CN106872558A - A kind of method of Nano Silver and silver ion in measure plant - Google Patents
A kind of method of Nano Silver and silver ion in measure plant Download PDFInfo
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- CN106872558A CN106872558A CN201710116531.0A CN201710116531A CN106872558A CN 106872558 A CN106872558 A CN 106872558A CN 201710116531 A CN201710116531 A CN 201710116531A CN 106872558 A CN106872558 A CN 106872558A
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Abstract
A kind of method of Nano Silver and silver ion in measure plant, first obtains the plant leaf blade of AgNP pollutions；Carried out extracting AgNP and Ag with weak base TMAH (TMAH) or mixed enzyme Macerozyme R 10 again+；Finally by individual particle inductivity coupled plasma mass spectrometry（spICP‑MS）Determine, realize granule density, particle size distribution and the Ag of AgNP particles in plant+Detection.The inventive method is based on the enzymes of Macerozyme R 10 and extracts individual particle inductivity coupled plasma mass spectrometry GC-MS, can quickly determine AgNP and Ag in plant sample+。
Description
Technical field
The invention belongs to Environmental Analytical Chemistry and field of ecology, it is related to a kind of extraction based on enzyme to couple individual particle-inductance
Nano Silver (AgNP) and silver ion (Ag in coupled plasma mass spectrometry (spICP-MS) technology quick detection plant+) side
Method.
Background technology
Because having excellent, wide spectrum anti-microbial property, AgNP has been widely used in medical nasal spray, textile, wound
In the consumer goods such as treatment bandage, mother and baby's articles for use, washing machine.Counted according to the inferior international scholar center of Wood Percival Lowell, existing market
On the about 410 kinds consumer goods contain AgNP, account for 25.2% (http of all nano material products://
www.nanotechproject.org/).AgNP is preparing, using will inevitably be entered into, discarded and cyclic process
In the middle of environment.Plant is important primary producer in terriecosystems, and more and more evidences show that AgNP can give birth to plant
Development long produces toxicity, including suppresses the sprouting of seed and root elongation, cause metabolic dysfunction, hinders photosynthesis etc..
Compared to AgNP phytotoxicity researchs, from cellular level the problems such as the plant internalization of understanding AgNP, accumulation, transhipment and conversion,
Directly constrain the understanding to AgNP toxicity mechanisms.Therefore, for conversion processes of the scientific evaluation AgNP in plant and enter
One step evaluates the exposure of AgNP, need to set up AgNP and Ag in effectively quick measure plant matrix+Method.
The analysis of nano particle is generally limited by extremely low ambient concentration and complex environment matrix in environmental sample.In recent years
Come cloud point extraction, field flow separation, hydrodynamic chromatography, Capillary Electrophoresis and ICP-MS on-line couplings be successfully used for AgNP and
Ag+Separation determination.But correlative study needs first to carry out the AgNP and Ag of complexity+Lock out operation, could then examine online
Survey.These not only cumbersome, labor intensives of operation and time, but also many manual operation errors can be introduced.It is worth noting that,
These methods fail to realize AgNP and Ag in complicated biological sample+Effective measure.TEM-EDS can be from the Electronic Speculum for photographing
The size information of nano particle in sample is read in photo, but it is not applied for environmental sample (the ng L of low concentration-1-μg
L-1) and the granule density information of nano particle can not be obtained.Synchrotron radiation correlation technique (XANES and μ-XRF etc.) can be analyzed
The combining form of AgNP in sample, there is provided atomic scale information, but this technology be limited to simultaneously Concentration Testing limit high and
Need the data handling procedure of prolonged data acquisition and complexity.Therefore, AgNP in a kind of quick measure biological sample is developed
And Ag+New method it is significant.
The content of the invention
The technical problem of solution：The present invention provides a kind of method for determining Nano Silver and silver ion in plant, the method
Based on Macerozyme R-10 enzyme extraction-individual particle inductivity coupled plasma mass spectrometry GC-MSs, plant can be quickly determined
AgNP and Ag in sample+。
Technical scheme：A kind of method of Nano Silver and silver ion in measure plant, step is：
(1) silver-colored Pollution Plant is obtained；
(2) carried out extracting AgNP and Ag with weak base TMAH or mixed enzyme Macerozyme R-10+；
(3) determined by individual particle-inductivity coupled plasma mass spectrometry (spICP-MS), realize AgNP particles in plant
Granule density, particle size distribution and Ag+Detection.
