CN101266231A - Pederin discrimination method - Google Patents
Pederin discrimination method Download PDFInfo
- Publication number
- CN101266231A CN101266231A CNA2008100362454A CN200810036245A CN101266231A CN 101266231 A CN101266231 A CN 101266231A CN A2008100362454 A CNA2008100362454 A CN A2008100362454A CN 200810036245 A CN200810036245 A CN 200810036245A CN 101266231 A CN101266231 A CN 101266231A
- Authority
- CN
- China
- Prior art keywords
- malicious
- ion peak
- ion
- pederin
- discrimination method
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Abstract
The invention discloses a discrimination method of Paederus comprising a sample processing and data capturing process, wherein the Paederus extract product is diluted by 100 times using 50% of HPLC methanol and the data capturing process comprises the following steps: respectively scanning the Paederus extract product diluent in anion mode and cation mode using the liquid chromagraphy/mass spectrometry to acquire parent ion peak value; performing the collision induced dissociation and scanning using trace sampling pump post-column injection system to acquire daughter ion peak data. The discrimination method of Paederus uses liquid chromagraphy/mass spectrometry to obtain the features that: excellent sensitivity, accurate discrimination result, efficiency and reliability, and convenient operation and provides convenience for the clinical research on the Paederus extract product.
Description
Technical field
The present invention relates to a kind of discrimination method of malicious pederin, specifically, relate to a kind of method of utilizing the liquid chromatograph mass spectrography method to differentiate malicious pederin.
Background technology
The poison rove beetle is a genus in the Staphylinidae, and known kind surpasses 250 kinds, originates from except that the Antarctica all over the world, and the Chinese malicious rove beetle of having reported has 21 kinds.The poison rove beetle mainly lives in moist farmland, vegetable plot, the weeds clump, is the common small insects in gully, farmland.The common malicious rove beetle of China has blue or green wing ant shape rove beetle (claiming fusiformis poison rove beetle, yellow sufficient ant shape rove beetle again), black sufficient ant shape rove beetle, the crinosity rove beetle, towel shape poison rove beetle, little red malicious rove beetle, ant attitude poison rove beetle, 7 kinds of Tibet poison rove beetles etc., it is the widest wherein to distribute with blue or green wing ant shape rove beetle, spreads all over areas such as each province and Taiwan.
Ovum, larva, pupa and the adult (each several part except that wing) of poison rove beetle all have toxin existence, and the toxin of adult is mainly derived from hemolymph, all contain toxin in female, the male worm body.The toxin of poison rove beetle is the chemical substance that polypide is used for resisting external invasion and attack.The venom complex chemical composition of poison rove beetle contains aldehydes, quinones, terpene etc., has its toxin of report to be mainly 3 kinds, is respectively malicious pederin, plan poison pederin and malicious rove beetle ketone.Wherein, the content of malicious pederin is the highest, and molecular weight is 503.65, is dissolved in organic solvent ethanol, ether, chloroform and benzene, and is water insoluble, and the malicious pederin of crystallization is 112 ℃ of fusings, and faintly acid is boiled 1h and is not destroyed in the water.
The inventor discovers that malicious rove beetle extract has the healing of the diabetes skin ulcer of promotion, suppresses the growth of HL-60 human leukemia cell line, suppresses growth of BEL-7402 human hepatoma cell strain and trichogenous medicinal efficacy, medically be with a wide range of applications, be expected to be used to prepare treatment diabetic skin telephium, the human leukemia for the treatment of some type and people's liver cancer and trichogenous pharmaceutical preparation.But at present the discrimination method of poisonous pederin not as yet makes the research clinically of malicious rove beetle extract have very big difficulty.
Summary of the invention
The discrimination method that the purpose of this invention is to provide a kind of malicious pederin is for the research clinically of malicious rove beetle extract provides convenient.
For solving the problems of the technologies described above, technical scheme of the present invention is as follows:
The discrimination method of malicious pederin of the present invention comprises sample preparation and data acquisition, it is characterized in that, described sample preparation is with 100 times of 50%HPLC level methyl alcohol dilutions with malicious rove beetle extract; It is at first to utilize liquid chromatograph-mass spectrometer to scan malicious rove beetle extract dilution respectively under negative ion mode and positive ion mode that described data are obtained, to obtain the parent ion peak data; Injection system carries out collision induced dissociation and scanning after adopting the micro-sampling pump plunger shaft then, to obtain the daughter ion peak data.
