CN102830192A - Method for simultaneously detecting nitrofurans raw drug residue in aquatic product - Google Patents
Method for simultaneously detecting nitrofurans raw drug residue in aquatic product Download PDFInfo
- Publication number
- CN102830192A CN102830192A CN2012102776380A CN201210277638A CN102830192A CN 102830192 A CN102830192 A CN 102830192A CN 2012102776380 A CN2012102776380 A CN 2012102776380A CN 201210277638 A CN201210277638 A CN 201210277638A CN 102830192 A CN102830192 A CN 102830192A
- Authority
- CN
- China
- Prior art keywords
- itrofurans
- former medicine
- aquatic products
- residual method
- simultaneously according
- 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
Images
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to a method for simultaneously detecting a nitrofurans raw drug residue in an aquatic product, comprising the following steps of: (1) preparing four types of mixed standard solutions; (2) preparing a sample; (3) extracting; (4) concentrating; (5) purifying; and (6) detecting. The method disclosed by the invention has the characteristics of high flexibility, simplicity and convenience in operation, low detection cost, accuracy and reliability; and the quantitative limit is 5 micrograms/kilogram and the recycling rate is more than 70%, so that the method is suitable for rapidly detecting imports and exports of the aquatic product.
Description
Technical field
The invention belongs to aquatic product quality safety detection field, particularly a kind ofly detect the residual method of the former medicine of itrofurans in the aquatic products simultaneously.
Background technology
The itrofurans medicine is the synthetic microbiotic with 5-nitrofuran structure; Mainly comprise furaltadone (Furaltadone; FTD), nitrofurazone (Nitrofurazone; NFZ), furantoin (Nitrofurantoin, NFT) and furazolidone (Furazolidone, FZD) these four kinds of medicines.Because this type microbiotic has good antibacterial and bactericidal effect, once be widely used in the prevention and treatment of aquatic animal disease.Correlative study has proved that the itrofurans medicine has teratogenesis, carcinogenic and mutagenic toxic and side effect.At present, European Union, the U.S., China and Japan and other countries have forbidden that the itrofurans medicine uses in the animal-breeding process, however because its result of treatment is remarkable, and still not having relevant alternative medicine, many raisers are still using the itrofurans medicine.Therefore, set up the detection method of measuring the itrofurans medicine and have crucial meaning.
Detect the residual main employing high performance liquid chromatography of itrofurans medicine, using high performance liquid chromatography tandem mass spectrum method and ELISA at present and measure, and detected object mainly concentrates on feed and breeding water, detects index and is mainly the nitrofuran metabolin.Because aquatic products are of a great variety; Matrix is complicated, and the method research that causes detecting the former medicine residue detection of nitrofuran in the aquatic products is also relatively deficienter, the residual method of the former medicine of nitrofuran in the existing liquid chromatographic detection aquatic products; It need adopt SPE to purify; In practical operation, increased needed time of pre-treatment and expense, the quantitative limit of this method is higher simultaneously; The quantitative limit of furaltadone, nitrofurazone, furantoin, furazolidone is respectively 25,10,15,10 μ g/kg, can't satisfy harsh day by day detection requirement.
Summary of the invention
Technical matters to be solved by this invention provides a kind of residual method of the former medicine of itrofurans in the aquatic products that detects simultaneously, and that this method has is highly sensitive, easy and simple to handle, it is with low cost to detect, characteristics accurately and reliably; Quantitative limit is 5 μ g/kg, and the recovery is suitable for aquatic products and imports and exports fast detecting greater than 70%.
A kind of residual method of the former medicine of itrofurans in the aquatic products that detects simultaneously of the present invention comprises:
(1) get furaltadone, nitrofurazone, furantoin and furazolidone and be dissolved in methyl alcohol, be mixed with the former medicine hybrid standard storing solution of 100 μ g/mL ,-10 ℃--preserve down for 20 ℃; The hybrid standard storing solution is diluted with methyl alcohol, be mixed with the hybrid standard intermediate liquid of 1 μ g/mL, again the hybrid standard intermediate liquid is mixed with standard operation solution with moving phase;
(2) the fish sample is scaled, is removed the peel along ridge and get muscle, pulverize-10 ℃--store down for 20 ℃, room temperature is thawed before measuring; Sample thief adds ethyl acetate, through vortex mixed, ultrasonic, centrifugal after the vortex mixed once more, gets upper solution and adds ethyl acetate again and repeat to extract once, merges extract;
(3) said extracted liquid is evaporated to dried, adds normal hexane, add moving phase behind the vortex, vortex mixed, centrifugal, after taking off layer solution and crossing the water film, last machine stratographic analysis promptly gets the concentration of the former medicine of itrofurans.
