CN108802223A - A kind of method of 9 plant growth regulators residual quantities in measurement melon and fruit - Google Patents
A kind of method of 9 plant growth regulators residual quantities in measurement melon and fruit Download PDFInfo
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Abstract
The invention discloses a kind of methods of 9 plant growth regulators residual quantities in measurement melon and fruit, and in particular to the residual detection field of medicine, this method comprises the following steps：Extractant is added in sample to be tested, homogenized obtains sample slurries.The sample slurries are subjected to centrifugal treating, obtain supernatant and residue.Supernatant is redissolved using pure water through rotating to after doing, obtains sample solution.Magnetic adsorbent is added in sample solution to detach magnetic adsorbent after centrifugation oscillation using magnetisable material, obtain magnetic adsorbent and extraction raffinate.The magnetic adsorbent of adsorbate is attached with through eluent, obtains the magnetic adsorbent of eluent and desorption.The eluent is blown to dry through nitrogen, and is redissolved using acetonitrile-aqueous formic acid, filter membrane, obtains machine analysis liquid.Upper machine analysis liquid is analyzed through high performance liquid chromatography tandem mass spectrometry to be measured, and the rate of recovery of each plant growth regulator is obtained.The present invention has the advantages that accuracy is high and easy to operate.
Description
Technical field
The present invention relates to the residual detection fields of medicine, specially a kind of to measure 9 plant growth regulators residual quantities in melon and fruit
Method.
Background technology
Plant growth regulator be it is a kind of it is artificial synthesized, have and natural phytohormone similar physiologic and biological effect
Active material has important adjusting and control action to the growth and development of plant.Because its effectiveness is high, dosage is small, residual hazard is few
Feature and be widely used, have become indispensable important measures in agricultural production.However, as PGRs is used increasingly
Increase and use scope is gradually expanded, safety is also increasingly taken seriously.Studies have shown that remaining in plant
PGRs can enter with food chain in human body, excessive edible immune function of human body to be caused to decline, skeleton bulking or even teratogenesis,
The consequences such as carcinogenic, cause gene mutation.
In recent years, mainly there are Capillary Electrophoresis-mass spectrography (CE-MS), enzyme about the remaining detection methods of PGRs both at home and abroad
Linked immunosorbent assay (ELISA), gas chromatography (GC), gas chromatography-mass spectrography (GC-MS), high performance liquid chromatography (HPLC) and
Liquid Chromatography-Tandem Mass Spectrometry (HPLC-MS/MS).The difficult point of the more residues detectons of PGRs is that its type is more, property phase in plant
Difference is larger, concentration is low and matrix interference is serious etc., for the more demanding of pre-treating method.Therefore, it establishes a set of efficient, accurate
The assay method of high plant growth regulator residual quantity is a problem to be solved.
Invention content
The present invention provides a kind of method of 9 plant growth regulators residual quantities in measurement melon and fruit, this method has standard
True property height and advantage easy to operate.
To achieve the above object, the present invention provides the following technical solutions：
A kind of method of 9 plant growth regulators residual quantities in measurement melon and fruit, this method comprises the following steps：
S01, extractant is added in sample to be tested, homogenized obtains sample slurries；
S02, the sample slurries are subjected to centrifugal treating, obtain supernatant and residue；
S03, the supernatant are redissolved using pure water through rotating to after doing, obtain sample solution；
S04, magnetic adsorbent is added in the sample solution, after centrifugation oscillation, using magnetisable material by sample solution
In be attached with plant growth regulator magnetic adsorbent separation, obtain the magnetic adsorbent for being attached with adsorbate and extraction raffinate；
S05, the magnetic adsorbent for being attached with adsorbate obtain the magnetic absorption of eluent and desorption through eluent
Agent；
Extraction raffinate discards；
S06, the eluent are blown to dry through nitrogen, and are redissolved using acetonitrile-aqueous formic acid, filter membrane obtains machine point
Analyse liquid；
S07, upper machine analysis liquid are analyzed through high performance liquid chromatography tandem mass spectrometry and are measured, and each plant growth regulator is obtained
The rate of recovery.
Preferably, the extractant is selected from：The mixed solution of formic acid and acetonitrile, acetonitrile, sodium hydroxide and acetonitrile it is mixed
Close any one in solution.Wherein, in the mixed solution of formic acid and acetonitrile, the mass fraction of formic acid is 1%~3%；Hydrogen
In the mixed solution of sodium oxide molybdena and acetonitrile, the mass fraction of sodium hydroxide is 1%.
