CN110095524A - A kind of triterpenoid saponin mass spectroscopy structural analytic method - Google Patents
A kind of triterpenoid saponin mass spectroscopy structural analytic method Download PDFInfo
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Abstract
The present invention relates to a kind of triterpenoid saponin mass spectroscopy structural analytic methods, sample is carried out to the second mass analysis under the positive ion mode under different impact energies, then the second mass analysis under the negative ion mode under different impact energies is carried out, the structure elucidation of the sugar chain composition and binding site of triterpene saponin componds is carried out using the second order ms daughter fragment ion feature that collisions different under cation and negative ion mode can induce.The precise Identification of triterpenoid saponin glycosyl and binding site can be fast implemented.
Description
Technical field
The invention belongs to separation analytical chemistry fields, and in particular to a kind of triterpenoid saponin mass spectroscopy structural analytic method.
Background technique
Disclosing the information of the background technology part, it is only intended to increase understanding of the overall background of the invention, without certainty
It is considered as recognizing or implying in any form that information composition has become existing skill well known to persons skilled in the art
Art.
Triterpene saponin componds are a kind of glycoside native compounds being made of triterpene parent nucleus and glycosyl, are had anti-swollen
Tumor, anti-aging, it is antifatigue, adjust immunity the effects of.This kind of compound is widely present among panax species, as ginseng,
American Ginseng etc..The sugar chain structure of triterpenoid saponin is sufficiently complex, including hexose, pentose, deoxidation hexose etc., common are Portugal
Grape sugar, galactolipin, rhamnose, arabinose, xylose, glucuronic acid and galacturonic acid etc..Common triterpenoid saponin has two
A sugar chain, the two sugar chains may be made of identical glycosyl, it is also possible to be made of different glycosyls, by monosaccharide, disaccharides, trisaccharide
Composition.Sugar chain has different binding sites on triterpene parent nucleus.There are a large amount of tetracyclic triterpene saponin(es in panax species, often
The binding site seen has C-3, C-6 and C-20, and the difference of binding site can cause its active dramatically different.
Sensitivity of mass spectrometry height, good resolution are used widely in the analysis of plant triterpene saponins compound.Liquid phase color
The separating capacity and mass-spectrometric technique combination for composing ultra high efficiency are that the high throughput analysis of triterpenoid saponin brings possibility.Inventors have found that
Structure is complicated for triterpenoid saponin, many kinds of, the standard specimen not usually being commercialized, and determines to its sugar chain composition and the accurate of binding site
Property brings difficulty.Common single collision energy, single ionic mode second mass analysis means tend not to accurately
Identify its binding site and sugar chain composition, and multi-stage ms are more demanding to compound concentration, limit its application.
Summary of the invention
For above-mentioned problems of the prior art, it is an object of the present invention to provide a kind of triterpenoid saponin mass spectrum knots
Structure analytic method.
In order to solve the above technical problems, the technical solution of the present invention is as follows:
A kind of triterpenoid saponin mass spectroscopy structural analytic method, specific steps are as follows: sample is first passed through into liquid chromatogram separation, will be divided
Sample from after carries out the second mass analysis under the positive ion mode under different impact energies, then carries out under different impact energies
Second mass analysis under negative ion mode, second order ms that can induce using collisions different under cation and negative ion mode
Fragment ion feature carries out the structure elucidation of the sugar chain composition and binding site of triterpene saponin compound.
By the mass spectral characteristic under negative ions mode, in conjunction with the second order ms cracking characteristic that collision can induce, to glycosyl
Structure and binding site carry out accurately it is qualitative.In the prior art, single collision energy, single ionic mode second level matter
Spectrum analysis means cannot accurately identify its binding site and sugar chain composition, and multi-stage ms are more demanding to compound concentration,
Accurate qualitative analysis cannot be carried out to the triterpenoid saponin of low concentration.The connection of positive ion mode and negative ion mode in the present invention
With respectively to the accurate structure elucidation of composition progress of binding site and sugar chain.In the present invention, by son under positive ion mode from
The energy resolution curve of son determines the binding site of sugar chain.Sugar is determined by the energy resolution curve of daughter ion under negative ion mode
The composition of glycosyl in chain.
