CN103399096B - Method for detecting content of malachite green and metabolin thereof in sediment of aquaculture environment - Google Patents
Method for detecting content of malachite green and metabolin thereof in sediment of aquaculture environment Download PDFInfo
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Abstract
The invention relates to a method for detecting content of malachite green and a metabolin thereof in sediment of aquaculture environment, and belongs to the technical field of environment quality and security detection. A sample liquid obtained by subjecting the sediment to acetonitrile ultrasonic extraction is quantitatively analyzed by liquid chromatography-tandem mass spectrometry after being purified by a multiwalled carbon nanotube solid phase extraction column. The method provided by the invention is convenient for operation, has relatively good repeatability, accurate determination and relatively high recovery rate, and can rapidly analyze the malachite green and the metabolin thereof in the sediment of the aquaculture environment.
Description
Technical field
The invention belongs to environment security detection technique field, relate to a kind of culture environment of aquatic products sediment Malachite Green and metabolite content detection method thereof.
Background technology
Malachite green belongs to a kind of industrial dye, in aquaculture through being commonly used to sterilization for the treatment of saprolegniasis, cheek mildew and environment etc.Research shows that malachite green and metabolic product procrypsis malachite green thereof eliminate slowly in animal body, residence time is long, and there is carcinogenic, teratogenesis and mutagenesis, American-European a lot of country and the Ministry of Agriculture of China No. 235 file of bulletin are all classified malachite green to ban use of medicine as, are defined in animal derived food and must not detect.In aquaculture at present, the phenomenon of Misuse malachite green has obtained effective control of government agencies at all levels, but still have minority fisherman using malachite green, added once and be widely used, component environment has been caused to pollution, cause residual malachite green class medicine in part breeding environment surface deposit, for follow-up aquaculture product quality is brought potential safety hazard, particularly absorption has the sediment of these materials can constantly slowly discharge to water body again, becomes the main source of secondary pollution.The aquatic products that in actual testing, often discovery is not used malachite green class medicine to carry out disease treatment yet also can detect the existence of malachite green and its residues.The existence of above situation makes troubles to a certain extent the work of tracing to the source of fish quality problem.Therefore monitor culture environment of aquatic products sediment Malachite Green and metabolite content thereof and there is positive effect to understanding breeding environment safety case, investigation aquatic products Malachite Green pollution source and fish quality supervision.At present less for the research of the trace malachite green and its residues detection method in culture environment of aquatic products bed mud.How by Sample Pretreatment Technique and instrument analysis technology effectively, set up that a kind of example enrichment degree is high, the recovery is high, the detection method of favorable reproducibility and the high culture environment of aquatic products sediment Malachite Green of degree of accuracy and metabolite residue amount thereof becomes the key of research.
Carbon nano-tube (Carbon nanotubes, CNTS) is as novel functional material, is widely applied at physics, chemistry and the field such as biological.CNTS has very strong adsorptive power, can be combined with object molecule by noncovalent interaction power such as π-π acting force, Van der Waals force and hydrophobic interactions, there is the advantages such as high-specific surface area, good chemistry, mechanical stability and thermal stability, can be used as good gas chromatography, Capillary Electrophoresis and performance liquid chromatographic column packing material, also can be used as sorbing material for being enriched with organic pollutants, metallic ion and biomacromolecule.The superperformance having due to CNTS material and cheap price advantage, the multi-walled carbon nano-tubes (MWCNTS) in existing bibliographical information CNTS is in the pre-treatment of the samples such as water body organophosphorus, organochlorine, agricultural chemicals and purifying at present.Multi-wall carbon nano-tube tube material is also had to positive meaning for detection of veterinary drugs in food work for expanding residue of veterinary drug pretreatment technology more.The detection technique of malachite green has liquid chromatography and liquid chromatograph mass spectrography.After propane sulfonic acid cation exchange solid-phase extraction column or aluminium oxide Solid-Phase Extraction column purification, realize liquid chromatography-fluoroscopic examination, the detection of liquid chromatography-ultraviolet and liquid chromatography-mass spectrometry analyzing and testing.Liquid phase chromatography complex pretreatment, consuming time, detect limit for height and be unfavorable for accurate quantitative analysis and qualitative analysis.Culture environment of aquatic products sediment matrix components complexity, and pollutant load is lower, if Impurity removal is bad in sample pretreatment process, directly understand the measurement result of interference Instrument, be therefore object of the present invention by carbon nanomaterial efficient and with low cost for Sample Pretreatment Technique.
