CN103399096A - 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 have carcinogenic, teratogenesis and mutagenesis, American-European a lot of country and China Ministry of Agriculture 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 the 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 to water body, slowly discharge again, becomes the main source of secondary pollution.In actual testing also often the discovery aquatic products that do not use malachite green class medicine to carry out disease treatment also can detect the existence of malachite green and its residues.The existence of above situation makes troubles for 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 have 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 the 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, at physics, chemistry and the field such as biological, is widely applied.CNTS has very strong adsorptive power, can pass through the noncovalent interaction power such as π-π acting force, Van der Waals force and hydrophobic interaction is combined with the object molecule, have high-specific surface area, good advantages such as 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 be used to being enriched with organic pollutants, metallic ion and biomacromolecule.The superperformance that has due to the 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.The multi-wall carbon nano-tube tube material is also had to positive meaning for detection of veterinary drugs in food work for expanding the 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 that liquid chromatography-fluoroscopic examination, liquid chromatography-ultraviolet detect and liquid chromatography-mass spectrometry analyzing and testing.The 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 in sample pretreatment process, Impurity removal is bad, directly understand the measurement result of interference Instrument, by carbon nanomaterial efficient and with low cost, for Sample Pretreatment Technique, be therefore purpose of the present invention.
Summary of the invention
The objective 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, with the qualitative analysis that quantitatively reaches that realizes culture environment of aquatic products sediment Malachite Green and metabolin thereof, measure.
The inventive method comprises the following steps successively: the extraction of ⑴ ultrasonic solvent, ⑵ solid phase extraction concentration purify, ⑶ liquid chromatography-Mass Spectrometer Method, and the method for operating of each step is as follows:
⑴ ultrasonic solvent extraction: accurately weighing 2.0～10.0g uniform deposition matter sample joins in polypropylene centrifuge tube, accurately add the 50ng deuterium for malachite green, 50ng deuterium for procrypsis malachite green and 10～12mL trifluoroacetic acid aqueous solution; The rear vortex vibration of jumping a queue mixes 2min, and 35～45 ℃ of temperature extraction 8min, then will extract sample centrifugal 6 min under rotating speed 5000 rpm under the 40KHz ul-trasonic irradiation, and supernatant is transferred in the 25mL color comparison tube; In residue, add again the extraction that repeats front after the 12mL acetonitrile and centrifugally operated once, merge supernatant and also with acetonitrile, be settled to 25mL, namely obtain sample liquid.
⑵ solid phase extraction concentration purifies: in 3mL tygon solid phase extraction column (the special-purpose void column of SPE) bottom, first place one deck aperture 20 μ m silica sand filter core sieve plates, 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 are 40～300m
2/ g), the above places one deck 20 μ m filter core sieve plates again, by regulate upper strata filter core sieve plate downwards the compression degree control the multi-walled carbon nano-tubes depth of packing at 0.5～0.6cm; The multi-walled carbon nano-tubes solid-phase extraction column that completes first, with methyl alcohol and pure water rinse activation, then accurately pipettes the 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.
⑶ liquid chromatography-Mass Spectrometer Method: efflux, after 45～55 ℃ of lower nitrogen dry up, fully dissolves with 2mL acetonitrile～5mmol/L ammonium acetate buffer solution (1+1, v/v); Get 1mL sample liquid, through 0.22 μ m organic phase filtering with microporous membrane, in sample flasket, under the liquid chromatography of having set-mass spectrum condition, detect, carry out qualitative and quantitative analysis.Chromatographic condition: C
18Chromatographic column (50mm * 2.1 mm, 1.7 μ m), 40 ℃ of column temperatures, sample size 10 μ L, mobile phase composition is with formic acid, to adjust 5mmol/L ammonium acetate buffer solution and the acetonitrile of pH to 4.5, flow velocity 0.3 mL/min, condition of gradient elution, 0～3.0min, 50%～5%(ammonium acetate buffer solution, the linear change), 3.0～5.0min, 5%～50%(ammonium acetate buffer solution, the linear change); Mass spectrum condition: electron spray positive ion mode (ESI
+), capillary voltage 3.50 kv, 120 ℃ of ion source temperatures, 380 ℃ of desolventizing temperature degree, desolventizing airshed 600 L/h, taper hole airshed 50 L/h, multiple-reaction monitoring (MRM) scan pattern; The malachite green quota ion is to 329.3/313.2, and qualitative ion pair 329.3/208.1, procrypsis malachite green quota ion be to 331.2/316.2, qualitative ion pair 331.2/239.2; The drawing standard working curve, calculate 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 the example enrichment purification as solid phase extraction column stuffing, good purification, reduced impurity effectively to measuring the interference of process; Adopt liquid chromatography-mass spectroscopy to carry out the monitoring of many reactive ions, have higher sensitivity and degree of accuracy; With existing measuring technology, compare, the present invention does not need commercialization PRS cation exchange or the aluminium oxide solid-phase extraction column that price is more expensive in detecting the 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.