Collision energy range under positive ion mode is 0-100eV, does not include 0eV, is divided into 5-10eV；
Collision energy range under negative ion mode is 0-100eV, does not include 0eV, is divided into 5-10eV.
In some embodiments, the number of impact energy selected element can be 4-7 under positive ion mode；In other implementation
In example, 4 impact energies, 40eV, 50eV, 60eV, 70eV；5 impact energies in yet other embodiments, respectively 45eV,
55eV,65eV,75eV,85eV；6 impact energies in yet other embodiments, respectively 40eV, 50eV, 60eV, 70eV,
80eV,90eV；7 impact energies in yet other embodiments, respectively 40eV, 45eV, 50eV, 55eV, 60eV, 65eV,
70eV。
The binding site information for obtaining sugar chain in the present invention in the positive-ion mode, carries out second level under different impact energies
Mass spectral analysis, and with the raising of impact energy, the signal of sugar chain is gradually appeared, and difference is obtained by the information of fragment ion
The information of the sugar chain of position.In the present invention under positive ion mode, with the increase of impact energy, sugar chain and triterpene parent nucleus junction
C-O key can be broken, can gradually appear sugar chain plus sodium peak, C-20 sugar chains add sodium peak to occur under low impact energy,
C-3 and C-6 sugar chain adds sodium peak to occur under high impact energy.
In some embodiments, 3-5 impact energy under negative ion mode；3 impact energies in yet other embodiments,
Respectively 65eV, 75eV, 85eV；4 impact energies, respectively 50eV, 60eV, 70eV, 80eV in yet other embodiments,；?
In other embodiment, 5 impact energies, respectively 55eV, 65eV, 75eV, 85eV, 95eV.
The sugar chain under positive ion mode is further analyzed in the negative ion mode in the present invention, obtains sugar chain
Composition.With the raising of energy, the glycosyl in sugar chain progressively disengages sugar chain, and can obtain specific glycosyl molecule.With touching
The increase for hitting energy, gradually appears the neutral loss of glycosyl, the composition of the glycosyl in sugar chain is determined according to the mass number of neutral loss.
Wherein, neutral loss 162 is hexose, neutral loss 146 is deoxidation hexose, neutral loss 132 is pentose, neutral loss
176 be hexose aldehydic acid.
In some embodiments, the second order ms daughter ion under positive ion mode is [M+Na]+。
In some embodiments, the second order ms daughter ion under negative ion mode is [M-H]-。
In some embodiments, sample material is American Ginseng, ginseng or Radix Notoginseng.
In some embodiments, liquid phase chromatogram condition: mobile phase A and B are respectively the aqueous solution containing 0.1% formic acid and contain
There is the acetonitrile solution of 0.1% formic acid.Flow velocity 0.3mL/min.Chromatographic column is reverse phase C18 chromatographic column.Column temperature 30-50 degree.When elution
Between be 20-50min.Elution start gradient is 5%B, and final gradient is 100%B, and maintains 3-5min using 100%B.
In some embodiments, pre-treatment is carried out before sample material is detected obtains sample, the preparation method of sample
For solid sample is prepared into powder, extractant is then added, the supernatant obtain after ultrasound, centrifugation is sample.
Extractant is the mixture of organic solvent and water in yet other embodiments, and the mass ratio of organic solvent and water is
5:1-3:1；Organic solvent is one of acetonitrile, methanol, ethyl alcohol in yet other embodiments,；1g in yet other embodiments,
The volume of the corresponding extractant of powder is 8-12mL；The ultrasonic time is 20-40min in yet other embodiments,；Other
In embodiment, the time of centrifugation is 7-13min.
Using single organic solvent and water as extractant in the present invention, and the supernatant after centrifugation is as extraction
Liquid, after liquid chromatogram, there are many contents are low and the high triterpenoid saponin of content, the present invention can be under low concentration for discovery
Triterpenoid saponin carries out accurate structural parsing.
