CN106546672A - A kind of residue analysis method of polyoxin on ginseng - Google Patents
A kind of residue analysis method of polyoxin on ginseng Download PDFInfo
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- CN106546672A CN106546672A CN201610907396.7A CN201610907396A CN106546672A CN 106546672 A CN106546672 A CN 106546672A CN 201610907396 A CN201610907396 A CN 201610907396A CN 106546672 A CN106546672 A CN 106546672A
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- 229930182764 Polyoxin Natural products 0.000 title claims abstract description 49
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- 239000000523 sample Substances 0.000 claims description 19
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- 239000007788 liquid Substances 0.000 claims description 13
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 12
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 11
- 238000002552 multiple reaction monitoring Methods 0.000 claims description 11
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- 239000012488 sample solution Substances 0.000 claims description 4
- 239000007864 aqueous solution Substances 0.000 claims description 3
- 238000000605 extraction Methods 0.000 claims description 3
- 239000012071 phase Substances 0.000 claims description 3
- 239000005695 Ammonium acetate Substances 0.000 claims description 2
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 claims description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 2
- 235000019257 ammonium acetate Nutrition 0.000 claims description 2
- 229940043376 ammonium acetate Drugs 0.000 claims description 2
- 238000003556 assay Methods 0.000 claims description 2
- 238000004364 calculation method Methods 0.000 claims description 2
- 238000000132 electrospray ionisation Methods 0.000 claims description 2
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- 229930182536 Antimycin Natural products 0.000 description 1
- 241000221198 Basidiomycota Species 0.000 description 1
- 241000219112 Cucumis Species 0.000 description 1
- 235000015510 Cucumis melo subsp melo Nutrition 0.000 description 1
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- 208000031888 Mycoses Diseases 0.000 description 1
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- 241000233679 Peronosporaceae Species 0.000 description 1
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- 235000014787 Vitis vinifera Nutrition 0.000 description 1
- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
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- 229940088710 antibiotic agent Drugs 0.000 description 1
- CQIUKKVOEOPUDV-IYSWYEEDSA-N antimycin Chemical compound OC1=C(C(O)=O)C(=O)C(C)=C2[C@H](C)[C@@H](C)OC=C21 CQIUKKVOEOPUDV-IYSWYEEDSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
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- 241000411851 herbal medicine Species 0.000 description 1
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- General Health & Medical Sciences (AREA)
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Abstract
The invention provides a kind of residue analysis method of polyoxin on ginseng, establishes residue analysis method of kind of the polyoxin on ginseng, the polyoxin remained in sample is extracted with the homogeneous of aqueous acid containing first, after crossing film, liquid chromatography mass mass spectrum（HPLCMS‑MS）Detection, sets up the calibration curve of correction, quantified by external standard method using the vehicle solution without agricultural chemicals to be measured.The simple organic reagent used of pre-treatment is less, and the rate of recovery meets residues of pesticides criterion calls, and method meets the requirement of current environmental protection.And the degree of accuracy of detection method, precision meet the technical requirements of retention analysis, detection lower bound reaches domestic and international technical regulation and limit standard requirement.Average recovery rate of the present invention is 74.6%～94.7%, average relative standard's deviation（RSD）For 1.3%～7.8%, detection limit is less than 0.06mg/kg, with easy to operate, quick, sensitivity it is high, characteristic is strong, the accurate advantage of reproducible, qualitative, quantitative.
Description
Technical field
The invention provides a kind of residue analysis method of polyoxin on ginseng, is to existing residue analysis method
Improvement, belong to Chinese herbal medicine chemical analysis technology field.
