WO2012033111A1 - 物質の構造解析方法 - Google Patents
物質の構造解析方法 Download PDFInfo
- Publication number
- WO2012033111A1 WO2012033111A1 PCT/JP2011/070322 JP2011070322W WO2012033111A1 WO 2012033111 A1 WO2012033111 A1 WO 2012033111A1 JP 2011070322 W JP2011070322 W JP 2011070322W WO 2012033111 A1 WO2012033111 A1 WO 2012033111A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- ion amount
- percentage
- value
- amount
- total ion
- Prior art date
Links
Images
Classifications
-
- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- H01J49/00—Particle spectrometers or separator tubes
- H01J49/0027—Methods for using particle spectrometers
- H01J49/0036—Step by step routines describing the handling of the data generated during a measurement
-
- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- H01J49/00—Particle spectrometers or separator tubes
- H01J49/0027—Methods for using particle spectrometers
- H01J49/0031—Step by step routines describing the use of the apparatus
-
- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- H01J49/00—Particle spectrometers or separator tubes
- H01J49/004—Combinations of spectrometers, tandem spectrometers, e.g. MS/MS, MSn
- H01J49/0045—Combinations of spectrometers, tandem spectrometers, e.g. MS/MS, MSn characterised by the fragmentation or other specific reaction
- H01J49/005—Combinations of spectrometers, tandem spectrometers, e.g. MS/MS, MSn characterised by the fragmentation or other specific reaction by collision with gas, e.g. by introducing gas or by accelerating ions with an electric field
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
- G01N30/7233—Mass spectrometers interfaced to liquid or supercritical fluid chromatograph
Definitions
- the present invention relates to a method for analyzing the structure of a target substance by measuring the target substance by triple quadrupole mass spectrometry and subjecting the obtained data to a predetermined deconvolution process.
- TQ-MS Triple-quadrupole mass spectrometry
- HPLC high performance liquid chromatography
- ESI electrospray ionization
- LC-MS liquid chromatograph mass spectrometry
- CID collision-induced dissociation
- affinity of the species for the stationary phase used in HPLC can be determined from the retention time of the given species in the chromatography column, and structural information can be obtained.
- this technique has a problem of providing only MS / MS information.
- quadrupole ion trap mass spectrometry enables multi-stage MS / MS analysis that provides detailed structural information (Non-Patent Documents 3 and 4). , 6-9).
- QIT-MS quadrupole ion trap mass spectrometry
- ERMS energy-resolved mass spectrometry
- MS n + 1 information information related to the fragmentation reaction of product ions that is not normally obtained in the CID process of QIT-MS. It has been clarified that it contains. However, since the QIT-MS instrument can be used to perform MS n experiments, such information may not be very important for QIT-MS analysis. On the other hand, since the TQ-MS apparatus cannot be used for the MS n experiment, the above-described information of MS n + 1 is extremely important for the TQ-MS analysis. Also, such MS n + 1 information is useful when the TQ-MS method is used in combination with HPLC for structure elucidation.
- An object of the present invention is a method for structural analysis of a substance using triple quadrupole mass spectrometry (TQ-MS), which enables acquisition of MS n information from data obtained by TQ-MS measurement. Is to provide a method.
- TQ-MS triple quadrupole mass spectrometry
- the present inventors have obtained the ratio of each product ion amount to the total ion amount with respect to the ERMS spectrum obtained in the TQ-MS measurement, and arbitrarily determined the ratio of the product ions. It was found that MS n information can be obtained by performing linear regression analysis on the sum of the combinations. The present invention has been completed based on such findings.
- the aspect of the present invention relates to the following.
- [1] (a) The target substance is subjected to triple quadrupole mass spectrometry (TQ-MS) measurement while changing the value of CID energy. (B) For each CID energy value, determine the percentage of the precursor ion amount relative to the total ion amount and the percentage of the specific m / z product ion amount relative to the total ion amount; (C) Each value of the percentage with respect to the total ion amount of the product ion amount of the specific m / z in each value of CID energy that gives each value of the percentage of the precursor ion amount with respect to the total ion amount obtained in the step (b) Extract (D) For each value of the percentage of the product ion amount of the specific m / z extracted in step (c) with respect to the total ion amount, the sum of all combinations is obtained, (E) A product that selects a function that can be linearly approximated from among functions in which the percentage value
- step (b) with the CID energy value on the X axis and the percentage value on the Y axis, the percentage of the precursor ion amount relative to the total ion amount and the total ion of the product ion amount of a specific m / z
- a graph is created by plotting the percentage against the amount.
