CN106568866A - Measuring method of desoxym equindox, 3-metyl-2-acetyl-quinoxaline and MQCA in trepang - Google Patents
Measuring method of desoxym equindox, 3-metyl-2-acetyl-quinoxaline and MQCA in trepang Download PDFInfo
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- CN106568866A CN106568866A CN201610988653.4A CN201610988653A CN106568866A CN 106568866 A CN106568866 A CN 106568866A CN 201610988653 A CN201610988653 A CN 201610988653A CN 106568866 A CN106568866 A CN 106568866A
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- mequindox
- mqca
- trepang
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- formic acid
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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
Abstract
The invention discloses a measuring method of desoxym equindox, 3-metyl-2-acetyl-quinoxaline and MQCA (3-methylquinoxaline-2-carboxylic acid) in a trepang through once-determination. The measuring method comprises selection and allocation of instruments and reagents, extraction and purification of a sample, specific chromatographic conditions and mass spectrum conditions. Compared with the prior art, the content of desoxym equindox, 3-metyl-2-acetyl-quinoxaline and MQCA (3-methylquinoxaline-2-carboxylic acid) in the trepang can be measured through one sample in once-determination, fast and accurate data support for conditions of abuse of antibiotics in the current trepang aquaculture industry is provided, and thus the measuring method of desoxym equindox, 3-metyl-2-acetyl-quinoxaline and MQCA in trepang is conducive to promotion of the healthy development of the trepang aquaculture industry.
Description
Technical field
The present invention relates to holothruian cultures technical field, takes off an oxygen mequindox, de- dioxy acetyl in specifically a kind of sea cucumber
First quinoline and MQCA（3- Jia based quinoxaline -2- carboxylic acids）Assay method.
Background technology
Modern farming increasingly tends to scale, intensive, using antibiotic, vitamin, hormone, metal trace element
Deng more becoming the guarantee requisite ring of animal husbandry development.Unfortunately, due to the shortage and economy profit of scientific knowledge
Beneficial orders about, the phenomenon generally existing of drug abuse in aquaculture.In China, situation is particularly acute.Abuse veterinary drug it is direct after
Fruit is the residual for causing veterinary drug in animal food, after intake human body, affects the health of the mankind.Veterinary drug in animal food
Remain the attention that the potential hazard to people more and more causes people.Only set up the medicament residue measure side of a set of simple and effective
Method, is the basis for efficiently controlling medicament residue generation.
Olaquindox（Olaquindox）It it is early 1970s by West Germany's Baeyer pharmaceutical factory also known as olaquindox, Olaquindox
The feed addictive of synthesis is developed, because it has broad-spectrum antimicrobial effect and growth promoting function, is widely used in livestock and poultry cultivation, antibacterial
Treatment and the control and prevention of disease of aquatic livestock.But a large amount of biological toxicity tests show：Excessive olaquindox can be residual in animal body
Stay, with obvious cumulative toxicity, the toxic and side effect such as carcinogenic, aberration inducing effect and DNA damage.In view of human health is caused
Olaquindox is classified as disabling medicine by harm, EU Committee EC2788/98 resolutions, and in domestic NY5070-2002 criterions quinoline is also specified
Ethanol disables fishing medicine.But olaquindox itself is unstable, in animal body can short time intracellular metabolite into ten multi-products, wherein 3-
Jia based quinoxaline -2- carboxylic acids（MQCA）It is one of major metabolite, it is relatively stable in vivo, is Codex Alimentary Commission
One of residual marker of identification, therefore generally using olaquindox metabolite MQCA as residual marker.China's agricultural in 2003
Portion defines the maximum residue limit of MQCA in muscle and liver organization and is respectively 4 μ g/kg and 50 μ g/kg.