The concentration of the standard operation solution in the said step (1) is respectively 0.010,0.015,0.020,0.025,0.05,0.1 and 0.25 μ g/mL.
Fish sample in the said step (2) is grass carp or large yellow croaker.
Moving phase in said step (1) and (3) is acetonitrile and the NaH of volume ratio 25:75
2PO
4The WS; Wherein, NaH
2PO
4The concentration of the WS is 0.05mol/L, pH=3.0.
Centrifugal speed in said step (2) and (3) is 4000r/min, and the centrifugal time is 5min.
Concentrating under reduced pressure in the said step (3) carries out under 30 ℃.
The thickness of the water film in the said step (3) is 0.45 μ m.
Chromatographiccondition in the said step (3) is: chromatographic column: ZORBAX SB-C18 post, specification are 250mm*4.6mm, and particle diameter is 5 μ m; Column temperature: 25 ℃; Detect wavelength: 365nm; The acetonitrile of moving phase: volume ratio 25:75 and NaH
2PO
4The WS; Wherein, NaH
2PO
4The concentration of the WS is 0.05mol/L, pH=3.0; Flow velocity: 1.0mL/min; Sample size: 50 μ L; Working time: 18.0min; Adopt gradient elution.
Beneficial effect
(1) the present invention have highly sensitive, easy and simple to handle, detect with low cost, characteristics accurately and reliably;
(2) the present invention has simplified the pre-treatment step of sample, has reduced the pre-treatment expense, and particularly the pre-treatment time with sample has shortened more than one times, and the quantitative limit of method also increases substantially;
(3) the present invention detects four kinds of residual quantitative limit of the former medicine of nitrofuran and is 5 μ g/kg, and the recovery is suitable for aquatic products and imports and exports fast detecting greater than 70%.
Description of drawings
Fig. 1 is 0.025 μ g/mL mixed standard solution chromatogram;
Fig. 2 is 5.0 μ g/kg grass carp sample mark-on chromatograms;
Fig. 3 is a grass carp dummy chromatogram.
Embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in the restriction scope of the present invention.Should be understood that in addition those skilled in the art can do various changes or modification to the present invention after the content of having read the present invention's instruction, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
1.0.05mol/L NaH
2PO
4(pH=3.0) solution allocation method: claim 7.8g NaH
2PO
42H
2O adds the 1000mL ultrapure water, regulates pH to 3.0 with phosphoric acid, is settled to 1L.
2. four kinds of mixed standard solution preparations: accurately take by weighing furaltadone, nitrofurazone, furantoin, each 10mg of furazolidone, to 100mL, be mixed with four kinds of former medicine hybrid standard storing solutions of 100 μ g/mL ,-20 ℃ of preservations with methanol constant volume.Get 1mL hybrid standard storing solution, to 100mL, be mixed with the hybrid standard intermediate liquid of 1 μ g/mL with methanol constant volume.Pipette the hybrid standard intermediate liquid, with moving phase (volume ratio: acetonitrile: 0.05M NaH
2PO
4=25:75, pH3.0) being mixed with concentration is 0.010,0.015,0.020,0.025,0.05,0.1,0.25 μ g/mL standard operation solution.
Specimen preparation: grass carp and large yellow croaker scale, remove the peel along ridge and get muscle, pulverizes through organizing comminutor, and-20 ℃ of storages, room temperature is thawed before measuring.
Sample pre-treatments:
1 extracts: accurately take by weighing sample (5.00 ± 0.05) g in the 50mL centrifuge tube, add 8mL ethyl acetate, vortex mixed 1min; Ultrasonic 5min; Vortex mixed 1min once more, the centrifugal 5min of 4000r/min is transferred to the upper strata in the brown heart bottle; Add 8mL ethyl acetate again and repeat to extract, merge extract.
2. concentrate: on Rotary Evaporators, be evaporated to dried extract in 30 ℃.
3. purify: add the 2mL normal hexane, add 1mL moving phase (volume ratio: acetonitrile: 0.05MNaH behind the vortex 30s
2PO
4=25:75 pH3.0), mixes 1min, changes in the 10mL centrifuge tube, and the centrifugal 5min of 4000r/min, after taking off layer and crossing 0.45 μ m water film, last machine analysis, the result sees Fig. 2 and Fig. 3.