- step (c) a graph is obtained by plotting each percentage of the total ion amount of the specific m / z product ion amount in the Y-axis direction with the percentage of the precursor ion amount with respect to the total ion amount as the X axis.
- the method according to [1] or [2] which is prepared.
- step (d) a graph is created by plotting the sum of the obtained combinations in the Y-axis direction with the percentage of the total amount of precursor ions with respect to the total ion amount as the X-axis. 3].
- step (e) The method according to any one of [1] to [4], wherein the linear approximation in step (e) is performed using the following formula (4): Where Syx represents the residual of the linear regression curve, x j and y j represent the coordinate elements of the data j, a j and b j represent the coordinate elements of the regression curve, and n ⁇ 2 It is a degree of freedom.
- Syx represents the residual of the linear regression curve
- x j and y j represent the coordinate elements of the data j
- a j and b j represent the coordinate elements of the regression curve
- n ⁇ 2 It is a degree of freedom.
- the method according to any one of [1] to [5] wherein MS n information is acquired in the step (e
- FIG. 1a shows a CID spectrum showing that fragment ions (f 1 -f 4 ) are generated from precursor ions X.
- FIG. 1b shows the ERMS spectrum observed for the dissociation of ion X, and the data can usually be analyzed by Boltzmann sigmoidal equations.
- FIG. 1c shows a graph (ratio plot) in which the ERMS of FIG. 1b is replotted with the precursor ion X as the x-axis and the individual product ions as the y-axis.
- FIG. 2a shows the ERMS spectrum of the sodium ester of the methyl ester of ganglioside GD1a (1).
- FIG. 1a shows a CID spectrum showing that fragment ions (f 1 -f 4 ) are generated from precursor ions X.
- FIG. 1b shows the ERMS spectrum observed for the dissociation of ion X, and the data can usually be analyzed by Boltzmann sigmoidal equations.
- FIG. 2b shows a ratio plot of the ERMS of FIG. 1a.
- FIG. 2c shows fragmentation of the precursor ion (m / z 1916.1). Dissociation corresponding to the cleavage of several glycosidic bonds was observed, with the most abundant ion being the y-ion with N-acetylgalactosaminyl cleavage.
- the ERMS of the dissociation of the ion X is shown, and one of the product ions (f 1 ) is further dissociated into f 3 and f 4 .
- FIG. 3a shows the dissociation skim.
- FIG. 3b shows the ERMS spectrum, and some product ion curves cannot be approximated by the Boltzmann sigmoid equation.
- FIG. 3c shows a ratio plot of ERMS, where the sum of the series of curves is a straight line.
- FIG. 6 shows an ERMS spectrum of [2 + Na] + obtained by QIT-MS.
- FIG. 4a shows the ERMS spectrum.
- FIG. 4b shows a ratio plot of the ERMS of FIG. 1a. Black circles indicate the sum of all product ions, and white square symbols indicate the sum of ions at m / z 466.2 and m / z 628.3.
- FIG. 4c shows the fragmentation of the precursor ion (m / z 646.3). Dissociation corresponding to galactosyl cleavage and dehydration occurred at the PA and N-acetyl groups was observed.
- FIG. 6 shows an ERMS spectrum of [2 + Na] + obtained by QIT-MS.
- FIG. 4a shows the ERMS spectrum.
- FIG. 4b shows a ratio plot of the ERMS of FIG. 1a. Black circles indicate the
- FIG. 6 shows an ERMS spectrum of [2 + Na] + obtained by TQ-MS.
- FIG. 5a shows the ERMS spectrum.
- FIG. 5b shows a ratio plot of the ERMS of FIG. 5a.
- FIG. 5c shows an arbitrary sum of the individual product ion intensities in the ratio plot.
- FIG. 5d shows the fragmentation of the precursor ion (m / z 646.3). Dissociation corresponding to galactosyl cleavage (f 2 ) and dehydration occurred at the PA and N-acetyl groups (f 1 and f 3 ) was observed. Furthermore, other fragments such as f 4 to f 6 that were not observed by QIT-MS were observed. The dissociation pathway of the sodium addition compound 2 obtained in TQ-MS is shown.
- the structure analysis method of the substance of the present invention is as follows: (a) TQ-MS measurement is performed on the target substance while changing the value of CID energy, and (b) the amount of precursor ion with respect to the total ion quantity at each CID energy value.
- Each of the CID energies that gives a percentage and a percentage of the product ion quantity of a specific m / z with respect to the total ion quantity, and gives each value of the percentage of the precursor ion quantity obtained in step (b) with respect to the total ion quantity.
