CN114113373B - System and method for measuring glyphosate, glufosinate and nitrite in urine - Google Patents
System and method for measuring glyphosate, glufosinate and nitrite in urine Download PDFInfo
- Publication number
- CN114113373B CN114113373B CN202111320527.9A CN202111320527A CN114113373B CN 114113373 B CN114113373 B CN 114113373B CN 202111320527 A CN202111320527 A CN 202111320527A CN 114113373 B CN114113373 B CN 114113373B
- Authority
- CN
- China
- Prior art keywords
- column
- detection unit
- urine
- glufosinate
- nitrite
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 210000002700 urine Anatomy 0.000 title claims abstract description 50
- IAJOBQBIJHVGMQ-UHFFFAOYSA-N 2-amino-4-[hydroxy(methyl)phosphoryl]butanoic acid Chemical compound CP(O)(=O)CCC(N)C(O)=O IAJOBQBIJHVGMQ-UHFFFAOYSA-N 0.000 title claims abstract description 40
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 title claims abstract description 40
- 239000005561 Glufosinate Substances 0.000 title claims abstract description 39
- 239000005562 Glyphosate Substances 0.000 title claims abstract description 39
- XDDAORKBJWWYJS-UHFFFAOYSA-N glyphosate Chemical compound OC(=O)CNCP(O)(O)=O XDDAORKBJWWYJS-UHFFFAOYSA-N 0.000 title claims abstract description 39
- 229940097068 glyphosate Drugs 0.000 title claims abstract description 39
- 238000000034 method Methods 0.000 title claims abstract description 22
- 238000001514 detection method Methods 0.000 claims abstract description 94
- 150000002500 ions Chemical class 0.000 claims abstract description 37
- 239000007788 liquid Substances 0.000 claims description 28
- 238000004458 analytical method Methods 0.000 claims description 18
- 239000003480 eluent Substances 0.000 claims description 12
- 238000010828 elution Methods 0.000 claims description 8
- 238000000926 separation method Methods 0.000 claims description 8
- 238000011084 recovery Methods 0.000 claims description 5
- 238000010829 isocratic elution Methods 0.000 claims description 3
- 239000003112 inhibitor Substances 0.000 claims 4
- 230000001681 protective effect Effects 0.000 claims 1
- 238000005342 ion exchange Methods 0.000 abstract description 7
- 239000012535 impurity Substances 0.000 abstract description 5
- 238000001212 derivatisation Methods 0.000 abstract description 3
- 230000006920 protein precipitation Effects 0.000 abstract description 2
- 230000035945 sensitivity Effects 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 9
- 239000013076 target substance Substances 0.000 description 8
- 238000004255 ion exchange chromatography Methods 0.000 description 5
- 238000010586 diagram Methods 0.000 description 4
- 239000002574 poison Substances 0.000 description 3
- 231100000614 poison Toxicity 0.000 description 3
- 238000011002 quantification Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- XYFCBTPGUUZFHI-UHFFFAOYSA-N Phosphine Chemical compound P XYFCBTPGUUZFHI-UHFFFAOYSA-N 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000000108 ultra-filtration Methods 0.000 description 2
- 206010003497 Asphyxia Diseases 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- QLULGSLAHXLKSR-UHFFFAOYSA-N azane;phosphane Chemical compound N.P QLULGSLAHXLKSR-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 229910052793 cadmium Inorganic materials 0.000 description 1
- BDOSMKKIYDKNTQ-UHFFFAOYSA-N cadmium atom Chemical compound [Cd] BDOSMKKIYDKNTQ-UHFFFAOYSA-N 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 239000000645 desinfectant Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 239000004009 herbicide Substances 0.000 description 1
- 150000008040 ionic compounds Chemical class 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 229910000073 phosphorus hydride Inorganic materials 0.000 description 1
- 238000007781 pre-processing Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
Classifications
-
- 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
- G01N30/08—Preparation using an enricher
-
- 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
- G01N30/24—Automatic injection systems
-
- 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
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Library & Information Science (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Description
技术领域Technical field
本发明属于尿液毒物检测，具体涉及一种离子交换色谱柱切换法同时测定尿液中的草甘膦、草铵膦和亚硝酸盐的系统和方法。The invention belongs to urine poison detection, and specifically relates to a system and method for simultaneously determining glyphosate, glufosinate and nitrite in urine using an ion exchange chromatography column switching method.