Described liquid phase chromatogram condition is: chromatographic column is that Shiseido C18/100 * 2.1mm/5u, moving phase are that volume ratio is that 70: 30 methyl alcohol one water and solution system, the flow velocity that contains the 1mM ammonium formate are 0.2mlmin
-1, column temperature is that 30 ℃, sample size are 10 μ l; Described mass spectrum condition is: Turbo Ionspray ion gun, triple quadrupole bar mass detector, spraying gas 13L/min, gas curtain gas 12L/min, spray voltage 5KV, focus voltage 400V, inlet voltage 10V, outlet voltage 10V, 450 ℃ of auxiliary heating actuator temperatures and quality of scanning scope are 400～800amu.
Described parent ion peak data are: chromatographic peak retention time t
R=4.0min, quasi-molecular ion peak [M-H]
-Be m/z=502.4, with the adduct ion peak [M+HCOO] of formate
-Be m/z=548.4, with the adduct ion peak [M+Na] of sodion
+Be m/z=526.4; Described daughter ion peak data is: m/z 470, m/z 438, m/z502.2, m/z 470, m/z 438, m/z 494.2, m/z 462.2, m/z 340.
The discrimination method of malicious pederin provided by the invention, owing to adopted the liquid chromatograph mass spectrography system, make this discrimination method have good sensitivity, identification result is accurate and effective and reliable, and easy to operate, for the research clinically of malicious rove beetle extract provides convenience.
Description of drawings
0～8min chromatogram that Fig. 1 scans under negative ion mode for malicious rove beetle extract dilution;
0～8min chromatogram that Fig. 2 scans under positive ion mode for malicious rove beetle extract dilution;
Fig. 3 is parent ion peak (t
R=4.0min) the mass spectrogram under negative ion mode;
Fig. 4 is parent ion peak (t
R=4.0min) the mass spectrogram under positive ion mode;
Fig. 5 is the daughter ion mass spectrogram of m/z=526.4 ion;
Fig. 6 is the daughter ion mass spectrogram of m/z=548.4 ion;
Fig. 7 is the daughter ion mass spectrogram of m/z=502.2 ion;
Fig. 8 is the chemical molecular structural formula of malicious pederin compound.
Embodiment
The invention will be further described below by embodiment, and its purpose only is better to understand content of the present invention and unrestricted protection scope of the present invention.
Embodiment
1, sample preparation
Malicious rove beetle extract is diluted 100 times with 50%HPLC level methyl alcohol.
2, data are obtained
A) the parent ion peak data obtains
Liquid phase chromatogram condition is: chromatographic column is that Shiseido C18/100 * 2.1mm/5u, moving phase are that volume ratio is that 70: 30 methyl alcohol one water and solution system, the flow velocity that contains the 1mM ammonium formate are 0.2mlmin
-1, column temperature is that 30 ℃, sample size are 10 μ l; The mass spectrum condition is: 450 ℃ of Turbo Ionspray ion gun, triple quadrupole bar mass detector, spraying gas 13L/min, gas curtain gas 12L/min, spray voltage 5KV, focus voltage 400V, inlet voltage 10V, outlet voltage 10V and auxiliary heating actuator temperatures; The mass spectrum working method is set at Q1 scanning, the quality of scanning scope is 400～800amu, the malicious rove beetle extract dilution of scanning under negative ion mode and positive ion mode obtains 0～8min chromatogram (seeing illustrated in figures 1 and 2) of scanning under different ionization patterns respectively, and visible two figure are at t
RThere is a total peak at=4.0min place.
By this total peak (t
R=4.0min) mass spectrogram under negative ion mode and positive ion mode (seeing Fig. 3 and shown in Figure 4) is as seen: the ion of m/z=502.4 is the quasi-molecular ion of malicious pederin compound molecule, can be expressed as [M-H]
-The ion of m/z=548.4 is the adduct ion of malicious pederin compound molecule and formate, can be expressed as [M+HCOO]
-The m/z=526.4 ion is the adduct ion of malicious pederin compound molecule and sodion, can be expressed as [M+Na]
+
B) the daughter ion peak data obtains
Injection system behind the employing micro-sampling pump plunger shaft carries out daughter ion scanning to m/z=526.4,548.4,502.2, and malicious rove beetle extract dilution sample introduction speed is 10ul/min, and collision gas is 7L/min.
