CN105929040A - Method for detecting citrinin in Guogong wine - Google Patents
Method for detecting citrinin in Guogong wine Download PDFInfo
- Publication number
- CN105929040A CN105929040A CN201610235099.2A CN201610235099A CN105929040A CN 105929040 A CN105929040 A CN 105929040A CN 201610235099 A CN201610235099 A CN 201610235099A CN 105929040 A CN105929040 A CN 105929040A
- Authority
- CN
- China
- Prior art keywords
- standard
- citrinin
- sample
- detection method
- ethanol
- 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.)
- Pending
Links
Classifications
-
- 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
-
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- 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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention provides a method for detecting citrinin in Guogong wine, and belongs to the technical field of chemical detection. The detection method has the advantages of high sensitivity and short detection time. The detection method comprises the following steps: (1) preparing a standard solution; (2) adding 0.5 mL of sample into a 25 mL volumetric flask, adding water to the scale, evenly shaking, filtering, making the solution go through a HLB solid phase extraction column at a flowing speed of 1 mL/min, washing the column by 5 mL of water, discarding, then washing the column by 5 mL of ethanol, collecting the eluent, blowing the eluent by nitrogen gas at a temperature of 40 DEG C to dry the eluent, precisely adding 1 mL of ethanol to dissolve the obtained solids, filtering the solution by an organic filter membrane with a thickness of 0.22 [mu]m to obtain a sample to be detected, loading the sample on an Agilent 1295-6550C triple quadrupole HPLC-MS, and analyzing the sample.
Description
Technical field
The present invention provides the detection method of a kind of citrinin, the detection method of citrinin in a kind of Guogong liquor；Belong to technical field of chemical detection.
Background technology
Citrinin (citrinin) is isolated from penicillium Penicillium citrinum first in 1931.Its molecular formula is C13H14O5, molecular weight is 250.25.Citrinin has certain nephrotoxicity and liver toxicity, energy teratogenesis, initiation tumor and Mutation induction.And Monas cuspurpureus Went is Chinese tradition medical material, it is to form by distinctive monascus ruber cultivation and fermentation in rice.Having the statin substance of suppression Biosynthesis of cholesterol because some monascus sp bacteria strain can produce, thus Hongqu is developed to state's Guogong liquor and the health food of blood fat reducing.It is found that in monascus product containing after citrinin first at nineteen ninety-five France scholar Blanc etc., the most using citrinin as the strict Con trolling index of food pollution.And in China, Chinese Pharmacopoeia not yet using citrinin as the Con trolling index containing Monas cuspurpureus Went Chinese patent medicine.
State's Guogong liquor XUEZHIKANG JIAONANG is documented and measures its citrinin content by RP-HPLC method, and in food, health food, also have the detection method of citrinin: thin layer chromatography, fluorescence spectrophotometry, high performance liquid chromatography etc..
Summary of the invention
For above-mentioned deficiency, it is an object of the invention to provide a kind of highly sensitive, the method for the detection method citrinin of citrinin in the Guogong liquor that analysis time is short.
For solving above-mentioned technical problem, the technical scheme that the present invention provides is as follows:
The detection method of citrinin in a kind of Guogong liquor, described detection method comprises the steps: 1) prepare standard solution；2) take sample 0.5ml, put in 25ml volumetric flask, be settled to scale with water, filter after shaking up, with 1ml/min flow velocity by HLB solid-phase extraction column, use 5ml water wash, discard, eluent is collected again with 5ml ethanol elution, eluent nitrogen at 40 DEG C is blown near dry, and the accurate 1ml ethanol that adds dissolves, with the 0.22 organic membrane filtration of μm, obtain testing sample, triple for Agilent1295-6550C on testing sample level Four bar liquid chromatography mass spectrometric combined instruments are detected.