背景技术Background technique
草甘膦（Glyphosate）、草铵膦（Glufosinate）、亚硝酸盐常被用于农药、消毒剂等各个方面，除了对环境产生一定的影响外，还会对人体产生一些影响。亚硝酸盐会使人体组织缺氧，严重时会使人窒息而亡。草甘膦和草铵膦为有机磷类除草剂，具有低毒、高效、范围广的特点，但是进入人体后会使人产生消化道、呼吸道、心尿管系统及神经系统等多脏器损伤的现象，严重时会使人死亡。Glyphosate, Glufosinate, and nitrite are often used in various aspects such as pesticides and disinfectants. In addition to having certain effects on the environment, they can also have some effects on the human body. Nitrite can deprive human tissues of oxygen, and in severe cases can cause suffocation and death. Glyphosate and glufosinate are organophosphorus herbicides with low toxicity, high efficiency and wide range. However, after entering the human body, they will cause damage to multiple organs such as the digestive tract, respiratory tract, cardio-urinary system and nervous system. phenomenon, which can lead to death in severe cases.
目前常用来检测这些毒物的方法有：分光光度法、液质-气质联用法（LC-GC）、镉柱法等。而由于草甘膦、草铵膦极性较强、易溶于水，缺少发光基团和荧光基团，所以用液质测定大多需要复杂的衍生化手段。Currently, the methods commonly used to detect these poisons include: spectrophotometry, liquid mass-mass spectrometry (LC-GC), cadmium column method, etc. Since glyphosate and glufosinate are highly polar, easily soluble in water, and lack luminescent and fluorescent groups, most liquid-mass determinations require complex derivatization methods.
草甘膦、草铵膦和亚硝酸盐在水里均为离子化合物，都能用离子色谱进行分离检测。而在唾液、尿液、血液等生物检材当中，尿液是最具有价值的。但是目前的LC、LC-MS、GC、GC-MS对尿液进行检测都需要进行前处理，需要将尿液中的蛋白质等大分子进行沉淀，将目标物分离进行检测。Glyphosate, glufosinate and nitrite are all ionic compounds in water and can be separated and detected by ion chromatography. Among biological test materials such as saliva, urine, and blood, urine is the most valuable. However, current LC, LC-MS, GC, and GC-MS detection of urine requires pre-processing. It is necessary to precipitate macromolecules such as proteins in urine and separate the target substances for detection.
发明内容Contents of the invention
发明目的：本发明的目的在于提供一种离子交换色谱柱切换联用法同时直接测定尿液中不同浓度的草甘膦、草铵膦和亚硝酸盐的系统；本发明的第二目的在于提供一种上述系统的检测方法。Purpose of the invention: The purpose of the present invention is to provide a system for directly measuring different concentrations of glyphosate, glufosinate and nitrite in urine at the same time using an ion exchange chromatography column switching combined method; the second purpose of the present invention is to provide a A detection method for the above system.