As seen from Figure 5, the daughter ion mass spectrum of m/z=526.4 ion has m/z=494.2, m/z=462.2 and m/z=340, and wherein m/z=494.2, m/z=462.2 are [M+Na]
+Lose CH
3The daughter ion of OH, m/z=340 are the daughter ion of the formation of carbonyl α side chain fracture.
As seen from Figure 6, the daughter ion mass spectrum of m/z=548.4 ion has m/z=502.2, m/z=470 and m/z=438, and wherein m/z=502.2 has proved m/z=548.4 ([M+HCOO]
-) and [M-H]
-Relation, m/z=470, m/z=438 are [M-H]
-+Lose CH
3The daughter ion of OH.
As seen from Figure 7, the daughter ion mass spectrum of m/z=502.2 ion has m/z=470 and m/z=438, and they are [M-H]
-+Lose CH
3The daughter ion of OH.
Comprehensive above-mentioned data analysis as seen, t
RIn the molecular structure of compounds of=4.0min chromatographic peak correspondence a plurality of CH are arranged
3The O group, the ESI mass spectrum of this compound easily forms adduct ion, molecular weight is 503, fits like a glove with the chemical molecular architectural feature of malicious pederin compound, and the discrimination method that it can be said that bright malicious pederin of the present invention can accurately identify the malicious pederin in the malicious rove beetle extract.
Claims (3)
1. the discrimination method of a malicious pederin comprises sample preparation and data acquisition, it is characterized in that, described sample preparation is with 100 times of 50%HPLC level methyl alcohol dilutions with malicious rove beetle extract; It is at first to utilize liquid chromatograph-mass spectrometer to scan malicious rove beetle extract dilution respectively under negative ion mode and positive ion mode that described data are obtained, to obtain the parent ion peak data; Injection system carries out collision induced dissociation and scanning after adopting the micro-sampling pump plunger shaft then, to obtain the daughter ion peak data.
2. the discrimination method of malicious pederin according to claim 1, it is characterized in that described liquid phase chromatogram condition is: chromatographic column is that Shiseido C18/100 * 2.1mm/5u, moving phase are that volume ratio is that 70: 30 methyl alcohol one water and solution system, the flow velocity that contains the 1mM ammonium formate are 0.2mlmin
-1, column temperature is that 30 ℃, sample size are 10 μ l; Described mass spectrum condition is: Turbo Ionspray ion gun, triple quadrupole bar mass detector, spraying gas 13L/min, gas curtain gas 12L/min, spray voltage 5KV, focus voltage 400V, inlet voltage 10V, outlet voltage 10V, 450 ℃ of auxiliary heating actuator temperatures and quality of scanning scope are 400～800amu.
3. the discrimination method of malicious pederin according to claim 1 is characterized in that, described parent ion peak data are: chromatographic peak retention time t
R=4.0min, quasi-molecular ion peak [M-H]
-Be m/z=502.4, with the adduct ion peak [M+HCOO] of formate
-Be m/z=548.4, with the adduct ion peak [M+Na] of sodion
+Be m/z=526.4; Described daughter ion peak data is: m/z 470, m/z 438, m/z 502.2, m/z 470, m/z 438, m/z494.2, m/z462.2, m/z340.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008100362454A CN101266231B (en) | 2008-04-18 | 2008-04-18 | Pederin discrimination method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008100362454A CN101266231B (en) | 2008-04-18 | 2008-04-18 | Pederin discrimination method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101266231A true CN101266231A (en) | 2008-09-17 |
| CN101266231B CN101266231B (en) | 2011-05-25 |
Family
ID=39988795
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2008100362454A Expired - Fee Related CN101266231B (en) | 2008-04-18 | 2008-04-18 | Pederin discrimination method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101266231B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101773101A (en) * | 2010-03-03 | 2010-07-14 | 中国农业科学院植物保护研究所 | Method for artificially and massively feeding natural enemy staphylinid |
-
2008
- 2008-04-18 CN CN2008100362454A patent/CN101266231B/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101773101A (en) * | 2010-03-03 | 2010-07-14 | 中国农业科学院植物保护研究所 | Method for artificially and massively feeding natural enemy staphylinid |