The accompanying drawing explanation
Accompanying drawing 1 is the standard solution chromatogram of 1.00 μ g/L;
Accompanying drawing 2 is the daughter ion scintigram of 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
⑴ ultrasonic solvent extraction: accurately weighing 2.0g uniform deposition matter sample joins in polypropylene centrifuge tube, the deuterium that adds 100ng/mL for malachite green and deuterium for procrypsis malachite green mixing inner mark solution 500 μ L and 10mL trifluoroacetic acid aqueous solution; The rear vortex vibration of jumping a queue mixes 2min, and 35 ℃ of temperature extraction 8min, then will extract sample centrifugal 6 min under rotating speed 5000 rpm under the 40KHz ul-trasonic irradiation, and supernatant is transferred in the 25mL color comparison tube; In residue, add again the extraction that repeats front after the 12mL acetonitrile and centrifugally operated once, merge supernatant and also with acetonitrile, be settled to 25mL.
⑵ solid phase extraction concentration purifies: in 3mL tygon solid phase extraction column (the special-purpose void column of SPE) bottom, first place one deck aperture 20 μ m silica sand filter core sieve plates, 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 are 40～300m
2/ g), the above places one deck 20 μ m filter core sieve plates again, by regulate upper strata filter core sieve plate downwards the compression degree control the multi-walled carbon nano-tubes depth of packing at 0.5cm; The multi-walled carbon nano-tubes solid-phase extraction column that completes first, with methyl alcohol and pure water rinse activation, then accurately pipettes the 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.
⑶ liquid chromatography-Mass Spectrometer Method: eluent, after 45 ℃ of lower nitrogen dry up, fully dissolves with 2mL acetonitrile-5mmol/L ammonium acetate buffer solution (1+1, v/v); Get 1mL sample liquid, through 0.22 μ m organic phase filtering with microporous membrane, in sample flasket, under the liquid chromatography of having set-mass spectrum condition, detect, carry out qualitative and quantitative analysis.Chromatographic condition: C
18Chromatographic column (50mm * 2.1 mm, 1.7 μ m), 40 ℃ of column temperatures, sample size 10 μ L, mobile phase composition is with formic acid, to adjust 5mmol/L ammonium acetate buffer solution and the acetonitrile of pH to 4.5, flow velocity 0.3 mL/min, condition of gradient elution, 0～3.0min, 50%～5%(ammonium acetate buffer solution, the linear change), 3.0～5.0min, 5%～50%(ammonium acetate buffer solution, the 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 ℃ of ion source temperatures, 380 ℃ of desolventizing temperature degree, desolventizing airshed 600 L/h, taper hole airshed 50 L/h, multiple-reaction monitoring (MRM) scan pattern; The 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); The drawing standard working curve, calculate malachite green and procrypsis malachite green content with inner mark method ration.
⑷ 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 internal standard compound deuterium is 5.00 μ g/L for malachite green and deuterium for procrypsis malachite green concentration, then the requirement according to above-mentioned steps ⑶ operates, and according to internal standard method, requires the drawing standard curve;
⑸ the mensuration of the method recovery
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, add respectively the mixed standard solution of malachite green and procrypsis malachite green, each interpolation level is done respectively 6 Duplicate Samples; According to step ⑴～⑷, carry out liquid chromatography-Mass Spectrometer Method, and compare with typical curve obtained above, by converting, finally obtain the concentration of sediment of pond Malachite Green to be measured and procrypsis malachite green.The method recovery is at 76～103%, RSD<15%.
Liquid chromatography-the Mass Spectrometer Method of embodiment 2 aquaculture pond bed mud Malachite Greens
⑴ ultrasonic solvent extraction: accurately weighing 5.0g uniform deposition matter sample joins in polypropylene centrifuge tube, the deuterium that adds 100ng/mL for malachite green and deuterium for procrypsis malachite green mixing inner mark solution 500 μ L and 11mL trifluoroacetic acid aqueous solution; The rear vortex vibration of jumping a queue mixes 2min, and 40 ℃ of temperature extraction 8min, then will extract sample centrifugal 6 min under rotating speed 5000 rpm under the 40KHz ul-trasonic irradiation, and supernatant is transferred in the 25mL color comparison tube; In residue, add again the extraction that repeats front after the 12mL acetonitrile and centrifugally operated once, merge supernatant and also with acetonitrile, be settled to 25mL.