Further, the detection method of citrinin in above-mentioned a kind of Guogong liquor, described standard solution of preparing comprises the steps:
1) preparation of standard solution
Standard reserving solution: precision weighs standard substance 5mg, puts in 5ml volumetric flask, adds ethanol and dissolves and to scale, and making into concentration is 1mg/ml；
Standard dilution: accurate draw standard solution 0.2ml, puts in the volumetric flask of 50ml, adds ethanol dilution to scale, and making into concentration is 2 μ g/ml1；
Standard working curve: take standard dilution liquid and add ethanol in right amount, be configured to respectively concentration be 0.01,2,5,5,20ng/ml standard working solution.
Further, the detection method of citrinin in above-mentioned a kind of Guogong liquor, described liquid chromatograph and Mass Spectrometry Conditions chromatographic column: Agilent poroshell 120EC-C18,3.0 × 50mm 2.7micron, 1.8 μm；Mobile phase A is 0.2% phosphoric acid, and Mobile phase B is methanol, uses gradient elution program as follows: 0-3min, A, 75-5%, B, 25-95%；3-5.5min, A, 5%, B, 95%；5.5.-5.6min, A, 75%, B, 25%；Flow velocity 0.2ml/min；Column temperature 40 DEG C；Sample size 2 μ l；Electrospray ionization source ESI, positive ion mode, selects multiple-reaction monitoring MRM, carrier gas temperature: 150 DEG C, carrier gas flux: 15L/min, nebulizer pressure 40psi, sheath temperature 300 DEG C, sheath throughput: 12L/min, capillary voltage 3500V, qualitative, quantitative ion: parent ion m/z=251.2, daughter ion m/z233.2 and m/z 205.2, collision energy is respectively 5V and 20V.
Establish the citrinin liquid chromatography tandom mass spectrometry determination containing Monas cuspurpureus Went state Guogong liquor herein, this method completes the analysis to this material in 6min and measures, in the case of ensureing that qualitative, quantitative is accurately, achieve high-sensitive analysis, and during extracting, decrease the use of the toxic reagents such as toluene, there is the features such as efficient, environmental protection, can be that the citrinin monitoring in state's Guogong liquor provides technical support.
Accompanying drawing explanation
Fig. 1 is 2 μ g/ml citrinin standard solution multiple-reaction monitoring figures.
Fig. 2 is the multiple-reaction monitoring figure of blank sample.
Detailed description of the invention
Below in conjunction with detailed description of the invention; the claim of the present invention is described in further detail; but not constituting any limitation of the invention, the amendment of any limited number of time made at the claims in the present invention protection domain, still within the claims of the present invention.
Embodiment 1
1 material
1.1 instrument
Agilent1295-6550C triple level Four bar liquid chromatography mass spectrometric combined instrument (U.S. Agilent), HLB-SPE post (60mg/3ml, Shanghai ANPEL Scientific Instrument Co., Ltd.), Agilent ZORBAX Eclipse Plus (2.1 × 50mm, 1.8 μm；U.S. Agilent), XA205DU electronic balance (Germany Mei Teletuo benefit company), ultrasonic cleaner.
1.2 state's Guogong liquor and reagent
State's Guogong liquor 4 batches, derives from market sale sample.Citrinin (lot number: 518-75-2-150403, standard substance are purchased from Pribolab, purity ＞ 98%), ethanol and methanol are chromatographically pure, and other reagent is analytical pure；The percentage concentration that the present invention relates to, is not particularly illustrated, and liquid is volumetric concentration, and solute is the finger mass concentration of solid；Experimental water is the deionized water filtered through ELGA PURELAB ClassicUVF cleaning system.
2 methods and result
The preparation of 2.1 standard solution
2.1.1 standard reserving solution: precision weighs standard substance 5mg, puts in 5ml volumetric flask, adds ethanol and dissolves and to scale, and making into concentration is 1mg/ml.
2.1.2 standard dilution: accurate draw standard solution 0.2ml, puts in the volumetric flask of 50ml, adds ethanol dilution to scale, and making into concentration is 2 μ g/ml, and testing result sees Fig. 1.
2.1.3 standard working curve: take standard dilution liquid and add ethanol in right amount, be configured to respectively concentration be 0.01,2,5,5,20ng/ml standard working solution.
