CN107941949A - The detection method of patulin and chloramphenicol in haw products - Google Patents
The detection method of patulin and chloramphenicol in haw products Download PDFInfo
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- CN107941949A CN107941949A CN201711209603.2A CN201711209603A CN107941949A CN 107941949 A CN107941949 A CN 107941949A CN 201711209603 A CN201711209603 A CN 201711209603A CN 107941949 A CN107941949 A CN 107941949A
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- mobile phase
- chloramphenicol
- patulin
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- haw
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- 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
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- 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
Abstract
The invention discloses the detection method of patulin and chloramphenicol in a kind of haw products, be measured using method associated with liquid chromatography mass, the chromatographic condition used for：Chromatographic column Agilent Eclipse Plus C18, 1.8 μm, 2.1 mm × 100mm；33~38 DEG C of column temperature, 0.2~0.5ml/min of flow velocity；Using 9mmol/L ammonium acetates as mobile phase A, using methanol as Mobile phase B, gradient elution, 1~2 μ l of sample size are carried out；Mass Spectrometry Conditions are：Using electron spray ionisation source, MRM modes detect, and dry temperature degree is 210 DEG C；Nebulizer pressure 20Psi；Dry gas stream 11~12L/min of speed, 350 DEG C of sheath stream temperature degree；Sheath stream gas velocity 10L/min；1000 V of spray nozzle voltage.The method of the invention have the advantages that it is easy to operate it is quick, accuracy is high, stability is good, suitable for quality control of the laboratory for haw products, this method can be detected in batches, and detection efficiency is high.
Description
Technical field
The present invention relates to food or medicine detection field, is specifically the detection side of patulin and chloramphenicol in haw products
Method.
Background technology
Patulin is also known as Penicillium patulum toxin, clavacin, coral mycotoxin of penicillium, it is one by fungies such as aspergillus and moulds
A kind of secondary metabolite produced, clavacin have the characteristics that the antibiotic of wide spectrum.Toxicology test shows that patulin has
Have an impact fertility, carcinogenic and immune etc. toxicological effect, while be also a kind of neurotoxin.Patulin has teratogenesis, to people
The harm of body is very big, causes the infringement of the systems such as breathing and uropoiesis, makes one neural paralysis, pulmonary edema, renal failure.
Chloramphenicol also known as chloramphenicol, be by chlorine Streptothrix produce it is a kind of have inhibit bacteria the wide spectrum of growth and resist
Rhzomorph, chloramphenicol have inhibitory action to gramnegative bacterium, gram-positive bacterium etc., available for treatment typhoid fever bacillary dysentery,
The diseases such as urinary tract infection, pertussis, pneumonia, septicemia, but have detrimental effect to human body.It has suppression to marrow hemopoiesis function
Effect, can cause agranulocytosis disease, alpastic anemia and hemolytic anemia, and lethal effect is produced to people.Chloramphenicol
Learn property relatively to stablize, be easy to remain in animal body, so as to cause directly to endanger, more seriously certain density medicine is residual
The drug resistance of pathogenic bacteria can be induced by staying.In addition, long-term micro intake chloramphenicol not only makes the generation such as Escherichia coli, salmonella resistance to
Pharmacological property, and body normal flora can be caused to lack of proper care, make the various diseases of people's easy infection, such as aplastic anemia and former baby
Syndrome can cause granulocyte and decrease of platelet, neonate, premature's Synthetic Grey disease etc..
Patulin is widely present in hawthorn, apple etc. and goes mouldy in fruit, and chloramphenicol is also included in disabling by the Ministry of Agriculture of China
Bee medicine list, can all add honey in most of haw productses at present and be made, therefore, detect patulin in haw products
It is necessary with the content of chloramphenicol.At present detection patulin and chloramphenicol method mainly have thin-layered chromatography,
Gas chromatography, high performance liquid chromatography, enzyme linked immunosorbent assay and high performance liquid chromatography mass spectrometry etc..
Hawthorn（Crataegus pinnatifida Bunge）Make with reducing blood lipid, blood pressure, cardiac stimulant, arrhythmia etc.