技术方案：本发明的一种测定尿液中草甘膦、草铵膦和亚硝酸盐的系统，包括用于高浓度检测的一维检测单元和用于低浓度检测的二维检测单元，所述一维检测单元和二维检测单元之间设有切换待测液体的第三六通阀；所述一维检测单元包括第一六通阀，所述第一六通阀上设有用于收集待测液体的定量环，所述第一六通阀的一端依次连接有第一保护柱、第一分析柱、第一抑制器和第一检测器；所述第一检测器与第三六通阀的一端连接；所述二维检测单元包括第二六通阀，所述第二六通阀上设有用于富集待测液体的富集柱，所述第二六通阀的一端与第三六通阀连接，所述第二六通阀的另一端依次连接有第二保护柱、第二分析柱、第二抑制器和第二检测器。Technical solution: A system for measuring glyphosate, glufosinate and nitrite in urine according to the present invention includes a one-dimensional detection unit for high-concentration detection and a two-dimensional detection unit for low-concentration detection. A third six-way valve for switching the liquid to be measured is provided between the one-dimensional detection unit and the two-dimensional detection unit; the one-dimensional detection unit includes a first six-way valve, and the first six-way valve is provided with a For the quantitative loop of the liquid to be measured, one end of the first six-way valve is connected in sequence with a first guard column, a first analysis column, a first suppressor and a first detector; the first detector and the third six-way valve One end of the valve is connected; the two-dimensional detection unit includes a second six-way valve, the second six-way valve is provided with an enrichment column for enriching the liquid to be tested, and one end of the second six-way valve is connected to the third Three six-way valves are connected, and the other end of the second six-way valve is connected in sequence with a second guard column, a second analysis column, a second suppressor and a second detector.
进一步的，所述第一保护柱和第一分析柱采用Ion Pac AG19色谱柱；所述第二保护柱和第二分析柱采用Ion Pac AG11色谱柱。Further, the first guard column and the first analysis column adopt Ion Pac AG19 chromatographic column; the second guard column and the second analysis column adopt Ion Pac AG11 chromatographic column.
进一步的，所述富集柱采用UATC-LP12富集柱。Further, the enrichment column uses a UATC-LP12 enrichment column.
进一步的，所述一维检测单元和二维检测单元还连接有用于通入淋洗液的第一泵和第二泵。Furthermore, the one-dimensional detection unit and the two-dimensional detection unit are also connected to a first pump and a second pump for passing eluent.
本发明还保护所述的测定尿液中草甘膦、草铵膦和亚硝酸盐的系统的检测方法，包括以下步骤：The invention also protects the detection method of the system for measuring glyphosate, glufosinate and nitrite in urine, which includes the following steps:
步骤一、在空白尿液中添加0.1~100 mg/L的目标离子分别对一维检测单元和二维检测单元进行检出限、定量限、线性和回收率的评价，确定一维检测单元和二维检测单元的检测浓度分界点；Step 1. Add 0.1~100 mg/L target ions to blank urine to evaluate the detection limit, quantification limit, linearity and recovery rate of the one-dimensional detection unit and the two-dimensional detection unit respectively, and determine the one-dimensional detection unit and The detection concentration cut-off point of the two-dimensional detection unit;
步骤二、通过自动进样器将待测液体装载在定量环中；Step 2: Load the liquid to be measured into the quantitative loop through the automatic sampler;
步骤三、当待测液体中目标离子的浓度高于浓度分界点时，在洗脱液流动相作用下待测液体进入第一保护柱和第一分析柱进行洗脱分离；分离后的液体依次进入第一抑制器和第一检测器进行检测；Step 3: When the concentration of the target ion in the liquid to be measured is higher than the concentration cut-off point, the liquid to be measured enters the first guard column and the first analysis column under the action of the eluent mobile phase for elution and separation; the separated liquids are sequentially Enter the first suppressor and first detector for detection;
步骤四、当待测液体中目标离子的浓度低于浓度分界点时，切换第三六通阀将定量环中的待测液流入到富集柱中进行目标离子的富集；Step 4: When the concentration of the target ions in the liquid to be measured is lower than the concentration cut-off point, switch the third six-way valve to flow the liquid to be measured in the quantitative loop into the enrichment column to enrich the target ions;
步骤五、富集柱中待测液体进入第二保护柱和第二分析柱进行洗脱分离，分离后的液体依次进入第二抑制器和第二检测器进行检测；Step 5: The liquid to be measured in the enrichment column enters the second guard column and the second analytical column for elution and separation, and the separated liquid enters the second suppressor and the second detector in sequence for detection;
其中，目标离子为草甘膦、草铵膦或亚硝酸盐。Among them, the target ions are glyphosate, glufosinate or nitrite.
