JP6020395B2 - Drug analysis method using chromatographic mass spectrometry - Google Patents

Description

  The present invention relates to a drug analysis method for identifying a drug using a gas chromatograph mass spectrometer or a liquid chromatograph mass spectrometer using a mass spectrometer capable of MS / MS measurement.

  The spread of illegal drugs such as narcotics and stimulants is a global problem. In Japan and other countries, illegal drugs with actions similar to stimulants and cannabis are regulated individually, that is, for each compound. There are many analogs that have changed, and it is difficult to regulate individual compounds. Under these circumstances, so-called comprehensive regulations are being implemented in Japan, the United Kingdom, and some states in the United States, which do not regulate illegal drugs individually, but regulate the main skeleton of compounds.

  For example, in Japan, the Ministry of Health, Labor and Welfare ministerial ordinance for comprehensively regulating synthetic cannabinoid compounds, which are cannabis-like compounds, under the Pharmaceutical Affairs Law was enforced in March 2013 (see Non-Patent Document 1). This ministerial ordinance consists of 772 compounds (1H-indol-3-yl) (naphthalen-1-yl) methanone containing naphthalene ring and indole ring (however, three compounds already regulated as narcotics) Are comprehensively regulated).

  Conventionally, identification of such illegal drugs and poisons has been carried out using a gas chromatograph mass spectrometer (GC-MS) or a liquid chromatograph mass spectrometer (LC-MS). Specifically, a mass spectrum obtained by measuring a target sample using GC-MS or LC-MS is converted into a mass spectrum obtained by measuring a known standard sample or a generally available database. It is common to identify a compound detected in a target sample by collating with a recorded mass spectrum and searching for a compound with a matching or similar spectrum pattern (see Non-Patent Document 2, etc.). .

  However, it is often difficult to obtain a standard sample for illegal drugs that have begun to be newly introduced due to partial modification of the chemical structure as described above. In addition, mass spectra corresponding to such drugs are not recorded in existing databases, and it is not easy to create a database in which mass spectra of all of the many compounds subject to comprehensive regulations are recorded. For this reason, it is quite difficult to identify all the illegal drugs subject to comprehensive regulation and to estimate their structures by the above-described conventional methods using GC-MS and LC-MS.

  As another method, an attempt to estimate the structure of an illegal drug by combining high-precision mass information obtained by a nuclear magnetic resonance (NMR) apparatus, a liquid chromatograph time-of-flight mass spectrometer (LC-TOFMS), or the like is also possible. However, such a method requires preparation of a plurality of devices, the device itself is expensive, and the measurement work and data analysis work are considerably troublesome.

“Regarding partial amendment of ministerial ordinances stipulating designated drugs prescribed in Article 2, Paragraph 14 of the Pharmaceutical Affairs Law and medical use prescribed in Article 76-4 of the Act” (Enforcement Notice), Ministry of Health, Labor and Welfare, [Search Date: August 14, 2013], Internet <URL: http://www.mhlw.go.jp/bunya/iyakuhin/yakubuturanyou/kanren-tuchi/yakuji/dl/H24-04.pdf> Hitoshi Dobashi, “SHIMADZU GC / MS Technical Report No.6“ Automatic Identification and Semiquantitative Analysis of Serum Psychiatric Drugs Using “GC / MS Forensic Toxicology Database” ”, Shimadzu Corporation, [Search Date: Heisei 25 August 14], Internet <URL: http://www.an.shimadzu.co.jp/gcms/support/lib/pdf/c146-0279.pdf>

  The present invention has been made to solve the above-mentioned problems, and the object of the present invention is to use a device that is easily available and relatively inexpensive, and no standard sample exists or information is recorded in a database. Provided is a drug analysis method capable of accurately identifying a specific drug of an untreated naphthoylindole system (that is, having (1H-indol-3-yl) (naphthalen-1-yl) methanone as a basic skeleton) There is to do.