Summary of the invention
The object of the invention is to overcome the deficiency of existing analytical approach, can find a kind of culture environment of aquatic products sediment Malachite Green quick, accurate and with low cost and the assay method of metabolite content thereof, measure with the quantitative and qualitative analysis that realizes culture environment of aquatic products sediment Malachite Green and metabolin thereof.
The inventive method comprises the following steps successively: (1) ultrasonic solvent extraction, (2) solid phase extraction concentration purify, (3) liquid chromatography-Mass Spectrometer Method, the method for operating of each step is as follows:
(1) ultrasonic solvent extraction: accurately weigh 2.0～10.0g uniform deposition matter sample and join in polypropylene centrifuge tube, accurately add the deuterated malachite green of 50ng, the deuterated procrypsis malachite green of 50ng and 10～12mL trifluoroacetic acid aqueous solution; After jumping a queue, vortex vibration mixes 2min, and under 40KHz ul-trasonic irradiation, 35～45 DEG C of temperature extraction 8min, then will extract sample centrifugal 6 min under rotating speed 5000 rpm, and supernatant is transferred in 25mL color comparison tube; In residue, add again the extraction that repeats after 12mL acetonitrile above and centrifugally operated once, merge supernatant and be settled to 25mL with acetonitrile, obtain sample liquid.
(2) solid phase extraction concentration purifies: first place one deck aperture 20 μ m silica sand filter core sieve plates in 3mL tygon solid phase extraction column (the special void column of SPE) bottom, add 20～60mg multi-wall carbon nano-tube tube material (diameter 40～60nm, length 5～15 μ m, purity > 95%, ash content≤2%, specific surface area is 40～300m
2/ g), place again one deck 20 μ m filter core sieve plates above, by regulate upper strata filter core sieve plate downwards compression degree control multi-walled carbon nano-tubes depth of packing at 0.5～0.6cm; The multi-walled carbon nano-tubes solid-phase extraction column completing first, with methyl alcohol and pure water rinse activation, then accurately pipettes 5mL sample liquid in solid-phase extraction column, and coutroi velocity, at 1.0mL/min, receives efflux; In solid-phase extraction column, add 5% formic acid acetonitrile 2～4mL to carry out drip washing again, merge efflux.
(3) liquid chromatography-Mass Spectrometer Method: efflux, after nitrogen dries up at 45～55 DEG C, uses 2mL acetonitrile～5mmol/L ammonium acetate buffer solution (1+1, v/v) fully to dissolve; Get 1mL sample liquid,, in sample flasket, under liquid chromatography-mass spectrum condition of having set, detect through 0.22 μ m organic phase filtering with microporous membrane, carry out qualitative and quantitative analysis.Chromatographic condition: C
18chromatographic column (50mm × 2.1 mm, 1.7 μ m), 40 DEG C of column temperatures, sample size 10 μ L, mobile phase composition is 5mmol/L ammonium acetate buffer solution and the acetonitrile of adjusting pH to 4.5 with formic acid, flow velocity 0.3 mL/min, condition of gradient elution, 0～3.0min, 50%～5%(ammonium acetate buffer solution, linear change), 3.0～5.0min, 5%～50%(ammonium acetate buffer solution, linear change); Mass spectrum condition: electron spray positive ion mode (ESI
+), capillary voltage 3.50 kv, 120 DEG C of ion source temperatures, 380 DEG C of desolventizing temperature degree, desolventizing airshed 600 L/h, taper hole airshed 50 L/h, multiple-reaction monitoring (MRM) scan pattern; Malachite green quota ion is to 329.3/313.2, qualitative ion pair 329.3/208.1, and procrypsis malachite green quota ion is to 331.2/316.2, qualitative ion pair 331.2/239.2; Drawing standard working curve, calculates malachite green and procrypsis malachite green content with inner mark method ration.
