CN108918503A - It is a kind of based on needle body SERS substrate to the detection method of pesticide residues in fruits - Google Patents
It is a kind of based on needle body SERS substrate to the detection method of pesticide residues in fruits Download PDFInfo
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- CN108918503A CN108918503A CN201810851306.6A CN201810851306A CN108918503A CN 108918503 A CN108918503 A CN 108918503A CN 201810851306 A CN201810851306 A CN 201810851306A CN 108918503 A CN108918503 A CN 108918503A
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- acupuncture
- needle
- gold nano
- detection method
- fruits
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/65—Raman scattering
- G01N21/658—Raman scattering enhancement Raman, e.g. surface plasmons
Abstract
The invention discloses a kind of based on needle body SERS substrate to the detection method of pesticide residues in fruits, includes the following steps:(1) acupuncture acupuncture needle is cleaned, is inserted into 3- aminopropyl-trimethoxy silane and impregnates, spontaneously dried after the completion of impregnating spare;(2) the stable gold nano grain of sodium citrate is prepared;(3) gold nano particle colloidal sols are prepared;(4) it after being impregnated in step (1) treated acupuncture acupuncture needle insertion gold nano particle colloidal sols, spontaneously dries spare;(5) by step (4) treated the acupuncture acupuncture needle 10-60min that is inserted into fruit to be measured that let the acupuncture needle remain at a certain point, SERS detection is carried out to acupuncture acupuncture needle using Raman spectrometer after extraction, the present invention has constructed a kind of highly sensitive plug-type SERS sampling substrate by uniformly modifying stable gold nano grain in metal surface, step is simple, it is convenient to operate, it is easy to carry, performance is stablized, with preferable reproducibility and stability, high sensitivity, detection precisely, can be widely applied to the detection of fruit internal pesticide residue.
Description
Technical field
The present invention relates to fruit the Detection Technologies of Pesticide Residues fields, more particularly to a kind of needle body SERS substrate that is based on is to fruit
The middle remaining detection method of pesticide.
Background technique
Surface enhanced Raman spectroscopy (Surface-Enhanced Raman Scattering, SERS) is that have high table
The vibrational spectrum technology of face detection sensitivity, can provide molecular characterization and material composition information.Since its superelevation enhances
The features such as effect can provide molecular fingerprint information, and testing conditions are mild, be not necessarily to sample pre-treatments, SERS technology is extensive
Ground is applied to environmental contaminants, biological medicine, the every field such as chemical reaction monitoring.However, current SERS detects only office
It is limited to the residual detection of agriculture of fruit surface, the pesticide residue of fruit internal still has very high harm to human body.Using common
SERS substrate can not carry out internal sampling and traditional technology can not achieve highly sensitive detection.Therefore, such as using SERS substrate
What realizes that the residual portable inspectiont of fruit internal agriculture is still a major challenge for researcher.
In view of this, the present invention is specifically proposed.
Summary of the invention
The object of the present invention is to provide a kind of based on needle body SERS substrate to the detection method of pesticide residues in fruits, is based on
The inspiration of traditional acupuncture acupuncture needle has constructed a kind of highly sensitive insertion by uniformly modifying stable gold nano grain in metal surface
The SERS of formula samples substrate, and step is simple, and it is convenient to operate, easy to carry, and performance is stablized, and has preferable reproducibility and stabilization
Property, high sensitivity, detection precisely, can be widely applied to the detection of fruit internal pesticide residue, have a extensive future, be conducive to push away
Wide application.
To achieve the goals above, a kind of inspection based on needle body SERS substrate to pesticide residues in fruits provided by the invention
Survey method, which is characterized in that include the following steps:
(1) acupuncture acupuncture needle is cleaned, is inserted into 3- aminopropyl-trimethoxy silane and impregnates, impregnated and complete
After spontaneously dry it is spare;
(2) the stable gold nano grain of sodium citrate is prepared;
(3) it after the stable gold nano grain eccentric cleaning of the sodium citrate prepared step (2), is concentrated as original content 10-
90% gold nano particle colloidal sols;
(4) it after being impregnated in step (1) treated acupuncture acupuncture needle insertion gold nano particle colloidal sols, spontaneously dries spare;
(5) step (4) treated acupuncture acupuncture needle is inserted into fruit to be measured, time of retaining needle 10-60min, after extraction
SERS detection is carried out to acupuncture acupuncture needle using Raman spectrometer.