4. chromatographic condition: the instrument chromatogram is Agilent 1200, and chromatographic column is ZORBAX SB-C18 post (250mm*4.6mm, 5 μ m), column temperature: 25 ℃, detect wavelength: 365nm, and moving phase is: volume ratio: acetonitrile: 0.05M NaH
2PO
4=25:75, pH3.0, flow velocity: 1.0mL/min; Sample size: 50 μ L; Be 18.0min working time.The gradient elution program is seen table 1.
Table 1 moving phase and gradient elution program
Embodiment 3
Chromatogram result: the standard items chromatogram of four kinds of former medicines of nitrofuran is seen accompanying drawing 1.The appearance time of furaltadone, nitrofurazone, furantoin and furazolidone is respectively: 3.922,5.294,6.678 and 8.785min.
The methodology checking:
Under the experiment condition that this method is confirmed; Using moving phase to be mixed with concentration is 0.010,0.015,0.020,0.025,0.05,0.1,0.25 μ g/mL standard operation solution; Last machine analysis is a horizontal ordinate with every kind of drug concentrations, and peak area is an ordinate drawing standard curve; Experimental result shows: in 0.01 ~ 0.25 μ g/mL scope; Former concentration of nitrofuran and peak area are good linear relationship, and the facies relationship number average is more than 0.998, the regression equation of four kinds of nitrofuran medicines and coefficient R
2See table 2.
The linear equation of four kinds of former medicines of nitrofuran of table 2 and detection by quantitative limit
In blank sample, add four kinds of medicines, the concentration of signal to noise ratio (S/N ratio) S/N >=10 o'clock is decided to be the detection by quantitative limit, and the quantitative limit of furaltadone, nitrofurazone, furantoin, furazolidone (LOQ) is 5.0 μ g/kg, and quantitative limit can satisfy daily detection requirement.With grass carp and large yellow croaker is research object, the recovery of investigation method and precision.In blank sample, add the former medicine standard solution of nitrofuran of 3 kinds of variable concentrations levels, record the recovery according to this method, each level repeats 5 times.The result sees table 3, and four kinds of former medicines are when addition 3 ~ 25 μ g/kg, and average recovery rate is that relative standard deviation is less than 13.5% between 70.9 ~ 116.8%.This shows that this method accuracy is high, favorable reproducibility.
Recovery of standard addition of table 3 this method and precision (n=5)
Claims (8)
1. one kind is detected the residual method of the former medicine of itrofurans in the aquatic products simultaneously, comprising:
(1) get furaltadone, nitrofurazone, furantoin and furazolidone and be dissolved in methyl alcohol, be mixed with the former medicine hybrid standard storing solution of 100 μ g/mL ,-10 ℃--preserve down for 20 ℃; The hybrid standard storing solution is diluted with methyl alcohol, be mixed with the hybrid standard intermediate liquid of 1 μ g/mL, again the hybrid standard intermediate liquid is mixed with standard operation solution with moving phase;
(2) the fish sample is scaled, is removed the peel along ridge and get muscle, pulverize-10 ℃--store down for 20 ℃, room temperature is thawed before measuring; Sample thief adds ethyl acetate, through vortex mixed, ultrasonic, centrifugal after the vortex mixed once more, gets upper solution and adds ethyl acetate again and repeat to extract once, merges extract;
(3) said extracted liquid is evaporated to dried, adds normal hexane, add moving phase behind the vortex, vortex mixed, centrifugal, after taking off layer solution and crossing the water film, last machine stratographic analysis promptly gets the concentration of the former medicine of itrofurans.
2. a kind of residual method of the former medicine of itrofurans in the aquatic products that detects simultaneously according to claim 1, it is characterized in that: the concentration of the standard operation solution in the said step (1) is respectively 0.010,0.015,0.020,0.025,0.05,0.1 and 0.25 μ g/mL.
3. a kind of residual method of the former medicine of itrofurans in the aquatic products that detects simultaneously according to claim 1 is characterized in that:
Fish sample in the said step (2) is grass carp or large yellow croaker.
4. a kind of residual method of the former medicine of itrofurans in the aquatic products that detects simultaneously according to claim 1 is characterized in that:
Moving phase in said step (1) and (3) is acetonitrile and the NaH of volume ratio 25:75
2PO
4The WS; Wherein, NaH
2PO
4The concentration of the WS is 0.05mol/L, pH=3.0.
5. a kind of residual method of the former medicine of itrofurans in the aquatic products that detects simultaneously according to claim 1 is characterized in that:
Centrifugal speed in said step (2) and (3) is 4000r/min, and the centrifugal time is 5min.
6. a kind of residual method of the former medicine of itrofurans in the aquatic products that detects simultaneously according to claim 1 is characterized in that:
Concentrating under reduced pressure in the said step (3) carries out under 30 ℃.