⑵ solid phase extraction concentration purifies: in 3mL tygon solid phase extraction column (the special-purpose void column of SPE) bottom, first place one deck aperture 20 μ m silica sand filter core sieve plates, 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 are 40～300m
2/ g), the above places one deck 20 μ m filter core sieve plates again, by regulate upper strata filter core sieve plate downwards the compression degree control the multi-walled carbon nano-tubes depth of packing at 0.6cm; The multi-walled carbon nano-tubes solid-phase extraction column that completes first, with methyl alcohol and pure water rinse activation, then accurately pipettes the 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.
⑶ liquid chromatography-Mass Spectrometer Method: eluent, after 50 ℃ of lower nitrogen dry up, fully dissolves with 2mL acetonitrile-5mmol/L ammonium acetate buffer solution (1+1, v/v); Get 1mL sample liquid, through 0.22 μ m organic phase filtering with microporous membrane, in sample flasket, under the liquid chromatography of having set-mass spectrum condition, detect, carry out qualitative and quantitative analysis.Chromatographic condition: C
18Chromatographic column (50mm * 2.1 mm, 1.7 μ m), 40 ℃ of column temperatures, sample size 10 μ L, mobile phase composition is with formic acid, to adjust 5 mmol/L ammonium acetate buffer solution and the acetonitriles of pH to 4.5, flow velocity 0.3 mL/min, condition of gradient elution, 0～3.0min, 50%～5%(ammonium acetate buffer solution, the linear change), 3.0～5.0min, 5%～50%(ammonium acetate buffer solution, the linear change); Mass spectrum condition: electron spray positive ion mode (ESI
+), capillary voltage 3.50 kv, 120 ℃ of ion source temperatures, 380 ℃ of desolventizing temperature degree, desolventizing airshed 600 L/h, taper hole airshed 50 L/h, multiple-reaction monitoring (MRM) scan pattern; The malachite green quota ion is to 329.3/313.2, and qualitative ion pair 329.3/208.1, procrypsis malachite green quota ion be to 331.2/316.2, qualitative ion pair 331.2/239.2; The drawing standard working curve, calculate malachite green and procrypsis malachite green content with inner mark method ration.
⑷ 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 internal standard compound deuterium is 5.00 μ g/L for malachite green and deuterium for procrypsis malachite green concentration, then the requirement according to above-mentioned steps ⑶ operates, and according to internal standard method, requires the drawing standard curve.
⑸ the mensuration of the method recovery
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, add respectively the mixed standard solution of malachite green and procrypsis malachite green, each interpolation level is done respectively 6 Duplicate Samples; According to step ⑴～⑷, carry out liquid chromatography-Mass Spectrometer Method, and compare with typical curve obtained above, by converting, finally obtain the concentration of sediment of pond Malachite Green to be measured and procrypsis malachite green.The method recovery is at 81～109%, RSD<15%.
Liquid chromatography-the Mass Spectrometer Method of embodiment 3 aquaculture pond bed mud Malachite Greens
⑴ ultrasonic solvent extraction: accurately weighing 10.0g uniform deposition matter sample joins in polypropylene centrifuge tube, the deuterium that adds 100ng/mL for malachite green and deuterium for procrypsis malachite green mixing inner mark solution 500 μ L and 12mL trifluoroacetic acid aqueous solution; The rear vortex vibration of jumping a queue mixes 2min, and 45 ℃ of temperature extraction 8min, then will extract sample centrifugal 6 min under rotating speed 5000 rpm under the 40KHz ul-trasonic irradiation, and supernatant is transferred in the 25mL color comparison tube; In residue, add again the extraction that repeats front after the 12mL acetonitrile and centrifugally operated once, merge supernatant and also with acetonitrile, be settled to 25mL.
⑵ solid phase extraction concentration purifies: in 3mL tygon solid phase extraction column (the special-purpose void column of SPE) bottom, first place one deck aperture 20 μ m silica sand filter core sieve plates, 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 are 40～300m
2/ g), the above places one deck 20 μ m filter core sieve plates again, by regulate upper strata filter core sieve plate downwards the compression degree control the multi-walled carbon nano-tubes depth of packing at 0.58cm; The multi-walled carbon nano-tubes solid-phase extraction column that completes first, with methyl alcohol and pure water rinse activation, then accurately pipettes the 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.