进一步的，所述步骤一中，检测浓度分界点为10mg/L。Further, in step one, the detection concentration cut-off point is 10 mg/L.
进一步的，所述步骤一中，一维检测单元中草甘膦、草铵膦、亚硝酸盐的检出限分别为：0.396 mg/L、5.88 mg/L、0.97 mg/L；定量限分别为1.32 mg/L、19.6 mg/L、3.25 mg/L；二维检测单元中草甘膦、草铵膦、亚硝酸盐的检出限分别为：0.05 mg/L、0.02 mg/L、0.03mg/L；定量限分别为：0.19 mg/L、0.06 mg/L、0.11mg/L。Further, in step one, the detection limits of glyphosate, glufosinate and nitrite in the one-dimensional detection unit are: 0.396 mg/L, 5.88 mg/L, and 0.97 mg/L respectively; the quantitation limits are respectively are 1.32 mg/L, 19.6 mg/L, and 3.25 mg/L; the detection limits of glyphosate, glufosinate, and nitrite in the two-dimensional detection unit are: 0.05 mg/L, 0.02 mg/L, and 0.03 respectively. mg/L; the limits of quantification are: 0.19 mg/L, 0.06 mg/L, and 0.11 mg/L.
进一步的，所述步骤三中，采用10~40 mmol/L的KOH溶液进行梯度洗脱，KOH溶液的流速为1.0mL/min，柱温为30℃。Further, in step three, a KOH solution of 10 to 40 mmol/L is used for gradient elution, the flow rate of the KOH solution is 1.0 mL/min, and the column temperature is 30°C.
进一步的，所述步骤四中，第三六通阀切换草铵膦、亚硝酸盐、草甘膦的时间分别为9.7~11.4 min、15~15.9 min、26.4~29 min。Further, in step four, the time for the third six-way valve to switch between glufosinate, nitrite, and glyphosate is 9.7~11.4 min, 15~15.9 min, and 26.4~29 min respectively.
进一步的，所述步骤五中，采用23 mmol/L的KOH溶液进行等度洗脱，KOH溶液的流速为1.0mL/min，柱温为30℃。Further, in step five, a 23 mmol/L KOH solution is used for isocratic elution, the flow rate of the KOH solution is 1.0 mL/min, and the column temperature is 30°C.
本发明的测试原理为，利用一维检测单元将尿液中的目标离子和其他杂质离子进行分离，检测尿液中较高浓度的草甘膦、草铵膦和亚硝酸盐；其次当尿液中目标物含量过低时可以通过富集柱进行目标离子的富集，再通过二维检测单元同时检测尿液中低含量的草甘膦、草铵膦、亚硝酸盐；通过离子交换柱切换联用法实现了尿液中不同浓度的草甘膦、草铵膦、亚硝酸盐的同时、快速、准确检测。The testing principle of the present invention is to use a one-dimensional detection unit to separate target ions and other impurity ions in urine, and detect higher concentrations of glyphosate, glufosinate and nitrite in urine; secondly, when urine When the content of the target substance is too low, the target ions can be enriched through the enrichment column, and then the low levels of glyphosate, glufosinate, and nitrite in the urine can be simultaneously detected through the two-dimensional detection unit; switching through the ion exchange column The combined method achieves simultaneous, rapid and accurate detection of different concentrations of glyphosate, glufosinate and nitrite in urine.