In order to solve the above problems, the present invention uses a chromatographic mass spectrometer capable of MS / MS measurement and uses (1H-indol-3-yl) (naphthalen-1-yl) methanone as a basic skeleton. A method for comprehensively identifying specific compounds to be analyzed, comprising:
a) Using a chromatographic mass spectrometer, the sample containing the compound to be identified is
a1) A naphthoyl produced by cleavage upon ionization is a precursor ion, a naphthalene from which the carbonyl group is eliminated from the naphthoyl is a product ion, and a plurality of MRMs according to the difference in mass of the third functional group bonded to the naphthalene ring A first MRM measurement using transition as a measurement condition, or a first product ion scan measurement using naphthoyl as a precursor ion corresponding to a difference in mass of the third functional group,
a2) The indole carbonyl generated by the cleavage during ionization is a precursor ion, the indole from which the carbonyl group is eliminated from the indole carbonyl is a product ion, and there are multiple numbers corresponding to the difference in mass of the second functional group bonded to the indole ring. A second MRM measurement using the MRM transition as a measurement condition, or a second product ion scan measurement using a precursor ion of indolecarbonyl corresponding to the difference in mass of the second functional group, and
a3) Precursor ion scan measurement using indole carbonyl in the state where the first functional group bonded to nitrogen of the indole ring is removed while the second functional group is added as a product ion,
A measurement execution step for repeatedly executing three types of MS / MS measurements,
b) Created based on data obtained by the first MRM measurement or the first product ion scan measurement, the second MRM measurement or the second product ion scan measurement, and the precursor ion scan measurement, respectively. A compound presence confirmation step for determining the presence of the specific compound having the basic skeleton by determining whether or not a peak is commonly present at the same retention time on the chromatogram corresponding to each type of measurement;
c) The type of the third functional group was obtained by the second MRM measurement or the second product ion scan measurement based on the data obtained by the first MRM measurement or the first product ion scan measurement. The type of the second functional group is estimated based on the data, the type of the first functional group is estimated based on the data obtained by the precursor ion scan measurement, and the compound presence confirmation step is performed based on the estimation results. A structure estimation step for estimating the chemical structure of a specific compound whose existence has been confirmed;
It is characterized by having.

  As a chromatograph mass spectrometer capable of MS / MS measurement, for example, a gas chromatograph or a liquid chromatograph and a tandem quadrupole mass spectrometer (also called a triple quadrupole mass spectrometer) are used. A combined device can be used.

  In the drug analysis method according to the present invention, the second and third functional groups include hydrogen. The first, second and third functional groups include at least a functional group (substituent) specified by the ministerial ordinance described in Non-Patent Document 1. Specifically, the first functional group includes at least a linear alkyl group (-CnH2n + 1) having 3 to 8 carbon atoms (n) and a linear alkenyl group having 5 carbon atoms (- C5H9) and the terminal carbon of a linear alkyl group having 3 to 5 carbon atoms, fluorine (F), chlorine (Cl), bromine (Br), iodine (I), cyano group (-CN) , A group in which any one of a hydroxyl group (—OH) or an acetoxy group (—OCOCH 3) is bonded. The second functional group includes at least two kinds of hydrogen and a methyl group (—CH 3). The third functional group includes at least a linear alkyl group having 1 to 6 carbon atoms (-CnH2n + 1), an alkoxy group having 1 or 2 carbon atoms (-OCnH2n + 1), fluorine (F ), Chlorine (Cl), bromine (Br), and iodine (I).

  In the drug analysis method according to the present invention, when a compound in a sample is introduced into a mass spectrometer and ionized, various ions generated by cleavage of molecular ions are used as precursor ions for MS / MS measurement. Therefore, as an ion source of the mass spectrometer, it is preferable to use an ion source that does not perform so-called soft ionization but easily causes cleavage, typically an electron ionization method. Further, when a liquid chromatograph and a mass spectrometer are combined, an electron ionization method cannot be used, but an ion source having an in-source decomposition function may be used instead.

  When a compound having (1H-indol-3-yl) (naphthalen-1-yl) methanone as a basic skeleton is introduced into a mass spectrometer, a partial bond is broken during ionization, and naphthoyl containing a naphthalene ring An ion or an indolecarbonyl ion containing an indole ring is generated. Of these, naphthoyl contains only the third functional group of the first to third functional groups. Accordingly, naphthoyl containing the third functional group is used as a precursor ion, naphthalene containing the third functional group is used as a product ion, and multiple MRM transitions according to the difference in mass of the third functional group are used as measurement conditions. When the reaction ion monitoring) measurement is performed, ions derived from the original compound are detected in the MRM transition corresponding to the type of the third functional group contained in naphthoyl and naphthalene.

  On the other hand, indolecarbonyl contains a first functional group and a second functional group, but the first functional group is eliminated during ionization. MRM measurement was performed using multiple MRM transitions according to the difference in mass of the second functional group, with indole carbonyl containing the second functional group as the precursor ion and indole containing the second functional group as the product ion. Then, the ion derived from the original compound is detected in the MRM transition according to the type of the second functional group contained in the indole carbonyl and indole.

