TWI334027B - Mass spectrometric analysis for detecting analytes in a solid-state sample - Google Patents

Description

1334027 IX. Description of the Invention: [Technical Field of the Invention] The present invention relates to an analysis method, and more particularly to a mass spectrometry method for detecting a test object in a solid sample. [Prior Art] Generally, when qualitative analysis is performed on a solid sample containing an unknown analyte, several instruments or classical analysis methods are used one after another. Each method is implemented in a different manner, and the manners of the test samples suitable for the method are different. Therefore, the solid sample is separately configured into an appropriate test sample according to various analytical methods to be performed. It is an extremely time consuming and cumbersome workflow. Since a larger number of analytical methods are suitable for the sample to be tested in the solution state (hereinafter referred to as "the solution to be tested"), a solid sample containing the unknown analyte is first dissolved in a suitable solvent to form a sample. The detection of solutions to facilitate the detection of a variety of analytical methods is often cited as the first most popular practice. However, the solvent used must be capable of dissolving the solid sample, but the risk is that there may be a chemical reaction between the solvent and the solid sample, otherwise the analysis results will be unreliable. Therefore, the preparation of the solution to be tested is not only difficult but also cumbersome and time consuming. Mass spectrometry (hereinafter referred to as "MS analysis") is the main technique for qualitative analysis. Some MS analysis [hereafter introduced in its free mode, desorption electrospray ionization (DESI) or direct atmospheric pressure chemical ionization (DAPCI)] is not required Under the premise of dissolving 5 1334027 of the solid sample, the solid sample is directly detected by an unknown sample, but the solid sample can not be transferred to other analytical methods. Therefore, when the results of these MS analyses are insufficient to identify the unambiguous identity of the unknown analytes, the operator is still inevitably preparing the solution to be tested. In addition, the electrospray-assisted laser desorption and deionization (eiectrospray_assisted laser des〇rpti〇ni〇nizati〇n, hereinafter abbreviated as ELDI) technology previously developed by the applicant of the present invention can be made by a pair of ingots. The medicine is irradiated with a laser beam, and the object to be tested is desorbed and combined with an electrically sprayed charged particle to be ionized, and then a mass spectrometry result is obtained; however, some are scattered ( A solid sample such as a powder (hereinafter referred to as a "powder sample") is not easily crushed into an ingot, and when a laser sample is directly fired by a laser beam, it will be scattered around, making most of the Powder samples are wasted, and only a small portion of the analytes are successfully ionized. Based on the above troubles, it is apparent that the mass spectrometry method required is that the solid sample used can also be used by other analytical methods, so that the solid sample can be used repeatedly and economically, especially when the amount is extremely limited; It also eliminates the inconvenience of dissolving the solid sample. SUMMARY OF THE INVENTION Accordingly, it is an object of the present invention to provide a mass spectrometry method for detecting a test object in a solid sample that meets the above needs. The method for mass spectrometry of the present invention for detecting a sample to be tested comprises: (a) providing a solid sample containing a plurality of analytes to be desorbed and freed for mass spectrometry; (b) providing An electric spraying unit, the electric spraying unit 6 1334027

a container having an electrosprayable medium, and a nozzle in fluid communication with the container, (c) providing a mass analyzer that is remote from the nozzle and is used for receiving and analyzing (d) providing a detector for detecting a signal generated by the object to be tested analyzed by the mass spectrometer, and (d) providing a detector for the object to be detected Generating a mass spectrometry result; (e) generating a potential difference between the nozzle and the mass analyzer, so that the nozzle extracts the electricity (4) medium and forms a plurality of tiny charged particles as charge donors, and the charged particles are The potential difference is moved along the charge transfer path toward the mass analyzer; and (1) using a laser beam to shoot the sample such that at least one analyte in the sample is desorbed and along a Flying along the flight path intersecting the charge transfer path, and causing the at least one object to be tested to be freed due to charge transfer after contacting at least one of the charged particles, and guided to the mass under the potential difference analysis Receiving and moving the mass analyzer; characterized in that:

The step (4) of the mass spectrometry method is to pressurize the test object and an additive to form the sample; wherein the additive is selected from a metal halide, a bromide planer, an iodized planer, or One of the combinations. In the present document, "subject" refers to any substance contained in the solid sample that can be desorbed by the shot of the laser beam. By the method of the present invention, the solid sample formed by pressing the test object with the additive can not only be scattered when receiving the laser beam, and the solid sample can be further Transfer to IR analysis 'Saves the amount of sample used and also eliminates the need to dissolve the sample. 7 1334027. [Embodiment] According to the method of the present invention described above, an object to be tested which is originally in a null state can be successfully fixed. Mass spectrometry was performed in the solid sample, and the solid sample was also used for IR spectral analysis. Preferably, the step (a) of the method of the present invention is to pressurize the object to be tested, an additive, and a carbon powder, so that the object to be tested can be easily removed by the aid of the carbon powder. In addition, the limitation of the carbon content is such that the solid sample can still have the transmittance allowed by IR spectrum analysis. On the other hand, the applicant also predicted that when the solid sample contains carbon powder, a laser beam with a lower radiation energy should be used. Further preferably, the method of the present invention further comprises a step (a') before the step (a), which is to atomize the analyte and the additive; for example, to grind the solution The particle size of the analyte and the additive is smaller to allow the analyte to be more evenly distributed, thereby increasing the chance that the analyte is shot by the laser beam. When the test object and the additive are mixed and ground to form a 'kun compound, the step (a) is continued to pressurize the mixture to form a solid sample in the form of a disk. Based on this practice, the weight ratio between the analyte and the additive should be substantially unlimited, as long as the laser beam can be fired onto the object to be tested and successfully desorbed for deuteration, ie, The more the amount of the test object is +, the closer the 疋 is to the surface of the solid sample, the higher the probability of being shot by the laser beam. Preferably, the weight ratio of the analyte to the additive is above the toilet; preferably, it is between 0. 01 and 1 GG, and more preferably, < 0.01 to 〇. In the embodiment of the method of the present invention, the weight ratio of the analyte to the 8 1334027 additive is 〇 〇1. Additives can be easily used to be chemically reacted by dust*" and the properties are stable and not easy to test with: the responders' such as metal toothings, desertification hammers, and bismuth telluride, which make the point lower than the compound It is easy to be compacted, and the sample with high light transmission is: After the eight methods, it is also advantageous for IR spectrum analysis. Preferably, the additives are fNarn^, and potassium bromide (KBr) is used. Sodium chloride), potassium chloride (KC1), potassium telluride (KI), sodium iodide _), or a combination of two. In the embodiment of the method of the present invention, the additive used is another worthwhile. When the step (4) is displayed in the positive mode (when the temperature is obtained, the (4) is obtained by (4) The analyte 1% proton (H) is bound, rather than the analyte ion formed by combining the positive ions (such as alkali metal ions such as +, K) in the additive, so the obtained mass spectrum is simpler and more It is conducive to improving the accuracy and accuracy of the analysis results. In addition, the laser beam of the (1) step is optional infrared laser, ultraviolet laser, milk laser, argon ion laser, krypton laser, carbon dioxide thunder. Shot, or Shiquanshi laser. The laser beam used in the embodiment of the method of the present invention is an infrared laser. The energy of the laser beam described herein means "radiation energy" ° The applicable energy of the laser beam of step t is based on the principle that the object to be tested can be taken off from the solid sample. Preferably, the energy of the laser beam is above 10 "J; more preferably, Is between 25 W and 9 1334027 ;; in the method of the invention The energy of the laser beam used in the embodiment is 6 mJ. In addition, the electric squirting medium in the method (8) of the present invention complicates the mass spectrometry analysis in order to avoid the infiltration of other positive ions such as bismuth, antimony and the like. Preferably, the aqueous solution containing a volatile liquid may be used, and the volatile liquid may be a combination of acetonitrile, acetone, an alcohol, or the like; more preferably an alcohol such as methanol. Further, in the step (6) In the positive mode, in order to improve the dissociation efficiency of the analyte to be desorbed, it is preferred that the electrospray medium in the step (8) is more containing an acid, and the acid can be used, for example, formic acid, acetic acid, or the like. An acetic acid, or a combination thereof, is used in the embodiment of the method of the present invention, and is an aqueous solution containing 5 Gvg of 1% methanol and g.1vq of 1% acetic acid. Others are used in the steps (4) to (1). The relevant components in the EFI unit, as well as the mass analyzer, the debt detector, and other related operating parameters, etc., are well known; reference may be made to the previously mentioned application number 〇95Η) 3439 National patent case In this case the text are incorporated by reference to the case of the embodiment literature. <Examples> The following is a description of how to carry out the mass spectrometry method of the present invention for measuring the analyte in the solid sample mouth by the embodiment; however, the examples are for illustrative purposes and should not be used. It is to be construed as limiting the implementation of the invention. In addition, each embodiment performs the steps (4) to (f) in the mass spectrometry method of the present invention for the measurement of the solid-state sample 丨+ (not shown) by using a mass spectrometer as shown in FIG. . The mass spectrometer comprises a carrier platform 2, an electric 10 1334027 spray unit 3, a laser device capable of emitting a laser light * μ, a cable mechanism 4 of the town 9 fruit 41, a mass analyzer 5 ' and a detection 6. J: in the middle, 此 τ τ 〒 该 'The loading platform 2 is used to put the solid sample i ^ on the top and the other parts of the function and setting details, is the method (4) ~ (f) steps of the present invention The statements in the same are the same, and they are expressed together. In each of the examples, the method of the present invention described below is carried out under the conditions of normal temperature and normal pressure, and the following method: (g) taking 1 gram of KBr as an additive, After mixing with a specific test substance of 1 gram, it is ground into a fine powder mixture by mortar and pestle; (a) The powder mixture is transferred to a tablet press at a pressure of 8 Torr. The mixture is solidified to obtain a solid sample in the form of a disk having a diameter of 13 mm; (b) an electric spraying unit 3 is provided, the electric spraying unit 3 having an electric spraying medium 31 a container 32, and a nozzle 33 in fluid communication with the container 32; (c) providing a mass analyzer 5 disposed away from the nozzle 33 for receiving and analyzing from the sample J a desorbed and free object to be tested; (d) providing a detector 6 for detecting a signal generated by the analyte analyzed by the mass analyzer 5 and generating a mass spectrometry The result 9 (e) causes a potential difference between the nozzle 33 and the mass analyzer 5 to cause the nozzle 33 to spray the electric spray medium 31, and a plurality of tiny charged particles acting as charge donors, the charged particles moving along the charge transfer path toward the mass analyzer 5 under the action of the potential difference 11 1334027; and (f) using a laser beam 41 To shoot the sample 丨 (each time a spot is formed at a different point on the surface of the solid sample 1 to thereby obtain a fresh spot/thumb), such that at least the sample 1 is desorbed and Flying along a flight path intersecting the charge transfer path, and causing the at least one analyte to contact at least one of the charged particles, (4) the charge is transferred and freed, and guided by the potential difference, Moving towards the mass analyzer 5 and being received by the mass analyzer 5. In each of the examples, the analysis results of the primary mass spectrometry and the secondary mass spectrometry are respectively obtained (the results of the analysis of the sample of the sample) are performed, and the primary/secondary mass spectrometry is set by the operation of the mass analyzer. . [Chemicals] The following examples of the present invention are prepared by using the following chemicals or equipment: L laser mechanism (source of laser beam): infrared laser (IR laser, hereinafter referred to as 1R laser) 'by Russia Manufactured by LOTIS TII, the $ is LS-2130SHP. The wavelength of the emitted laser beam is 1〇64 nm 'The frequency is 2 Hz' The pulse length is 9 ns, and the energy per radiant is 6 mJ 〇2. The quality of the knives is arbitrarily (including enough): Ion trap Mass analyzer by Germany