Further include step S01.1 preferably, between step S01 and step S02, the step S01.1 includes：
(1) sample slurries obtain first-time filtrate and filter residue through filtering；
(2) Wagner's reagent is added in filter residue, and is stirred continuously, after 30~50min, coloured filter is rinsed with clear water
Remaining Wagner's reagent on slag obtains the coloured filter residue that dyeing is completed；
(3) coloured filter residue is redissolved using extractant, and calcium sulfate or aluminum sulfate is added, stirring uses first to dissolving
Acid for adjusting pH is to 2.5~3.5, and after reacting 30~50min, centrifugal filtration obtains secondary filtrate and residue；
(4) first-time filtrate and secondary filtrate are mixed, obtains mixing filtrate；
(5) chitosan is added in obtained mixing filtrate, after stirring 5~10min, stands up to mixed solution clarification,
Filtering, obtains filtrate three times and filters out slag；
(6) step (3)~(5) are repeated until filtering out slag lighter to colourless, and merge the filtrate being obtained by filtration every time,
Weak aqua ammonia is added in filtrate after merging and adjusts pH to 6.8~7.2 to get to the supernatant in step S02.
Preferably, in step (3), pH value 2.8.
As further preferred, in step (6), pH value 7.0.
Preferably, in step S04, the magnetic adsorbent be selected from magnetic graphene with it is compound made of beta-cyclodextrin
Composite material, Fe made of material, chitosan and ferroso-ferric oxide3O4Dopamine, Fe3O4- SiO2- chitosan graphite oxides
Alkene, Fe3O4Nitrogen-doped graphene, Fe3O4At least one of beta cyclodextrin-carboxymethyl cellulose.
Preferably, above-mentioned magnetic graphene and the preparation method of composite material made of beta-cyclodextrin include following step
Suddenly：
(a) magnetic graphene is even added in water, is ultrasonically treated, and concentrated ammonia liquor is added, β-is added after mixing
Cyclodextrin reacts 230~260min under 55~68 DEG C of water bath condition, after the completion of reaction, obtains solidliquid mixture；
(b) after the solidliquid mixture is cooled to room temperature, solid phase is detached to get to magnetic graphene using magnetisable material
With composite material made of beta-cyclodextrin.
Preferably, the preparation method of composite material made of above-mentioned chitosan and ferroso-ferric oxide includes the following steps：
(i) the full and uniform mixing of ferroso-ferric oxide, chitosan is taken, is heated under the conditions of 90~100 DEG C of oil bath, and constantly stir
10~15 hours are mixed to get to composite material made of chitosan and ferroso-ferric oxide.
Preferably, in step S05, eluant, eluent is arbitrary in methanol, ethyl acetate, acetonitrile and dichloromethane
It is a kind of.
Preferably, in step S05, it is attached with the magnetic adsorbent of adsorbate through eluant, eluent in triplicate, and merge
The solution obtained three times is the eluent in step S05.
Beneficial effects of the present invention are：
Magnetic solid phases abstraction technique based on magnetic graphene is a kind of novel sample treatment skill developed in recent years
Art, it combine magnetic extracting operation it is simple, it is time saving quickly and graphene has the specific surface area of super large, preferable chemical stabilization
Property and the characteristics of powerful π-π interactions is generated to organic compound, made some progress in terms of sample pre-treatments.
Also, nano-modified material is prepared using beta-cyclodextrin modified magnetic graphene, not only remains beta-cyclodextrin and magnetic graphite
The proper property of both alkene also plays enhancing magnetic graphene water solubility, prevents the effect of its self aggregation.Therefore, of the invention
The scheme provided composite material adsorbed target determinand made of above-mentioned magnetic graphene and beta-cyclodextrin, not only operates
Simpler, accuracy is also very high.
Specific implementation mode
Embodiment 1
The present embodiment provides a kind of technical solutions：
A kind of method of 9 plant growth regulators residual quantities in measurement melon and fruit, which is characterized in that include the following steps：
S01, extractant is added in sample to be tested, homogenized obtains sample slurries.
S02, sample slurries obtain first-time filtrate and filter residue through filtering.
S03, Wagner's reagent is added in filter residue obtained above, and is stirred continuously, after 45min, rinsed with clear water
Remaining Wagner's reagent on coloured filter residue obtains the coloured filter residue that dyeing is completed.Due to being between each cell of melon and fruit
There is pectin to be tightly bonded, is difficult to separate the cell of sample to be tested by mechanical homogenate therefore.Therefore, in this step
In rapid, the cell of melon and fruit is dyed, to facilitate experimenter to judge in subsequent processing, whether melon and fruit cell separates.
S04, coloured filter residue is redissolved using extractant, and calcium sulfate or aluminum sulfate is added, stirring uses first to dissolving
Acid for adjusting pH is to 2.8, and after reacting 40min, centrifugal filtration obtains secondary filtrate and residue.In this step, since system is in
Acidity, and calcium sulfate or aluminum sulfate are added in system, it is equivalent in solution system and generates dehydrating agent sulfuric acid.Pectin is de-
Under the action of aqua, hydrated film is lost, also, under acid condition, it is suppressed that the dissociation of carboxyl in pectin.Meanwhile methoxyl group
Between form hydrogen bond, therefore more and more pectin molecule glue connect webbed structure, to which flocculation is at gel.When pectin quilt
It is in loose condition (of surface) between melon and fruit cell after removing, so that the plant growth regulator remained into the cell can quickly be precipitated,
And it is precipitated more complete.