Above-mentioned triterpenoid saponin mass spectroscopy structural analytic method in Panax Chinese medicine triterpenoid saponin it is qualitative in application.
In some embodiments, Panax Chinese medicine includes ginseng, American Ginseng and Radix Notoginseng.
Beneficial effects of the present invention:
Second order ms feature under cation and negative ion mode that the present invention passes through different impact energies realizes triterpenoid saponin
Sugar chain composition and binding site structure elucidation；
It is that one of acetonitrile, methanol, ethyl alcohol extract that the present invention, which extracts liquid and preparation method thereof using organic solvent, first benefit
Extracting solution is separated with liquid chromatogram, then carries out second mass analysis, improves the detection and analysis of triterpenoid saponin
Sensitivity and accuracy；
The method of the present invention is easy to operate, quick, accuracy is high.
Detailed description of the invention
The Figure of description for constituting a part of the invention is used to provide further understanding of the present application, and of the invention shows
Examples and descriptions thereof are used to explain the present invention for meaning property, does not constitute improper limitations of the present invention.
Fig. 1 is the energy resolution curve of the daughter ion of triterpenoid saponin Re under 1 positive ion mode of embodiment；
Fig. 2 is the energy resolution curve of the daughter ion of triterpenoid saponin Re under 1 negative ion mode of embodiment.
Fig. 3 is triterpenoid saponin Rb under 2 positive ion mode of embodiment2Daughter ion energy resolution curve；
Fig. 4 is triterpenoid saponin Rb under 2 negative ion mode of embodiment2Daughter ion energy resolution curve.
Fig. 5 is triterpenoid saponin Rb under 3 positive ion mode of embodiment1Daughter ion energy resolution curve；
Fig. 6 is triterpenoid saponin Rb under 3 negative ion mode of embodiment1Daughter ion energy resolution curve.
Specific embodiment
It is noted that following detailed description is all illustrative, it is intended to provide further instruction to the present invention.Unless another
It indicates, all technical and scientific terms used herein has usual with general technical staff of the technical field of the invention
The identical meanings of understanding.
It should be noted that term used herein above is merely to describe specific embodiment, and be not intended to restricted root
According to the illustrative embodiments of the application.As used herein, unless the context clearly indicates otherwise, otherwise singular
Also it is intended to include plural form, additionally, it should be understood that, when in the present specification using term "comprising" and/or " packet
Include " when, indicate existing characteristics, step, operation, device, component and/or their combination.Below with reference to embodiment to this hair
Bright further explanation
Embodiment 1
Triterpenoid saponin is qualitative in American Ginseng:
(1) American Ginseng extracting solution is prepared
American Ginseng powder 1g is added to methanol/water (v/v, 4:1) solution of 10mL, ultrasonic 30min is centrifuged 10min, takes
Clear liquid.
(2) second mass analysis
Supernatant is subjected to LC-MS analysis (UHPLC-Q-TOF-MS), chromatographic column is Acquity BEH C18
(2.1 × 100mm, 1.7 μm, Waters), mobile phase A and B are respectively the aqueous solution containing 0.1% formic acid and 0.1% formic acid
Acetonitrile solution, 40 DEG C of column temperature, gradient 0min, 5%B；10min, 50%B；25min, 100%B；30min, 100%B.
Mass spectrum dries gas 8L/min, dry 200 DEG C, atomization gas 2bar of temperature degree, and capillary voltage 3500V in the positive-ion mode is born
3000V under ion mode, 80 μ s, RF 750Vpp of collision cell transmission time, isolation width are 4, mass number range 50-1200.
Carry out the second mass analysis of different impact energies, the impact energy under positive ion mode be 40eV, 50eV, 60eV,
70eV.Impact energy under negative ion mode is 50eV, 60eV, 70eV, 80eV.