Manufactured by BrUker Dalt〇n, model Esquire Plus 3000. The scanning rate is one second each time (2 〇〇 ms/scan), and the scanning result of 5 knives is continuously recorded, and the mass spectrum is presented in an averaging mode. 12 1334027 3. Electrospray medium: a 50% by weight aqueous solution of decyl alcohol containing 0.1% by weight of acetic acid. Among them, methanol and acetic acid were manufactured by Sigma-Aldrich, USA, as HPLC grade. 4. Test object: a. Blood matrix (hemin) - molecular weight 652.0, manufactured by Sigma-Aldrich, USA, model number 16009-13-5. b. - 2'-deoxyadenosine monohydrate - molecular weight 269.26, manufactured by Sigma, USA, model #16373-93-6. c. 2-Aminobenzimidazole--Molecular weight: 133,15, manufactured by Tokyo Chemical Industry Co., Ltd., Japan, model number A0850. d. 3-aminoquinoline--molecular weight: 144.17, manufactured by Tokyo Chemical Industry Co., Ltd., model A0418 〇e. 1,12-dodecanediamine (l,12 -dodecanediamine) - Molecular Weight: 200.36, manufactured by Tokyo Chemical Industry Co., Ltd., Japan, model number D0091. f. cytochrome c - molecular weight 12230, manufactured by Sigma-Aldrich, USA, HPLC grade. Unless otherwise stated, each embodiment is carried out in a positive mode at normal temperature and pressure, and the electrospray medium used is an aqueous solution containing % vol% methanol and 0.1 vol% acetic acid; the laser beam is The radiati〇n energy is an infrared laser of 6 mJ; the solid sample is translucent. 13 [Examples 1 to 6] The types of analytes selected for each example and the obtained mass spectrum map numbers are not listed in Table 1. It is worth mentioning that, due to the molecular weight of the analyte other than cytochrome All are deleted below, based on the applicant's practical experience 'except for the analyte ions formed by cytochrome will have a multi-valent positive charge', other analyte ions should only carry a positive valence charge; and each charge A contribution to a proton (H+).