- each value of the percentage of the product ion amount of the specific m / z with respect to the total ion amount is extracted, and (d) the total ion amount of the product ion amount of the specific m / z extracted in the step (c)
- the sum of all combinations is determined, (e) the percentage value of the precursor ion quantity to the total ion quantity is x, Selecting a function that can be linearly approximated among the functions having the sum of the obtained combinations as y, and analyzing the structure of the target substance from the m / z values of the product ions that give the selected functions. It is characterized by.
- CID energy generally indicates energy applied when CID (collision induced dissociation) occurs, and specifically, an AC electric field of a certain frequency for vibrating ions. Indicates voltage.
- Precursor ion means an ion corresponding to a target substance as a precursor
- product ion means a fragment ion having each m / z obtained by performing TQ-MS measurement on the precursor ion. means.
- m / z indicates a ratio of mass number (m) to electric charge (z).
- MS n information means information on the structure of the precursor ion obtained from the m / z value, signal intensity, CID energy, etc. of the product ion generated by the precursor ion being decomposed in multiple stages.
- TQ-MS measurement is performed while changing the value of CID energy until a product ion having a specific m / z is generated from a precursor ion corresponding to the target substance.
- the TQ-MS measurement is performed while changing the value of the CID energy until the intensity of the precursor ion becomes almost zero.
- the target substance to be subjected to the structural analysis of the present invention is not particularly limited as long as it can be used for TQ-MS measurement, but is preferably a sugar chain.
- the sugar chain may be obtained from a living tissue or cell, or may be obtained from a substance bound to a synthesized protein, or may be subjected to acid hydrolysis or enzymatic decomposition, HPLC, etc. Those separated and purified by the above can be used. A chemically synthesized sugar chain can also be used.
- step (b) for each value of the changed CID energy, the percentage of the precursor ion amount with respect to the total ion amount and the percentage with respect to the total ion amount of the product ion amount of a specific m / z are obtained. Specifically, based on the TQ-MS measurement data obtained in step (a), the percentage of the precursor ion intensity with respect to the total ion intensity at each value of CID energy and the product ion intensity of a specific m / z Obtain the percentage of total ionic strength. That is, at the time when TQ-MS measurement is started by applying CID energy, the amount of precursor ions corresponds to the total amount of ions, and the percentage value is 100%.
- step (b) the CID energy is taken as the X-axis and the percentage as the Y-axis, the percentage of the precursor ion amount obtained as described above with respect to the total ion amount, and the total ion amount of the product ion amount of a specific m / z
- a graph may be generated by plotting the percentage with respect to the ERMS spectrum for the precursor ion and the product ion having each m / z value.
- the ERMS spectrum includes a plurality of data points whose parameters are CID energy and the percentage of each ion amount.
- step (c) the total ion of the product ion amount of the specific m / z in each value of CID energy giving each value of the percentage of the precursor ion amount obtained in the step (b) with respect to the total ion amount. Extract each value as a percentage of the quantity.
- step (c) plotting each value of the percentage of the product ion amount of the specific m / z extracted with respect to the total ion amount, where the percentage of precursor ions is the X axis and the percentage of product ions is the Y axis.
- step (d) the sum of all combinations is obtained for each value of the percentage of the product ion amount of the specific m / z extracted in step (c) with respect to the total ion amount.
- a graph may be created by plotting the sum of the obtained combinations in the Y-axis direction with the percentage of the precursor ion amount relative to the total ion amount as the X-axis.
- the graph of FIG. The graph thus obtained includes a plurality of curves indicating the sum of the percentages for each combination of product ions having a specific m / z value.
- step (e) a function that can be linearly approximated among the functions in which the percentage value of the precursor ion amount with respect to the total ion amount is x and the sum of the combinations obtained in step (d) is y.
- step (e) a function that can be linearly approximated among the functions in which the percentage value of the precursor ion amount with respect to the total ion amount is x and the sum of the combinations obtained in step (d) is y.
- step (d) when a graph is created in step (d), it is only necessary to select a curve that can be linearly approximated among a plurality of curves included in the graph.
- linear approximation in the step (d) can be performed by using a generally known least square method, the following formula (4) or formula (5).
- Syx indicates the residual of the linear regression curve
- x j and y j indicate the coordinate elements of the data j
- a j and b j indicate the coordinate elements of the regression curve.
- n-2 is the degree of freedom.
- the Syx value as the threshold value can be appropriately determined according to data obtained in the TQ-MS.
- x j and y j indicate coordinate elements of data j, and p and q indicate the average of the data coordinates, respectively.
- the R 2 value as the threshold value can be appropriately determined according to the data obtained in the TQ-MS.