2.2 sample pre-treatments
Precision measures sample 0.5ml, puts in 25ml volumetric flask, is settled to scale with water, filters after shaking up, stand-by.
2.3 sample purification
By above-mentioned sample pre-treatments liquid, with about 1ml/min flow velocity by HLB solid-phase extraction column (activating with 3ml ethanol and 3ml water respectively before use), use 5ml water wash, discard, collecting eluent with 5ml ethanol elution again, eluent nitrogen at 40 DEG C is blown near dry, and the accurate 1ml ethanol that adds dissolves, with the 0.22 organic membrane filtration of μm, obtain testing sample.
2.4 liquid chromatograph and Mass Spectrometry Conditions
Chromatographic column: Agilent ZORBAX Eclipse Plus post (2.1 × 50mm, 1.8 μm)；Mobile phase A is 0.2% phosphoric acid, and Mobile phase B is methanol, uses gradient elution program, is shown in Table 1.Flow velocity 0.2ml/min；Column temperature 40 DEG C；Sample size 2 μ l；Electrospray ionization source (ESI), positive ion mode, select multiple-reaction monitoring (MRM), carrier gas temperature: 150 DEG C, carrier gas flux: 15L/min, nebulizer pressure 40psi, sheath temperature 300 DEG C, sheath throughput: 12L/min, capillary voltage 3500V, qualitative, quantitative ion: parent ion m/z=251.2 (M+1), daughter ion m/z233.2 and m/z 205.2, collision energy is respectively 5V and 20V, Frag and is 380V.
Table 1 gradient elution program
Tab 1 Gradient elution program
The mensuration of 2.5 actual samples
In this test, substitute into the regression equation calculation of standard series, result state Guogong liquor 4 batches with quota ion peak area, all do not detect citrinin.
The accuracy of 2.4 methods
Carrying out recovery of standard addition test in not containing the blank sample of two kinds of substrate of citrinin after measured, spiked levels is 5ng/ml.Parallel assay 6 times, the average recovery of standard addition liquid preparation scope obtained is 95%～50%, sees Fig. 2.
Technical scheme use water (containing 0.2% phosphoric acid)+methanol that the present invention provides=75+25 makees flowing phase, flow velocity 0.3ml/min, the chromatographic peak of citrinin is symmetrical, tandem mass spectrum detector is used to detect, owing to tandem mass spectrum has in separation, high resolution and highly sensitive characteristic, multiple-reaction monitoring pattern (MRM) and parent ion and two corresponding daughter ions are coordinated to be monitored, reach the most qualitative and quantitative purpose, simplify the step that flowing is selected mutually, so this method has selected above-mentioned flowing phase and ratio thereof.
nullAfter using HLB solid-phase extraction column that the citrinin in sample is purified for understanding this method，The matrix interference situation to Mass Spectrometer Method when measuring，Select a sample not detecting citrinin to carry out extraction by method and make matrix liquid，And add standard series，External standard standard series is carried out mass spectroscopy simultaneously，Regression analysis is carried out with its concentration according to the quota ion peak area that retention time is corresponding，Matrix liquid to add the regression equation of standard series be Y=3 298.522531X+4 528.423 351，R=0.999 4，Compare with external standard regression equation Y=3 329.186799X+4 586.551 631 (r=0.999 4)，Both are without substantial differences，Show that HLB solid-phase extraction column has obvious purification to sample substrate.
Establish the citrinin liquid chromatography tandom mass spectrometry determination containing Monas cuspurpureus Went state Guogong liquor herein, this method completes the analysis to this material in 6min and measures, in the case of ensureing that qualitative, quantitative is accurately, achieve high-sensitive analysis, and during extracting, decrease the use of the toxic reagents such as toluene, there is the features such as efficient, environmental protection, can be that the citrinin monitoring in state's Guogong liquor provides technical support.