Mequindox（Mequindox, MEQ）It is tool that China voluntarily develops also known as LIJUNJING, Shu quinoxaline compounds
There is a class novel chiral synthon of independent intellectual property right.Due to broad spectrum antibiotic activity, being widely used on China's veterinary clinic.But
It is that production practices show, mequindox toxicity is bigger than normal, dosage is higher than that 3～5 times of therapeutic dose or long-time can draw when continuously using
Play poisoning even dead.Clinical toxicology research also points out it to cause gene mutation, chromosome aberration, with hepatotoxicity,
Adrenal gland toxicity and two generation breeding toxicities.
To mequindox, the metabolin in hepatomicrosome has carried out relative quantification to the Liu Zhaoying of Hua Zhong Agriculture University, it is believed that N
The de- oxygen mequindoxes of 4- are one of the main metabolites of mequindox in pig, chicken and rat liver microsomes.Central China agricultural
The Yin Fujun of university has then carried out absorption distribution, excretion and metabolism research of the tritium mark mequindox in rat, chicken and pig body, most
De- dioxy mequindox is determined eventually（3- methyl -2- acetyl group-quinoxaline）It is mequindox in these three animal bodies
Residual marker.
Still it is both needed to lead to without the method for determining three kinds of residual markers in sea cucumber simultaneously, and existing method both at home and abroad at present
Cross three kinds of different methods to be determined respectively, there is no carries out the elder generation that three kinds of residual markers are once determined by a sample
Example.
The content of the invention
The purpose of the present invention is open a kind of by once determining a de- oxygen mequindox in sea cucumber, de- dioxy mequindox
And MQCA（3- Jia based quinoxaline -2- carboxylic acids）Assay method, including instrument and the selection of reagent and allotment, sample extraction with it is net
Change, specific chromatographic condition and Mass Spectrometry Conditions.
Compared with prior art, the present invention can once be determined by a sample and take off in sea cucumber an oxygen mequindox, take off two
Oxygen mequindox and MQCA（3- Jia based quinoxaline -2- carboxylic acids）Content, and then to current holothruian cultures industry abuse of antibiotics
Situation provide fast and accurately data support, may advantageously facilitate the sound development of holothruian cultures industry.
Specific embodiment
The assay method of an oxygen mequindox, de- dioxy mequindox and MQCA is taken off in sea cucumber, it is characterised in that：
A) instrument and reagent：
Ultra Performance Liquid Chromatography-tandem mass spectrometer (Quattro Premier XE, Waters, USA), ultra-pure water instrument
(Milli-Q Gradient, Millipore, France), supercentrifuge (TGL-10C, Town in Shanghai booth scientific instrument
Factory), ultrasonic cleaner (KQ-600E, Kunshan Ultrasonic Instruments Co., Ltd.), Nitrogen evaporator (N-EVAPTM112,
Organomation Associates, USA), Full-automatic solid phase extraction instrument (ASPEC XL4+, GILSON, France)；Methyl alcohol
(chromatographically pure), acetonitrile (chromatographically pure), formic acid (top pure grade), hydrochloric acid（Top pure grade）, ethyl acetate（Chromatographically pure）, formic acid（Top grade
It is pure）, sodium acetate（Top pure grade）, solid phase extraction column (MAX 60mg 3mL, Waters, USA)；Mequindox metabolin：MQCA
（German Dr.）Purity>98%, an oxygen mequindox and de- dioxy mequindox are taken off by Wuhan Green's Baeyer biotechnology company