Beneficial effect of the present invention: the present invention has set up the detection method of culture environment of aquatic products sediment Malachite Green and metabolite content thereof; Adopt cheap multi-wall carbon nano-tube tube material to carry out example enrichment purification as solid phase extraction column stuffing, good purification, has reduced impurity effectively to measuring the interference of process; Adopt liquid chromatography-mass spectroscopy to carry out the monitoring of many reactive ions, there is higher sensitivity and degree of accuracy; Compared with existing measuring technology, the present invention does not need commercialization PRS cation exchange or the aluminium oxide solid-phase extraction column that price is more expensive in detection malachite green and its residues process, method can be carried out qualitative and quantitative measurement simultaneously, operation is simple, consuming cost is low, the malachite green and its residues content in can express-analysis culture environment of aquatic products sediment.
Brief description of the drawings
Accompanying drawing 1 is the standard solution chromatogram of 1.00 μ g/L;
The daughter ion scintigram that accompanying drawing 2 is malachite green;
Accompanying drawing 3 is the daughter ion scintigram of procrypsis malachite green.
Embodiment
Below in conjunction with drawings and Examples, technical scheme of the present invention is further described.
Liquid chromatography-the Mass Spectrometer Method of embodiment 1 aquaculture pond bed mud Malachite Green
(1) ultrasonic solvent extraction: accurately weigh 2.0g uniform deposition matter sample and join in polypropylene centrifuge tube, add deuterated malachite green and deuterated procrypsis malachite green mixing inner mark solution 500 μ L and the 10mL trifluoroacetic acid aqueous solution of 100ng/mL; After jumping a queue, vortex vibration mixes 2min, and under 40KHz ul-trasonic irradiation, 35 DEG C of temperature extraction 8min, then will extract sample centrifugal 6 min under rotating speed 5000 rpm, and supernatant is transferred in 25mL color comparison tube; In residue, add again the extraction that repeats after 12mL acetonitrile above and centrifugally operated once, merge supernatant and be settled to 25mL with acetonitrile.
(2) solid phase extraction concentration purifies: first place one deck aperture 20 μ m silica sand filter core sieve plates in 3mL tygon solid phase extraction column (the special void column of SPE) bottom, add 20mg multi-wall carbon nano-tube tube material (diameter 40～60nm, length 5～15 μ m, purity > 95%, ash content≤2%, specific surface area is 40～300m
2/ g), place again one deck 20 μ m filter core sieve plates above, by regulate upper strata filter core sieve plate downwards compression degree control multi-walled carbon nano-tubes depth of packing at 0.5cm; The multi-walled carbon nano-tubes solid-phase extraction column completing first, with methyl alcohol and pure water rinse activation, then accurately pipettes 5mL sample liquid in solid-phase extraction column, and coutroi velocity, at 1.0mL/min, receives efflux; In solid-phase extraction column, add 5% formic acid acetonitrile 2mL to carry out drip washing again, merge efflux.
(3) liquid chromatography-Mass Spectrometer Method: eluent, after nitrogen dries up at 45 DEG C, uses 2mL acetonitrile-5mmol/L ammonium acetate buffer solution (1+1, v/v) fully to dissolve; Get 1mL sample liquid,, in sample flasket, under liquid chromatography-mass spectrum condition of having set, detect through 0.22 μ m organic phase filtering with microporous membrane, carry out qualitative and quantitative analysis.Chromatographic condition: C
18chromatographic column (50mm × 2.1 mm, 1.7 μ m), 40 DEG C of column temperatures, sample size 10 μ L, mobile phase composition is 5mmol/L ammonium acetate buffer solution and the acetonitrile of adjusting pH to 4.5 with formic acid, flow velocity 0.3 mL/min, condition of gradient elution, 0～3.0min, 50%～5%(ammonium acetate buffer solution, linear change), 3.0～5.0min, 5%～50%(ammonium acetate buffer solution, linear change), obtain the chromatogram that appearance time is moderate, peak shape is sharp-pointed, degree of separation is good, appearance time malachite green 1.06min, procrypsis malachite green 3.01min(is shown in accompanying drawing 1); Mass spectrum condition: electron spray positive ion mode (ESI
+), capillary voltage 3.50 kv, 120 DEG C of ion source temperatures, 380 DEG C of desolventizing temperature degree, desolventizing airshed 600 L/h, taper hole airshed 50 L/h, multiple-reaction monitoring (MRM) scan pattern; Malachite green quota ion is to 329.3/313.2, and qualitative ion pair 329.3/208.1(is shown in accompanying drawing 2), procrypsis malachite green quota ion is to 331.2/316.2, qualitative ion pair 331.2/239.2(is shown in accompanying drawing 3); Drawing standard working curve, calculates malachite green and procrypsis malachite green content with inner mark method ration.