7. a kind of residual method of the former medicine of itrofurans in the aquatic products that detects simultaneously according to claim 1 is characterized in that:
The thickness of the water film in the said step (3) is 0.45 μ m.
8. a kind of residual method of the former medicine of itrofurans in the aquatic products that detects simultaneously according to claim 1 is characterized in that:
Chromatographiccondition in the said step (3) is: chromatographic column: ZORBAX SB-C18 post, specification are 250mm*4.6mm, and particle diameter is 5 μ m; Column temperature: 25 ℃; Detect wavelength: 365nm; The acetonitrile of moving phase: volume ratio 25:75 and NaH
2PO
4The WS; Wherein, NaH
2PO
4The concentration of the WS is 0.05mol/L, pH=3.0; Flow velocity: 1.0mL/min; Sample size: 50 μ L; Working time: 18.0min; Adopt gradient elution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012102776380A CN102830192A (en) | 2012-08-06 | 2012-08-06 | Method for simultaneously detecting nitrofurans raw drug residue in aquatic product |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012102776380A CN102830192A (en) | 2012-08-06 | 2012-08-06 | Method for simultaneously detecting nitrofurans raw drug residue in aquatic product |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102830192A true CN102830192A (en) | 2012-12-19 |
Family
ID=47333400
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012102776380A Pending CN102830192A (en) | 2012-08-06 | 2012-08-06 | Method for simultaneously detecting nitrofurans raw drug residue in aquatic product |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102830192A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103293039A (en) * | 2013-05-30 | 2013-09-11 | 福建出入境检验检疫局检验检疫技术中心 | Method for quickly obtaining positive material containing AOZ (3-amino-2-oxazolidinone) minced shrimp natural matrix in laboratory |
| CN103926340A (en) * | 2014-04-04 | 2014-07-16 | 中国检验检疫科学研究院 | Method for measuring nitrofuran antibiotics in cosmetics |
| CN109580793A (en) * | 2017-09-28 | 2019-04-05 | 中国水产科学研究院东海水产研究所 | Multiple types residue of veterinary drug high throughput quick screening method in a kind of aquatic products |
| CN109632427A (en) * | 2019-01-28 | 2019-04-16 | 广州食源生物科技有限公司 | A kind of pre-treating method that Nitrofuran metatolites quickly detect and its detection method |
| CN115372528A (en) * | 2022-09-20 | 2022-11-22 | 北京云鹏鹏程医药科技有限公司 | Detection method for simultaneously determining multiple impurities in nitrofurantoin |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SU1748057A1 (en) * | 1990-06-04 | 1992-07-15 | Курский Государственный Медицинский Институт | Method of determination of 5-nitrofurane derivatives and semi-products |
| CN101625339A (en) * | 2008-07-07 | 2010-01-13 | 吴光红 | Method for detecting residue of seven synthetic antibacterial agents in aquatic products |
| CN101949897A (en) * | 2010-06-30 | 2011-01-19 | 北京六角体科技发展有限公司 | Method for detecting pre-treatment of nitrofurans metabolite |
| CN102621252A (en) * | 2012-03-19 | 2012-08-01 | 福建中检华日食品安全检测有限公司 | Pretreatment method for detecting nitrofuran metabolites in aquatic product |
-
2012
- 2012-08-06 CN CN2012102776380A patent/CN102830192A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SU1748057A1 (en) * | 1990-06-04 | 1992-07-15 | Курский Государственный Медицинский Институт | Method of determination of 5-nitrofurane derivatives and semi-products |
| CN101625339A (en) * | 2008-07-07 | 2010-01-13 | 吴光红 | Method for detecting residue of seven synthetic antibacterial agents in aquatic products |
| CN101949897A (en) * | 2010-06-30 | 2011-01-19 | 北京六角体科技发展有限公司 | Method for detecting pre-treatment of nitrofurans metabolite |
| CN102621252A (en) * | 2012-03-19 | 2012-08-01 | 福建中检华日食品安全检测有限公司 | Pretreatment method for detecting nitrofuran metabolites in aquatic product |
Non-Patent Citations (9)
| Title |
|---|
| M. VASS 等: "Nitrofuran antibiotics: a review on the application, prohibition and residual analysis", 《VETERINARNI MEDICINA》 * |
| NICKOS A. BOTSOGLOU: "Determination of Furazolidone in Eggs by High-Performance Liquid", 《JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY》 * |