其中，一维检测单元的色谱柱采用Ion Pac AG19，二维检测单元的色谱柱采用IonPac AG11，这是因为草甘膦、草铵膦、亚硝酸盐和尿液中的其他杂质能够在Ion Pac AG19、Ion Pac AG11上完全分离，并且分离度和峰形较好；三种目标物与尿液中的氯离子、硫酸根离子、尿素等杂质首先在Ion Pac AG19分离柱上进行预分离，检测较高浓度的目标物；随后，三种目标物进入二维检测单元，经过富集柱富集后在Ion Pac AG11分离柱上进行再一次分离，检测较低浓度的目标物。Among them, the chromatographic column of the one-dimensional detection unit uses Ion Pac AG19, and the chromatographic column of the two-dimensional detection unit uses IonPac AG11. This is because glyphosate, glufosinate, nitrite and other impurities in urine can be detected in Ion Pac AG19 and Ion Pac AG11 are completely separated with good resolution and peak shape; the three target substances and chloride ions, sulfate ions, urea and other impurities in urine are first pre-separated on the Ion Pac AG19 separation column and detected. Higher concentration target substances; then, the three target substances enter the two-dimensional detection unit, and are separated again on the Ion Pac AG11 separation column after being enriched by the enrichment column to detect lower concentration target substances.
有益效果：与现有技术相比，本发明的具有如下显著优点：本发明与现有的气质联用、液质联用方法相比，该离子色谱法无需尿液的蛋白沉淀衍生化等复杂的前处理，可以直接进样，并且能够同时测定草甘膦、草铵膦和亚硝酸盐等三种强极性毒物，方法具有快速、操作简单、灵敏度高的优点；同时，本发明还可以高效测定不同浓度的草甘膦、草铵膦和亚硝酸盐，目标离子的检测浓度区间在0.1~100mg/L。Beneficial effects: Compared with the existing technology, the present invention has the following significant advantages: compared with the existing GC-MS and LC-MS methods, the ion chromatography method does not require complexities such as urine protein precipitation and derivatization. The pretreatment can directly inject samples, and can simultaneously measure three highly polar poisons such as glyphosate, glufosinate and nitrite. The method has the advantages of rapidity, simple operation and high sensitivity; at the same time, the present invention can also Efficiently determine different concentrations of glyphosate, glufosinate and nitrite, and the detection concentration range of target ions is 0.1~100mg/L.
附图说明Description of drawings
图1为本发明系统进样过程示意图；Figure 1 is a schematic diagram of the sampling process of the system of the present invention;
图2为本发明系统一维离子交换的示意图；Figure 2 is a schematic diagram of one-dimensional ion exchange in the system of the present invention;
图3为本发明系统二维离子富集的示意图；Figure 3 is a schematic diagram of two-dimensional ion enrichment of the system of the present invention;
图4为本发明系统二维离子交换的示意图；Figure 4 is a schematic diagram of two-dimensional ion exchange in the system of the present invention;
图5为尿液的第一维离子交换色谱图；Figure 5 shows the first-dimensional ion exchange chromatogram of urine;
图6为尿液的第二维离子交换色谱图。Figure 6 shows the second-dimensional ion exchange chromatogram of urine.
具体实施方式Detailed ways
下面结合实施例对本发明的技术方案做进一步详细说明。The technical solution of the present invention will be further described in detail below with reference to the examples.
参见图1-4的一种测定尿液中草甘膦、草铵膦和亚硝酸盐的系统，包括用于高浓度检测的一维检测单元和用于低浓度检测的二维检测单元；一维检测单元和二维检测单元之间设有切换待测液体的第三六通阀4；其中，一维检测单元包括第一六通阀3、定量环6、第一保护柱7、第一分析柱8、第一抑制器9和第一检测器10；定量环6设置在第一六通阀3上，一维检测单元的一侧设有第一泵1，第一泵1外接洗脱液；第一六通阀3依次与第一保护柱7、第一分析柱8、第一抑制器9和第一检测器10连接；自动进样器16与第一六通阀3连接；二维检测单元包括第二六通阀5、富集柱11、第二保护柱12、第二分析柱13、第二抑制器14和第二检测器15；富集柱11设置在第二六通阀5上，第二六通阀5的一侧连接第二泵2，第二泵2外接洗脱液，第二六通阀5的一端与第三六通阀4连接，第二六通阀5的另一端依次连接有第二保护柱12、第二分析柱13、第二抑制器14和第二检测器15。