Background technology
Polyoxin be by three doctor Lang of Japan Chemical research institute Suzuki et al. in 1963 from Kumamoto county Su Jun mills
Soil in a kind of actinomyces for isolating, be a kind of broad spectrum activity antimicrobial fungicides, with Uptake and translocation effect.Nineteen eighty-two
Rise to go into operation in China and apply, be mainly used in preventing and treating wheat powdery mildew, black fleck disease of ginseng, Alternaria alternate, cucumber downy mildew are melon
Various fungal diseases such as droop, rice sheath blight disease, alternaria leaf spot of apple, strawberry and grape grey mould, the withered sheath disease of forest.Its
Mechanism of action is unique, can interfere with the biosynthesis of cell membrane chitin, the ascus of cell membrane is constituted with chitin as matrix
Some fungies of Gammaproteobacteria, Basidiomycetes and deuteromycetes, germ tube, mycelium are understood local bulkiness, rupture, are overflowed carefully after contacting medicament
Intracellular inclusion, causes germ dead, so as to reach bactericidal effect.
Used as classical bactericide, polyoxin series bactericidal agent since the advent of the world is fast in countries in the world pesticide market portion
Speed expands, and extensively applies.As people are environmentally safe and the concern of food security, all kinds of agricultural chemicals are in surrounding medium and various works
Method for detecting residue in thing becomes pesticide field study hotspot.Therefore, how efficiently and rapidly to determine polyoxin class sterilized
Method for detecting residue in agent surrounding medium and various crops also becomes the hot issue of the medicament.However, due to medicament itself
Feature, although also having residue analysis method of the document report with regard to such agricultural chemicals in the surrounding mediums such as soil both at home and abroad, for example
Become pesticide science worker for many years using C18 reverse-phase chromatographic columns and Diode Array Detector product and the residue limits in medicinal material
The straight problem for making great efforts to solve.In recent years, my device, high performance liquid chromatography, Capillary Electrophoresis-electrogenerated chemiluminescence method (CE-ECL)
Deng, but these methods are applied to that the generally existing rate of recovery in ginseng matrix is low, and the phenomenon such as poor reproducibility does not possess reference, because
This, the polyoxin series bactericidal agent determination of residual amount in ginseng becomes the difficult point in current residual analysis.Polyoxin series bactericidal agent
As a kind of bactericide of high-efficiency broad spectrum, it is widely used in the agricultural production processes such as veterinary antibiotics, medicinal material, it is how efficiently quick
Which is determined in crop, fruit state in ginseng production process, ginseng agricultural university amount prevents and treats black fleck disease of ginseng using polyoxin series bactericidal agent
Etc. disease, in order to avoid polyoxin becomes the external technology barriers for limiting China's ginseng outlet, and solve China's ginseng outlet
Fall into a passive position in polyoxin residue problem situation, for polyoxin series bactericidal agent, successfully filter out one it is simple to operate,
The rate of recovery is high, the residue analysis method of favorable reproducibility is imperative.The foundation of this residue analysis method not only can make China
The residual detection technique level of ginseng agriculture, standard and external Complete Synchronization, while meeting with during ginseng exit procedure can also be avoided external
Green barrier.
The content of the invention：
The invention discloses a kind of residue analysis method of polyoxin on ginseng, the method is mainly used in determining people
The method of the polyoxin content remained in the complex matrices Chinese medicine such as ginseng, American Ginseng, method have stronger reappearance, reclaim
Rate can reach more than 80%.
A kind of residue analysis method of the polyoxin of the present invention on ginseng, comprises the following steps：
1st, in sample, polyoxin B is extracted with aqueous formic acid：
(1) fresh ginseng：Weigh fresh ginseng sample 5.0g to be put in beaker, add 0.3% aqueous formic acid 20mL, adjust pH
Scope 2-5, is homogenized about 2min, and reduce pressure in the Buchner funnel for be covered with filter aid suction filtration, and filtrate is all collected in tool plug graduated cylinder
In, portion filtrate is taken in centrifuge tube, 12000rpm centrifugation 5min cross 0.22 μm of filter membrane, treat that liquid chromatograph-mass spectrometer is examined
Survey；
(2) ginseng dry powder：Ginseng dry powder sample 5.0g is weighed in conical flask with stopper, 0.3% aqueous formic acid is added
20mL, regulation pH scopes 2-5, mechanical shaking extraction 1h, reduce pressure in the Buchner funnel for be covered with filter aid suction filtration, and filtrate is all collected
In tool plug graduated cylinder, portion filtrate is taken in centrifuge tube, 12000rpm centrifugation 5min cross 0.22 μm of filter membrane, treat liquid chromatogram-matter
Spectrum combined instrument detection；
2nd, analysis test：
(1) instrument condition：
Chromatographic column：Agilent Zorbax RRHD Eclipse Plus C18 3.0×100mm 1.8μm；
Mobile phase：Ammonium acetate 2.5mmol/L+0.05% aqueous formic acids: methanol elution gradient；
Gradient elution program：
Flow velocity：0.3mL/min；
Column temperature：30℃；
Sample size：10μl.