Claims (4)
1. the detection method of citrinin in a Guogong liquor, it is characterised in that described detection
Method comprises the steps: 1) prepare standard solution；2) take sample 0.5ml, put 25ml
In volumetric flask, it is settled to scale with water, filters after shaking up, pass through with 1ml/min flow velocity
HLB solid-phase extraction column, uses 5ml water wash, discards, then washes with the collection of 5ml ethanol elution
De-liquid, eluent nitrogen at 40 DEG C is blown near dry, and the accurate 1ml ethanol that adds dissolves, and uses
The 0.22 organic membrane filtration of μm, obtains testing sample, by Agilent1295-on testing sample
6550C triple level Four bar liquid chromatography mass spectrometric combined instrument detects.
The detection method of citrinin in a kind of Guogong liquor the most according to claim 1, its
Being characterised by, described standard solution of preparing comprises the steps:
1) preparation of standard solution
Standard reserving solution: precision weighs standard substance 5mg, puts in 5ml volumetric flask, adds second
Alcohol dissolves and to scale, and making into concentration is 1mg/ml；
Standard dilution: accurate absorption standard solution 0.2ml, puts in the volumetric flask of 50ml,
Adding ethanol dilution to scale, making into concentration is 2 μ g/ml1；
Standard working curve: take standard dilution liquid and add ethanol in right amount, be configured to concentration respectively
Be 0.01,2,5,5,20ng/ml standard working solution.
The detection method of citrinin in a kind of Guogong liquor the most according to claim 1, its
It is characterised by, described liquid chromatograph and Mass Spectrometry Conditions chromatographic column: Agilent poroshell
120EC-C18,3.0 × 50mm 2.7micron, 1.8 μm；Mobile phase A is 0.2% phosphorus
Acid, Mobile phase B is methanol, uses gradient elution program as follows: 0-3min, A, 75-
5%, B, 25-95%；3-5.5min, A, 5%, B, 95%；5.5.-5.6min, A,
75%, B, 25%；Flow velocity 0.2ml/min；Column temperature 40 DEG C；Sample size 2 μ l；Electron spray
Source ESI, positive ion mode, selection multiple-reaction monitoring MRM, carrier gas temperature: 150 DEG C,
Carrier gas flux: 15L/min, nebulizer pressure 40psi, sheath temperature 300 DEG C, sheath air-flow
Amount: 12L/min, capillary voltage 3500V, qualitative, quantitative ion: parent ion m/
Z=251.2, daughter ion m/z233.2 and m/z 205.2, collision energy be respectively 5V and
20V。
The detection method Fructus Citri tangerinae of citrinin in a kind of Guogong liquor the most according to claim 3,
It is characterized in that, respectively with 3ml methanol and 3 before described HLB solid-phase extraction column use
Ml water activates.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610235099.2A CN105929040A (en) | 2016-04-15 | 2016-04-15 | Method for detecting citrinin in Guogong wine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610235099.2A CN105929040A (en) | 2016-04-15 | 2016-04-15 | Method for detecting citrinin in Guogong wine |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105929040A true CN105929040A (en) | 2016-09-07 |
Family
ID=56838249
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610235099.2A Pending CN105929040A (en) | 2016-04-15 | 2016-04-15 | Method for detecting citrinin in Guogong wine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105929040A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102175788A (en) * | 2010-12-27 | 2011-09-07 | 晨光生物科技集团股份有限公司 | Method for detecting contents of citrinin in monascus by liquid chromatograph/mass spectrometer (LC/MS) |
| CN102590366A (en) * | 2011-01-05 | 2012-07-18 | 中国医学科学院药用植物研究所 | Detection method for citrinin in Chinese herbal medicines of different matrixes |
| KR101494131B1 (en) * | 2014-02-27 | 2015-02-16 | 숭실대학교산학협력단 | Detection method of the mycotoxin citrinin using silver substrates and Raman spectroscopy |
-
2016
- 2016-04-15 CN CN201610235099.2A patent/CN105929040A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102175788A (en) * | 2010-12-27 | 2011-09-07 | 晨光生物科技集团股份有限公司 | Method for detecting contents of citrinin in monascus by liquid chromatograph/mass spectrometer (LC/MS) |