- R 2 0.90 is preferably used as the threshold value.
- a curve with a value of R 2 of 0.90 or more is selected by linear approximation.
- the structure of the target substance is analyzed from the m / z value of the product ion giving each function (curve in the graph) selected by performing linear approximation. Specifically, by comparing m / z values of product ions related to combinations that give a function that can be linearly approximated, it is possible to analyze in what order each product ion is generated from the precursor ions. .
- the sample Gibili antigen trisaccharide
- Hex represents hexose (hexose sugar)
- HexNAc represents N-acetylhexosamine
- Ganglioside GD1a (1) was purchased from Seikagaku Corporation (Tokyo, Japan) and esterified.
- Galili antigen trisaccharide (2) was purchased from Carbohydrate Synthesis Ltd. (Oxford, UK) and previously reported procedures (Hase, S .; Ikenaka, T .; Matsushima, Y. Biochem. Biophys. Res. Commun. 1978, 85, 257-263.).
- the experiment conducted using the API Q-Star pulsar i Q-TOF MS apparatus (controlled by Analyst QS) as TQ-MS measurement is as follows. Data collection was done in positive mode with full scans between m / z 100 and 2000. A sample (1 pmol / ⁇ L) was dissolved in methanol and introduced into the ion source using a coated nanospray tip (New Objective, Inc. MA, USA) at a flow rate of 150 nL / min. The standard ion spray voltage for nanospray was 1600V. Nitrogen gas was used as curtain gas (15 psi) and collision gas.
- the first and second declustering potentials, ie DP1 and DP2 were 50V and 15V, respectively, and the focusing potential was 250V.
- MS / MS spectra were acquired using selection of precursor ions by quadrupole in low resolution mode (less than unit resolution). As a result, all the parent isotope groups were sent to the collision cell, and all fragment isotope groups were obtained. All spectra reported here are averages of 1.0 minute scans.
- rel C represents the ion current (%) of a predetermined ion among the observed ions, and is expressed as a percentage of the total ion current.
- C Ii represents the ionic current observed at the focal point, and C Ip represents the ionic current of the precursor ion.
- x j and y j indicate coordinate elements of the data j.
- a j and b j indicate the coordinate elements of the regression curve.
- n-2 is the degree of freedom.
- Syx ⁇ 0 there is a strong relationship between ions.
- Syx 2 was used as the threshold value.
- the dissociation reaction of the ganglioside GD1a (compound 1) methyl ester can be considered a typical example of what follows a “simple” fragmentation process (FIG. 2). Observed that under CID conditions, a number of fragment ions (m / z 1610.9, 1245.8, 940.6, 778.6, and 693.2) were formed from sodium chloride 1 (m / z 1916.1: [M + Na + ] + ). It was done. Each series of data points was well fitted using the Boltzmann sigmoid equation. The precursor (precursor) -product (product) relationship (ratio plot) shows that the ratio of fragment ions to precursor is constant over a wide range of CID energies, as shown in FIG. 2b.
- Table 1 shows combinations of product ions (fragment ions) constituting a curve selected by linear approximation among the curves shown in FIG. 5c.
- product ions indicated by circles are elements constituting each curve, and Syx values corresponding to each curve are shown.
- a series of fragment ions f 1 to f 4 [m / z 304 (f 4 ), 466 (f 3 ), 484 (f 2 ), 628 (f 1 )] showed a certain relationship.
- the ions f 2 and f 6 are compounds with a PA residue, and f 1 , f 3 and f 4 are dehydration ions between the PA residue and the N-acetyl group. It was. Hexose-derived C ion species (f 5 ) was also observed as a small signal. Information on the imidazoline structure under CID conditions has been reported in the past (Kurimoto, A .; Kanie, O. Rapid Commun. Mass Spectrom. 2007, 21, 2770-2778.). All fragment ions corresponded to the structure of Compound 2, but only limited information was available from these m / z values alone. The information obtained from the above analysis provided further details as follows.
- the precursor ion (2 + Na + ) was fragmented into product ions f 1 , f 2 , f 5 and f 6 through cleavage of two glycosidic bonds and dehydration of the reducing end.
- f 2 further generated f 3 and f 4 through dehydration and glycosyl cleavage.
- the ion f 1 was converted to f 4 by two glycosyl cleavages.