The results obtained in the respective examples are respectively explained. [Example 1] The structure of the blood matrix is also as follows:

The blood matrix of g and 〇.1 g of the solid sample of Example 1 (containing 〇〇〇1 14 1334027 KB r) was dark brown. As far as the results of Fig. 2(a) are concerned, only the ion peak with a w々 value of 6ΐ62 is present, which is the analyte ion (AH+ 'A table to be tested) formed after the blood matrix is connected to the proton. The generated signal. Referring to Fig. 2(b), the main ion peaks (indicated as 557 ι, 498, and 418.6) indicate that the structure of the blood matrix has been broken during secondary mass spectrometry, and m々 The ion of the analyte represented by the ion peak of 557.1 is the +1 valent ion formed by the structure of the blood matrix breaking [C2〇2H3], and the content of the +1 valent ion is also the most; the m々 value of 498.1 is the blood matrix. The +1 valent ion formed after the structure of [C4〇4H6] is broken; and the 々 value of 418 6 represents the +1 valent ion formed after the structure of the blood matrix breaks [0^()04%4] 〇 [Example 2] The structure of the water-reoxygen gland with one water 2·-deoxygen gland as the test substance is as follows:

In Fig. 3(a), the two ion peaks with a claw value of 252 〇 and I% 主要 are mainly present; 252 of them represent a water 2·-deoxygen gland after losing one water molecule and attaching a proton The ions formed, while the 136.0 represents a water 2, and the deoxygen gland breaks the +1 valent ion formed by the structure of [C:5〇3H8]. This indicates that the energy of the laser beam has broken the structure of some of the analyte molecules. The amount of analyte ions with m/ζ value of 252.0 is still the majority. However, in Figure 3(b), only the ion peak signal with a value of 136 々 is seen, and the ion peak signal with m/ζ value of 252.0 seems to have disappeared. [Example 3] 2-Amino-benzimidazole as a test substance The structural formula of 2-aminobenzimidazole is as follows:

NΗ / ~NH2 is produced in Figure 4(a), which is similar to Figure 2 (4) of Example 1, which only produces the analyte ion (10) value of 134.0 formed by the 2-aminobenzopyrene attached to the proton. Signal. In Fig. 4(8), the μ value which is not in Fig. 4(a) is 1〇5" (the ion represented by it is 2-aminobenzene and the saliva breaks off the structure of [C2H4].

+1 > (Bei Ions) and 91.1 (2-Amino benzopyrene cleavage off the +1 ionic ion formed by the structure of [(:3116]), and the _ value is 134 〇 It seems to have disappeared. [Example 4] The structural formula of 3-aminoquinoline as the test substance 3-aminoquinoline is as follows: In Figure 5 (4), the & 3_amine base is attached to the proton The formed analyte 16 1334027 is the most intense ion peak, and the ion represented by the ion peak signal with a m々 value of 176.9 may be 3 aminoquinoline combined with a methanol molecule. The ions formed after a proton, or formed by impurities. In Figure 5(b), the intensity of the ion peak with a m々 value of 145.0 is too low to be observed, and the ion peak intensity in the figure is The higher value is 118 〇, and the ion represented by it is the +1 valent ion formed by the structure of 3_aminoquinoline cleavage [C^2], and the ion represented by the w々 value of 9L1 is It is a +1 valent ion formed by the structure of [C4H5] after the cleavage of [C4H5]. [Example 5] The structure of M2-dodecanediamine with ι, ΐ2·dodecanediamine as the analyte Such as Shown: H2NCH2(CH2)1〇CH2NH2 In Figure 6(4), the analyte ion (m/z value 201.1) formed by the 142·12-burned diamine attached to the -f is the highest intensity ion. Peak, and the ion peak signal with m/z value of 184.2, the representative ion may be the amine group at the two ends of the structure of u2_ dodecanediamine. The amine group is removed (νη^ combines to form a cyclic molecule). After that, the secondary amine group thereon is reconnected with ions formed by protons. In Fig. 6(b), the ion peak with m々 value of 201.1 seems to have disappeared, and the peak value of the ion peak intensity in the figure is the highest. 184.1. [Example 6] using cytochrome c as the analyte in Fig. 7, for example, w々 value is 767 6q value is 16), 817.00 value is 15), 875.3Q value is 14), 942 1q Ion peaks with values of 13), 1020.0 U values of 12) 'and 1128.9 U values of u). These 17 1334027 12228±2, compared with the original manufacturer's 12230, are very qualitative. The molecular weight obtained after the § ten nose is the molecular weight value of the provided cytochrome C standard. It is apparent that Example 6 demonstrates that the method of the present invention can indeed successfully desorb the analyte therein from the solid sample containing KBr and the analyte and perform mass spectrometry.

The mass spectrometry method for the analyte to be detected by the above embodiments is used to detect the fact that a solid sample has the following advantages. The shell test result has a credibility. The results of the spectral analysis are in full compliance with the test object used. Obviously, the results of the test are actually comparable to those of the tester. In addition, in all the mass spectra obtained in the examples, there is no such thing as Ακ+ When the resulting signal 'confirmed that the fine version is an additive, the result of the method of the present invention is not