Preferably, the cleaning process of acupuncture acupuncture needle is in the step (1):First through acetone soak and ultrasound 20-40min,
Simultaneously ultrasound 20-40min is impregnated with ethyl alcohol again.
Preferably, the preparation method of the stable gold nano grain of sodium citrate is in the step (2):
(1) HAuCl that volume ratio is 1: 99 is added into the three-necked flask with magneton4Solution and ultrapure water, the HAuCl4
The mass fraction of solution is 1%;
(2) three-necked flask is placed in oil bath or heating mantle and is heated, twice of HAuCl is added in when boiling4Liquor capacity
Sodium citrate solution, the mass fraction of the sodium citrate solution are 1%, keep boiling and uniform stirring 20-40min, then
Stop heating, it is spare.
Preferably, it is impregnated in acupuncture acupuncture needle insertion 3- aminopropyl-trimethoxy silane in the step (1), impregnates temperature
Degree is room temperature, soaking time 0.5-1h.
Preferably, it being impregnated in acupuncture acupuncture needle insertion gold nano particle colloidal sols in the step (4), soaking temperature is 3-5 DEG C,
Soaking time is 4-10h.
Preferably, the applicable fruit type to be measured of the detection method includes apple, peach, Lee, jujube, chestnut, pears, plum, apricot,
Hazel, persimmon, melon, hawthorn, mulberry fruit, premium, cherry, tangerine, shaddock, mandarin orange, orange, lichee, longan, loquat, red bayberry and olive.
It is provided by the invention it is a kind of based on needle body SERS substrate to the detection method of pesticide residues in fruits, have has as follows
Beneficial effect.
1. the present invention is based on the inspiration of traditional acupuncture acupuncture needle, by uniformly modifying stable gold nano grain in metal surface
A kind of highly sensitive plug-type SERS sampling substrate is constructed, step is simple, and it is convenient to operate, and easy to carry, performance is stablized, and has
Preferable reproducibility and stability, high sensitivity, detection are accurate.
2. the composite can be widely applied to the detection of fruit internal pesticide residue, be conducive to be applied to actual production
In practical, and commercialization transformation is carried out, had broad application prospects, is conducive to promote and apply.
3. acupuncture acupuncture needle of the present invention passes through the processing of Chemical Felter APTMS, so that gold nano grain tends to intensive list
Layer arrangement, ensures that the reproducibility of Detecting Pesticide.
4. between the gold nano grain on acupuncture acupuncture needle of the present invention surface, there are local surface coupled plasma and gold nanos
Existing surface plasma polarization, increases SERS signal jointly, further improves pesticide between particle and acupuncture acupuncture needle
The sensitivity of residue detection.
Detailed description of the invention
Fig. 1 is the scanning electron microscope (SEM) photograph that embodiment 1 modifies the functionalization acupuncture acupuncture needle SERS substrate obtained after gold nano grain;
Fig. 2 be embodiment 2 using functionalization acupuncture acupuncture needle SERS substrate to using same concentration different batches thiram molten
The peach that liquid impregnates carries out the result figure of SERS detection;
Fig. 3 be embodiment 3 using functionalization acupuncture acupuncture needle SERS substrate to use various concentration thiram solution impregnate
The result figure of peach progress SERS detection.
Specific embodiment
The present invention will be further described with attached drawing combined with specific embodiments below, to help the contents of the present invention are understood.