  Further, indolecarbonyl ions in which both the second functional group and the first functional group are added without ionization of the first functional group during ionization are also generated during ionization. When cleaved by dissociation or the like, the first functional group is preferentially eliminated. The mass of indole carbonyl after the first functional group is eliminated depends on the type of the second functional group, but since there are at most two kinds of the second functional group, the mass after the first functional group is eliminated. The mass-to-charge ratio of the indolecarbonyl ion, that is, the product ion, is one of two. Therefore, when a precursor ion scan measurement is performed in which these two mass-to-charge ratios are determined as product ions, the precursor ion spectrum has a peak at a position corresponding to the mass of indolecarbonyl before the first functional group is eliminated. Appears. The type of the first functional group can be specified from the difference between the detected mass and the mass set for the product ion.

  In the measurement execution step in the drug analysis method according to the present invention, for example, three types of MS / MS measurements determined in advance as described above are executed on the sample. When the target compound is contained in the sample, a chromatogram obtained corresponding to one MRM transition in the first MRM measurement, a chromatogram obtained corresponding to one MRM transition in the second MRM measurement, and A peak appears at the same retention time of the chromatogram obtained corresponding to one product ion in the precursor ion scan measurement. Therefore, in the compound presence confirmation step, it is determined whether or not the specific compound having the basic skeleton exists by determining whether or not these three types of common peaks exist at the same retention time on the chromatogram. To do. If no peaks are obtained at the same retention time, it means that at least one of the functional groups is missing.

  In the case of a specific compound, in the structure estimation step, the type of the third functional group is estimated based on which MRM transition in the first MRM measurement the peak is detected, and which MRM in the second MRM measurement is estimated. The type of the second functional group is estimated based on whether the peak is detected in the transition, and further, based on the position (mass-to-charge ratio) where the peak is detected on the map of the precursor ion obtained by the precursor ion scan measurement. Estimate the type of one functional group. Then, by combining these estimation results, the chemical structure of the specific compound whose existence has been confirmed is estimated, and the target compound is identified.

  Both the first and second MRM measurements may be product ion scan measurements. That is, instead of the first MRM measurement, the first product ion scan measurement using naphthoyl as a precursor ion according to the difference in the mass of the third functional group may be performed, and instead of the second MRM measurement, What is necessary is just to perform the 2nd product ion scan measurement which used indolecarbonyl as a precursor ion according to the difference in the mass of a 2nd functional group.

  In addition, in the drug analysis method according to the present invention, although it is possible to identify a large number of compounds having different combinations of three types of functional groups, mass spectra are recorded in existing drug databases for some drugs, It is more efficient to identify such drugs by database search. In addition, by using database search, it is possible to identify drugs other than naphthoylindoles.

Therefore, in the drug analysis method according to the present invention, preferably,
In the measurement execution step, a normal scan measurement that does not intentionally cleave ions derived from the sample is repeatedly performed in combination with the three types of MS / MS measurements,
A database utilization identification step of identifying a known compound by collating a mass spectrum created based on the data obtained by the normal scan measurement with a drug database;
In the structure estimation step, structure estimation for an unknown compound that is not recorded in the drug database may be performed.

  Here, “normal scan measurement that does not intentionally cleave ions from the sample” means that, for example, unintentional cleavage of ions at the ion source by the electron ionization method is allowed, and intentional cleavage in a collision cell or the like. A scan measurement that is performed without any manipulation. For example, in a tandem quadrupole mass spectrometer, no voltage is applied to the collision cell, and mass separation is not performed in the first quadrupole mass filter, but only in the second quadrupole mass filter. What is necessary is just to carry out scan measurement for scanning.

  According to the preferred method described above, since the compounds recorded in the drug database are identified by database search based on mass spectrum, naphthoylindole drugs can be efficiently identified and recorded in the database. Drugs other than naphthoylindoles can also be identified.

  According to the drug analysis method of the present invention, even when a standard sample is not available and a database for collating mass spectra or the like is not prepared, the gas chromatograph tandem quadrupole mass spectrometer is Using only such a relatively inexpensive device, the chemical structure of a naphthoylindole drug that is the subject of comprehensive regulation can be estimated. In addition, since the apparatus itself is easily available and various settings and operations for measurement are simple, it is convenient to use widely while suppressing costs.