Above-mentioned TMAH extracts AgNP and Ag+The step of be：Step 1) 0.1g plant leaf blades are weighed, it is placed in after being cut into small pieces
In 50mL beakers, it is marked with the AgNP standard liquids of 80nm so that the concentration of AgNP is 28,000NPs in final solution
mL-1；Step 2) to the TMAH that 10mL 1wt.% are added in the plant leaf blade of mark completion, water bath sonicator 24h dispersion tissues simultaneously prevent
Only AgNP reunites, and the tissue suspension after ultrasound is moved into the filter membrane that aperture is 1 μm is crossed after centrifuge tube centrifugation；Step 3) utilize
The detection of spICP-MS instruments, model：NexION 300, PerkinElmer, USA；Instrument measuring parameter is：ICP-MS is operated
Condition：Flow rate of carrier gas 1L min-1；Auxiliary gas flow speed 1.2L min-1；Plasma (orifice) gas flow velocity 18L min-1；ICP radio frequency lines
Rate energy 1600W；Dummy run phase voltage -2850V；Stage pulse voltage 1000V；Pond enters voltage -6V；Pond goes out voltage -6V；Pond bar
Offset -14；Sampling centrum is nickel；Filtering centrum is nickel；Sampling system is that rotary spray chamber is Meinhard atomizers；spICP-MS
Method parameter is：Analytical element Ag；Density 10.5gcm-3；Quality 106.9amu；Holdup time 0.05ms；Stabilization time 0ms.
Above-mentioned Macerozyme R-10 enzymes extract AgNP and Ag+The step of be：Step 1) 0.1g plant leaf blades are weighed, cut
In 8mL after into fritter, 2mmolL-1, it is homogenized in the citrate buffer solutions of pH 6.0, afterwards by the AgNP standard liquids of 80nm
It is tagged in the plant substrates solution, AgNP ultimate densities are 28,000NPs mL-1；Step 2) added in homogenate
Macerozyme R-10, Macerozyme R-10 are 1 with the mass ratio of plant leaf blade:3,37 DEG C of heating water bath 36h are carried
Take, extraction terminates rear sample and stands, and takes supernatant and crosses the filter membrane that aperture is 1 μm；Step 3) using the detection of spICP-MS instruments, instrument
Device model：NexION 300,PerkinElmer,USA；Instrument measuring parameter is ICP-MS operating conditions：Flow rate of carrier gas 1L
min-1；Auxiliary gas flow speed 1.2Lmin-1；Plasma (orifice) gas flow velocity 18Lmin-1；ICP radio frequency energies 1600W；Mould
Intend stage voltage -2850V；Stage pulse voltage 1000V；Pond enters voltage -6V；Pond goes out voltage -6V；Pond bar offsets -14；Sampling vertebra
Body is nickel；Filtering centrum is nickel；Sampling system is that rotary spray chamber is Meinhard atomizers；SpICP-MS method parameters are：Point
Analysis elements A g；Density 10.5gcm-3；Quality 106.9amu；Holdup time 0.05ms；Stabilization time 0ms.
Above-mentioned AgNP particle diameters are 86nm.
Above-mentioned plant is paddy rice.
Know-why：TMAH is a kind of water-soluble highly basic compound, metal ion that can be in stablizing solution.It can be used for
The pre-treatment of trace element detection in biological sample.Different from strong acid decomposition, TMAH will not cause the mineralising of organic matter, but energy
It is enough that organically combine state metal is discharged into solution.Under this background, TMAH is to realize extracting nanometer from biological sample
Grain and corresponding metal ion, and not on nano particle produce influence provide may.
The main constituents of plant cell are some fibre element, pectin and polysaccharide etc..Macerozyme R-10 are many
The mixture of kind of enzyme, comprising cellulase, hemicellulase and pectase, with clearing up plant tissue cell and release wherein
Nano particle potentiality.Macerozyme R-10 during Digestion of Plant Samples, will not cause nano particle dissolving and
The change (see Fig. 1) of particle size.
The operation principle of spICP-MS is：As only Ag in sample+In the presence of, solution system is homogeneous, what detector was produced
Signal is a continual and steady signal (background signal).And when nano particle is contained in sample, single nano particle enters
Enter to plasma and be ionized, produce " ion cloud "；" ion cloud " enter level Four bar mass analyzer, detector detect from
Subsignal, produces a pulse signal；Nano particle is bigger, and the atomicity for containing is more, " ion cloud " intermediate ion of generation
Quantity is also more, and the corresponding pulse signal for producing is also stronger；According to signal intensity and the relation of quality, letter can will be obtained
Number be converted into granular mass, in the case of known to grain shape, formula (m=ρ V) according to quality and volume and density and
Volume and particle diameter relationSo as to obtain the particle diameter of particle.The background signal that detects is corresponding to be exactly solution
Ag+Concentration information.SpICP-MS fundamental diagrams refer to Fig. 2.