(3) triterpenoid saponin is qualitative:
By the energy resolution curve of daughter ion under cation and negative ion mode, qualitative 35 kinds of triterpenes out in American Ginseng
Saponin component, and its glycosyl connection site and glycosyl composition has been determined.By taking Re as an example.As depicted in figs. 1 and 2, Re just from
Under subpattern, the signal of 203 daughter ion of sugar chain occurs at low energies, and the signal of 349 daughter ion of sugar chain goes out at a high energy
It is existing, illustrate sugar chain 203 at C-20, and sugar chain 349 is at C-6.In the negative ion mode, there is 783 ([M-H- of daughter ion
162]-)、799([M-H-146]-)、637([M-H-162-146]-)、475([637-162]-).Illustrate that sugar chain 203 is one six
Carbon sugar.Sugar chain 349 is a deoxidation hexose and a hexose.Wherein M refers to the molecular weight of Re, 783 ([M-H-
132]-) it is the daughter ion that one hexose of sugar chain neutral loss of Re obtains, 799 ([M-H-146]-) lost for the sugar chain neutrality of Re
Lose the daughter ion that a deoxidation hexose obtains, 637 ([M-H-162-146]-) be Re one hexose of sugar chain neutral loss
The daughter ion obtained with a deoxidation hexose, 475 ([637-162]-) one hexose of neutral loss obtains again later
Daughter ion.
Embodiment 2
Triterpenoid saponin is qualitative in ginseng:
(1) Ginseng extract is prepared
Ginseng Root Powder 1g is added to methanol/water (v/v, 4:1) solution of 11mL, ultrasonic 35min is centrifuged 12min, takes supernatant
Liquid.
(2) second mass analysis
Supernatant is subjected to LC-MS analysis (UHPLC-Q-TOF-MS), chromatographic column ZORBAX-SB-AQ
C18 (2.1 × 100mm, 1.8 μm, Agient), mobile phase A and B are respectively the aqueous solution containing 0.1% formic acid and 0.1% formic acid
Acetonitrile solution, 45 DEG C of column temperature, gradient 0min, 5%B；6min, 40%B；20min, 100%B；25min, 100%B.
Mass spectrum dries gas 8L/min, dry 200 DEG C, atomization gas 2bar of temperature degree, and capillary voltage 3500V in the positive-ion mode is born
3000V under ion mode, 80 μ s, RF 750Vpp of collision cell transmission time, isolation width are 4, mass number range 50-1200.
Carry out the second mass analysis of different impact energies, the impact energy under positive ion mode be 45eV, 55eV, 65eV,
75eV,85eV.Impact energy under negative ion mode is 55eV, 65eV, 75eV, 85eV, 95eV.
(3) triterpenoid saponin is qualitative:
By the energy resolution curve of daughter ion under cation and negative ion mode, qualitative 50 kinds of triterpene soaps out in ginseng
Methods of glycosides, and its glycosyl connection site and glycosyl composition has been determined.With Rb2For.As shown in Figure 3 and Figure 4, Rb2In cation
Under mode, the signal of 355 daughter ion of sugar chain occurs at low energies, and the signal of 365 daughter ion of sugar chain occurs at a high energy,
Illustrate sugar chain 355 at C-20, and sugar chain 365 is at C-3.In the negative ion mode, there is 945 ([M-H- of daughter ion
132]-)、915([M-H-162]-)、783([M-H-162-132]-)、621([783-162]-)、459([621-162]-).Explanation
Sugar chain 365 is two hexoses.Sugar chain 355 is a pentose and a hexose.Wherein M refers to Rb2Molecular weight,
945([M-H-132]-) it is Rb2The obtained daughter ion of one pentose of sugar chain neutral loss, 915 ([M-H-162]-) it is Rb2
The obtained daughter ion of one hexose of sugar chain neutral loss, 783 ([M-H-162-132]-) it is Rb2Sugar chain neutral loss one
The daughter ion that a pentose and a hexose obtain, 621 ([783-162]-) it is that one hexose of neutral loss obtains again later
The daughter ion arrived, 459 ([621-162]-) it is the daughter ion that one hexose of neutral loss obtains again later.