BRIEF DESCRIPTION OF THE DRAWINGS The schematic block diagram of one Example of GC-MS / MS system which implements the drug analysis method based on this invention. The flowchart which shows the procedure which implements a comprehensive drug identification process in the GC-MS / MS system of a present Example. The figure which shows the chemical structure of the naphthol indole type | system | group drug which is an identification object. The figure which shows an example of the mass spectrum obtained with respect to the naphthoyl indole type | system | group drug (JWH-020). Explanatory drawing of the detection object fragment | piece in 1st MRM measurement. The figure which shows the correspondence of MRM transition in 1st MRM measurement, and functional group R3 of estimation object. Explanatory drawing of the detection object fragment | piece in 2nd MRM measurement. The figure which shows the correspondence of the MRM transition in 2nd MRM measurement, and the functional group R2 of estimation object. Explanatory drawing of the detection object fragment | piece in precursor ion scan measurement. The figure which shows the correspondence of the precursor ion in the precursor ion scan measurement, and the functional group R1 of estimation object. The figure which shows an example of the chromatogram which is a measurement result, and the peak detection result on this chromatogram. The figure which shows the verification result with respect to the standard sample of 20 types of naphthoyl indole type drugs.

  An embodiment of a drug analysis method according to the present invention will be described with reference to the accompanying drawings. FIG. 1 is a schematic configuration diagram of an embodiment of a GC-MS / MS system for implementing a drug analysis method according to the present invention.

  As shown in FIG. 1, the GC-MS / MS system includes a GC unit 1 and an MS / MS unit 2. The GC unit 1 includes a sample vaporizing chamber 10 that vaporizes a small amount of liquid sample, a column 12 that separates sample components in a time direction, and a column oven 11 that controls the temperature of the column 12. On the other hand, the MS / MS unit 2 includes an ion source 21 that ionizes a sample to be measured by an electron ionization (EI) method in an analysis chamber 20 that is evacuated by a vacuum pump (not shown), and four rods each. A front-stage quadrupole mass filter 22 and a rear-stage quadrupole mass filter 25 composed of electrodes, a collision cell 23 in which a multipole ion guide 24 is disposed, and a detection signal corresponding to the amount of ions by detecting ions Is provided.

  The analysis control unit 33 has a function of controlling the operations of the GC unit 1 and the MS / MS unit 2 under the instruction of the central control unit 34. The central control unit 34 to which the input unit 36 and the display unit 37 are connected is responsible for overall control of the entire system, in addition to the user interface through these. The storage device included in the central control unit 34 stores a comprehensive drug identification control program 35 for performing characteristic control for performing a comprehensive drug identification process to be described later. By controlling each unit through the analysis control unit 33 according to the program 35, measurement and data processing necessary to comprehensively identify the drug are executed. At this time, the detection signal (ion intensity signal) from the detector 26 is input to the data processing unit 30, and the comprehensive drug identification data processing unit 31 included in the data processing unit 30 is stored in the drug database 32. By executing data processing using the information, the various drugs contained in the sample are comprehensively identified.

  The central control unit 34 and the data processing unit 30 can be embodied by executing a dedicated control / processing software installed in the computer using a personal computer as hardware. In this case, the input unit 36 is a keyboard or a pointing device (such as a mouse), and the display unit 37 is a display monitor.

A basic MS / MS measurement operation in the GC-MS / MS system of this embodiment will be schematically described.
When a small amount of sample liquid is dropped into the sample vaporizing chamber 10, the sample liquid is vaporized in a short time, and the compound in the sample is sent into the column 12 on a carrier gas such as helium. While passing through the column 12, each compound in the sample reaches the outlet of the column 12 with a different delay. The column oven 11 is heated according to a predetermined temperature profile. The ion source 21 sequentially ionizes the compounds in the gas supplied from the column 12 outlet. Since the ion source 21 is an EI ion source, a part of the bond of the compound is cleaved (that is, cleavage occurs) at the time of ionization, and ions of various fragments derived from one compound are generated.

  The analysis controller 33 applies a voltage that allows ions having a specific mass-to-charge ratio to pass through the rod electrodes of the front-stage quadrupole mass filter 22 and the rear-stage quadrupole mass filter 25. Thereby, ions having a specific mass-to-charge ratio among various ions derived from the compound pass through the front quadrupole mass filter 22 and are introduced into the collision cell 23. A collision-induced dissociation gas is introduced into the collision cell 23, and ions introduced into the collision cell 23 come into contact with the gas and are cleaved.