Beneficial effect：With existing AgNP and Ag+Analysis method compare, this method has advantages below：
1) AgNP particles, Ag in complicated plant substrates are realized+Quick measure.In separate unit spICP-MS, same sample
The detection of the granule density, particle diameter distribution and deliquescent metal ion of sample can simultaneously be completed；
2) test limit can as little as ng L-1, the promising approach as the detection of environmental sample nano particle；
3) it is simple to operate, quick, can batch quantity analysis sample；
4) sample determination loss amount is few.
Brief description of the drawings
Fig. 1 is influence figures of the TMAH and Macerozyme R-10 to AgNP grain diameters；
Fig. 2 is AgNP granule densities, particle diameter distribution and the dissolubility Ag that spICP-MS completes sample simultaneously+Detection figure；
Fig. 3 is droplet measurement limit figures of the spICP-MS to AgNP；
Fig. 4 is the primary signal figure that spICP-MS is detected to AgNP in plant substrates, and a and b, c, d are respectively plants in figure
Unmarked AgNP and mark 30nm, 50nm and 100nm AgNP (label concentration 50,000NPs mL in matrix-1) instrument signal
Figure.
Fig. 5 is the graph of a relation of the AgNP granule densities that instrument is detected and AgNP theoretical particles concentration.
Specific embodiment
The present invention extracts AgNP particles in plant substrates by contrasting two kinds of extractants of TMAH and Macerozyme R-10,
The plasma mass detection method of joint individual particle, it is found that the extraction effect of macerozyme R-10 is more reliable and stable.Contrast two
Plant extracting method to find, Macerozyme R-10 extracting methods are not notable to AgNP grain diameter influences, there is no dissolution and reunite now
As occurring, average grain diameter is at 85.5 ± 1.1nm (Fig. 1), and the average recovery rate of granule density reaches 97.9 ± 4.9% (tables 3).
And TMAH weak base extracting methods cause the dissolution and reunion of AgNP particles, average grain diameter at 38.2 ± 4.2nm (Fig. 1), and
The average recovery rate of granule density is only 60.7 ± 8.5% (tables 3), it can be seen that, the extraction effect of Macerozyme R-10 is more
It is stabilization, measured result is more accurate.
AgNP and Ag in plant substrates is proposed in the present invention+Effective extraction and on-line determination, including sample simultaneously in
The extraction of AgNP, AgNP particles and Ag+Measure.
Embodiment 1
TMAH extracts AgNP and Ag+The step of be：Step 1) 0.1g plant leaf blades are weighed, 50mL burnings are placed in after being cut into small pieces
In cup, it is marked with the AgNP standard liquids of 80nm so that the concentration of AgNP is 28,000NPs mL in final solution-1；Step
The TMAH of 10mL 1wt.% is added in the rapid plant leaf blade for 2) completing to mark, water bath sonicator 24h dispersion tissues simultaneously prevent AgNP
Reunite, the tissue suspension after ultrasound is moved into the filter membrane that aperture is 1 μm is crossed after centrifuge tube centrifugation；Step 3) utilize spICP-MS
Instrument detects (parameter setting is shown in Table 1, the results are shown in Table 3 and Fig. 1).
Embodiment 2
Macerozyme R-10 enzymes extract AgNP and Ag+The step of be：Step 1) 0.1g plant leaf blades are weighed, it is cut into small
In 8mL after block, 2mmolL-1, it is homogenized in the citrate buffer solutions of pH 6.0, the AgNP standard liquids of 80nm are marked afterwards
To in the plant substrates solution so that AgNP ultimate densities are 28,000NPs mL in final solution-1；Step 2) in homogenate
It is 1 with the mass ratio of plant leaf blade to add Macerozyme R-10, Macerozyme R-10:3,37 DEG C of heating water bath 36h enter
Row is extracted, and extraction terminates rear sample and stands, and takes supernatant and crosses the filter membrane that aperture is 1 μm；Step 3) using the inspection of spICP-MS instruments
Survey, (parameter setting is shown in Table 1, the results are shown in Table 3 and Fig. 1).
Embodiment 3
A, cut 0.1g plant leaf blades (growth period is surrounding) into pieces (0.5 × 0.5cm2), be transferred to 15mL plastics from
In heart pipe.In centrifuge tube add 8mL citrate buffer solutions (2mM, pH 6.0), then with tissue refiner (28,000rpm,
360W；DY89-1, NingBo XinZhi Biology Science Co., Ltd) homogenate.