Embodiment 1
It is a kind of based on needle body SERS substrate to the detection method of pesticide residues in fruits:
(1) acupuncture acupuncture needle is cleaned, cleaning process is first then to be impregnated with ethyl alcohol through acetone soak and ultrasound 20min
And ultrasound 40min.It is inserted into 3- aminopropyl-trimethoxy silane (APTMS) and impregnates, soaking temperature is room temperature, is impregnated
Time is 0.5h, is spontaneously dried after the completion of impregnating spare;
(2) the stable gold nano grain of sodium citrate is prepared:To addition 1ml in 250ml three-necked flask (with magneton)
HAuCl4Solution (mass fraction 1%) and 99ml ultrapure water;Three-necked flask is placed in oil bath or heating mantle and is heated, is boiled
When be added 2ml sodium citrate solution (mass fraction 1%), keep boiling and uniform stirring 40min;Then stop heating,
It is spare;
(3) it by after the stable gold nano grain eccentric cleaning of sodium citrate, is concentrated as the gold nano grain of original content 10%
Colloidal sol;
(4) it being impregnated in the acupuncture acupuncture needle insertion gold nano particle colloidal sols by APTMS modified, soaking temperature is 5 DEG C,
Soaking time is 4h;
The scanning electron microscope (SEM) photograph of the functionalization acupuncture acupuncture needle SERS substrate obtained after modification gold nano grain is as shown in Figure 1, table
Bright gold nano grain has established structure by the stability that this method uniform can modify on acupuncture acupuncture needle surface, for postorder detection
Basis.
(5) the functionalization acupuncture acupuncture needle SERS substrate of preparation is inserted into from the peach that food market is bought, time of retaining needle is
60min carries out SERS detection to acupuncture acupuncture needle using Raman spectrometer after extraction.
Embodiment 2
It is a kind of based on needle body SERS substrate to the detection method of pesticide residues in fruits:
(1) acupuncture acupuncture needle is cleaned, cleaning process is first then to be impregnated with ethyl alcohol through acetone soak and ultrasound 40min
And ultrasound 20min.It is inserted into 3- aminopropyl-trimethoxy silane (APTMS) and impregnates, soaking temperature is room temperature, is impregnated
Time is 1h, is spontaneously dried after the completion of impregnating spare;
(2) the stable gold nano grain of sodium citrate is prepared:To addition 1ml in 250ml three-necked flask (with magneton)
HAuCl4Solution (mass fraction 1%) and 99ml ultrapure water;Three-necked flask is placed in oil bath or heating mantle and is heated, is boiled
When be added 2ml sodium citrate solution (mass fraction 1%), keep boiling and uniform stirring 20min;Then stop heating,
It is spare;
(3) it by after the stable gold nano grain eccentric cleaning of sodium citrate, is concentrated as the gold nano grain of original content 90%
Colloidal sol;
(4) it being impregnated in the acupuncture acupuncture needle insertion gold nano particle colloidal sols by APTMS modified, soaking temperature is 3 DEG C,
Soaking time is 10h;
(5) the functionalization acupuncture acupuncture needle SERS substrate of preparation is inserted into from the peach that food market is bought, time of retaining needle is
10min carries out SERS detection to acupuncture acupuncture needle using Raman spectrometer after extraction.
Repetitive test:
It is beautiful to same concentration different batches good fortune is used using the functionalization acupuncture acupuncture needle SERS substrate prepared by the above method
The peach that double solution impregnate carries out SERS detection, and totally 10 batches, the pre-treating method of peach are:Peach to be measured is impregnated respectively
In the thiram solution of same concentration different batches, take out, peach surface clear water is rinsed and wipe after 30min it is dry, to go
Except peach thiram solution remained on surface, to reduce test error.Testing result is as shown in Fig. 2, show that the substrate can be real
The detection of existing high duplication is 8.54% using the thiram RSD that the detection method can detecte.
Embodiment 3
It is a kind of based on needle body SERS substrate to the detection method of pesticide residues in fruits:
(1) acupuncture acupuncture needle is cleaned, cleaning process is first then to be impregnated with ethyl alcohol through acetone soak and ultrasound 30min
And ultrasound 30min.It is inserted into 3- aminopropyl-trimethoxy silane (APTMS) and impregnates, soaking temperature is room temperature, is impregnated
Time is 0.75h, is spontaneously dried after the completion of impregnating spare;
(2) the stable gold nano grain of sodium citrate is prepared:To addition 1ml in 250ml three-necked flask (with magneton)
HAuCl4Solution (mass fraction 1%) and 99ml ultrapure water;Three-necked flask is placed in oil bath or heating mantle and is heated, is boiled
When be added 2ml sodium citrate solution (mass fraction 1%), keep boiling and uniform stirring 30min;Then stop heating,
It is spare;
(3) it by after the stable gold nano grain eccentric cleaning of sodium citrate, is concentrated as the gold nano grain of original content 50%
Colloidal sol;
(4) it being impregnated in the acupuncture acupuncture needle insertion gold nano particle colloidal sols by APTMS modified, soaking temperature is 4 DEG C,
Soaking time is 7h;
(5) the functionalization acupuncture acupuncture needle SERS substrate of preparation is inserted into from the peach that food market is bought, time of retaining needle is
35min carries out SERS detection to acupuncture acupuncture needle using Raman spectrometer after extraction.