  Various product ions generated by the cleavage are introduced into the rear quadrupole mass filter 25 while being converged by the ion guide 24, and product ions having a specific mass-to-charge ratio pass through the rear quadrupole mass filter 25 to detect the detector 26. To reach. A detection signal from the detector 26 is input to the data processing unit 30. The data processing unit 30 creates a mass spectrum, a mass chromatogram, and the like, and performs compound identification as described later.

  Similar to general GC-MS / MS, in the system of the present embodiment, MRM measurement, product ion scan measurement, precursor ion scan measurement, and neutral loss scan measurement are prepared as MS / MS measurement modes. Further, Q1 scan measurement, Q3 scan measurement, Q1-SIM measurement, and Q3-SIM measurement are prepared as normal measurements that do not involve ion cleavage in the collision cell 23. In the comprehensive drug identification process described below, MRM measurement and precursor ion scan measurement are used as MS / MS measurement, and Q3 scan measurement is used as normal measurement.

Hereinafter, a comprehensive naphthoylindole drug identification process performed using the system of this example will be described. FIG. 2 is a flowchart showing the identification procedure.
FIG. 3 shows the chemical structure of the naphthoylindole drug to be identified. As shown in the figure, this compound has (1H-indol-3-yl) (naphthalen-1-yl) methanone as a basic skeleton. A functional group R1 (first functional group) is bonded to the 1-position of the indole ring, a functional group R2 (second functional group) is bonded to the 2-position of the indole ring, and a functional group is bonded to the 4-position of the naphthalene ring. R3 (third functional group) is bonded. In the ministerial ordinance described in Non-Patent Document 1, the types of functional groups R1 and R3 in the compounds included in the designated drug are indicated, and compounds in which the functional group R2 is substituted with a methyl group instead of hydrogen are also included in the designated drug. ing.
Therefore, in order to comprehensively identify this naphthoylindole drug, the following two are required.
[A] Confirmation of (1H-indol-3-yl) (naphthalen-1-yl) methanone as a basic skeleton.
[B] Judgment whether or not the three types of functional groups R1, R2, and R3 correspond to those specified by the ministerial ordinance.

  FIG. 4 is an example of a mass spectrum obtained by scanning (Q3 scan) measurement of a naphthoylindole compound having functional groups R1: C7H15, R2: H, R3: H. When ionization is performed by the electron ionization method, partial bonds in molecular ions are cleaved by excessive energy given from thermal electrons, and peaks derived from various fragments having a mass to charge ratio smaller than that of molecular ions are observed. Since some of the ions generated by unintentional cleavage in the ion source contain only one type of functional group, the type of functional group can be specified by using MS / MS measurement using such ions as precursor ions. Is possible. Therefore, here, the following three types of MS / MS measurements are performed.

[First MRM measurement]
As shown in FIGS. 4 and 5 (a), the naphthoyl produced upon ionization contains only the functional group R3 among the three types of functional groups. Further, as shown in FIG. When target-induced collision-induced dissociation is performed, naphthalene from which the carbonyl group is eliminated is observed with the functional group R3 remaining. In this case, both naphthoyl and naphthalene contain a functional group R3, and the mass-to-charge ratio differs between naphthoyl and naphthalene depending on the type of the functional group R3. Therefore, the first MS / MS measurement is MRM measurement in which MRM transitions are set so that naphthoyl is a precursor ion, naphthalene is a product ion, and the masses of 13 kinds of functional groups R3 added to naphthalene are respectively corresponded. .

  FIG. 6 shows the relationship between the MRM transition and the type of the functional group R3 in the first MRM measurement. That is, if ions are observed in any one of these 13 types of MRM transitions, it can be said that the functional group corresponding to the MRM transition is likely to be the functional group R3.

[Second MRM measurement]
Of the three types of functional groups, the functional group R1 was eliminated during ionization, and when collision-induced dissociation was performed targeting indolecarbonyl containing only the functional group R2, the functional group R2 remained as shown in FIG. An indole from which the carbonyl group is eliminated is observed. In this case, both the indole carbonyl and the indole from which the carbonyl group is eliminated contain the functional group R2, and the mass-to-charge ratio differs between the indole carbonyl and the indole depending on the type of the functional group R2. Therefore, the indole carbonyl from which the functional group R1 is eliminated is a precursor ion, and the indole from which the functional group R1 is eliminated is a product ion, which corresponds to the masses of the two types of functional groups R2 attached to the 2-position of the indole ring. The MRM measurement in which the MRM transition was set as described above was taken as the second MS / MS measurement.