(4) Specification Curve of Increasing
With acetonitrile-5mmol/L ammonium acetate buffer solution (1+1, v/v) compound concentration is respectively malachite green and the procrypsis malachite green mixed standard solution of 0.02 μ g/L, 0.05 μ g/L, 0.10 μ g/L, 0.25 μ g/L, 1.00 μ g/L, 2.00 μ g/L and 5.00 μ g/L, the deuterated malachite green of internal standard compound and deuterated procrypsis malachite green concentration are 5.00 μ g/L, then operate according to above-mentioned steps requirement (3), require drawing standard curve according to internal standard method;
(5) the mensuration of the method recovery
The mixed standard solution that adds respectively malachite green and procrypsis malachite green by the concentration of 0.10 μ g/kg, 0.20 μ g/kg, 0.50 μ g/kg, 1.00 μ g/kg and 2.00 μ g/kg in sediment of pond, each interpolation level is done respectively 6 Duplicate Samples; According to step (1)～(4) carry out liquid chromatography-Mass Spectrometer Method, and with typical curve comparison obtained above, finally obtain the concentration of sediment of pond Malachite Green to be measured and procrypsis malachite green by converting.The method recovery is at 76～103%, RSD < 15%.
Liquid chromatography-the Mass Spectrometer Method of embodiment 2 aquaculture pond bed mud Malachite Greens
(1) ultrasonic solvent extraction: accurately weigh 5.0g uniform deposition matter sample and join in polypropylene centrifuge tube, add deuterated malachite green and deuterated procrypsis malachite green mixing inner mark solution 500 μ L and the 11mL trifluoroacetic acid aqueous solution of 100ng/mL; After jumping a queue, vortex vibration mixes 2min, and under 40KHz ul-trasonic irradiation, 40 DEG C of temperature extraction 8min, then will extract sample centrifugal 6 min under rotating speed 5000 rpm, and supernatant is transferred in 25mL color comparison tube; In residue, add again the extraction that repeats after 12mL acetonitrile above and centrifugally operated once, merge supernatant and be settled to 25mL with acetonitrile.
(2) solid phase extraction concentration purifies: first place one deck aperture 20 μ m silica sand filter core sieve plates in 3mL tygon solid phase extraction column (the special void column of SPE) bottom, add 40mg multi-wall carbon nano-tube tube material (diameter 40～60nm, length 5～15 μ m, purity > 95%, ash content≤2%, specific surface area is 40～300m
2/ g), place again one deck 20 μ m filter core sieve plates above, by regulate upper strata filter core sieve plate downwards compression degree control multi-walled carbon nano-tubes depth of packing at 0.6cm; The multi-walled carbon nano-tubes solid-phase extraction column completing first, with methyl alcohol and pure water rinse activation, then accurately pipettes 5mL sample liquid in solid-phase extraction column, and coutroi velocity, at 1.0mL/min, receives efflux; In solid-phase extraction column, add 5% formic acid acetonitrile 3mL to carry out drip washing again, merge efflux.