(2) mass spectral analysis condition：
Ionization source module：Electro-spray ionization；
Ionization source polarity：Positive ion mode (ESI+)；
Scan mode：Multiple-reaction monitoring (MRM)；
Atomization gas：Nitrogen；
Atomization gas pressure：45psi；
Capillary voltage：4000V；
It is dried temperature degree：400℃；
Dry gas stream speed：12L/min；
Polyoxin：Quota ion：508.1/490.1；
Qualitative ion：508.1/366；
Fragmentation voltage in source：120V；
Collision gas energy：508.1/490.1(10V)；
508.1/366(20V)。
3rd, calibration curve, quantified by external standard method polyoxin B are set up with matrix；
(1) the foundation of calibration curve：
Polyoxin standard items 0.0345g is accurately weighed, and 10ml is settled to 10% methanol aqueous solution, is configured to
The standard mother liquor of 1000mg/kg, then this standard mother liquor is diluted to into 0.05,0.1,0.2,0.5,1.0, the mark of 2.0mg/kg series
Quasi- solution, determines according to above-mentioned liquid-phase condition, and 5 μ l of sample size obtain polyoxin B MRM spectrograms 1, with sample introduction concentration as horizontal seat
Mark, peak area is ordinate, draws calibration curve, and linear regression equation is Y=12216x-149.29, and coefficient correlation is r=
0.9994, the results are shown in Table 1
The calibration curve of 1 polyoxin B of table
(2) minimum detection limit：
Under these conditions, the limit of identification of polyoxin B is 1.05 × 10-1ng。
(3) concentration limit
Tested according to TIANZHU XINGNAO Capsul, under above-mentioned instrument condition, polyoxin is dry in soil, plant, fresh ginseng and ginseng
Concentration limit in powder is 0.2mg/kg.
(4) relative retention time
Relative retention time of the polyoxin B on liquid chromatograph-mass spectrometer is 1.69min or so.
(5) TIANZHU XINGNAO Capsul and relative standard deviation：
With fresh ginseng and ginseng dry powder as matrix, addition polyoxin B be 0.2,1, tri- levels of 5mg/kg, be repeated 5 times,
According to step 1 and the step 2 assay method rate of recovery, 2-3 is the results are shown in Table, typical MRM spectrograms are shown in Fig. 2-4.Polyoxin is in fresh people
Ginseng and ginseng dry powder in average recovery rate difference 74.7～94.7% and 74.6～86.0%, the coefficient of variation be respectively 1.3～
4.2% and 4.6～7.8%.
4th, result is calculated
The residual quantity of test portion Pesticides is counted with mass fraction w, and numerical value is represented with milligrams per kilogram (mg/kg), by formula (1)
Calculate：
In formula：
The residual quantity mass fraction of w --- test portion Pesticides, unit are milligrams per kilogram (mg/kg)；
ρ s --- sample solution mass concentration, unit are milligrams per liter (mg/L)；
V0 --- the final constant volume of sample solution, unit are milliliter (ml)；
M --- sample mass, unit are gram (g)；
Result of calculation retains 2 significant digits significant digits.