  FIG. 8 shows the relationship between the MRM transition and the type of the functional group R2 in this MRM measurement mode. That is, if ions are observed in one of these two types of MRM transitions, it can be said that there is a high possibility that the functional group corresponding to the MRM transition is the functional group R2.

[Precursor ion scan measurement]
As described above, the functional group R1 that modifies the nitrogen of the indole ring is relatively easily released upon ionization, but the functional group R1 is not eliminated on the mass spectrum (that is, as-added) indolecarbonyl. Is also observed. When this indolecarbonyl is cleaved by collision-induced dissociation, the functional group R1 is eliminated (see FIG. 9). Therefore, the difference between the mass-to-charge ratio of the precursor ion before the collision-induced dissociation and the mass-to-charge ratio of the product ion after the collision-induced dissociation corresponds to the functional group R1. At this time, indolecarbonyl also contains functional group R2, but only two types of functional group R2 (hydrogen or methyl group) need be known, so the mass-to-charge ratio of indolecarbonyl from which functional group R1 is eliminated is the functional group. There are at most two types depending on the type of R2. Therefore, a precursor ion scan measurement was carried out by using a precursor ion scan measurement in which two kinds of mass-to-charge ratios in which either hydrogen or a methyl group was added to the position of the functional group R2 in the indole carbonyl from which the R1 functional group was eliminated were respectively set as product ions. / MS measurement.

  As described above, since the difference between the mass-to-charge ratio of the ions and the mass-to-charge ratio of the product ions observed by the precursor ion scan measurement should correspond to the type of the functional group R1, a peak is observed on the precursor ion spectrum. The type of the functional group R1 can be estimated from the difference between the mass-to-charge ratio of ions to be set and the set mass-to-charge ratio of product ions. FIG. 10 shows the relationship between the mass-to-charge ratio of the precursor ion and the functional group R1.

  Since the measurement conditions for the three types of MS / MS measurement described above (measurement mode, mass-to-charge ratio to be detected, etc.) are determined in advance, such measurement conditions may be stored in the comprehensive drug identification control program 35. it can.

  When identifying an illegal drug in a sample, the measurer first sets measurement conditions from the input unit 36 (step S1). Since basic measurement conditions for MS / MS measurement are determined, other measurement conditions such as GC separation conditions, scan speed at the time of scan measurement, collision energy, and the like may be set as necessary. Setting of such measurement conditions can be omitted by using all default values or using a predetermined method file.

  When the measurement is started by the measurer's instruction, the analysis control unit 33 controls the GC unit 1 and the MS / MS unit 2 based on the instruction from the central control unit 34, and from the measurement start time to the measurement end time, The first MRM measurement including the 13 types of MRM transitions, the second MRM measurement including the 2 types of MRM transitions, and the precursor ion scan measurement for the two types of product ions are repeatedly collected (step S2). ). Data obtained by each measurement is stored in a memory in the data processing unit 30. When the measurement end time is reached (Yes in step S3), the measurement is ended, and subsequently, the comprehensive drug identification data processing unit 31 executes data processing (step S4).

  First, the comprehensive drug identification data processing unit 31 creates a mass chromatogram for each MRM transition of the MRM measurement, and creates a total ion chromatogram for each product ion of the precursor ion scan measurement (step S5). Then, peak detection is performed on each of the chromatograms, and one of the 13 mass chromatograms obtained by the first MRM measurement and one of the two mass chromatograms obtained by the second MRM measurement. In any one and three chromatograms of any one of the two total ion chromatograms obtained by the precursor ion scan measurement, a peak appearing at the same retention time is searched (step S6).

  The presence of peaks appearing at the same retention time in the three chromatograms means that the compound appearing at the retention time has naphthoyl and indolecarbonyl in its chemical structure, which is (1H-indole It means having -3-yl) (naphthalen-1-yl) methanone as a basic skeleton. Therefore, when the presence of a common peak is not confirmed at the same retention time (No in step S7), it is determined that the drug is not at least a naphthoylindole drug (step S8).

