sample target cleaning solution, wherein, the semiconductor nanometer material is ZnO, (Bi.sub.2O.sub.3).sub.0.07(CoO).sub.0.03(ZnO).sub.0.9, BN, AIN, TiO.sub.2, or Ga.sub.2O.sub.3.

2. The mass calibration kit according to claim 1, wherein a solvent of the semiconductor nanometer material suspension is isopropanol.

3. The mass calibration kit according to claim 1, wherein the free fatty acid standard solution comprises nine types of free fatty acids in total, namely, a free fatty acid C6:0, a free fatty acid C8:0, a free fatty acid C10:0, a free fatty acid C12:0, a free fatty acid C14:0, a free fatty acid C16:0, a free fatty acid C18:0, a free fatty acid C20:0, and a free fatty acid C22:0, and the nine types of free fatty acids have a same amount of substance.

4. The mass calibration kit according to claim 1, wherein a solvent of the free fatty acid standard solution is normal hexane.

5. The mass calibration kit according to claim 1, wherein components of the MALDI sample target cleaning solution are acetone whose volume concentration is 50% and normal hexane whose volume concentration is 50%.

6. A calibration method of the mass calibration kit for a low-mass area of a high-resolution mass spectrometer in a negative ion mode according to claim 1, comprising the following steps: (1) cleaning a MALDI mass spectrometer sample target by using a MALDI sample target cleaning solution, adjusting a sample target voltage, a hexapole voltage, an ion extraction voltage, and a slit voltage in a mass spectrometer ion source, so that a voltage difference between the sample target and the slit is 20 volts; (2) dripping 1 microliter of semiconductor nanometer material suspension onto a surface of e sample target, keeping the sample target at a room temperature, and after a solvent in the semiconductor nanometer material suspension is completely volatilized and dried, obtaining a sample target whose surface is covered by a semiconductor nanometer material; (3) taking 1 microliter of free fatty acid standard solution to drip it onto a surface of the semiconductor nanometer material on the sample target of step (2), after a solvent in the free fatty acid standard solution is completely volatilized and dried, placing the sample target into a mass spectrometer, and performing mass calibration in a correction mode of the mass spectrometer, wherein a calibration coefficient obtained after the mass spectrometer is corrected may be automatically used to correct a result of sample mass spectrometric detection; wherein the process of the sample mass spectrometric detection comprises: taking 1 microliter of a sample solution to drip it onto the surface of the semiconductor nanometer material of the sample target, naturally drying it, and placing the sample target into the mass spectrometer to perform the sample mass spectrometric detection.

7. The calibration method according to claim 6, wherein the mass calibration is real-time calibration or off-line calibration.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is a diagram of mass-spectral peaks that are generated by nine types of free fatty acids C6:0, C8:0, C10:0, C12:0, C14:0, C16:0, C18:0, C20:0, and C22:0 which are uniformly different from each other by 28 Da.

(2) FIG. 2 is a diagram of a sample mass spectrum in Embodiment 1.

(3) FIG. 3 is a diagram of a sample mass spectrum in Embodiment 2.

DESCRIPTION OF THE EMBODIMENTS

(4) In order to provide a better understanding of the present invention, the embodiments will be described in the following to further elaborate the content of the present invention, but the content of the present invention is not limited by the following embodiments.

(5) In the following embodiments, a calibration method for a low-mass area of a high-resolution mass spectrometer in a negative ion mode is used and includes the following steps:

(6) (1) cleaning a MALDI mass spectrometer sample target by using a MALDI sample target cleaning solution, adjusting a sample target voltage, a hexapole voltage, an ion extraction voltage, and a slit voltage in a mass spectrometer ion source, so that a voltage difference between the sample target and the slit is 20 volts;

(7) (2) dripping 1 microliter of semiconductor nanometer material suspension onto a surface of the sample target, keeping the sample target at a room temperature, and after a solvent in the semiconductor nanometer material suspension is completely volatilized and dried, obtaining a sample target whose surface is covered by a semiconductor nanometer material;

(8) (3) taking 1 microliter of free fatty acid standard solution to drip it onto a surface of the semiconductor nanometer material on the sample target of step (2), after a solvent in the free fatty acid standard solution is completely volatilized and dried, placing the sample target into a mass spectrometer, and performing mass calibration in a correction mode of the mass spectrometer, to obtain a diagram of mass-spectral peaks that are generated by nine types of free fatty acids C6:0, C8:0, C10:0, C12:0, C14:0, C16:0, C18:0, C20:0, and C22:0 which are uniformly different from each other by 28 Da, as shown in FIG. 1.