Landscapes
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
Description
〔1〕 (a)目的物質について、CIDエネルギーの値を変化させて三連四重極質量分析(TQ-MS)測定を行い、
(b)各CIDエネルギーの値において、プレカーサーイオン量の総イオン量に対する百分率と、特定のm/zのプロダクトイオン量の総イオン量に対する百分率とを求め、
(c)工程(b)において求めたプレカーサーイオン量の総イオン量に対する百分率の各値を与えるCIDエネルギーの各値における、前記特定のm/zのプロダクトイオン量の総イオン量に対する百分率の各値を抽出し、
(d)工程(c)において抽出した前記特定のm/zのプロダクトイオン量の総イオン量に対する百分率の各値について、全ての組合せの和を求め、
(e)プレカーサーイオン量の総イオン量に対する百分率の値をxとし、かつ前記求めた各組合せの和の値をyとする関数のうち直線近似できるものを選択し、選択した各関数を与えるプロダクトイオンのm/z値から目的物質の構造を解析することを含む、
物質の構造解析方法。
〔2〕 工程(b)において、CIDエネルギーの値をX軸とし、かつ百分率の値をY軸として、プレカーサーイオン量の総イオン量に対する百分率と、特定のm/zのプロダクトイオン量の総イオン量に対する百分率とをプロットしてグラフを作成する、〔1〕に記載の方法。
〔3〕 工程(c)において、プレカーサーイオン量の総イオン量に対する百分率をX軸として、前記特定のm/zのプロダクトイオン量の総イオン量に対する各百分率をY軸方向にプロットしてグラフを作成する、〔1〕又は〔2〕に記載の方法。
〔4〕 工程(d)において、プレカーサーイオン量の総イオン量に対する百分率をX軸として、前記求めた各組合せの和の値をY軸方向にプロットしてグラフを作成する、〔1〕~〔3〕のいずれかに記載の方法。
〔5〕 工程(e)における直線近似を以下の式(4)を用いて行う、〔1〕~〔4〕のいずれかに記載の方法:
〔6〕 工程(e)において、MSn情報を取得する、〔1〕~〔5〕のいずれかに記載の方法。
〔7〕 前記目的物質が糖鎖である、〔1〕~〔6〕のいずれかに記載の方法。
(材料)
ガングリオシドGD1a(1)は、生化学工業株式会社(日本、東京)から購入し、エステル化した。ガリリ抗原三糖(2)は、Carbohydrate Synthesis Ltd.(オックスフォード、イギリス)から購入し、過去に報告されている手順(Hase, S.; Ikenaka, T.; Matsushima, Y. Biochem. Biophys. Res. Commun. 1978, 85, 257-263.)に従い、ピリジルアミノ化した。
試料は、ポジティブモードで、Pico View ESI (nanospray) source (New Objective, Inc. MA、米国)を装備した、四重極イオントラップ型質量分析装置Esquire 3000 plus (Bruker Daltonics GmbH, Bremen, Germany)、及びAPI Q-Star pulsar i (Applied Biosystems/SCIEX, ON、カナダ)を用いて分析した。
ERMSのグラフを得るために、以下の方程式(1)を用いた。イオン“IP”が一連のプロダクトイオンI1,I2,I3,…Iiを生成する場合、個々のイオンについての相対イオン電流を、以下の方程式(1)で定義した。この方程式(1)を用いることにより、本発明のERMSスペクトル(図5a)が得られる。
(広範囲のCIDエネルギーに対して得られるプレカーサーに対するフラグメントイオンの比率間の直線的な関係)
まず、CID条件下でのプレカーサーイオンXのMS/MS分析を検討する(図1a)。得られたスペクトル(図1a)は、イオンの個々のシグナルの強度を示す。未同定の化合物の構造を決定することを目的として、比較のためにそれらシグナルの比率を得るかもしれない。一方、CID条件下におけるイオンXのERMS分析(図1b)は、通常のMS/MS分析によって得ることができる情報に加え、プレカーサーイオンの分裂に必要な活性化エネルギーに関係する情報を提供する。この事実から、衝突セルで起きている化学反応に関係する情報を抽出できるかもしれない。直線関係にある一連の曲線が第二のプロットにおいて見られ、プレカーサーイオンの各百分率に対する個々のイオンの百分率を示している(図1c)。このような場合に、適用されたCIDエネルギーに関わらず、プレカーサーに対するフラグメントイオンの強度の比率が一定であるという重要な情報を得ることができる。この結果から、異なるCIDエネルギーにおいて得られたMS/MSスペクトル同士の比較が可能であることが示され、複雑なグリカンの構造決定における断片ピークの比率の比較のために極めて重要な基準が得られる(Takegawa, Y.; Deguchi, K.; Ito, S.; Yoshioka, S.; Sano, A.; Yoshinari, K.; Kobayashi, K.; Nakagawa, H.; Monde, K.; Nishimura, S.-I. Anal. Chem. 2004, 76, 7294-7303、Kameyama, A.; Kikuchi, N.; Nakaya, S.; Ito, H.; Sato, T.; Shikanai, T.; Takahashi, Y.; Takahashi, K.; Narimatsu, H. Anal. Chem. 2005, 77, 4719-4725、Ashline, D.; Singh, S.; Hanneman, A.; Reinhold, V. Anal. Chem. 2005, 77, 6250-6262)。過去の報告において検体の大部分について観察されたように、異なるCIDエネルギーに対するこのような直線性が、この種のERMSスペクトル(図1b)において一般に見受けられたことは注目すべきである(非特許文献10~13)。
まれに、図3に示すような複雑なERMSスペクトルが得られる。このようなデータの取扱いは、近似の困難性ゆえに、ERMSの定量的な分析において問題となっており、これらのスペクトル自身は「構造指紋(structural fingerprints)」と考えることができるものの、それらはそのような定量的な分析に利用されていない(Daikoku, S.; Kurimoto, A.; Mutsuga, S.; Ako, T.; Kanemitsu, T.; Shioiri, Y.; Ohtake, A.; Kato, R.; Saotome, C.; Ohtsuka, I.; Koroghi, S.; Sarkar, S. K.; Tobe, A.; Adachi, S.; Suzuki, K.; Kanie, O. Carbohydr. Res. 2009, 344, 384-394.)。このような場合には、プレカーサーに対するフラグメントイオンの比率のいくつかは直線性を示し、その他は直線性を示さず、シグナル強度のいくつか(f1、f3及びf4)の合計(f1’)が直線になる(図3c)。フラグメントイオンf3及びf4は、f1の曲線が点線(f1’)から離れた地点において、f1から生成したものと考えられた。この結果から、フラグメントイオンの断片化反応は直線離脱の地点に対応するCIDエネルギーにおいて起こり始めることが示された。