  On the other hand, if the presence of a common peak is confirmed at the same retention time (Yes in step S7), it can be estimated that the drug is a naphthoylindole-based drug, so the 13 obtained in the first MRM measurement In the mass chromatogram, the type of the functional group R3 is determined from the MRM transition in which the common peak is observed. From the MRM transition in which the common peak is observed in the two mass chromatograms obtained in the second MRM measurement. Of the two total ion chromatograms obtained by the precursor ion scan measurement, the type of the functional group R2, and the position of the peak on the chromatogram where the common peak is observed, that is, the mass charge of the detected precursor ion The type of the functional group R1 is estimated from the ratio (step S9). The chemical structure of the detected naphthoylindole drug is estimated from the functional groups R1, R2, and R3 thus estimated, and the compound is identified (step S10). When the drug is identified, the identification result is displayed on the display unit 37 when the drug is not identified, and presented to the measurer (step S11).

  In the above-described processing, the drug database 32 is not used when identifying the compound in the sample, but naphthoylindole drugs whose mass spectrum is recorded in the drug database 32 are conventionally performed. It is more efficient to identify compounds by such simple database search. Therefore, normal scan measurement (for example, Q3 scan measurement) is repeatedly executed together with the above-described MRM measurement and precursor ion scan measurement, and mass spectra are sequentially obtained. On the total ion chromatogram based on the data obtained by the scan measurement. By comparing the mass spectrum of the detected appearance position of the peak with the mass spectrum in the drug database 32, identification of the first-stage compound is performed, and subsequently, the identification process after step S5 described above is performed. The naphthoylindole drug that was not found in the identification of the compound in the first step (missed from the database search) may be identified. Thereby, not only the identification efficiency of a naphthoyl indole type | system | group drug improves but the drug and poison other than a naphthoyl indole type | system | group can also be identified.

  Further, both the first and second MRM measurements in the above description may be product scan measurements. That is, instead of the first MRM measurement, a product ion scan measurement using naphthoyl as a precursor ion corresponding to the difference in mass of the functional group R3 may be performed. Instead of the second MRM measurement, the functional group R2 What is necessary is just to perform the product ion scan measurement which used indolecarbonyl as a precursor ion according to the difference in mass. However, there are 13 types of functional groups R3 and 2 types of functional groups R2, and when data is acquired by performing a total of 15 types of product ion scan measurement, the sensitivity is inferior to MRM measurement. Therefore, in the above example, MRM measurement capable of detecting ions with higher sensitivity is used.

An application example of the comprehensive drug identification process described above will be specifically described.
The equipment used for the measurement is as follows.
GC-MS / MS: Shimadzu GCMS-TQ8030
Column: Resxi Rxi-5Sil MS (length 30m, inner diameter 0.25mm, df = 0.25μm)
A standard sample of a compound which is an illegal drug having 20 types of (1H-indol-3-yl) (naphthalen-1-yl) methanone having different combinations of functional groups R1, R2 and R3 as a basic skeleton was measured. The types of the compounds and the functional groups R1, R2, and R3 are as shown in FIG.

  As a typical example, it is a measurement result of common name AM2232 (5- [3- (1-naphthoyl) 1-1H-indol-1-yl] pentanenitrile) which is R1: C4H8CN, R2: H, R3: H. A portion of the chromatogram and peak detection results are shown in FIG. The peak observed in the mass chromatogram corresponding to the MRM transition of m / z 144> m / z 116 in the second MRM measurement and the MRM transition of m / z 155> m / z 127 in the first MRM measurement The peak observed in the corresponding mass chromatogram and the peak observed in the chromatogram where the product ion is m / z 144 and the precursor ion is m / z 225 have the same retention time. Therefore, since the retention times of these peaks coincide, the compound detected at the retention time has (1H-indol-3-yl) (naphthalen-1-yl) methanone as a basic skeleton. I can confirm that.

  Further, since the MRM transition of the first MRM measurement in which the common peak is detected is m / z 155> m / z 127, it can be estimated that the functional group R3 is H (see FIG. 6). Since the MRM transition of the second MRM measurement in which the peak is detected is m / z 144> m / z 116, it can be estimated that the functional group R2 is also H (see FIG. 8). Since the precursor ion of the detected precursor ion scan measurement is m / z 225, it can be estimated that the functional group R1 is H and C4H8CN. This is consistent with the chemical structure of AM2232 and it can be concluded that AM2232 can be identified.