(3) liquid chromatography-Mass Spectrometer Method: eluent, after nitrogen dries up at 50 DEG C, uses 2mL acetonitrile-5mmol/L ammonium acetate buffer solution (1+1, v/v) fully to dissolve; Get 1mL sample liquid,, in sample flasket, under liquid chromatography-mass spectrum condition of having set, detect through 0.22 μ m organic phase filtering with microporous membrane, carry out qualitative and quantitative analysis.Chromatographic condition: C
18chromatographic column (50mm × 2.1 mm, 1.7 μ m), 40 DEG C of column temperatures, sample size 10 μ L, mobile phase composition is 5 mmol/L ammonium acetate buffer solution and the acetonitriles of adjusting pH to 4.5 with formic acid, flow velocity 0.3 mL/min, condition of gradient elution, 0～3.0min, 50%～5%(ammonium acetate buffer solution, linear change), 3.0～5.0min, 5%～50%(ammonium acetate buffer solution, linear change); Mass spectrum condition: electron spray positive ion mode (ESI
+), capillary voltage 3.50 kv, 120 DEG C of ion source temperatures, 380 DEG C of desolventizing temperature degree, desolventizing airshed 600 L/h, taper hole airshed 50 L/h, multiple-reaction monitoring (MRM) scan pattern; Malachite green quota ion is to 329.3/313.2, qualitative ion pair 329.3/208.1, and procrypsis malachite green quota ion is to 331.2/316.2, qualitative ion pair 331.2/239.2; Drawing standard working curve, calculates malachite green and procrypsis malachite green content with inner mark method ration.
(4) Specification Curve of Increasing
With acetonitrile-5mmol/L ammonium acetate buffer solution (1+1, v/v) compound concentration is respectively malachite green and the procrypsis malachite green mixed standard solution of 0.02 μ g/L, 0.05 μ g/L, 0.10 μ g/L, 0.25 μ g/L, 1.00 μ g/L, 2.00 μ g/L and 5.00 μ g/L, the deuterated malachite green of internal standard compound and deuterated procrypsis malachite green concentration are 5.00 μ g/L, then operate according to above-mentioned steps requirement (3), require drawing standard curve according to internal standard method.
(5) the mensuration of the method recovery
The mixed standard solution that adds respectively malachite green and procrypsis malachite green by the concentration of 0.10 μ g/kg, 0.20 μ g/kg, 0.50 μ g/kg, 1.00 μ g/kg and 2.00 μ g/kg in sediment of pond, each interpolation level is done respectively 6 Duplicate Samples; According to step (1)～(4) carry out liquid chromatography-Mass Spectrometer Method, and with typical curve comparison obtained above, finally obtain the concentration of sediment of pond Malachite Green to be measured and procrypsis malachite green by converting.The method recovery is at 81～109%, RSD < 15%.
Liquid chromatography-the Mass Spectrometer Method of embodiment 3 aquaculture pond bed mud Malachite Greens
(1) ultrasonic solvent extraction: accurately weigh 10.0g uniform deposition matter sample and join in polypropylene centrifuge tube, add deuterated malachite green and deuterated procrypsis malachite green mixing inner mark solution 500 μ L and the 12mL trifluoroacetic acid aqueous solution of 100ng/mL; After jumping a queue, vortex vibration mixes 2min, and under 40KHz ul-trasonic irradiation, 45 DEG C of temperature extraction 8min, then will extract sample centrifugal 6 min under rotating speed 5000 rpm, and supernatant is transferred in 25mL color comparison tube; In residue, add again the extraction that repeats after 12mL acetonitrile above and centrifugally operated once, merge supernatant and be settled to 25mL with acetonitrile.
(2) solid phase extraction concentration purifies: first place one deck aperture 20 μ m silica sand filter core sieve plates in 3mL tygon solid phase extraction column (the special void column of SPE) bottom, add 60mg multi-wall carbon nano-tube tube material (diameter 40～60nm, length 5～15 μ m, purity > 95%, ash content≤2%, specific surface area is 40～300m
2/ g), place again one deck 20 μ m filter core sieve plates above, by regulate upper strata filter core sieve plate downwards compression degree control multi-walled carbon nano-tubes depth of packing at 0.58cm; The multi-walled carbon nano-tubes solid-phase extraction column completing first, with methyl alcohol and pure water rinse activation, then accurately pipettes 5mL sample liquid in solid-phase extraction column, and coutroi velocity, at 1.0mL/min, receives efflux; In solid-phase extraction column, add 5% formic acid acetonitrile 4mL to carry out drip washing again, merge efflux.