S05, first-time filtrate and secondary filtrate are mixed, obtains mixing filtrate.
S06, chitosan is added in obtained mixing filtrate, after stirring 8min, stands until mixed solution clarification, mistake
Filter, obtains filtrate three times and filters out slag.
S07, step S04~S06 is repeated until filtering out slag lighter to colourless, and merge the filter being obtained by filtration every time
Weak aqua ammonia is added in filtrate after merging and adjusts pH to 7 to get to the supernatant in step S02 for liquid.
Due to heavy metal component, the pigment composition in the adsorbable filtrate of chitosan, when multiple cells are viscous by pectin
When connecting together, the iodine in cell is not easy to be adsorbed by chitosan, so that filter residue is in aubergine.Pass through repetitive operation step
S04~S06 may make pectin to be completely removed so that melon and fruit cell is in be completely dispersed state, remaining to be more conducive to
Plant growth substance is precipitated.
S08, the sample slurries are subjected to centrifugal treating, obtain supernatant and residue.
S09, the supernatant are redissolved using pure water through rotating to after doing, obtain sample solution.
S10, composite material made of magnetic graphene and beta-cyclodextrin, centrifugation oscillation are added in the sample solution
Afterwards, it will be answered made of the magnetic graphene for being attached with plant growth regulator in sample solution and beta-cyclodextrin using magnetisable material
Condensation material detaches, and obtains composite material and extraction raffinate made of the magnetic graphene for being attached with adsorbate and beta-cyclodextrin.
Composite material is eluted through methanol made of S11, the magnetic graphene for being attached with adsorbate and beta-cyclodextrin, is washed
Composite material made of the magnetic graphene and beta-cyclodextrin of de- liquid and desorption.Obtained extraction raffinate discards.Meanwhile formaldehyde can be used
The chitosan filtered out in above-mentioned steps is eluted, a small amount of plant growth regulator adsorbed elutes.
S12, the eluent are blown to dry through nitrogen, and are redissolved using acetonitrile-aqueous formic acid, filter membrane obtains machine point
Analyse liquid；
S13, upper machine analysis liquid are analyzed through high performance liquid chromatography tandem mass spectrometry and are measured, and each plant growth regulator is obtained
The rate of recovery.
Embodiment 2
The present embodiment provides a kind of preparation method of composite material made of magnetic graphene and beta-cyclodextrin, the preparation sides
Method includes the following steps：
(a) magnetic graphene is even added in water, is ultrasonically treated, and concentrated ammonia liquor is added, β-is added after mixing
Cyclodextrin reacts 240min under 60 DEG C of water bath condition, after the completion of reaction, obtains solidliquid mixture；
(b) after the solidliquid mixture is cooled to room temperature, solid phase is detached to get to magnetic graphene using magnetisable material
With composite material made of beta-cyclodextrin.
Embodiment 3
The present embodiment provides a kind of preparation method of composite material made of chitosan and ferroso-ferric oxide, the preparation methods
Include the following steps：
(i) the full and uniform mixing of ferroso-ferric oxide, chitosan is taken, is heated under the conditions of 95 DEG C of oil bath, and be stirred continuously 12
Hour to get to composite material made of chitosan and ferroso-ferric oxide.
Embodiment 4
Select the blank samples such as tomato, watermelon and strawberry to be added recovery experiment, each pitch-based sphere do 6 it is parallel,
It the results are shown in Table 4.The average recovery rate of 9 kinds of PGRs be 71.7%~101%, relative standard deviation (RSD) be 0.9%~
10.9%, there is good accuracy and precision, meet multi-residue analysis requirement.Concrete outcome is as follows：
Claims (10)
1. a kind of method measuring 9 plant growth regulators residual quantities in melon and fruit, which is characterized in that include the following steps：
S01, extractant is added in sample to be tested, homogenized obtains sample slurries；
S02, the sample slurries are subjected to centrifugal treating, obtain supernatant and residue；
S03, the supernatant are redissolved using pure water through rotating to after doing, obtain sample solution；
S04, magnetic adsorbent is added in the sample solution, it, will be attached in sample solution using magnetisable material after centrifuging oscillation
The magnetic adsorbent separation for plant growth regulator, obtains the magnetic adsorbent for being attached with adsorbate and extraction raffinate；
S05, the magnetic adsorbent for being attached with adsorbate obtain the magnetic adsorbent of eluent and desorption through eluent；
Extraction raffinate discards；
S06, the eluent are blown to dry through nitrogen, and are redissolved using acetonitrile-aqueous formic acid, filter membrane, obtain machine analysis liquid；
S07, upper machine analysis liquid are analyzed through high performance liquid chromatography tandem mass spectrometry and are measured, and the recycling of each plant growth regulator is obtained
Rate.