(9) Preparation of the foregoing semiconductor nanometer material suspension includes: weighing 10 mg of semiconductor nanometer particles (Bi.sub.2O.sub.3).sub.0.07(CoO).sub.0.03(ZnO).sub.0.9, dissolving them in 1 mL of isopropanol, and performing ultrasonic oscillation for 1 minute to uniformly disperse the nanometer particles.

(10) Preparation of the foregoing free fatty acid standard solution includes: taking a free fatty acid C6:0, a free fatty acid C8:0, a free fatty acid C10:0, a free fatty acid C12:0, a free fatty acid C14:0, a free fatty acid C16:0, a free fatty acid C18:0, a free fatty acid C20:0, and a free fatty acid C22:0 that have a same amount of substance, and using normal hexane as a solvent to obtain a free fatty acid standard solution whose solution concentration is 5 mg/mL.

(11) Components of the foregoing MALDI sample target cleaning solution are acetone whose volume concentration is 50% and normal hexane whose volume concentration is 50%.

(12) The foregoing mass calibration is real-time calibration or off-line correction.

(13) Embodiment 1

(14) Mass spectrometric detection of oestrogen diethylstilbestrol includes the following specific operation steps:

(15) (1) preparing a sample solution: weighing 100 mg of diethylstilbestrol and dissolving it in 1 mL of ethanol;

(16) (2) transferring 1 microliter of the sample solution onto a surface of a semiconductor nanometer material that covers the foregoing sample target, and naturally drying it;

(17) (3) adjusting a sample target, a hexapole, an extraction plate, and a slit voltage, so that a voltage difference between the sample target and the slit is 20 volts; placing the sample target into the mass spectrometer to perform mass-spectral detection, and after correcting a detection result by using a diagram of mass-spectral peaks of C6:0, C8:0, C10:0, C12:0, C14:0, C16:0, C18:0, C20:0, and C22:0, a diagram of a sample mass spectrum is obtained, as shown in FIG. 2.

(18) A combination of results of FIG. 1 and FIG. 2 indicates that: the mass spectrum has a signal free of background interference, uniform mass distribution, high accuracy, high resolution, and a stable property. After mass calibration, accurate measurement can be performed on measuring the mass of a small-molecule compound (such as oestrogen diethylstilbestrol), and a relative error between an actual detected value and a theoretical value is less than 6 ppm.

(19) Embodiment 2

(20) Mass spectrometric detection of phytohormone gibberellin includes the following specific operation steps:

(21) (1) preparing a sample solution: weighing 100 mg of gibberellin and dissolving it in 1 mL of ethanol;

(22) (2) transferring 1 microliter of the sample solution onto a surface of a semiconductor nanometer material that covers the foregoing sample target, and naturally drying it;

(23) (3) adjusting a sample target, a hexapole, an extraction plate, and a slit voltage, so that a voltage difference between the sample target and the slit is 20 volts; placing the sample target into the mass spectrometer to perform mass-spectral detection, and after correcting a detection result by using a diagram of mass-spectral peaks of C6:0, C8:0, C10:0, C12:0, C14:0, C16:0, C18:0, C20:0, and C22:0, a diagram of a sample mass spectrum is obtained, as shown in FIG. 3.

(24) A combination of results of FIG. 1 and FIG. 3 indicates that: the mass spectrum has a signal free of background interference, uniform mass distribution, high accuracy, high resolution, and a stable property. After mass calibration, accurate measurement can be performed on measuring the mass of a small-molecule compound (such as gibberellin), and a relative error between an actual detected value and a theoretical value is less than 6 ppm.

(25) Apparently, the aforementioned embodiments are merely used as examples for describing the present invention more clearly, and are not used to limit the method for implementation. To those having ordinary skill in the art, various modifications and variations can be made based on the above description. All possible implementations could not and need not be exhaustively listed here. Therefore, all the obvious modifications and variations derived froze here still fall within the protective scope of the present invention.