Embodiment 3
Triterpenoid saponin is qualitative in Radix Notoginseng:
(1) Radix Notoginseng extracting solution is prepared
Notoginseng Root 1g is added to methanol/water (v/v, 4:1) solution of 9mL, ultrasonic 26min is centrifuged 8min, takes supernatant
Liquid.
(2) second mass analysis
Supernatant is subjected to LC-MS analysis (UHPLC-Q-TOF-MS), chromatographic column ZORBAX-SB-AQ
C18 (2.1 × 100mm, 1.8 μm, Agient), mobile phase A and B are respectively the aqueous solution containing 0.1% formic acid and 0.1% formic acid
Acetonitrile solution, 40 DEG C of column temperature, gradient 0min, 5%B；7min, 50%B；21min, 100%B；25min, 100%B.
Mass spectrum dries gas 8L/min, dry 200 DEG C, atomization gas 2bar of temperature degree, and capillary voltage 3500V in the positive-ion mode is born
3000V under ion mode, 80 μ s, RF 750Vpp of collision cell transmission time, isolation width are 4, mass number range 50-1200.Into
The second mass analysis of the different impact energies of row, the impact energy under positive ion mode is 40eV, 50eV, 60eV, 70eV, 80eV,
90eV.Impact energy under negative ion mode is 65eV, 75eV, 85eV.
(3) triterpenoid saponin is qualitative:
By the energy resolution curve of daughter ion under cation and negative ion mode, qualitative 20 kinds of triterpene soaps out in Radix Notoginseng
Methods of glycosides, and its glycosyl connection site and glycosyl composition has been determined.With Rb1For.As shown in Figure 5 and Figure 6, Rb1In cation
Under mode, only there is the signal of 365 daughter ion of sugar chain, relative intensity is always 100% when impact energy is more than or equal to 60eV.?
Under negative ion mode, there is daughter ion 945 ([M-H-162]-)、783([M-H-162-162]-)、621([783-162]-)、459
([621-162]-).Illustrate that sugar chain 365 is two hexoses.And C-20 and C-3 be all two hexoses sugar chain.Its
Middle M refers to Rb1Molecular weight, 945 ([M-H-162]-) it is Rb1The obtained daughter ion of one hexose of sugar chain neutral loss,
783([M-H-162-162]-) it is Rb1The obtained daughter ion of two hexoses of sugar chain neutral loss, 621 ([783-162]-) be
The daughter ion that one hexose of neutral loss obtains again later, 459 ([621-162]-) be after one six carbon of neutral loss again
The daughter ion that sugar obtains.
Embodiment 4
Triterpenoid saponin is qualitative in American Ginseng:
(1) American Ginseng extracting solution is prepared
American Ginseng powder 1g is added to methanol/water (v/v, 4:1) solution of 8mL, ultrasonic 25min is centrifuged 13min, takes
Clear liquid.
(2) second mass analysis
Supernatant is subjected to LC-MS analysis (UHPLC-Q-TOF-MS), chromatographic column is Acquity BEH C18
(2.1 × 100mm, 1.7 μm, Waters), mobile phase A and B are respectively the aqueous solution containing 0.1% formic acid and 0.1% formic acid
Acetonitrile solution, 40 DEG C of column temperature, gradient 0min, 5%B；10min, 50%B；25min, 100%B；30min, 100%B.
Mass spectrum dries gas 8L/min, dry 200 DEG C, atomization gas 2bar of temperature degree, and capillary voltage 3500V in the positive-ion mode is born
3000V under ion mode, 80 μ s, RF 750Vpp of collision cell transmission time, isolation width are 4, mass number range 50-1200.Into
The second mass analysis of the different impact energies of row, the impact energy under positive ion mode is 40eV, 45eV, 50eV, 55eV, 60eV,
65eV,70eV.Impact energy under negative ion mode is 50eV, 60eV, 70eV, 80eV.