(3) liquid chromatography-Mass Spectrometer Method: eluent, after nitrogen dries up at 55 DEG C, uses 2mL acetonitrile-5mmol/L ammonium acetate buffer solution (1+1, v/v) fully to dissolve; Get 1mL sample liquid,, in sample flasket, under liquid chromatography-mass spectrum condition of having set, detect through 0.22 μ m organic phase filtering with microporous membrane, carry out qualitative and quantitative analysis.Chromatographic condition: C
18chromatographic column (50mm × 2.1 mm, 1.7 μ m), 40 DEG C of column temperatures, sample size 10 μ L, mobile phase composition is 5 mmol/L ammonium acetate buffer solution and the acetonitriles of adjusting pH to 4.5 with formic acid, flow velocity 0.3 mL/min, condition of gradient elution, 0～3.0min, 50%～5%(ammonium acetate buffer solution, linear change), 3.0～5.0min, 5%～50%(ammonium acetate buffer solution, linear change); Mass spectrum condition: electron spray positive ion mode (ESI
+), capillary voltage 3.50 kv, 120 DEG C of ion source temperatures, 380 DEG C of desolventizing temperature degree, desolventizing airshed 600 L/h, taper hole airshed 50 L/h, multiple-reaction monitoring (MRM) scan pattern; Malachite green quota ion is to 329.3/313.2, qualitative ion pair 329.3/208.1, and procrypsis malachite green quota ion is to 331.2/316.2, qualitative ion pair 331.2/239.2; Drawing standard working curve, calculates malachite green and procrypsis malachite green content with inner mark method ration.
(4) Specification Curve of Increasing
With acetonitrile-5mmol/L ammonium acetate buffer solution (1+1, v/v) compound concentration is respectively malachite green and the procrypsis malachite green mixed standard solution of 0.02 μ g/L, 0.05 μ g/L, 0.10 μ g/L, 0.25 μ g/L, 1.00 μ g/L, 2.00 μ g/L and 5.00 μ g/L, the deuterated malachite green of internal standard compound and deuterated procrypsis malachite green concentration are 5.00 μ g/L, then operate according to above-mentioned steps requirement (3), require drawing standard curve according to internal standard method.
(5) the mensuration of the method recovery
The mixed standard solution that adds respectively malachite green and procrypsis malachite green by the concentration of 0.10 μ g/kg, 0.20 μ g/kg, 0.50 μ g/kg, 1.00 μ g/kg and 2.00 μ g/kg in sediment of pond, each interpolation level is done respectively 6 Duplicate Samples; According to step (1)～(4) carry out liquid chromatography-Mass Spectrometer Method, and with typical curve comparison obtained above, finally obtain the concentration of sediment of pond Malachite Green to be measured and procrypsis malachite green by converting.The method recovery is at 87～101%, RSD < 15%.
Above-described embodiment is to explanation of the present invention, is not limitation of the invention, any scheme after simple change of the present invention is all belonged to protection scope of the present invention.
Claims (2)
1. culture environment of aquatic products sediment Malachite Green and metabolite content detection method thereof, is characterized in that the method comprises that ultrasonic solvent extraction, solid phase extraction concentration purify and these three steps of liquid chromatography-Mass Spectrometer Method, wherein:
Described ultrasonic solvent extraction, operation by the following step: accurately weigh 2.0～10.0g uniform deposition matter sample and join in polypropylene centrifuge tube, accurately add the deuterated malachite green of 50ng, the deuterated procrypsis malachite green of 50ng and 10～12mL trifluoroacetic acid aqueous solution; After jumping a queue, vortex vibration mixes 2min, and under 40KHz ul-trasonic irradiation, 35～45 DEG C of temperature extraction 8min, then will extract sample centrifugal 6 min under rotating speed 5000 rpm, and supernatant is transferred in 25mL color comparison tube; In residue, add again the extraction that repeats after 12mL acetonitrile above and centrifugally operated once, merge supernatant and be settled to 25mL with acetonitrile, obtain sample liquid;