上述の通り、選択したフラグメントイオンの強度の合計を調査することによって断片化経路を得る分析方法の潜在的有用性が示されたので、次に、TQ-MSを用いて、イオン[2 + Na]+のERMSスペクトルを調査した。TQ-MSにより得られたERMSスペクトルのプロファイルは、QIT-MSにより得られたものと全く異なっており、また、より複雑であった(図5a)。個々のフラグメントイオンの「生成曲線」の間の明らかな差異に加えて、QIT-MS実験では観察されなかったm/z 304.1及び322.2のイオンが観測された。また、QIT-MSの実験において観察されたように、m/z 628.3 (f1) 及び 466.2 (f3)のイオンは強力な関係を有しているものと考えられた。比率プロットは、また、フラグメントイオンの多くがそれらのプレカーサーと非直線関係を有することを示した(図5b)。一対を超えるフラグメントイオンが互いに関係しているものと考えられたので、個々のフラグメントイオンの組合せの合計を調査した。任意の組合せ、即ち、全体のイオン(m)からのイオン(l)のC(m,l)を考慮し、比率プロットにおいて線形回帰曲線の残差(Syx)の標準偏差を調査した(図5c)。Syx値が小さいほど、選択される対のイオンの間の関係はより強いと考えられる。図5cに表される曲線のうち、直線近似により選択された曲線を構成するプロダクトイオン(フラグメントイオン)の組合せを表1に示す。表中、丸印で示されるプロダクトイオンが各曲線を構成する要素であり、各曲線に対応するSyx値が示されている。例えば、一連のフラグメントイオンf1~f4[m/z 304 (f4), 466 (f3), 484 (f2), 628 (f1)]は、一定の関係を示した。
表1の結果から、具体的には、以下の情報が得られる。
・f1=[2+Na+](プレカーサー:646)-H2O(18)
・f2=[2+Na+]-Hex(162) f3= f1-Hex、かつ、f3= f2-H2O
・f4= f1-2Hex、かつ、f4= f3-Hex
なお、「Hex」はヘキソース(六炭糖)を示し、括弧内の数値はm/z値ないし分子量を示す。上記情報から、図6に示す通り、[2+Na+](プレカーサーイオン;試料ガリリ抗原三糖)の解離経路に関する情報を取得することができた。
Claims (7)
- (a)目的物質について、CIDエネルギーの値を変化させて三連四重極質量分析(TQ-MS)測定を行い、
(b)各CIDエネルギーの値において、プレカーサーイオン量の総イオン量に対する百分率と、特定のm/zのプロダクトイオン量の総イオン量に対する百分率とを求め、
(c)工程(b)において求めたプレカーサーイオン量の総イオン量に対する百分率の各値を与えるCIDエネルギーの各値における、前記特定のm/zのプロダクトイオン量の総イオン量に対する百分率の各値を抽出し、
(d)工程(c)において抽出した前記特定のm/zのプロダクトイオン量の総イオン量に対する百分率の各値について、全ての組合せの和を求め、
(e)プレカーサーイオン量の総イオン量に対する百分率の値をxとし、かつ前記求めた各組合せの和の値をyとする関数のうち直線近似できるものを選択し、選択した各関数を与えるプロダクトイオンのm/z値から目的物質の構造を解析することを含む、
物質の構造解析方法。 - 工程(b)において、CIDエネルギーの値をX軸とし、かつ百分率の値をY軸として、プレカーサーイオン量の総イオン量に対する百分率と、特定のm/zのプロダクトイオン量の総イオン量に対する百分率とをプロットしてグラフを作成する、請求項1に記載の方法。
- 工程(c)において、プレカーサーイオン量の総イオン量に対する百分率をX軸として、前記特定のm/zのプロダクトイオン量の総イオン量に対する各百分率をY軸方向にプロットしてグラフを作成する、請求項1又は2に記載の方法。
- 工程(d)において、プレカーサーイオン量の総イオン量に対する百分率をX軸として、前記求めた各組合せの和の値をY軸方向にプロットしてグラフを作成する、請求項1~3のいずれか1項に記載の方法。
- 工程(e)において、MSn情報を取得する、請求項1~5のいずれか1項に記載の方法。
- 前記目的物質が糖鎖である、請求項1~6のいずれか1項に記載の方法。
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US13/818,141 US8575543B2 (en) | 2010-09-07 | 2011-09-07 | Method for analyzing structure of substance |
DE112011102604T DE112011102604T5 (de) | 2010-09-07 | 2011-09-07 | Verfahren zur Analyse einer Stoffstruktur |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2010-199676 | 2010-09-07 | ||
JP2010199676A JP5441064B2 (ja) | 2010-09-07 | 2010-09-07 | 物質の構造解析方法 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2012033111A1 true WO2012033111A1 (ja) | 2012-03-15 |
Family
ID=45810708
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2011/070322 WO2012033111A1 (ja) | 2010-09-07 | 2011-09-07 | 物質の構造解析方法 |
Country Status (4)
Country | Link |
---|---|
US (1) | US8575543B2 (ja) |
JP (1) | JP5441064B2 (ja) |
DE (1) | DE112011102604T5 (ja) |