With, while be also spleen benefiting and stimulating the appetite, relieving dyspepsia, the good medicine of phlegm reduction of blood circulation promoting, the disease such as, hernia, blood stasis full to chest diaphragm spleen, amenorrhoea has very
The effect of good.Flavone compound Vitexin in hawthorn, is a kind of stronger medicine of antitumaous effect, its extract is to suppressing body
Interior growth of cancer cells, propagation and infiltration metastasis have certain effect.Haw products is not only tasty, suitable for people of all ages, and has
There is the effect of certain health care appetizing, liked be subject to numerous people, hawthorn is as China a kind of special food, nutrition and medicine
With fruit, there is very big potentiality to be exploited.
The content of the invention
The object of the present invention is to provide the detection method of patulin and chloramphenicol in a kind of haw products, is detection platform
The detection method that more accuracy are high, precision is good, stability is good, fast and convenient is provided.
The present invention is achieved through the following technical solutions：
The detection method of patulin and chloramphenicol in haw products, its detection method comprise the following steps：
（1）The preparation of sample solution and vehicle solution：Haw products and bare substrate 5~6g of sample are taken respectively, are smashed, essence
It is close weighed；25~30ml of methanol is added, is ultrasonically treated 30~40 minutes, shakes up, is filtered；It is accurate respectively to draw subsequent filtrate 1ml, add
10 times of methanol dilution, shakes up, and crosses 0.2μM filter membranes, you can obtain testing sample solution and vehicle solution；
（2）The preparation of mixed reference substance solution：Precision weighs patulin and chloramphenicol reference substance respectively, adds vehicle solution
Dilution, be made patulin and chloramphenicol mass concentration be 1 μ g/ml mixed solution, obtain mixing reference substance storing solution；Essence
It is close to measure above-mentioned mixing reference substance storing solution, add vehicle solution and dilute 10 times, up to mixed reference substance solution；
（3）Carry out liquid chromatography-tandem mass spectrometry analysis：Accurate respectively to draw mixed reference substance solution and sample solution, injection is high
Effect liquid phase chromatogram-tandem mass spectrum combined instrument, is measured according to certain chromatographic condition and Mass Spectrometry Conditions.
The supersound process condition is：300~350W of power, 40~50kHz of frequency.
The chromatographic condition is：Chromatographic column Agilent Eclipse Plus C18 , 1.8 μm, 2.1 mm × 100mm；Column
33~38 DEG C of temperature, 0.2~0.5ml/min of flow velocity；Using 9mmol/L ammonium acetates as mobile phase A, using methanol as Mobile phase B, ladder is carried out
Degree elution, 1~2 μ l of sample size.
In the chromatographic condition, the program of gradient elution is：0~1min, mobile phase A from 5% to 70%, Mobile phase B from
5% to 30%；1~6min, mobile phase A is from 70% to 40%, and Mobile phase B is from 30% to 60%；6~8min, mobile phase A from 40% to
95%, Mobile phase B is from 60% to 5%；8~15min, mobile phase A holding 95% is constant, and Mobile phase B holding 5% is constant；Wherein, percentage
Number represents the volume accounting of mobile phase A and Mobile phase B.
The Mass Spectrometry Conditions are：Using electron spray ionisation source, MRM modes detect, and dry temperature degree is 210 DEG C；Atomizer
Pressure 20Psi；Dry gas stream speed 11L/min, sheath stream temperature degree：350℃；Sheath stream gas velocity 10L/min；Spray nozzle voltage 1000
V。
The haw products includes haw jelly, haw flakes, hawthorn bar, hawthorn haw sheet, hawthorn conserve, preserved haw and mountain
Short, bristly hair or beard ball etc..
The measure of the method for the invention detection limit：Respectively it is accurate measure above-mentioned 10 μ l of mixed reference substance solution, 100 μ l,
500 μ l, add blank mechanism solution and are diluted to 1000 μ l, measured by the method for the invention sample introduction, patulin is calculated
Detection limit with chloramphenicol is respectively 4 ng/g and 1 μ g/g.