| CN101773101B (en) * | 2010-03-03 | 2012-07-04 | 中国农业科学院植物保护研究所 | Method for artificially and massively feeding natural enemy staphylinid |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101266231B (en) | 2011-05-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhang et al. | Comparison of phenolic compounds extracted from Diaphragma juglandis fructus, walnut pellicle, and flowers of Juglans regia using methanol, ultrasonic wave, and enzyme assisted-extraction | |
| Kujala et al. | Betalains and phenolics in red beetroot (Beta vulgaris) peel extracts: extraction and characterisation | |
| Tang et al. | Rapid and simple method for screening of natural antioxidants from Chinese herb Flos Lonicerae Japonicae by DPPH‐HPLC‐DAD‐TOF/MS | |
| He et al. | Cimicifuga species identification by high performance liquid chromatography–photodiode array/mass spectrometric/evaporative light scattering detection for quality control of black cohosh products | |
| Xu et al. | Chemistry, bioactivity and quality control of Dendrobium, a commonly used tonic herb in traditional Chinese medicine | |
| Khatib et al. | An insight on the alkaloid content of Capparis spinosa L. root by HPLC-DAD-MS, MS/MS and 1H qNMR | |
| Su et al. | Online identification of the antioxidant constituents of traditional Chinese medicine formula Chaihu-Shu-Gan-San by LC–LTQ-Orbitrap mass spectrometry and microplate spectrophotometer | |
| Maldini et al. | Phenolic compounds from Byrsonima crassifolia L. bark: Phytochemical investigation and quantitative analysis by LC-ESI MS/MS | |
| Ma et al. | Metabolic profiling of Actaea species extracts using high performance liquid chromatography coupled with electrospray ionization time-of-flight mass spectrometry | |
| Xu et al. | A rapid method for simultaneous determination of triterpenoid saponins in Pulsatilla turczaninovii using microwave-assisted extraction and high performance liquid chromatography–tandem mass spectrometry | |
| El-sayed et al. | UPLC-ESI-MS/MS profile of the ethyl acetate fraction of aerial parts of Bougainvillea'Scarlett O'Hara'cultivated in Egypt | |
| Huang et al. | Multicomponent assessment and ginsenoside conversions of Panax quinquefolium L. roots before and after steaming by HPLC-MSn | |
| Sun et al. | Simultaneous detection of flavonoids and phenolic acids in Herba Lysimachiae and Herba Desmodii Styracifolii using liquid chromatography tandem mass spectrometry | |
| Qiao et al. | Comprehensive chemical analysis of triterpenoids and polysaccharides in the medicinal mushroom Antrodia cinnamomea | |
| Ling et al. | High performance liquid chromatography coupled to electrospray ionization and quadrupole time-of-flight–mass spectrometry as a powerful analytical strategy for systematic analysis and improved characterization of the major bioactive constituents from Radix Dipsaci | |
| CN102352402B (en) | Method for preparing red ginseng saponins Rg3 group and Rh2 group mixed saponins | |
| Li et al. | The strategy for establishment of the multiple reaction monitoring based characteristic chemical profile of triterpenes in Alismatis rhizoma using two combined tandem mass spectrometers | |
| Vijendra et al. | Product enhancement of triterpenoid saponins in cell suspension cultures of Leucas aspera Spreng | |
| Xie et al. | Rapid identification and determination of 11 polyphenols in Herba lycopi by HPLC–MS/MS with multiple reactions monitoring mode (MRM) | |
| Montoro et al. | Determination of steroidal glycosides in Yucca gloriosa flowers by LC/MS/MS | |
| Fan et al. | HPLC–DAD–MS/MS identification and HPLC–ABTS·+ on-line antioxidant activity evaluation of bioactive compounds in liquorice (Glycyrrhiza uralensis Fisch.) extract | |
| Yang et al. | Simultaneous determination of 15 components in Radix Glehniae by high performance liquid chromatography–electrospray ionization tandem mass spectrometry | |
| Shi et al. | Chemical profiling of jinqi jiangtang tablets by HPLC-ESI-Q-TOF/MS | |
| CN101266231B (en) | Pederin discrimination method | |
| Wu et al. | New approach to the characterization and quantification of Antrodia cinnamomea benzenoid components utilizing HPLC-PDA, qNMR and HPLC-tandem MS: Comparing the wild fruiting bodies and its artificial cultivated commercial products |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20110525 Termination date: 20170418 |