(3) triterpenoid saponin is qualitative:
By the energy resolution curve of daughter ion under cation and negative ion mode, qualitative 35 kinds of triterpenes out in American Ginseng
Saponin component, and its glycosyl connection site and glycosyl composition has been determined.With Rb2For.Rb2In the positive-ion mode, sugar chain
355 signal occurs at low energies, and the signal of sugar chain 365 occurs at a high energy, illustrate sugar chain 355 at C-20, and
Sugar chain 365 is at C-3.In the negative ion mode, there is daughter ion 945 ([M-H-132]-)、915([M-H-162]-)、783
([M-H-162-132]-)、621([783-162]-)、459([621-162]-).Illustrate that sugar chain 365 is two hexoses.Sugar chain
355 be a pentose and a hexose.Wherein M refers to Rb2Molecular weight.
Comparative example 1
Triterpenoid saponin is qualitative in American Ginseng:
(1) American Ginseng extracting solution is prepared
American Ginseng powder 1g is added to methanol/water (v/v, 4:1) solution of 10mL, ultrasonic 30min is centrifuged 10min, takes
Clear liquid.
(2) second mass analysis
Supernatant is subjected to LC-MS analysis (UHPLC-Q-TOF-MS), chromatographic column is Acquity BEH C18
(2.1 × 100mm, 1.7 μm, Waters), mobile phase A and B are respectively the aqueous solution containing 0.1% formic acid and 0.1% formic acid
Acetonitrile solution, 40 DEG C of column temperature, gradient 0min, 5%B；10min, 50%B；25min, 100%B；30min, 100%B.
Mass spectrum dries gas 8L/min, dry 200 DEG C, atomization gas 2bar of temperature degree, and capillary voltage 3500V in the positive-ion mode is born
3000V under ion mode, 80 μ s, RF 750Vpp of collision cell transmission time, isolation width are 4, mass number range 50-1200.Into
The second mass analysis of the different impact energies of row, the impact energy under positive ion mode is 70eV, 80eV, 90eV, 100eV.Anion
Impact energy under mode is 40eV, 50eV, 60eV.
(3) triterpenoid saponin is qualitative:
By the energy resolution curve of daughter ion under cation and negative ion mode, there was only 15 kinds of triterpene soaps in American Ginseng
Methods of glycosides has determined its glycosyl connection site and glycosyl composition.Since impact energy selection does not conform under cation and negative ion mode
It is suitable, it not can determine that its glycosyl connection site and sugar chain form by 20 kinds of triterpenoid saponins.Impact energy is too under positive ion mode
Height, so that the discrimination of sugar chain daughter ion is deteriorated.Such as Re, the phase of sugar chain 349 and sugar chain 203 under 70eV-100eV impact energy
It is relatively high to intensity, it cannot effectively distinguish it and be broken successive problem, so not can determine that the position that sugar chain is connect with aglycon.
Impact energy under negative ion mode is too low, cannot obtain daughter fragment ion data abundant.Such as Rb2, three sugar of neutral loss
Daughter ion 621 and neutral loss four sugared daughter ions 459 do not obtained under 40-60eV impact energy, cannot effectively determine
The composition of its sugar chain.
Comparative example 2
Triterpenoid saponin is qualitative in American Ginseng:
(1) American Ginseng extracting solution is prepared
American Ginseng powder 1g is added to methanol/water (v/v, 4:1) solution of 10mL, ultrasonic 30min is centrifuged 10min, takes
Clear liquid.
(2) second mass analysis
Supernatant is subjected to LC-MS analysis (UHPLC-Q-TOF-MS), chromatographic column is Acquity BEH C18
(2.1 × 100mm, 1.7 μm, Waters), mobile phase A and B are respectively the aqueous solution containing 0.1% formic acid and 0.1% formic acid
Acetonitrile solution, 40 DEG C of column temperature, gradient 0min, 5%B；10min, 50%B；25min, 100%B；30min, 100%B.
