CN101266231A - Pederin discrimination method - Google Patents

Pederin discrimination method Download PDF

Info

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
ion peak
discrimination
pederin
Prior art date
Application number
CNA2008100362454A
Other languages
Chinese (zh)
Other versions
CN101266231B (en
Inventor
万平
李利珍
赵梅君
胡佳耀
Original Assignee
上海师范大学
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by 上海师范大学 filed Critical 上海师范大学
Priority to CN2008100362454A priority Critical patent/CN101266231B/en
Publication of CN101266231A publication Critical patent/CN101266231A/en
Application granted granted Critical
Publication of CN101266231B publication Critical patent/CN101266231B/en

Links

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

A kind of discrimination method of malicious pederin
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.
CN2008100362454A 2008-04-18 2008-04-18 Pederin discrimination method CN101266231B (en)

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 CN101266231B (en) 2008-04-18 2008-04-18 Pederin discrimination method

Country Status (1)

Country Link
CN (1) CN101266231B (en)

Cited By (1)

* Cited by examiner, † Cited by third party
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

Cited By (2)

* Cited by examiner, † Cited by third party
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
Wang et al. Stability of curcumin in buffer solutions and characterization of its degradation products
Guo et al. Simultaneous qualitative and quantitative analysis of triterpenic acids, saponins and flavonoids in the leaves of two Ziziphus species by HPLC–PDA–MS/ELSD
Hu et al. Multi-component HPLC fingerprinting of Radix Salviae Miltiorrhizae and its LC-MS-MS identification
De Leo et al. Profiling the chemical content of Opuntia ficus-indica flowers by HPLC–PDA-ESI-MS and GC/EIMS analyses
Aberham et al. Analysis of iridoids, secoiridoids and xanthones in Centaurium erythraea, Frasera caroliniensis and Gentiana lutea using LC–MS and RP-HPLC
Vinholes et al. In vitro studies to assess the antidiabetic, anti-cholinesterase and antioxidant potential of Spergularia rubra
Hayes et al. Steroidal saponins from the roots of Asparagus racemosus
Quirantes-Piné et al. High-performance liquid chromatography with diode array detection coupled to electrospray time-of-flight and ion-trap tandem mass spectrometry to identify phenolic compounds from a lemon verbena extract
Zhao et al. Identification of monofloral honeys using HPLC–ECD and chemometrics
Savarese et al. Characterization of phenolic extracts from olives (Olea europaea cv. Pisciottana) by electrospray ionization mass spectrometry
Wang et al. Rapid characterization of ginsenosides in the roots and rhizomes of Panax ginseng by UPLC-DAD-QTOF-MS/MS and simultaneous determination of 19 ginsenosides by HPLC-ESI-MS
Li et al. Systematic chemical profiling of Citrus grandis ‘Tomentosa’by ultra-fast liquid chromatography/diode-array detector/quadrupole time-of-flight tandem mass spectrometry
Chang et al. Quantification of (+)-catechin and (−)-epicatechin in coconut water by LC–MS
Jaiswal et al. Profiling the chlorogenic acids of Rudbeckia hirta, Helianthus tuberosus, Carlina acaulis and Symphyotrichum novae‐angliae leaves by LC‐MSn
Lin et al. Identification of the phenolic components of chrysanthemum flower (Chrysanthemum morifolium Ramat)
KR20050025588A (en) An hydroxytyrosol-rich composition from olive vegetation water and method of use thereof
He et al. A sensitive LC–MS/MS method for simultaneous determination of six flavonoids in rat plasma: Application to a pharmacokinetic study of total flavonoids from mulberry leaves
Liau et al. LC-APCI-MS method for detection and analysis of tryptanthrin, indigo, and indirubin in Daqingye and Banlangen
Miles et al. Isolation of pectenotoxin-2 from Dinophysis acuta and its conversion to pectenotoxin-2 seco acid, and preliminary assessment of their acute toxicities
Wang et al. A strategy for identification and structural characterization of compounds from Gardenia jasminoides by integrating macroporous resin column chromatography and liquid chromatography-tandem mass spectrometry combined with ion-mobility spectrometry
Kao et al. Determination of flavonoids and saponins in Gynostemma pentaphyllum (Thunb.) Makino by liquid chromatography–mass spectrometry
Kim et al. Antioxidant activities and polyphenol content of Morus alba leaf extracts collected from varying regions
Abas et al. Chemical characterization and antioxidant activity of three medicinal Apiaceae species
Avula et al. Quantitative determination of flavonoids and cycloartanol glycosides from aerial parts of Sutherlandia frutescens (L.) R. BR. by using LC-UV/ELSD methods and confirmation by using LC–MS method
Hadjmohammadi et al. Hollow fibre-based liquid phase microextraction combined with high-performance liquid chromatography for the analysis of flavonoids in Echinophora platyloba DC. and Mentha piperita

Legal Events

Date Code Title Description
PB01 Publication
C06 Publication
SE01 Entry into force of request for substantive examination
C10 Entry into substantive examination
GR01 Patent grant
C14 Grant of patent or utility model
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20110525

Termination date: 20170418

CF01 Termination of patent right due to non-payment of annual fee