WO (1) | WO2012033111A1 (ja) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115184530A (zh) * | 2022-08-19 | 2022-10-14 | 北京中医药大学 | 一种多级在线能量分辨质谱法及其在结构精准鉴定中的应用 |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8653448B1 (en) | 2012-09-07 | 2014-02-18 | Riken | Method for analyzing glycan structure |
WO2015021134A2 (en) * | 2013-08-06 | 2015-02-12 | University Of South Florida (A Florida Non-Profit Corporation) | Target binding molecules identified by kinetic target-guided synthesis |
US9466474B2 (en) * | 2013-08-08 | 2016-10-11 | Shimadzu Corporation | Triple quadrupole mass spectrometer |
CN108140060B (zh) | 2015-05-29 | 2022-06-28 | 沃特世科技公司 | 用于处理质谱数据的技术 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2010133707A (ja) * | 2008-06-17 | 2010-06-17 | Mitsubishi Chemicals Corp | 糖鎖構造解析方法 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1971998B1 (en) * | 2006-01-11 | 2019-05-08 | DH Technologies Development Pte. Ltd. | Fragmenting ions in mass spectrometry |
-
2010
- 2010-09-07 JP JP2010199676A patent/JP5441064B2/ja not_active Expired - Fee Related
-
2011
- 2011-09-07 WO PCT/JP2011/070322 patent/WO2012033111A1/ja active Application Filing
- 2011-09-07 DE DE112011102604T patent/DE112011102604T5/de not_active Ceased
- 2011-09-07 US US13/818,141 patent/US8575543B2/en not_active Expired - Fee Related
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2010133707A (ja) * | 2008-06-17 | 2010-06-17 | Mitsubishi Chemicals Corp | 糖鎖構造解析方法 |
Non-Patent Citations (1)
Title |
---|
OSAMU KANIE ET AL.: "Multi-stage mass spectrometric information obtained by deconvolution of energy-resolved spectra acquired by triple-quadrupole mass spectrometry", RAPID COMMUNUNICATIONS IN MASS SPECTROMETRY, vol. 25, no. IS.11, 15 June 2011 (2011-06-15), pages 1617 - 1624 * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115184530A (zh) * | 2022-08-19 | 2022-10-14 | 北京中医药大学 | 一种多级在线能量分辨质谱法及其在结构精准鉴定中的应用 |
CN115184530B (zh) * | 2022-08-19 | 2023-11-24 | 北京中医药大学 | 一种多级在线能量分辨质谱法及其在结构精准鉴定中的应用 |
Also Published As
Publication number | Publication date |
---|---|
US8575543B2 (en) | 2013-11-05 |
JP2012058002A (ja) | 2012-03-22 |
DE112011102604T5 (de) | 2013-05-23 |
JP5441064B2 (ja) | 2014-03-12 |
US20130153760A1 (en) | 2013-06-20 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Steen et al. | Quadrupole time‐of‐flight versus triple‐quadrupole mass spectrometry for the determination of phosphopeptides by precursor ion scanning | |
JP5111399B2 (ja) | 質量分析のための化学ノイズの低減 | |
US7189964B2 (en) | System and method for isotopic signature and mass analysis | |
Niessen et al. | Introduction to mass spectrometry, a tutorial | |
JP5441064B2 (ja) | 物質の構造解析方法 | |
JP7233436B2 (ja) | 糖タンパク質のための分析方法 | |
Satomi et al. | Site‐specific carbohydrate profiling of human transferrin by nano‐flow liquid chromatography/electrospray ionization mass spectrometry | |
Przybylski et al. | Discrimination of cyclic and linear oligosaccharides by tandem mass spectrometry using collision‐induced dissociation (CID), pulsed‐Q‐dissociation (PQD) and the higher‐energy C‐trap dissociation modes | |