| XUAN-YUN HUANG 等: "Determination of Nitrofurans Residue in Fishery Products Using High Performance Liquid Chromatography", 《ADVANCED MATERIALS RESEARCH》 * |
| 于慧娟 等: "高效液相色谱法测定水产品中呋喃唑酮的残留量", 《色谱》 * |
| 吴富忠 等: "水产品中呋喃唑酮、呋喃西林药物残留的HPLC法测定", 《中国卫生检验杂质》 * |
| 朱筱玲: "同位素内标法对水产品中硝基呋喃代谢物液相色谱-串联质谱法测定", 《江西科学》 * |
| 王媛 等: "高效液相色谱法检测水产品中硝基呋喃类代谢物残留量", 《分析实验室》 * |
| 葛宝坤 等: "测定鸡肉、水产品中四种硝基呋喃类药物残留量的固相萃取-液相色谱法", 《分析测试学报》 * |
| 邢丽红 等: "液相色谱-串联质谱（ＬＣ-ＭＳ／ＭＳ）法快速测定水产品中硝基呋喃类代谢物", 《环境化学》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103293039A (en) * | 2013-05-30 | 2013-09-11 | 福建出入境检验检疫局检验检疫技术中心 | Method for quickly obtaining positive material containing AOZ (3-amino-2-oxazolidinone) minced shrimp natural matrix in laboratory |
| CN103926340A (en) * | 2014-04-04 | 2014-07-16 | 中国检验检疫科学研究院 | Method for measuring nitrofuran antibiotics in cosmetics |
| CN109580793A (en) * | 2017-09-28 | 2019-04-05 | 中国水产科学研究院东海水产研究所 | Multiple types residue of veterinary drug high throughput quick screening method in a kind of aquatic products |
| CN109632427A (en) * | 2019-01-28 | 2019-04-16 | 广州食源生物科技有限公司 | A kind of pre-treating method that Nitrofuran metatolites quickly detect and its detection method |
| CN115372528A (en) * | 2022-09-20 | 2022-11-22 | 北京云鹏鹏程医药科技有限公司 | Detection method for simultaneously determining multiple impurities in nitrofurantoin |
| CN115372528B (en) * | 2022-09-20 | 2023-06-23 | 北京云鹏鹏程医药科技有限公司 | Detection method for simultaneously measuring various impurities in nitrofurantoin |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP2842957B1 (en) | Method for extracting and separating ginkgolides | |
| CN102830192A (en) | Method for simultaneously detecting nitrofurans raw drug residue in aquatic product | |
| CN103175932B (en) | Method for determining four hormones in rubber tree through high-efficiency liquid chromatography | |
| CN104569187B (en) | The detection method of whitening and speckle dispelling composition and device in cosmetics | |
| CN107561172B (en) | Method for simultaneously detecting content of multiple vitamins in nutrient soft capsule | |
| CN101721516B (en) | Preparation method of gardenia extract | |
| CN104922196A (en) | General ohwia flavone extract preparation method and general ohwia flavone extract quality detecting method | |
| CN104147054A (en) | Ginkgo biloba leaf extract as well as preparation method and application thereof | |
| CN108614052B (en) | Method for simultaneously detecting gelsemine, gelsemine and gelsemine in biological sample based on two-dimensional liquid chromatography | |
| CN104829666A (en) | Method for preparing high purity baicalin from radix scutellariae | |
| CN111410643B (en) | Preparation and application of novel cinnamoyl ester catechin and four novel phenylpropanoid flavan alkaloids | |
| CN105954432B (en) | A kind of detection method of ginkolide B content | |
| CN111595972A (en) | Preparation method and quality detection method of lithocarpus polystachyus rehd total flavone extract | |
| CN109265494B (en) | Method for extracting kaempferol glucoside compounds from camellia reticulata | |
| CN107202840A (en) | A kind of method for detecting 4 kinds of barbiturates downerns of fresh milk | |
| CN101639467A (en) | Method for determining amount of residual florfenicol in aquatic product by using high-efficiency liquid chromatogram fluorescence method | |
| CN114315806B (en) | Preparation method of ester catechin-theanine adduct | |
| CN103239487A (en) | Bilobalide injection and content determination method | |
| CN103145729B (en) | Bilobalide B compound and preparation method thereof | |
| CN113968785B (en) | Oxygen-containing benzoic acid in purslane and extraction and separation method and application thereof | |
| CN102491898B (en) | Method for simultaneously extracting chlorogenic acid and sunflower protein from sunflower seed meals | |
| CN103323551A (en) | Method for detecting content of medlar acid | |
| CN1994995A (en) | Method for extracting and purifying sequoyitol | |
| CN104198623A (en) | Method for detecting menbutone residue in pig tissue | |
| CN104892620B (en) | A kind of preparation method of high-purity karanjin |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20121219 |