  FIG. 12 shows the mass-to-charge ratio of ions detected when illegal drugs having 20 types of (1H-indol-3-yl) (naphthalen-1-yl) methanone including AM2232 as a basic skeleton are measured. Show. For all these compounds, ions corresponding to actual functional groups have been detected, and conversely, the three types of functional groups R1, R2, and R3 can be identified from the mass-to-charge ratio of the detected ions. I can confirm that.

  From the above results, according to the drug analyzer according to the present invention, from the measurement results using GC-MS / MS, (1H-indol-3-yl) (naphthalen-1-yl) methanone is used as the basic skeleton. It was verified that comprehensive identification of naphthoylindole illegal drugs is possible.

Note that the above embodiment is merely an example of the present invention, and it is obvious that changes and modifications can be appropriately made within the scope of the gist of the present invention.
For example, in the above embodiment, the gas chromatograph is used to separate the compounds in the sample, but a liquid chromatograph may be used. However, in order to ionize the compound in the sample solution eluted from the column of the liquid chromatograph, the atmospheric pressure ionization method is generally used. In the atmospheric pressure ionization method, ions are not easily cleaved at the ionization stage. Therefore, an ion source having an in-source decomposition function may be used so that fragment ions similar to those obtained by electron ionization can be obtained.

DESCRIPTION OF SYMBOLS 1 ... GC unit 10 ... Sample vaporization chamber 11 ... Column oven 12 ... Column 2 ... MS / MS unit 20 ... Analysis chamber 21 ... Ion source 22 ... Pre-stage quadrupole mass filter 23 ... Collision cell 24 ... Multipole ion guide 25 ... latter-stage quadrupole mass filter 26 ... detector 30 ... data processing unit 31 ... comprehensive drug identification data processing unit 32 ... drug database 33 ... analysis control unit 34 ... central control unit 35 ... comprehensive drug identification control program 36 ... Input unit 37 ... Display unit

Claims (2)

A drug analysis method for comprehensively identifying specific compounds having (1H-indol-3-yl) (naphthalen-1-yl) methanone as a basic skeleton using a chromatographic mass spectrometer capable of MS / MS measurement There,
a) Using a chromatographic mass spectrometer, the sample containing the compound to be identified is
a1) A naphthoyl produced by cleavage upon ionization is a precursor ion, a naphthalene from which the carbonyl group is eliminated from the naphthoyl is a product ion, and a plurality of MRMs according to the difference in mass of the third functional group bonded to the naphthalene ring A first MRM measurement using transition as a measurement condition, or a first product ion scan measurement using naphthoyl as a precursor ion corresponding to a difference in mass of the third functional group,
a2) The indole carbonyl generated by the cleavage during ionization is a precursor ion, the indole from which the carbonyl group is eliminated from the indole carbonyl is a product ion, and there are multiple numbers corresponding to the difference in mass of the second functional group bonded to the indole ring. A second MRM measurement using the MRM transition as a measurement condition, or a second product ion scan measurement using a precursor ion of indolecarbonyl corresponding to the difference in mass of the second functional group, and
a3) Precursor ion scan measurement using indole carbonyl in the state where the first functional group bonded to nitrogen of the indole ring is removed while the second functional group is added as a product ion,
A measurement execution step for repeatedly executing three types of MS / MS measurements,
b) Created based on data obtained by the first MRM measurement or the first product ion scan measurement, the second MRM measurement or the second product ion scan measurement, and the precursor ion scan measurement, respectively. A compound presence confirmation step for determining the presence of the specific compound having the basic skeleton by determining whether or not a peak is commonly present at the same retention time on the chromatogram corresponding to each type of measurement;
c) The type of the third functional group was obtained by the second MRM measurement or the second product ion scan measurement based on the data obtained by the first MRM measurement or the first product ion scan measurement. The type of the second functional group is estimated based on the data, the type of the first functional group is estimated based on the data obtained by the precursor ion scan measurement, and the compound presence confirmation step is performed based on the estimation results. A structure estimation step for estimating the chemical structure of a specific compound whose existence has been confirmed;
A drug analysis method using chromatographic mass spectrometry, comprising: The drug analysis method according to claim 1,
In the measurement execution step, a normal scan measurement that does not intentionally cleave ions derived from the sample is repeatedly executed together with the three types of MS / MS measurements,
A database utilization identification step of identifying a known compound by collating a mass spectrum created based on data obtained by the normal scan measurement with a drug database;
A drug analysis method using chromatographic mass spectrometry, wherein in the structure estimation step, structure estimation is performed for an unknown compound that is not recorded in the drug database.

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