The simpleness of the map and the ease of interpretation will affect the simplicity of the mass spectrum due to the presence of unloading, which is also in line with the requirements of mass spectrometry in the field of mass spectrometry. 2. Saving the amount of use and increasing the chemical points. (IV) Convenience of the industry: For each of the examples, the waiting object only uses G.GG1 g, and it is uniformly present in the solid sample to which it belongs. The method of the invention requires only a small amount of the object to be tested to successfully achieve the purpose of the pre-measurement; in addition, if β deliberately presses the object to be tested on the surface of the solid sample, then the beam is applied: Yes - the method can be used ' ^ The demand for the object to be tested will be lower' and the IU sample can be reused for ir photodegradation, which can save the amount of the analyte and increase the chemical analysis Convenience. Because 18 丄 this field is extremely small, it is suitable for the mass spectrometry analysis by the method of the present invention, which also represents the use of Α π water. The moon-issuing method is extremely advantageous for criminal identification. According to the above description, the method LV _y of the present invention is performed by pressing an object to be tested (4) into an (-) sample with an additive, and then applying a laser beam thereto. Shooting for dislocation, and then the desorbed analyte is ionized by electro-spraying. It has been proved that:

High mass spectrometry results. Furthermore, the method of the present invention is more advantageous in that the method of the present invention has the advantages of "pumping test =::degree" and the map is simple and convenient for interpretation"; "saving the test to be tested: the convenience of increasing the convenience of chemical analysis work", and the like It facilitates the chemical analysis of unknowns and the conduct of criminal identification. The above-mentioned ones are only the preferred embodiments of the present invention, and the scope of the present invention cannot be limited thereto, that is, the application according to the present invention: the simple equivalent of the content of the invention and the description of the invention Variations and modifications are still within the scope of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a mass spectrometer used in the mass spectrometry method for testing a test object in a solid sample, illustrating the relevant positions of the components required for the method of the present invention; Is a mass spectrum containing t 2 (4 and 2 (8), respectively illustrating the results of the method of the present invention, wherein Figure 2 (4) is performed -: under-mass spectrometry, and Figure 2 (b) is subjected to secondary mass spectrometry; Is a mass spectrum containing 3 (hook and 3 (b), respectively illustrating the results of the method of the present invention carried out in Example 2, wherein Figure 3 (a) is carried out - mass spectrometry 19 1334027, Figure 3 (b) is carried out Secondary mass spectrometry; Figure 4 is a mass spectrum containing 4(a) and 4(b), respectively illustrating the results of Example 3 of the method of the present invention, wherein Figure 4(a) is a mass spectrometry analysis, Figure 4 (b) is a secondary mass spectrometry; FIG. 5 is a mass spectrum comprising 5 (a) and 5 (b), respectively illustrating the results of the method of the present invention performed in Example 4, wherein FIG. 5(a) is performed once Mass spectrometry, Figure 5 (b) is a secondary mass spectrometry analysis; Figure 6 is a mass spectrum containing 6 (a) and 6 (b), respectively illustrating the present invention The results of Example 5 were carried out, wherein FIG. 6(a) is performed once for mass spectrometry, FIG. 6(b) is subjected to secondary mass spectrometry; and FIG. 7 is a mass spectrum showing that the method of the present invention is carried out in Example 6. result.

20 1334027 [Description of main component symbols] 2 Carrier platform 4 Laser mechanism 3 Electrospray unit 41 Laser beam 31 Electrospray media 5 Mass analyzer 32 Container 6 Detector 33 Nozzle 21

Claims (1)