B, by above-mentioned homogenate stand 1h after, take supernatant as plant background matrix.By 80nm AgNP particles and Ag+
Mark into plant substrates solution.
1. mark AgNP particle diameters are 80nm, and granule density is 80,000NPs mL-1。
2. Ag is marked+Concentration is 0.261 μ g L-1。
C, Macerozyme R-10 are with plant sample in (1) according to 1:3 (mass ratioes) mix, and plant is cleared up at 37 DEG C
Sample 36h.
D, will be cleared up in (2) sample stand 1h, take supernatant and cross the filter membrane that aperture is 1 μm, then the survey for carrying out spICP-MS
It is fixed that (parameter setting is shown in Table 1, the results are shown in Table 4).
Embodiment 4
Referring to embodiment 1, difference is to mark in step b AgNP conditions to be replaced by mark 30nm AgNP particles.Mark
AgNP particle diameters are 30nm, and granule density is respectively 35,000NPs mL-1(parameter setting is shown in Table 1, the results are shown in Table 4).
The Instrument measuring condition of table 1
Table 2 is the measure (by taking 80nm AgNP as an example) of spICP-MS granule density test limits
Influence of table 3 TMAH and Macerozyme R-10, the two kinds of extractants to concentrations of nanoparticles
Table 4 is AgNP and Ag in plant substrates solution+Detection
ND：Do not detect
As can be seen from Table 4, Macerozyme R-10 enzymes extraction-individual particle inductivity coupled plasma mass spectrometry combination skill
Art can quickly determine AgNP and Ag in plant sample+.AgNP grain diameter evaluated errors<2nm, AgNP the granule density rate of recovery
It is 99.6-103.7%, Ag+The rate of recovery is 98.2 ± 2.9%.
Claims (5)
1. a kind of method for determining Nano Silver and silver ion in plant, it is characterised in that step is：
(1) silver-colored Pollution Plant is obtained；
(2) carried out extracting AgNP and Ag with weak base TMAH or mixed enzyme Macerozyme R-10+；
(3) determined by individual particle-inductivity coupled plasma mass spectrometry (spICP-MS), realize the particle of AgNP particles in plant
Concentration, particle size distribution and Ag+Detection.
2. it is according to claim 1 determine plant in Nano Silver and silver ion method, it is characterised in that the TMAH
Extract AgNP and Ag+The step of be：Step 1) 0.1g plant leaf blades are weighed, it is placed in after being cut into small pieces in 50mL beakers, use 80nm
AgNP standard liquids be marked so that in final solution the concentration of AgNP be 28,000NPsmL-1；Step 2) to mark
The TMAH of 10mL 1wt.% is added in the plant leaf blade of completion, water bath sonicator 24h dispersion tissues simultaneously prevent AgNP from reuniting, will be super
Tissue suspension after sound crosses the filter membrane that aperture is 1 μm after being moved into centrifuge tube centrifugation；Step 3) using the detection of spICP-MS instruments,
Model：NexION 300, PerkinElmer, USA；Instrument measuring parameter is：ICP-MS operating conditions：Flow rate of carrier gas 1L
min-1；Auxiliary gas flow speed 1.2L min-1；Plasma (orifice) gas flow velocity 18L min-1；ICP radio frequency energies 1600W；Simulation
Stage voltage -2850V；Stage pulse voltage 1000V；Pond enters voltage -6V；Pond goes out voltage -6V；Pond bar offsets -14；Sampling centrum
It is nickel；Filtering centrum is nickel；Sampling system is that rotary spray chamber is Meinhard atomizers；SpICP-MS method parameters are：Analysis
Elements A g；Density 10.5gcm-3；Quality 106.9amu；Holdup time 0.05ms；Stabilization time 0ms.