Sensitivity test:
Using the functionalization acupuncture acupuncture needle SERS substrate prepared by the above method to use various concentration thiram solution soak
The peach of bubble carries out SERS detection, and the pre-treating method of peach is:Peach to be measured is separately immersed in the thiram of various concentration
In solution, take out, peach surface clear water is rinsed and wipe after 30min it is dry, to remove peach thiram solution remained on surface,
Reduce test error.Testing result is as shown in figure 3, be 1nM, table using the thiram minimum concentration that the detection method can detecte
The highly sensitive detection of thiram may be implemented in the bright substrate.
The present invention is based on the inspirations of traditional acupuncture acupuncture needle, by uniformly modifying stable gold nano grain structure in metal surface
Built a kind of highly sensitive plug-type SERS sampling substrate, step is simple, and it is convenient to operate, and easy to carry, performance is stablized, have compared with
Good reproducibility and stability, high sensitivity, detection precisely, can be widely applied to the detection of fruit internal pesticide residue, such as apple
Fruit, peach, Lee, jujube, chestnut, pears, plum, apricot, hazel, persimmon, melon, hawthorn, mulberry fruit, premium, cherry, tangerine, shaddock, mandarin orange, orange, lichee, longan, Pi
Rake, red bayberry and olive.Be conducive to be applied to actual production and it is practical in, and carry out commercialization transformation, have wide
Application prospect, be conducive to promote and apply.Acupuncture acupuncture needle of the present invention passes through the processing of Chemical Felter APTMS, so that gold nano
Particle tends to intensive monolayer alignment, ensures that the reproducibility of Detecting Pesticide.The gold on acupuncture acupuncture needle of the present invention surface
Between nano particle there are surface existing between local surface coupled plasma and gold nano grain and acupuncture acupuncture needle etc. from
Daughter polarization, increases SERS signal jointly, further improves the sensitivity of Detecting Pesticide.
Specific case used herein elaborates inventive concept, the explanation of above example is only intended to
Help understands core of the invention thought.It should be pointed out that for those skilled in the art, not departing from this
Under the premise of inventive concept, any obvious modification, equivalent replacement or the other improvements made should be included in the present invention
Protection scope within.
Claims (6)
1. it is a kind of based on needle body SERS substrate to the detection method of pesticide residues in fruits, which is characterized in that include the following steps:
(1) acupuncture acupuncture needle is cleaned, is inserted into 3- aminopropyl-trimethoxy silane and impregnates, after the completion of impregnating certainly
Right drying for standby;
(2) the stable gold nano grain of sodium citrate is prepared;
(3) it after the stable gold nano grain eccentric cleaning of the sodium citrate prepared step (2), is concentrated as original content 10-90%
Gold nano particle colloidal sols;
(4) it after being impregnated in step (1) treated acupuncture acupuncture needle insertion gold nano particle colloidal sols, spontaneously dries spare;
(5) step (4) treated acupuncture acupuncture needle is inserted into fruit to be measured, time of retaining needle 10-60min is utilized after extraction
Raman spectrometer carries out SERS detection to acupuncture acupuncture needle.
2. it is according to claim 1 it is a kind of based on needle body SERS substrate to the detection method of pesticide residues in fruits, it is special
Sign is that the cleaning process of acupuncture acupuncture needle is in the step (1):First through acetone soak and ultrasound 20-40min, then use ethyl alcohol
Impregnate simultaneously ultrasound 20-40min.