US7158903B2 (en) | Methods for quantitative analysis by tandem mass spectrometry | |
Konda et al. | Assignment of the stereochemistry and anomeric configuration of sugars within oligosaccharides via overlapping disaccharide ladders using MSn | |
JP2021183977A (ja) | 最適化された標的を絞った分析 | |
Dudley et al. | Study of the mass spectrometric fragmentation of pseudouridine: comparison of fragmentation data obtained by matrix‐assisted laser desorption/ionisation post‐source decay, electrospray ion trap multistage mass spectrometry, and by a method utilising electrospray quadrupole time‐of‐flight tandem mass spectrometry and in‐source fragmentation | |
US20090216705A1 (en) | Method for predicting sugar chain structure | |
Cooks et al. | Direct analysis of complex mixtures by mass spectrometry | |
Zhang et al. | Hybrid triple quadrupole‐linear ion trap mass spectrometry in fragmentation mechanism studies: application to structure elucidation of buspirone and one of its metabolites | |
Harvey | Postsource decay fragmentation of N-linked carbohydrates from ovalbumin and related glycoproteins | |
Straub et al. | ‘Metabolic mapping’of drugs: rapid screening techniques for xenobiotic metabolites with ms/ms techniques | |
US20040180446A1 (en) | Methods and devices for identifying biopolymers using mass spectroscopy | |
Froesch et al. | Sialylation analysis of O‐glycosylated sialylated peptides from urine of patients suffering from Schindler's disease by Fourier transform ion cyclotron resonance mass spectrometry and sustained off‐resonance irradiation collision‐induced dissociation | |
Jovanović et al. | Negative ion MALDI‐TOF MS, ISD and PSD of neutral underivatized oligosaccharides without anionic dopant strategies, using 2, 5‐DHAP as a matrix | |
Xue et al. | Determination of linkage position and anomeric configuration in Hex‐Fuc disaccharides using electrospray ionization tandem mass spectrometry | |
Carlesso et al. | Stereochemical differentiation of four monosaccharides using transition metal complexes by electrospray ionization/ion-trap mass spectrometry | |
Daikoku et al. | Analysis of a series of isomeric oligosaccharides by energy‐resolved mass spectrometry: a challenge on homobranched trisaccharides | |
Daikoku et al. | Discrimination of 16 structural isomers of fucosyl galactoside based on energy-resolved mass spectrometry | |
Moneti et al. | Do collisions inside the collision cell play a relevant role in CID‐LIFT experiments? |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 11823585 Country of ref document: EP Kind code of ref document: A1 |
|
WWE | Wipo information: entry into national phase |
Ref document number: 13818141 Country of ref document: US |
|
WWE | Wipo information: entry into national phase |
Ref document number: 1120111026048 Country of ref document: DE Ref document number: 112011102604 Country of ref document: DE |
|
122 | Ep: pct application non-entry in european phase |
Ref document number: 11823585 Country of ref document: EP Kind code of ref document: A1 |