The beneficial effects of the invention are as follows：
1st, the detection method of patulin and chloramphenicol in haw products of the present invention, can make up in the prior art for hawthorn
The deficiency of Part Quality Control, the method is easy to operate, and detection efficiency is high, can detect the content of patulin and chloramphenicol at the same time,
Reduce testing cost.
2nd, the detection method of patulin and chloramphenicol in haw products of the present invention, is combined using liquid chromatography mass
Method be measured, accuracy is high, and stability is good, suitable for quality control of the laboratory for haw products, can batch into
Row detection, easy to operate, detection limit is low.
Brief description of the drawings
Fig. 1 is the LC-MS/MS TIC figures for mixing reference substance.
Fig. 2 is the LC-MS/MS MRM figures for mixing reference substance.
Embodiment
The present invention is introduced in order to more detailed, with reference to embodiment, the present invention will be further described.
The quick high separating liquid phase chromatographic systems of Agilent 1290, Agilent 6490Triple Quad LC/MS/MS
Liquid chromatography/mass spectrometry combined instrument, distribution electrospray ionization source（ESI）；Chromatographic column is Agilent Eclipse Plus C18（100×
2.1mm ,1.8μm）, methanol is chromatographically pure, and ammonium acetate is chromatographically pure, and water is high purity water.
Embodiment 1
The detection method of patulin and chloramphenicol in haw products, its detection method comprise the following steps：
（1）The preparation of sample solution and vehicle solution：Haw products and bare substrate sample 5g are taken respectively, are smashed, it is accurate
It is weighed；Methanol 30ml is added, is ultrasonically treated 40 minutes, shakes up, is filtered；It is accurate respectively to draw subsequent filtrate 1ml, add methanol dilution 10
Times, shake up, cross 0.2μM filter membranes, you can obtain testing sample solution and vehicle solution；
（2）The preparation of mixed reference substance solution：Precision weighs patulin and chloramphenicol reference substance respectively, adds vehicle solution
Dilution, be made patulin and chloramphenicol mass concentration be 1 μ g/ml mixed solution, obtain mixing reference substance storing solution；Essence
It is close to measure above-mentioned mixing reference substance storing solution, add vehicle solution and dilute 10 times, up to mixed reference substance solution；
（3）Carry out liquid chromatography-tandem mass spectrometry analysis：Accurate respectively to draw mixed reference substance solution and sample solution, injection is high
Effect liquid phase chromatogram-tandem mass spectrum combined instrument, is measured according to certain chromatographic condition and Mass Spectrometry Conditions.
The supersound process condition is：Power 300W, frequency 40kHz.
The chromatographic condition is：Chromatographic column Agilent Eclipse Plus C18 , 1.8 μm, 2.1 mm × 100mm；Column
36 DEG C of temperature, flow velocity 0.4ml/min；Using 9mmol/L ammonium acetates as mobile phase A, using methanol as Mobile phase B, gradient elution is carried out, into
2 μ l of sample amount.
In the chromatographic condition, the program of gradient elution is：0~1min, mobile phase A from 5% to 70%, Mobile phase B from
5% to 30%；1~6min, mobile phase A is from 70% to 40%, and Mobile phase B is from 30% to 60%；6~8min, mobile phase A from 40% to
95%, Mobile phase B is from 60% to 5%；8~15min, mobile phase A holding 95% is constant, and Mobile phase B holding 5% is constant；Wherein, percentage
Number represents the volume accounting of mobile phase A and Mobile phase B.