The TIANZHU XINGNAO Capsul of polyoxin in 2 fresh ginseng of table
The TIANZHU XINGNAO Capsul of polyoxin in 3 ginseng dry powder of table
The positive effect of the present invention is：Residue analysis method of kind of the polyoxin on ginseng is established, with containing formic acid
Aqueous solution homogeneous extract the polyoxin that remains in sample, after crossing film, liquid chromatography mass-mass spectrum (HPLCMS-MS) detection,
The calibration curve of correction, quantified by external standard method are set up using the vehicle solution without agricultural chemicals to be measured.Have used by pre-treatment is simple
Machine reagent is less, and the rate of recovery meets residues of pesticides criterion calls, and method meets the requirement of current environmental protection.And detection method
The degree of accuracy, precision meet the technical requirements of retention analysis, detection lower bound reaches domestic and international technical regulation and limit standard will
Ask.Average recovery rate of the present invention is 74.6%～94.7%, and average relative standard's deviation (RSD) is 1.3%～7.8%, detection
Limit less than 0.06mg/kg, with easy to operate, quick, sensitivity it is high, characteristic is strong, reproducible, qualitative, quantitative is accurately excellent
Point.
Description of the drawings：
Fig. 1 is polyoxin B MRM spectrograms；
Fig. 2 is ginseng blank spectrogram；
Fig. 3 is that ginseng adds spectrogram；
Fig. 4 is ginseng sample spectra figure.
Fig. 5：Plant not acid adding pitch-based sphere 1mg/kg spectrograms；
Fig. 6：Plant acid adding pitch-based sphere 1mg/kg spectrograms；
Fig. 7：Soil not acid adding pitch-based sphere 1mg/kg spectrograms；
Fig. 8：Soil:Acid adding pitch-based sphere 1mg/kg spectrograms；
Fig. 9：Fresh ginseng not acid adding pitch-based sphere 5mg/kg spectrograms；
Figure 10：Fresh ginseng acid adding pitch-based sphere 5mg/kg spectrograms；
Figure 11：Dry ginseng's not acid adding pitch-based sphere 5mg/kg spectrograms；
Figure 12：Dry ginseng's acid adding pitch-based sphere 5mg/kg spectrograms.
Specific embodiment：
The two of three rooms in Tonghua, Jilin Province fair An County Yulin town Zhu Xian villages and Jilin Province Baishan Fusong County Songjianghe Area town
Year two places residue test result, 10% Wettable polyoxin powder, 100～150mg/kg of dosage (amount of formulation 666.7～
1000 times of liquid), in black fleck disease of ginseng initial stage (high more than the 15cm of ginseng plant) application method：Cauline leaf spray-on process.Harvest time ginseng
In plant, the residual quantity of polyoxin is 0.201～0.404mg/kg；In harvest time soil, the residual quantity of polyoxin is 0.234
～1.109mg/kg；In harvest time fresh ginseng, the residual quantity of polyoxin is respectively less than 0.2mg/kg；It is many in harvest time ginseng dry powder
The residual quantity of antimycin is respectively less than 0.2mg/kg.
Embodiment 1：The measure of polyoxin in ginseng plant
Weigh ginseng Plant samples 5.0g to be put in beaker, add 0.3% aqueous formic acid 20mL, adjust pH scopes 2-5,
Homogenate about 2min, reduce pressure in the Buchner funnel for be covered with filter aid suction filtration, filtrate is all collected in tool plug graduated cylinder, drawing section
Divide filtrate in centrifuge tube, 12000rpm centrifugation 5min cross 0.22 μm of filter membrane, treat that liquid chromatograph-mass spectrometer is detected, MRM
Spectrogram is shown in Fig. 5-Fig. 6, can be seen that the rate of recovery is significantly improved after acid adding from Fig. 5, Fig. 6.
The measurement result of polyoxin in 1 ginseng plant of embodiment
Embodiment 2：The measure of polyoxin in ginseng soil
Soil sample 5.0g is weighed in conical flask with stopper, 0.3% aqueous formic acid 20mL is added, pH scopes 2-5, vibration is adjusted
1h is extracted, reduce pressure in the Buchner funnel for be covered with filter aid suction filtration, filtrate is all collected in tool plug graduated cylinder, draws part and filter
In centrifuge tube, 12000rpm centrifugation 5min cross 0.22 μm of filter membrane, treat that liquid chromatograph-mass spectrometer is detected, MRM spectrograms liquid
See Fig. 7-Fig. 8, can be seen that the rate of recovery is significantly improved after acid adding from Fig. 7, Fig. 8.