| CN102590366A (en) * | 2011-01-05 | 2012-07-18 | 中国医学科学院药用植物研究所 | Detection method for citrinin in Chinese herbal medicines of different matrixes |
| KR101494131B1 (en) * | 2014-02-27 | 2015-02-16 | 숭실대학교산학협력단 | Detection method of the mycotoxin citrinin using silver substrates and Raman spectroscopy |
Non-Patent Citations (3)
| Title |
|---|
| UTE MEISTER: "New method of citrinin determination by HPLC after polyamide column clean-up", 《EUR FOOD RES TECHNOL》 * |
| ZHENG HAN 等: "Multianalysis of 35 Mycotoxins in Traditional Chinese Medicines by Ultra-High-Performance Liquid Chromatography−Tandem Mass Spectrometry Coupled with Accelerated Solvent Extraction", 《JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY》 * |
| 潘振球 等: "液相色谱-串联质谱法检测食品中的桔霉素", 《中国卫生检验杂志》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102854271B (en) | Method for measuring residues of three phenoxy carboxylic acid pesticides in tobacco and tobacco products | |
| CN106093261B (en) | A kind of method that starch syrup is mixed in discriminating honey | |
| CN103344725B (en) | Method for simultaneously and quantitatively detecting content of ethyl carbamate and phthalate in distilled white wine | |
| CN106814153A (en) | The liquid chromatography mass screening technique of the double target spot anticoagulating active materials based on fibrin ferment and clotting factor | |
| CN112684030A (en) | Method for detecting perfluoroalkanoic acid compound in aquatic product by enrichment purification-liquid chromatography tandem mass spectrometry and application | |
| CN103308641A (en) | High performance liquid chromatography-tandem mass spectrometry measuring method of three amide herbicides in tobacco and tobacco products | |
| CN109682897A (en) | A kind of method of a variety of incretion interferents in while determination of the environment water sample | |
| CN105784858B (en) | Method for measuring PPCPs in environmental soil | |
| CN106645518B (en) | The measuring method of chloramphenicol residue in a kind of propolis virgin rubber | |
| RU2425380C1 (en) | Method of quantitative analysis of urine dimethyl terephthalate by liquid chromatography | |
| CN102980968A (en) | Liquid chromatogram tandem mass spectrum measuring method for creatinine in urine | |
| CN107422053B (en) | Method for detecting trichloroacetic acid in cosmetics by liquid chromatography-mass spectrometry | |
| CN108593790A (en) | Detect serum 24,25 simultaneously（OH）The method of 2D and 25OHD | |
| CN102944635B (en) | Method for determining tris (2,3-dibromopropyl) phosphate content of water | |
| CN105651904B (en) | The detection method of citrinin in a kind of Zhibituo Tabiet | |
| CN105929040A (en) | Method for detecting citrinin in Guogong wine | |
| CN105929038A (en) | Method for detecting citrinin in red yeast rice pigment powder | |
| CN105911178A (en) | Detection method of citrinin in Xuezhikang capsules | |
| CN105911180A (en) | Method for detecting citrinin in Guogong medicinal wine by solid-phase extraction-liquid chromatography mass spectrometry | |
| CN105911179A (en) | Method for detecting citrinin in monascus pigment powder by solid-phase extraction-liquid chromatography mass spectrometry | |
| CN105929037A (en) | Method using solid extraction-(LC-MS) to detect citrinin in Zhibituo tablets | |
| CN105866307A (en) | Method for detecting citrinin in Xuezhikang capsule through solid phase extraction-liquid-mass chromatography | |
| CN111337611A (en) | Method for detecting malachite green, leucomalachite green, crystal violet and leucocyte crystal violet in aquatic products | |
| CN107300597B (en) | The detection method of 2-PHENYLBENZIMIDAZOLE-5-SULFONIC ACID in a kind of soil | |
| CN109324139A (en) | Ribosylzeatin liquid-liquid extraction-liquid chromatography-tandem mass spectrometry measuring method in a kind of tobacco leaf |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20160907 |
|
| RJ01 | Rejection of invention patent application after publication |