Mass spectrum dries gas 8L/min, dry 200 DEG C, atomization gas 2bar of temperature degree, and capillary voltage 3500V in the positive-ion mode is born
3000V under ion mode, 80 μ s, RF 750Vpp of collision cell transmission time, isolation width are 4, mass number range 50-1200.Into
Second mass analysis under the positive ion mode of the different impact energies of row, impact energy 50eV, 60eV, 70eV, 80eV, 90eV.
(3) triterpenoid saponin is qualitative:
By the energy resolution curve of daughter ion under positive ion mode, during American Ginseng triterpenoid saponin structure elucidation,
The sugar chain of triterpenoid saponins and the connection site of aglycon can only be identified, cannot it is qualitative go out sugar chain in specific glycosyl group
At.
The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention, for the skill of this field
For art personnel, the invention may be variously modified and varied.All within the spirits and principles of the present invention, made any to repair
Change, equivalent replacement, improvement etc., should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of triterpenoid saponin mass spectroscopy structural analytic method, it is characterised in that: specific steps are as follows:
Sample is first passed through into liquid chromatogram separation, the sample after separation is carried out to two under the positive ion mode under different impact energies
Grade mass spectral analysis, then carry out the second mass analysis under the negative ion mode under different impact energies, using cation and bear from
The second level daughter ion feature that different collisions can induce under subpattern carries out the sugar chain composition and binding site of triterpene saponin compound
Structure elucidation.
2. according to the method described in claim 1, it is characterized by: the number of impact energy selected element can be under positive ion mode
4-7；
Preferably, 4 impact energies, 40eV, 50eV, 60eV, 70eV；
Preferably, 5 impact energies, 45eV, 55eV, 65eV, 75eV, 85eV；
Preferably, 6 impact energies, respectively 40eV, 50eV, 60eV, 70eV, 80eV, 90eV；
Preferably, 7 impact energies, respectively 40eV, 45eV, 50eV, 55eV, 60eV, 65eV, 70eV.
3. according to the method described in claim 1, it is characterized by: 3-5 impact energy under negative ion mode；
Preferably, 3 impact energies, respectively 65eV, 75eV, 85eV；
Preferably, 4 impact energies, respectively 50eV, 60eV, 70eV, 80eV；
Preferably, 5 impact energies, respectively 55eV, 65eV, 75eV, 85eV, 95eV.
4. according to the method described in claim 1, it is characterized by: the second order ms daughter ion under positive ion mode is [M+Na]+。
5. according to the method described in claim 1, it is characterized by: the second order ms daughter ion under negative ion mode is [M-H]-。
6. according to the method described in claim 1, it is characterized by: sample material is American Ginseng, ginseng or Radix Notoginseng.
7. according to the method described in claim 1, it is characterized by: liquid phase chromatogram condition: mobile phase A and B respectively contain
The aqueous solution of 0.1% formic acid and acetonitrile solution containing 0.1% formic acid.Flow velocity 0.3mL/min.Chromatographic column is reverse phase C18 chromatography
Column.30-50 DEG C of column temperature.Elution time is 20-50min.Elution start gradient is 5%B, and final gradient is 100%B, and is used
100%B maintains 3-5min.
8. according to the method described in claim 1, it is characterized by: sample material, which carries out pre-treatment before being detected, obtains sample
Product, the preparation method of sample are that solid sample is prepared into powder, and extractant dissolution is then added, after progress ultrasound, centrifugation
To supernatant be sample；
Preferably, the ultrasonic time is 20-40min；
Preferably, the time of centrifugation is 7-13min；
Preferably, the volume of the corresponding extractant of 1g powder is 8-12mL.
9. according to the method described in claim 8, it is characterized by: extractant is the mixture of organic solvent and water, You Jirong
The mass ratio of agent and water is 5:1-3:1；
Preferably, organic solvent is one of acetonitrile, methanol, ethyl alcohol.
10. application of the described in any item methods of claim 1-9 in Panax Chinese medicine in triterpenoid saponin structure elucidation；
Preferably, Panax Chinese medicine includes ginseng, American Ginseng and Radix Notoginseng.
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