1334027 -----_______ Patent application No. 096115324 added, amended the description of the non-lined statement replacement page year > month (. 曰correct winter ten, the scope of patent application: 'fixed spoon = used to measure a solid Mass spectrometric analysis method of the analyte in the sample, (for 3 solid samples to be desorbed and freed to be analyzed by mass spectrometry ^ test object; (b) providing an electrospray unit, the electrospray ^ 70 has - a container accommodating an electrospray medium and a nozzle in fluid communication with the container; (c) providing a mass analyzer disposed away from the nozzle for receiving and analyzing the sample a detector that is desorbed and free; (4) providing a detector that analyzes the signal generated by the object to be tested by using a (four) (four) amount analyzer, and produces a mass spectrometry result; (e) a potential difference is generated between the nozzle and the mass analyzer, so that the nozzle sprays the electrospray medium and forms: a small charged particle as a charge donor, the charged particles move in the separator; and (f ) using a path toward the mass score ~ from ( Shooting the sample with each shot first beam such that at least one of the analytes is desorbed and flies along a flight path that intersects the charge transfer path and causes the at least one object to be tested to = at least one of the contacts will be guided by the free difference due to the charge transfer, moving towards the mass analyzer and being received by the mass knives; characterized in that: (4) the step of the Ζ spectrum analysis method is The sample to be tested and the metal toothing of the additive; wherein the additive is selected from the group consisting of a metal tooth, an evolution planer, an iodized planer, or the like. 〉 Item σ The quality of the male and female ploughing, the scallop-knife analysis method, wherein the (4) step of the analysis method is to add, correct, and correct the test object, the additive, and the patent application No. 096115324 The specification of the line replaces the stomach, and the carbon powder is pressed and fixed. The date of correction is August, 1999. 3. According to the mass spectrometry method described in the first item of the patent scope, wherein the mass spectrometry method further includes An implementation before Ding 4 (a) (a,) a step of micronizing the analyte and the additive. (In accordance with the mass spectrometry method described in the scope of the patent, wherein the metal halide is potassium chloride, sodium chloride , a combination of gasification, decommissioning, sodium, or a combination thereof. 8.5. According to the method of mass spectrometry described in claim 3, wherein the step (8) is to be mixed After the sample and the additive are ground to form a mixture, the mixture is pressed in the step (4), and the formed sample is in the form of a disk. wherein the sample is in accordance with the fifth item of the patent application. The mass spectrometry method has a weight ratio of the analyte to the additive of 0.001 or more. Wherein, the waiting for, the waiting for, the (7) 7 · according to the scope of the patent application 帛 6 items of the mass spectrometry method and the weight ratio of the additive is between 〇〇ι~ι〇〇8. The mass ratio method of the mass spectrometry method described in the seventh aspect is the ratio of the weight of the analyte to the additive is 〇〇1~"5 9. According to the mass spectrometry method described in the application patent _ 1, the step: the laser beam is an infrared laser , ultraviolet laser: nitrogen two-wind ion laser, krypton laser, carbon dioxide laser, or Shiquan stone laser. 10. The laser beam according to the first step of the patent application scope is red. 11. The mass spectrometry method according to claim 9 of the patent application, wherein the (1) external laser or ultraviolet laser. The mass spectrometry method according to the above, wherein the energy of the laser beam in the step of replacing the page of the syllabus of the sorcerer is replaced by 1 (23) 23 1334027, the patent application No. 096115324, the correction of the unlined specification. ) ^ Above. LjE Date_ "August August (f) 12. According to the mass spectrometry method described in the scope of claim patent, wherein the energy of the laser beam in the step is between 25 and ι〇〇13. The mass spectrometer, 〒, 〒, 歹 (the energy of the laser beam in step 0 is 6 mJ.) 14. According to the mass spectrometry method described in the scope of the patent application, 兮The electrospray medium in the step is an aqueous solution containing a volatile liquid Γ W 1 5. According to the mass spectrometry method described in claim 14 of the patent application, the '1, 5⁄4 volatile liquid is acetonitrile, acetone, alcohol , or a combination thereof, etc. 16. According to the mass spectrometry method of claim 15 of the patent application, the T-volatile liquid is an alcohol. 17. The mass spectrometry method according to claim 16, wherein哕 The volatile liquid is methanol. / 18. The mass spectrometry method according to the scope of claim [01], wherein the electrospray medium in the step (b) of Hunchun is more containing an acid. X 19. According to the scope of the patent application 16 mass spectrometry methods, The electrospray medium contains an alcohol and further contains citric acid, acetic acid, trifluoroacetic acid, or a combination thereof. 20. The mass spectrometry method according to claim 19, wherein the electrospray The sprinkling medium is an aqueous solution containing methanol and acetic acid.