⑶ liquid chromatography-Mass Spectrometer Method: eluent, after 55 ℃ of lower nitrogen dry up, fully dissolves with 2mL acetonitrile-5mmol/L ammonium acetate buffer solution (1+1, v/v); Get 1mL sample liquid, through 0.22 μ m organic phase filtering with microporous membrane, in sample flasket, under the liquid chromatography of having set-mass spectrum condition, detect, carry out qualitative and quantitative analysis.Chromatographic condition: C
18Chromatographic column (50mm * 2.1 mm, 1.7 μ m), 40 ℃ of column temperatures, sample size 10 μ L, mobile phase composition is with formic acid, to adjust 5 mmol/L ammonium acetate buffer solution and the acetonitriles of pH to 4.5, flow velocity 0.3 mL/min, condition of gradient elution, 0～3.0min, 50%～5%(ammonium acetate buffer solution, the linear change), 3.0～5.0min, 5%～50%(ammonium acetate buffer solution, the linear change); Mass spectrum condition: electron spray positive ion mode (ESI
+), capillary voltage 3.50 kv, 120 ℃ of ion source temperatures, 380 ℃ of desolventizing temperature degree, desolventizing airshed 600 L/h, taper hole airshed 50 L/h, multiple-reaction monitoring (MRM) scan pattern; The malachite green quota ion is to 329.3/313.2, and qualitative ion pair 329.3/208.1, procrypsis malachite green quota ion be to 331.2/316.2, qualitative ion pair 331.2/239.2; The drawing standard working curve, calculate malachite green and procrypsis malachite green content with inner mark method ration.
⑷ 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 internal standard compound deuterium is 5.00 μ g/L for malachite green and deuterium for procrypsis malachite green concentration, then the requirement according to above-mentioned steps ⑶ operates, and according to internal standard method, requires the drawing standard curve.
⑸ the mensuration of the method recovery
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, add respectively the mixed standard solution of malachite green and procrypsis malachite green, each interpolation level is done respectively 6 Duplicate Samples; According to step ⑴～⑷, carry out liquid chromatography-Mass Spectrometer Method, and compare with typical curve obtained above, by converting, finally obtain the concentration of sediment of pond Malachite Green to be measured and procrypsis malachite green.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:
The extraction of described ultrasonic solvent, operation by the following step: accurately weighing 2.0～10.0g uniform deposition matter sample joins in polypropylene centrifuge tube, accurately add the 50ng deuterium for malachite green, 50ng deuterium for procrypsis malachite green and 10～12mL trifluoroacetic acid aqueous solution; The rear vortex vibration of jumping a queue mixes 2min, and 35～45 ℃ of temperature extraction 8min, then will extract sample centrifugal 6 min under rotating speed 5000 rpm under the 40KHz ul-trasonic irradiation, and supernatant is transferred in the 25mL color comparison tube; In residue, add again the extraction that repeats front after the 12mL acetonitrile and centrifugally operated once, merge supernatant and also with acetonitrile, be settled to 25mL, namely 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, the above places one deck 20 μ m filter core sieve plates again, by regulate upper strata filter core sieve plate downwards the compression degree control the multi-walled carbon nano-tubes depth of packing at 0.5～0.6cm; The multi-walled carbon nano-tubes solid-phase extraction column that completes first, with 5mL methyl alcohol and 5mL pure water rinse activation, discards efflux; Then accurately pipette the 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, operation by the following step: efflux, after 45～55 ℃ of lower nitrogen dry up, fully dissolves with 2mL initial composition mobile phase; Get 1mL sample liquid, through 0.22 μ m organic phase filtering with microporous membrane, in sample flasket, under the liquid chromatography of having set-mass spectrum condition, detect, carry out qualitative and quantitative analysis;
Chromatographic condition: C
1840 ℃ of chromatographic column column temperatures, sample size 10 μ L, mobile phase composition are with formic acid, to adjust 5mmol/L ammonium acetate buffer solution and the acetonitrile of pH to 4.5, flow velocity 0.3 mL/min;
In condition of gradient elution: 0～3.0min, the ammonium acetate buffer solution volumetric concentration is down to 5% by 50% linearity; 3.0 in～5.0min, the 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 ℃ of ion source temperatures, 380 ℃ of desolventizing temperature degree, desolventizing airshed 600 L/h, taper hole airshed 50 L/h, multiple-reaction monitoring scan pattern; The malachite green quota ion is to 329.3/313.2, and qualitative ion pair 329.3/208.1, procrypsis malachite green quota ion be to 331.2/316.2, qualitative ion pair 331.2/239.2; The drawing standard working curve, calculate 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, and length is 5～15 μ m, purity>95%, ash content≤2%, specific surface area are 40～300m
2/ g.
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| 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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| 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 |
| CN105301161A (en) * | 2015-10-27 | 2016-02-03 | 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 |
| CN110006880A (en) * | 2019-03-20 | 2019-07-12 | 浙江农林大学 | A kind of develop the color for direct-reading quickly detects compound and the application of malachite green |
| 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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