The Mass Spectrometry Conditions are：Using electron spray ionisation source, MRM modes detect, and dry temperature degree is 210 DEG C；Atomizer
Pressure 20Psi；Dry gas stream speed 11L/min, sheath stream temperature degree：350℃；Sheath stream gas velocity 10L/min；Spray nozzle voltage 1000
V。
Embodiment 2
The detection method of patulin and chloramphenicol in haw products, its detection method comprise the following steps：
（1）The preparation of sample solution and vehicle solution：Haw products and bare substrate sample 5g are taken respectively, are smashed, it is accurate
It is weighed；Methanol 30ml is added, is ultrasonically treated 30 minutes, shakes up, is filtered；It is accurate respectively to draw subsequent filtrate 1ml, add methanol dilution 10
Times, shake up, cross 0.2μM filter membranes, you can obtain testing sample solution and vehicle solution；
（2）The preparation of mixed reference substance solution：Precision weighs patulin and chloramphenicol reference substance respectively, adds vehicle solution
Dilution, be made patulin and chloramphenicol mass concentration be 1 μ g/ml mixed solution, obtain mixing reference substance storing solution；Essence
It is close to measure above-mentioned mixing reference substance storing solution, add vehicle solution and dilute 10 times, up to mixed reference substance solution；
（3）Carry out liquid chromatography-tandem mass spectrometry analysis：Accurate respectively to draw mixed reference substance solution and sample solution, injection is high
Effect liquid phase chromatogram-tandem mass spectrum combined instrument, is measured according to certain chromatographic condition and Mass Spectrometry Conditions.
The supersound process condition is：Power 350W, frequency 40kHz.
The chromatographic condition is：Chromatographic column Agilent Eclipse Plus C18 , 1.8 μm, 2.1 mm × 100mm；Column
35 DEG C of temperature, flow velocity 0.5ml/min；Using 9mmol/L ammonium acetates as mobile phase A, using methanol as Mobile phase B, gradient elution is carried out, into
2 μ l of sample amount.
In the chromatographic condition, the program of gradient elution is：0~1min, mobile phase A from 5% to 70%, Mobile phase B from
5% to 30%；1~6min, mobile phase A is from 70% to 40%, and Mobile phase B is from 30% to 60%；6~8min, mobile phase A from 40% to
95%, Mobile phase B is from 60% to 5%；8~15min, mobile phase A holding 95% is constant, and Mobile phase B holding 5% is constant；Wherein, percentage
Number represents the volume accounting of mobile phase A and Mobile phase B.
The Mass Spectrometry Conditions are：Using electron spray ionisation source, MRM modes detect, and dry temperature degree is 210 DEG C；Atomizer
Pressure 20Psi；Dry gas stream speed 11L/min, sheath stream temperature degree：350℃；Sheath stream gas velocity 10L/min；Spray nozzle voltage 1000
V。
The mass spectrometry parameters of patulin and chloramphenicol are as shown in the table：
Claims (6)
1. the detection method of patulin and chloramphenicol in haw products, it is characterised in that its detection method comprises the following steps：
（1）The preparation of sample solution and vehicle solution：Haw products and bare substrate 5~6g of sample are taken respectively, are smashed, essence
It is close weighed；25~30ml of methanol is added, is ultrasonically treated 30~40 minutes, shakes up, is filtered；It is accurate respectively to draw subsequent filtrate 1ml, add
10 times of methanol dilution, shakes up, and crosses 0.2μM filter membranes, you can obtain testing sample solution and vehicle solution；
（2）The preparation of mixed reference substance solution：Precision weighs patulin and chloramphenicol reference substance respectively, adds vehicle solution
Dilution, be made patulin and chloramphenicol mass concentration be 1 μ g/ml mixed solution, obtain mixing reference substance storing solution；Essence
It is close to measure above-mentioned mixing reference substance storing solution, add vehicle solution and dilute 10 times, up to mixed reference substance solution；
（3）Carry out liquid chromatography-tandem mass spectrometry analysis：Accurate respectively to draw mixed reference substance solution and sample solution, injection is high
Effect liquid phase chromatogram-tandem mass spectrum combined instrument, is measured according to certain chromatographic condition and Mass Spectrometry Conditions.
2. the detection method of patulin and chloramphenicol in haw products according to claim 1, it is characterised in that described super
Sonication condition is：300~350W of power, 40~50kHz of frequency.