2. a kind of method measuring 9 plant growth regulators residual quantities in melon and fruit according to claim 1, feature exist
In the extractant is selected from：The mixed solution of formic acid and acetonitrile, acetonitrile, sodium hydroxide and acetonitrile mixed solution in it is arbitrary
It is a kind of；
Wherein, in the mixed solution of formic acid and acetonitrile, the mass fraction of formic acid is 1%~3%；Sodium hydroxide and acetonitrile it is mixed
It closes in solution, the mass fraction of sodium hydroxide is 1%.
3. a kind of method measuring 9 plant growth regulators residual quantities in melon and fruit according to claim 1, feature exist
In further including step S01.1 between step S01 and step S02, the step S01.1 includes：
(1) sample slurries obtain first-time filtrate and filter residue through filtering；
(2) Wagner's reagent is added in filter residue, and is stirred continuously, after 30~50min, is rinsed on coloured filter residue with clear water
Remaining Wagner's reagent obtains the coloured filter residue that dyeing is completed；
(3) coloured filter residue is redissolved using extractant, and calcium sulfate or aluminum sulfate is added, stirring uses formic acid tune to dissolving
PH to 2.5~3.5 is saved, after reacting 30~50min, centrifugal filtration obtains secondary filtrate and residue；
(4) first-time filtrate and secondary filtrate are mixed, obtains mixing filtrate；
(5) chitosan is added in obtained mixing filtrate, after stirring 5~10min, stands up to mixed solution clarification, filtering,
It obtains filtrate three times and filters out slag；
(6) step (3)~(5) are repeated until filtering out slag lighter to colourless, and merges the filtrate being obtained by filtration every time, closed
Weak aqua ammonia is added in filtrate after and and adjusts pH to 6.8~7.2 to get to the supernatant in step S02.
4. a kind of method measuring 9 plant growth regulators residual quantities in melon and fruit according to claim 3, feature exist
In, in step (3), pH value 2.8.
5. a kind of method measuring 9 plant growth regulators residual quantities in melon and fruit according to claim 3, feature exist
In, in step (6), pH value 7.0.
6. a kind of method measuring 9 plant growth regulators residual quantities in melon and fruit according to claim 1, feature exist
In, in step S04, the magnetic adsorbent be selected from composite material, chitosan made of magnetic graphene and beta-cyclodextrin and
Composite material, Fe made of ferroso-ferric oxide3O4Dopamine, Fe3O4- SiO2- chitosans graphene oxide, Fe3O4N doping
Graphene, Fe3O4At least one of beta cyclodextrin-carboxymethyl cellulose.
7. a kind of method measuring 9 plant growth regulators residual quantities in melon and fruit according to claim 6, feature exist
In magnetic graphene and the preparation method of composite material made of beta-cyclodextrin include the following steps：
(a) magnetic graphene is even added in water, is ultrasonically treated, and concentrated ammonia liquor is added, β-ring paste is added after mixing
Essence reacts 230~260min under 55~68 DEG C of water bath condition, after the completion of reaction, obtains solidliquid mixture；
(b) after the solidliquid mixture is cooled to room temperature, solid phase is detached to get to magnetic graphene and β-using magnetisable material
Composite material made of cyclodextrin.
8. a kind of method measuring 9 plant growth regulators residual quantities in melon and fruit according to claim 6, feature exist
In the preparation method of composite material made of chitosan and ferroso-ferric oxide includes the following steps：
(i) the full and uniform mixing of ferroso-ferric oxide, chitosan is taken, is heated under the conditions of 90~100 DEG C of oil bath, and be stirred continuously 10
~15 hours to get to composite material made of chitosan and ferroso-ferric oxide.
9. a kind of method measuring 9 plant growth regulators residual quantities in melon and fruit according to claim 1, feature exist
In, in step S05, any one of eluant, eluent in methanol, ethyl acetate, acetonitrile and dichloromethane.
10. a kind of method measuring 9 plant growth regulators residual quantities in melon and fruit according to claim 1, feature
Be, in step S05, be attached with the magnetic adsorbent of adsorbate through eluant, eluent in triplicate, and merge obtain three times it is molten
Liquid is the eluent in step S05.
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| CN111089917A (en) * | 2019-12-23 | 2020-05-01 | 浙江省食品药品检验研究院 | Method for measuring residual quantity of plant growth regulator in melons and fruits |
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| CN112816635B (en) * | 2020-12-31 | 2023-08-18 | 绿城农科检测技术有限公司 | Method for determining fatty acid in black fungus |
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