3. it is according to claim 1 or 2 it is a kind of based on needle body SERS substrate to the detection method of pesticide residues in fruits,
It is characterized in that, the preparation method of the stable gold nano grain of sodium citrate is in the step (2):
(1) HAuCl that volume ratio is 1: 99 is added into the three-necked flask with magneton4Solution and ultrapure water, the HAuCl4Solution
Mass fraction be 1%;
(2) three-necked flask is placed in oil bath or heating mantle and is heated, twice of HAuCl is added in when boiling4The citric acid of liquor capacity
Sodium solution, the mass fraction of the sodium citrate solution are 1%, keep boiling and uniform stirring 20-40min, then stop adding
Heat, it is spare.
4. it is according to claim 3 it is a kind of based on needle body SERS substrate to the detection method of pesticide residues in fruits, it is special
Sign is, impregnates in acupuncture acupuncture needle insertion 3- aminopropyl-trimethoxy silane in the step (1), and soaking temperature is room temperature,
Soaking time is 0.5-1h.
5. it is according to claim 4 it is a kind of based on needle body SERS substrate to the detection method of pesticide residues in fruits, it is special
Sign is, impregnates in acupuncture acupuncture needle insertion gold nano particle colloidal sols in the step (4), soaking temperature is 3-5 DEG C, soaking time
For 4-10h.
6. it is according to claim 5 it is a kind of based on needle body SERS substrate to the detection method of pesticide residues in fruits, it is special
Sign is, the applicable fruit type to be measured of the detection method includes apple, peach, Lee, jujube, chestnut, pears, plum, apricot, hazel, persimmon, melon,
Hawthorn, mulberry fruit, premium, cherry, tangerine, shaddock, mandarin orange, orange, lichee, longan, loquat, red bayberry and olive.
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Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1356543A (en) * | 2001-12-10 | 2002-07-03 | 中国科学院长春应用化学研究所 | Process for preparing film electrode of gold-monocrystal nano island array |
CN103105386A (en) * | 2013-01-29 | 2013-05-15 | 福州大学 | Method for detecting malachite green in water body and aquatic products |
CN106990092A (en) * | 2017-04-17 | 2017-07-28 | 中国科学院合肥物质科学研究院 | A kind of preparation method of the highly sensitive portable SERS substrates based on acupuncture acupuncture needle |
CN107044974A (en) * | 2017-03-28 | 2017-08-15 | 北京工业大学 | 2,4 dichlorphenoxyacetic acid method for detecting residue in a kind of fruits and vegetables |
CN107328750A (en) * | 2017-06-19 | 2017-11-07 | 吉林大学 | A kind of high activity, surface enhanced Raman scattering substrate of high homogeneity and preparation method thereof |
CN107727637A (en) * | 2017-10-17 | 2018-02-23 | 云南技师学院 | A kind of Practice for Pesticide Residue in Agricultural Products detection method |
-
2018
- 2018-07-27 CN CN201810851306.6A patent/CN108918503A/en active Pending
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1356543A (en) * | 2001-12-10 | 2002-07-03 | 中国科学院长春应用化学研究所 | Process for preparing film electrode of gold-monocrystal nano island array |
CN103105386A (en) * | 2013-01-29 | 2013-05-15 | 福州大学 | Method for detecting malachite green in water body and aquatic products |
CN107044974A (en) * | 2017-03-28 | 2017-08-15 | 北京工业大学 | 2,4 dichlorphenoxyacetic acid method for detecting residue in a kind of fruits and vegetables |
CN106990092A (en) * | 2017-04-17 | 2017-07-28 | 中国科学院合肥物质科学研究院 | A kind of preparation method of the highly sensitive portable SERS substrates based on acupuncture acupuncture needle |
CN107328750A (en) * | 2017-06-19 | 2017-11-07 | 吉林大学 | A kind of high activity, surface enhanced Raman scattering substrate of high homogeneity and preparation method thereof |
CN107727637A (en) * | 2017-10-17 | 2018-02-23 | 云南技师学院 | A kind of Practice for Pesticide Residue in Agricultural Products detection method |
Non-Patent Citations (1)
Title |
---|
周宥辰等: "湿化学镀SPR 金基底及其性能表征", 《电化学》 * |
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