3. the detection method of patulin and chloramphenicol in haw products according to claim 1, it is characterised in that the color
Spectral condition is：Chromatographic column Agilent Eclipse Plus C18 , 1.8 μm, 2.1 mm × 100mm；33~38 DEG C of column temperature, flow velocity
0.2~0.5ml/min；Using 9mmol/L ammonium acetates as mobile phase A, using methanol as Mobile phase B, gradient elution, sample size 1 are carried out
~2 μ l.
4. the detection method of patulin and chloramphenicol in haw products according to claim 1, it is characterised in that described
In chromatographic condition, the program of gradient elution is：0~1min, mobile phase A is from 5% to 70%, and Mobile phase B is from 5% to 30%；1~
6min, mobile phase A is from 70% to 40%, and Mobile phase B is from 30% to 60%；6~8min, mobile phase A from 40% to 95%, Mobile phase B from
60% to 5%；8~15min, mobile phase A holding 95% is constant, and Mobile phase B holding 5% is constant；Wherein, percentage represents mobile phase A
With the volume accounting of Mobile phase B.
5. the detection method of patulin and chloramphenicol in haw products according to claim 1, it is characterised in that the matter
Spectral condition is：Using electron spray ionisation source, MRM modes detect, and dry temperature degree is 210 DEG C；Nebulizer pressure 20Psi；It is dry
Gas velocity 11L/min, sheath stream temperature degree：350℃；Sheath stream gas velocity 10L/min；1000 V of spray nozzle voltage.
6. the detection method of patulin and chloramphenicol in haw products according to claim 1, it is characterised in that the mountain
Short, bristly hair or beard product includes haw jelly, haw flakes, hawthorn bar, hawthorn haw sheet, hawthorn conserve, preserved haw and Crataegi pill etc..
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| Application Number | Priority Date | Filing Date | Title |
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| CN201711209603.2A CN107941949B (en) | 2017-11-27 | 2017-11-27 | Detection method of patulin and chloramphenicol in hawthorn product |
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| CN201711209603.2A CN107941949B (en) | 2017-11-27 | 2017-11-27 | Detection method of patulin and chloramphenicol in hawthorn product |
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| CN107941949A true CN107941949A (en) | 2018-04-20 |
| CN107941949B CN107941949B (en) | 2021-01-15 |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110470772A (en) * | 2019-09-19 | 2019-11-19 | 厦门鉴科检测技术有限公司 | The detection method of patulin in the application of WATERS HILIC chromatographic column and food |
| CN111239288A (en) * | 2020-02-26 | 2020-06-05 | 欧陆分析技术服务(苏州)有限公司 | Method for rapidly detecting patulin in apples and apple juice |
| CN111289637A (en) * | 2020-01-19 | 2020-06-16 | 中国计量科学研究院 | Method for detecting patulin in apple juice |
| CN111693639A (en) * | 2020-06-12 | 2020-09-22 | 扬州大学 | Confirmation analysis method for detecting penicillin G residue in poultry tissue, poultry egg or pork |
| CN112763591A (en) * | 2020-12-15 | 2021-05-07 | 上海明捷医药科技有限公司 | Method for determining penicillin content in protein solution |
-
2017
- 2017-11-27 CN CN201711209603.2A patent/CN107941949B/en active Active
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110470772A (en) * | 2019-09-19 | 2019-11-19 | 厦门鉴科检测技术有限公司 | The detection method of patulin in the application of WATERS HILIC chromatographic column and food |
| CN111289637A (en) * | 2020-01-19 | 2020-06-16 | 中国计量科学研究院 | Method for detecting patulin in apple juice |
| CN111239288A (en) * | 2020-02-26 | 2020-06-05 | 欧陆分析技术服务(苏州)有限公司 | Method for rapidly detecting patulin in apples and apple juice |
| CN111693639A (en) * | 2020-06-12 | 2020-09-22 | 扬州大学 | Confirmation analysis method for detecting penicillin G residue in poultry tissue, poultry egg or pork |
| CN112763591A (en) * | 2020-12-15 | 2021-05-07 | 上海明捷医药科技有限公司 | Method for determining penicillin content in protein solution |
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| Publication number | Publication date |
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| CN107941949B (en) | 2021-01-15 |
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