The measurement result of polyoxin in 2 soil of embodiment
Embodiment 3：The measure of polyoxin in fresh ginseng
Weigh fresh ginseng sample 5.0g to be put in beaker, add 0.3% aqueous formic acid 20mL, adjust pH scopes 2-5, it is even
Slurry about 2min, reduce pressure in the Buchner funnel for be covered with filter aid suction filtration, filtrate is all collected in tool plug graduated cylinder, draws part
In centrifuge tube, 12000rpm centrifugation 5min cross 0.22 μm of filter membrane, treat that liquid chromatograph-mass spectrometer is detected filtrate, MRM spectrums
Figure is shown in Fig. 9-Figure 10, can be seen that the rate of recovery is significantly improved after acid adding from Fig. 9, Figure 10.
The measurement result of polyoxin in 2 soil of embodiment
Embodiment 4：The measure of polyoxin in ginseng dry powder
Ginseng dry powder sample 5.0g is weighed in conical flask with stopper, 0.3% aqueous formic acid 20mL is added, pH scopes are adjusted
2-5, mechanical shaking extraction 1h, reduce pressure in the Buchner funnel for be covered with filter aid suction filtration, filtrate is all collected in tool plug graduated cylinder, is inhaled
Portion filtrate is taken in centrifuge tube, 12000rpm centrifugation 5min cross 0.22 μm of filter membrane, treat that liquid chromatograph-mass spectrometer is detected,
MRM spectrograms are shown in figure, from Figure 11, Figure 12,11- Figure 12 can be seen that the rate of recovery is significantly improved after acid adding..
The measurement result of polyoxin in 4 ginseng dry powder of embodiment
Claims (1)
1. a kind of residue analysis method of polyoxin on ginseng, comprises the following steps：
1) in sample, polyoxin B is extracted with aqueous formic acid：
(1) fresh ginseng：
Fresh ginseng sample 5.0g is weighed, 0.3% aqueous formic acid 20mL is added, pH scopes 2-5 are adjusted, about 2min is homogenized, in paving
Reduce pressure in having the Buchner funnel of filter aid suction filtration, after filtrate is all collected, draws portion filtrate 12000rpm centrifugation 5min, mistake
0.22 μm of filter membrane, treats that liquid chromatograph-mass spectrometer is detected；
(2) ginseng dry powder：
Weigh ginseng dry powder sample 5.0g, add 0.3% aqueous formic acid 20mL, adjust pH scopes 2-5, mechanical shaking extraction 1h,
Reduce pressure in being covered with the Buchner funnel of filter aid suction filtration, after filtrate is all collected, draws portion filtrate 12000rpm centrifugation 5min,
0.22 μm of filter membrane is crossed, treats that liquid chromatograph-mass spectrometer is detected；
2) analysis test：
(1) instrument condition：
Chromatographic column：Agilent Zorbax RRHD Eclipse Plus C18 3.0×100mm 1.8μm；
Mobile phase：Ammonium acetate 2.5mmol/L+0.05% aqueous formic acids: methanol elution gradient；
Gradient elution program：
Flow velocity：0.3mL/min；Column temperature：30℃；Sample size：10μl；
(2) mass spectral analysis condition：
Ionization source module：Electro-spray ionization；
Ionization source polarity：Positive ion mode (ESI+)；
Scan mode：Multiple-reaction monitoring (MRM)；
Atomization gas：Nitrogen；
Atomization gas pressure：45psi；
Capillary voltage：4000V；
It is dried temperature degree：400℃；
Dry gas stream speed：12L/min；
Polyoxin：Quota ion：508.1/490.1；Qualitative ion：508.1/366；Fragmentation voltage in source：120V；Collision gas
Energy：508.1/490.1(10V)；508.1/366(20V)；
3) calibration curve, quantified by external standard method polyoxin B are set up with matrix；
(1) the foundation of calibration curve：
Polyoxin standard items 0.0345g is accurately weighed, and 10ml is settled to 10% methanol aqueous solution, is configured to 1000mg/kg
Standard mother liquor, then this standard mother liquor is diluted to into 0.05,0.1,0.2,0.5,1.0, the standard liquid of 2.0mg/kg series, press
Determine according to above-mentioned liquid-phase condition, 5 μ l of sample size obtain chromatogram, and with sample introduction concentration as abscissa, peak area is ordinate, is painted
Calibration curve processed, linear regression equation are Y=12216x-149.29, and coefficient correlation is r=0.9994.