Synthesis, purity>90%；Mixed standard solution：Appropriate mequindox metabolin standard items are weighed, is dissolved with acetonitrile, be made into 100
Mg/L Standard Stock solutions；
B) sample extraction and purification：
5.00g sea cucumber samples are weighed, the mol/L hydrochloric acid of 15mL 2, ultrasonic extraction 1h, 10000 turns/min centrifugations at 4 DEG C is added
10min.MAX posts with 3mL methyl alcohol and 3mL 2mol/L hydrochloric acid activations, add the supernatant in centrifuge tube in advance, use 3mL sodium acetates
Methanol solution drip washing, the 6mL wash-outs of the ethyl acetate containing 2% formic acid are collected.Nitrogen is dried up at 40 DEG C, adds 1mL to contain 0.1% formic acid
Acetonitrile-aqueous solution（70:30, v/v）1mL constant volumes, Ultra Performance Liquid Chromatography tandem mass spectrometry is determined；Wherein take off an oxygen acetyl first
Quinoline, de- dioxy mequindox and MQCA（3- Jia based quinoxaline -2- carboxylic acids）Quantitative limit be：2μg/kg；
C) chromatographic condition：
Chromatographic column：ACQUITYTM UPLC BEHC18(1.7μm, 2.1mm i.d.×100mm)；Chromatogram column temperature：40℃；Stream
Dynamic phase：Acetonitrile (A) and containing 0.5% formic acid water (B)；Flow velocity：0.25mL/min；Sample size：10μL；Column temperature：40℃；Three kinds of second
Acyl first quinoline metabolin liquid phase gradient elution program is as follows：
D) Mass Spectrometry Conditions：
Compound concentration is 1 μ g/mL, tri- kinds of mequindox metabolins and tunes liquid automatically, using peristaltic pump input mode (10 μ L/
Min), tuning automatically, the Mass Spectrometry Conditions of optimization are as follows：Ionization mode：Cation (ESI+)；Ionization voltage：2.50 kV；
Taper hole voltage：30 V；Ion source temperature：110℃；Taper hole blowback gas velocity：50 L/h；Desolvation temperature：350℃；Precipitation
Agent gas velocity：700 L/h；Argon gas flow velocity：0.11 mL/min；Other specification is as follows, and * represents quota ion：
Claims (1)
1. the assay method of an oxygen mequindox, de- dioxy mequindox and MQCA is taken off in sea cucumber, it is characterised in that：
A) instrument and reagent：
Ultra Performance Liquid Chromatography-tandem mass spectrometer (Quattro Premier XE, Waters, USA), ultra-pure water instrument
(Milli-Q Gradient, Millipore, France), supercentrifuge (TGL-10C, Town in Shanghai booth scientific instrument
Factory), ultrasonic cleaner (KQ-600E, Kunshan Ultrasonic Instruments Co., Ltd.), Nitrogen evaporator (N-EVAPTM112,
Organomation Associates, USA), Full-automatic solid phase extraction instrument (ASPEC XL4+, GILSON, France)；Methyl alcohol
(chromatographically pure), acetonitrile (chromatographically pure), formic acid (top pure grade), hydrochloric acid（Top pure grade）, ethyl acetate（Chromatographically pure）, formic acid（Top grade
It is pure）, sodium acetate（Top pure grade）, solid phase extraction column (MAX 60mg 3mL, Waters, USA)；Mequindox metabolin：MQCA
（German Dr.）Purity>98%, an oxygen mequindox and de- dioxy mequindox are taken off by Wuhan Green's Baeyer biotechnology company
Synthesis, purity>90%；Mixed standard solution：Appropriate mequindox metabolin standard items are weighed, is dissolved with acetonitrile, be made into 100
Mg/L Standard Stock solutions；
B) sample extraction and purification：
5.00g sea cucumber samples are weighed, the mol/L hydrochloric acid of 15mL 2, ultrasonic extraction 1h, 10000 turns/min centrifugations at 4 DEG C is added
10min；Solid phase extraction column with 3mL methyl alcohol and 3mL 2mol/L hydrochloric acid activations, adds the supernatant in centrifuge tube in advance, uses
3mL sodium acetate methanol solution drip washing, the 6mL wash-outs of the ethyl acetate containing 2% formic acid are collected；Nitrogen is dried up at 40 DEG C, adds 1mL to contain