Described solid phase extraction concentration purifies, operation by the following step: first place one deck aperture 20 μ m silica sand filter core sieve plates in 3mL tygon solid phase extraction column bottom, add 20～60mg multi-wall carbon nano-tube tube material, place again one deck 20 μ m filter core sieve plates above, by regulate upper strata filter core sieve plate downwards compression degree control multi-walled carbon nano-tubes depth of packing at 0.5～0.6cm; The multi-walled carbon nano-tubes solid-phase extraction column completing first, with 5mL methyl alcohol and 5mL pure water rinse activation, discards efflux; Then accurately pipette 5mL sample liquid in solid-phase extraction column, coutroi velocity, at 1.0mL/min, receives efflux; In solid-phase extraction column, add 5% formic acid acetonitrile 2～4mL to carry out drip washing again, merge efflux;
Described liquid chromatography-Mass Spectrometer Method, by the following step operation: efflux, after nitrogen dries up at 45～55 DEG C, is used 2mL acetonitrile-5mmol/L ammonium acetate buffer solution, 1+1, v/v fully dissolves; Get 1mL sample liquid,, in sample flasket, under liquid chromatography-mass spectrum condition of having set, detect through 0.22 μ m organic phase filtering with microporous membrane, carry out qualitative and quantitative analysis;
Chromatographic condition: C
1840 DEG C of chromatographic column column temperatures, sample size 10 μ L, mobile phase composition is 5mmol/L ammonium acetate buffer solution and the acetonitrile of adjusting pH to 4.5 with formic acid, flow velocity 0.3 mL/min;
In condition of gradient elution: 0～3.0min, ammonium acetate buffer solution volumetric concentration is down to 5% by 50% linearity; In 3.0～5.0min, ammonium acetate buffer solution volumetric concentration rises to 50% by 5% linearity;
Mass spectrum condition: electron spray positive ion mode, capillary voltage 3.50 kv, 120 DEG C of ion source temperatures, 380 DEG C of desolventizing temperature degree, desolventizing airshed 600 L/h, taper hole airshed 50 L/h, multiple-reaction monitoring scan pattern; Malachite green quota ion is to 329.3/313.2, qualitative ion pair 329.3/208.1, and procrypsis malachite green quota ion is to 331.2/316.2, qualitative ion pair 331.2/239.2; Drawing standard working curve, calculates malachite green and procrypsis malachite green content with inner mark method ration.
2. the detection method of a kind of culture environment of aquatic products sediment Malachite Green content according to claim 1, it is characterized in that: described multi-walled carbon nano-tubes material diameter is 40～60nm, length is 5～15 μ m, purity > 95%, ash content≤2%, specific surface area is 40～300m
2/ g.
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| CN104730157A (en) * | 2013-12-23 | 2015-06-24 | 天津优标技术检测服务有限公司 | Method for detecting residues of malachite green in health products by using UPLC-MS |
| CN105301161B (en) * | 2015-10-27 | 2017-05-10 | 四川出入境检验检疫局检验检疫技术中心 | Pre-column electrochemical derivatization one-time total quantity measuring method for malachite green and leucomalachite green in aquatic products |
| CN107462640A (en) * | 2017-06-14 | 2017-12-12 | 大连市水产技术推广总站 | The method for determining breeding water Malachite Green and crystal violet residual quantity simultaneously |
| CN109541088A (en) * | 2018-11-28 | 2019-03-29 | 北京农业质量标准与检测技术研究中心 | A kind of purification method of patulin, solid-phase extraction column and its application |
| CN109633045A (en) * | 2018-12-20 | 2019-04-16 | 广州广电计量检测股份有限公司 | A kind of method that Liquid Chromatography-Tandem Mass Spectrometry measures 6 kinds of pigment residue amounts in aquatic products simultaneously |
| CN110006880B (en) * | 2019-03-20 | 2021-05-25 | 浙江农林大学 | Compound for direct-reading chromogenic rapid detection of malachite green and application |
| CN110376307A (en) * | 2019-08-13 | 2019-10-25 | 深圳市深大检测有限公司 | The method for detecting residue of aquatic products Malachite Green |
| CN116908338A (en) * | 2023-09-11 | 2023-10-20 | 中国热带农业科学院三亚研究院 | Method for simultaneously measuring 110 pesticide residues in various tropical fruits |
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| CN102974129A (en) * | 2012-12-10 | 2013-03-20 | 浙江省海洋水产研究所 | Multi-stage combined type multi-wall carbon nano tube solid phase extraction column |
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