(2) minimum detection limit：
Under these conditions, the limit of identification of polyoxin B is 1.05 × 10-1ng；
(3) concentration limit
Tested according to TIANZHU XINGNAO Capsul, under above-mentioned instrument condition, polyoxin is in soil, plant, fresh ginseng and ginseng dry powder
Concentration limit be 0.2mg/kg；
(4) relative retention time
Relative retention time of the polyoxin B on liquid chromatograph-mass spectrometer is 1.69min；
(5) TIANZHU XINGNAO Capsul and relative standard deviation：
With fresh ginseng and ginseng dry powder as matrix, addition polyoxin B be 0.2,1, tri- levels of 5mg/kg, be repeated 5 times, according to
Above-mentioned pre-treating method and the instrument condition assay method rate of recovery.Average recovery of the polyoxin in fresh ginseng and ginseng dry powder
Rate difference 74.7～94.7% and 74.6～86.0%, the coefficient of variation is respectively 1.3～4.2% and 4.6～7.8%；
4) result is calculated
The residual quantity of test portion Pesticides is counted with mass fraction w, and numerical value is represented with milligrams per kilogram (mg/kg), based on formula (1)
Calculate：
In formula：
The residual quantity mass fraction of w --- test portion Pesticides, unit are milligrams per kilogram (mg/kg)；
ρ s --- sample solution mass concentration, unit are milligrams per liter (mg/L)；
V0 --- the final constant volume of sample solution, unit are milliliter (ml)；
M --- sample mass, unit are gram (g)；
Result of calculation retains 2 significant digits significant digits.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107153101A (en) * | 2017-04-24 | 2017-09-12 | 上海市农业科学院 | A kind of detection method of plant-derived feed polyoxin |
| CN109001328A (en) * | 2018-08-17 | 2018-12-14 | 山东省农业科学院农业质量标准与检测技术研究所 | A kind of LC-MS/MS detection method of polyoxin B |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103776929A (en) * | 2014-01-23 | 2014-05-07 | 陕西绿盾生物制品有限责任公司 | Method for detecting polyoxins B by high performance liquid chromatography method |
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| CN103776929A (en) * | 2014-01-23 | 2014-05-07 | 陕西绿盾生物制品有限责任公司 | Method for detecting polyoxins B by high performance liquid chromatography method |
Non-Patent Citations (6)
| Title |
|---|
| JOHN P. DANIELS等: "Residual Efficacy of Fungicides for Controlling Brown Patch on Creeping Bentgrass Fairways", 《PLANT DISEASE》 * |
| KAKEN 制药株式会社: "多氧霉素B在烟草上的残留分析", 《中国烟草学报》 * |
| LI SUN等: "Microbiological assay for quantitative determination of polyoxin B", 《PROCESS BIOCHEMISTRY》 * |
| 于福利等: "多抗霉素B的高效液相色谱分析方法", 《农药》 * |
| 陶宁: "多抗霉素在烟田生态系统中的残留降解及生态效应", 《中国优秀硕士学位论文全文数据库》 * |
| 陶宁等: "稻田土壤及水中多抗霉素残留量的分析与检测方法研究", 《分析测试学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107153101A (en) * | 2017-04-24 | 2017-09-12 | 上海市农业科学院 | A kind of detection method of plant-derived feed polyoxin |
| CN109001328A (en) * | 2018-08-17 | 2018-12-14 | 山东省农业科学院农业质量标准与检测技术研究所 | A kind of LC-MS/MS detection method of polyoxin B |
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