The acetonitrile-aqueous solution of 0.1% formic acid（70:30, v/v）1mL constant volumes, Ultra Performance Liquid Chromatography tandem mass spectrometry is determined；Wherein take off
One oxygen mequindox, de- dioxy mequindox and MQCA（3- Jia based quinoxaline -2- carboxylic acids）Quantitative limit be：2μg/kg；
C) chromatographic condition：
Chromatographic column：ACQUITYTM UPLC BEHC18(1.7μm, 2.1mm i.d.×100mm)；Chromatogram column temperature：40℃；Stream
Dynamic phase：Acetonitrile (A) and containing 0.5% formic acid water (B)；Flow velocity：0.25mL/min；Sample size：10μL；Column temperature：40℃；Three kinds of second
Acyl first quinoline metabolin liquid phase gradient elution program is as follows：
D) Mass Spectrometry Conditions：
Compound concentration is 1 μ g/mL, tri- kinds of mequindox metabolins and tunes liquid automatically, using peristaltic pump input mode (10 μ L/
Min), tuning automatically, the Mass Spectrometry Conditions of optimization are as follows：Ionization mode：Cation (ESI+)；Ionization voltage：2.50 kV；
Taper hole voltage：30 V；Ion source temperature：110℃；Taper hole blowback gas velocity：50 L/h；Desolvation temperature：350℃；Precipitation
Agent gas velocity：700 L/h；Argon gas flow velocity：0.11 mL/min；Other specification is as follows：*Represent quota ion
。
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| CN201610988653.4A CN106568866A (en) | 2016-11-10 | 2016-11-10 | Measuring method of desoxym equindox, 3-metyl-2-acetyl-quinoxaline and MQCA in trepang |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108918748A (en) * | 2018-07-25 | 2018-11-30 | 北京出入境检验检疫局检验检疫技术中心 | Olaquindox metabolite and the method and its sample-pretreating method of carbadox metabolite content in a kind of detection animal muscle tissue |
| CN109164185A (en) * | 2018-10-10 | 2019-01-08 | 福建省农业科学院农业质量标准与检测技术研究所 | The method of mequindox and its major metabolite in HPLC Simultaneous Determination aquatic products |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2006105699A (en) * | 2004-10-01 | 2006-04-20 | Maruzen Pharmaceut Co Ltd | Quantitatively determining method of peptide |
| CN101648917A (en) * | 2009-07-14 | 2010-02-17 | 华南农业大学 | Mequindox metabolic product and preparation method and application thereof |
-
2016
- 2016-11-10 CN CN201610988653.4A patent/CN106568866A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2006105699A (en) * | 2004-10-01 | 2006-04-20 | Maruzen Pharmaceut Co Ltd | Quantitatively determining method of peptide |
| CN101648917A (en) * | 2009-07-14 | 2010-02-17 | 华南农业大学 | Mequindox metabolic product and preparation method and application thereof |
Non-Patent Citations (5)
| Title |
|---|
| 刘慧慧等: "HPLC-MS-MS法分析乙酰甲喹在海参中的主要代谢物", 《质谱学报》 * |
| 刘迎春等: "高效液相色谱-电喷雾离子阱飞行时间质谱法鉴定乙酰甲喹及其代谢物", 《分析化学研究报告》 * |
| 宫向红等: "高效液相色谱法测定水产品中乙酰甲喹残留量", 《中国渔业质量与标准》 * |
| 徐英江等: "超高效液相色谱-串联质谱测定水产品中的卡巴氧、喹烯酮、乙酰甲喹及其代谢物", 《分析测试学报》 * |
| 汤有志等: "脱二氧乙酰甲喹及N4-脱一氧乙酰甲喹的合成及结构表征", 《动物医学进展》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108918748A (en) * | 2018-07-25 | 2018-11-30 | 北京出入境检验检疫局检验检疫技术中心 | Olaquindox metabolite and the method and its sample-pretreating method of carbadox metabolite content in a kind of detection animal muscle tissue |
| CN109164185A (en) * | 2018-10-10 | 2019-01-08 | 福建省农业科学院农业质量标准与检测技术研究所 | The method of mequindox and its major metabolite in HPLC Simultaneous Determination aquatic products |
| CN109164185B (en) * | 2018-10-10 | 2021-08-10 | 福建省农业科学院农业质量标准与检测技术研究所 | Method for simultaneously and quantitatively determining mequindox and main metabolites thereof in aquatic products by HPLC |
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Application publication date: 20170419 |