3. it is according to claim 1 determine plant in Nano Silver and silver ion method, it is characterised in that it is described
Macerozyme R-10 enzymes extract AgNP and Ag+The step of be：Step 1) weigh 0.1g plant leaf blades, after being cut into small pieces
8mL, 2mmolL-1, it is homogenized in the citrate buffer solutions of pH 6.0, the AgNP standard liquids of 80nm are tagged to the plant afterwards
In thing matrix solution, AgNP ultimate densities are 28,000NPs mL-1；Step 2) to addition Macerozyme R-10 in homogenate,
Macerozyme R-10 are 1 with the mass ratio of plant leaf blade:3,37 DEG C of heating water bath 36h are extracted, and extraction terminates rear sample
Stand, take supernatant and cross the filter membrane that aperture is 1 μm；Step 3) using the detection of spICP-MS instruments, INSTRUMENT MODEL is：NexION
300,PerkinElmer,USA；Instrument measuring parameter is ICP-MS operating conditions：Flow rate of carrier gas 1Lmin-1；Auxiliary gas flow
Fast 1.2Lmin-1；Plasma (orifice) gas flow velocity 18Lmin-1；ICP radio frequency energies 1600W；Dummy run phase voltage-
2850V；Stage pulse voltage 1000V；Pond enters voltage -6V；Pond goes out voltage -6V；Pond bar offsets -14；Sampling centrum is nickel；Filtering
Centrum is nickel；Sampling system is that rotary spray chamber is Meinhard atomizers；SpICP-MS method parameters are：Analytical element Ag；It is close
Degree 10.5gcm-3；Quality 106.9amu；Holdup time 0.05ms；Stabilization time 0ms.
4. it is according to claim 1 determine plant in Nano Silver and silver ion method, it is characterised in that the AgNP
Particle diameter is 86nm.
5. according to any method for determining Nano Silver and silver ion in plant of Claims 1 to 4, it is characterised in that described
Plant is paddy rice.
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Cited By (8)
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| CN109060933A (en) * | 2018-09-14 | 2018-12-21 | 中南大学 | The detection method of silver nano-grain content in soil |
| CN110153440A (en) * | 2019-05-14 | 2019-08-23 | 桂林理工大学 | A kind of aspergillus japonicus fermentation liquid green prepares the method and application of nano silver |
| CN110608981A (en) * | 2019-09-09 | 2019-12-24 | 中国科学院高能物理研究所 | Method for measuring mercury sulfide nanoparticles in plants |
| CN111105979A (en) * | 2018-10-26 | 2020-05-05 | 安捷伦科技有限公司 | Automatic detection of nanoparticles using single particle inductively coupled plasma mass spectrometry (SP-ICP-MS) |
| CN111537595A (en) * | 2020-04-28 | 2020-08-14 | 浙江方圆检测集团股份有限公司 | Characterization method for nano silver migration in antibacterial product |
| CN112415079A (en) * | 2019-08-22 | 2021-02-26 | 四川大学 | Double-parameter self-verification homogeneous immunoassay method for single-particle inductively coupled plasma mass spectrometry |
| CN113777149A (en) * | 2021-09-24 | 2021-12-10 | 中国科学院南京土壤研究所 | Method for simultaneously detecting particle size distribution and concentration of mercury nanoparticles in environmental soil |
| WO2023095020A1 (en) * | 2021-11-26 | 2023-06-01 | Ecole Polytechnique Federale De Lausanne (Epfl) | Single-particle inductively-coupled plasma mass spectrometry particle sizing and counting method, system, computer program and computer-readable data carrier |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109060933A (en) * | 2018-09-14 | 2018-12-21 | 中南大学 | The detection method of silver nano-grain content in soil |
| CN111105979A (en) * | 2018-10-26 | 2020-05-05 | 安捷伦科技有限公司 | Automatic detection of nanoparticles using single particle inductively coupled plasma mass spectrometry (SP-ICP-MS) |
| CN110153440A (en) * | 2019-05-14 | 2019-08-23 | 桂林理工大学 | A kind of aspergillus japonicus fermentation liquid green prepares the method and application of nano silver |
| CN112415079A (en) * | 2019-08-22 | 2021-02-26 | 四川大学 | Double-parameter self-verification homogeneous immunoassay method for single-particle inductively coupled plasma mass spectrometry |
| CN112415079B (en) * | 2019-08-22 | 2021-10-22 | 四川大学 | Double-parameter self-verification homogeneous immunoassay method for single-particle inductively coupled plasma mass spectrometry |
| CN110608981A (en) * | 2019-09-09 | 2019-12-24 | 中国科学院高能物理研究所 | Method for measuring mercury sulfide nanoparticles in plants |
| CN111537595A (en) * | 2020-04-28 | 2020-08-14 | 浙江方圆检测集团股份有限公司 | Characterization method for nano silver migration in antibacterial product |
| CN113777149A (en) * | 2021-09-24 | 2021-12-10 | 中国科学院南京土壤研究所 | Method for simultaneously detecting particle size distribution and concentration of mercury nanoparticles in environmental soil |
| WO2023095020A1 (en) * | 2021-11-26 | 2023-06-01 | Ecole Polytechnique Federale De Lausanne (Epfl) | Single-particle inductively-coupled plasma mass spectrometry particle sizing and counting method, system, computer program and computer-readable data carrier |
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Application publication date: 20170620 |