Referring to Figures 1-4, a system for measuring glyphosate, glufosinate and nitrite in urine includes a one-dimensional detection unit for high-concentration detection and a two-dimensional detection unit for low-concentration detection; 1. A third six-way valve 4 for switching the liquid to be measured is provided between the one-dimensional detection unit and the two-dimensional detection unit; wherein, the one-dimensional detection unit includes a first six-way valve 3, a quantitative loop 6, a first guard column 7, a first Analytical column 8, first suppressor 9 and first detector 10; quantitative loop 6 is set on the first six-way valve 3, and a first pump 1 is provided on one side of the one-dimensional detection unit, and the first pump 1 is connected to an external eluent liquid; the first six-way valve 3 is connected to the first guard column 7, the first analysis column 8, the first suppressor 9 and the first detector 10 in sequence; the automatic sampler 16 is connected to the first six-way valve 3; two The dimensional detection unit includes a second six-way valve 5, a enrichment column 11, a second guard column 12, a second analysis column 13, a second suppressor 14 and a second detector 15; the enrichment column 11 is arranged in the second six-way On the valve 5, one side of the second six-way valve 5 is connected to the second pump 2, the second pump 2 is externally connected to the eluent, one end of the second six-way valve 5 is connected to the third six-way valve 4, and the second six-way valve 5 is connected to the third six-way valve 4. The other end of 5 is connected in sequence with a second guard column 12, a second analysis column 13, a second suppressor 14 and a second detector 15.
第一保护柱7和第一分析柱8采用Ion Pac AG19色谱柱；所述第二保护柱12和第二分析柱13采用Ion Pac AG11色谱柱；富集柱11采用UATC-LP12富集柱；第一抑制器9为AERS500；第二抑制器14为AERS 500 mm；第一检测器10和第二检测器15为抑制性电导检测器。The first guard column 7 and the first analysis column 8 adopt Ion Pac AG19 chromatographic column; the second guard column 12 and the second analysis column 13 adopt Ion Pac AG11 chromatographic column; the enrichment column 11 adopts UATC-LP12 enrichment column; The first suppressor 9 is AERS 500; the second suppressor 14 is AERS 500 mm; the first detector 10 and the second detector 15 are suppressive conductivity detectors.
尿液中草甘膦、草铵膦、亚硝酸盐的离子色谱法分析检测步骤如下：The steps for ion chromatography analysis and detection of glyphosate, glufosinate and nitrite in urine are as follows:
步骤一、将待测尿液进行稀释除去大颗粒固体进样：取待测尿液1mL加入15mL试管中，加入19mL去离子水稀释，以7830 r/min的速度离心10min后，取上清液放入超滤管中。超滤管再以7830r/min的速度离心10min后，取上清液过0.22μm尼龙滤膜除去固体颗粒，直接进样；进样过程具体参见图1，实线表示连通的状态，虚线表示断开的状态，自动进样器16将待测尿液通过第一六通阀3输送到定量环6中，使待测尿液进入一维检测单元；Step 1. Dilute the urine to be tested to remove large solid particles and inject: add 1 mL of urine to be tested into a 15 mL test tube, add 19 mL of deionized water to dilute, centrifuge at 7830 r/min for 10 minutes, and take the supernatant Put into ultrafiltration tube. After the ultrafiltration tube was centrifuged at a speed of 7830r/min for 10 minutes, the supernatant was passed through a 0.22μm nylon filter to remove solid particles, and the sample was injected directly. For details on the injection process, see Figure 1. The solid line indicates the connected state, and the dotted line indicates disconnection. In the open state, the autosampler 16 transports the urine to be tested into the quantitative loop 6 through the first six-way valve 3, so that the urine to be tested enters the one-dimensional detection unit;
步骤二、参见图2，自动进样器16的进样端断开，第一泵1将洗脱液送入定量环6中，随后混合的待测液体进入第一保护柱7和第一分析柱8，将目标离子和杂质离子进行分离；淋洗液为10~40 mol/L 的KOH溶液梯度淋洗，具体为：0~5 min，10 mmol/L；5~25 min，10~40mmol/L；25~27 min，40 mmol/L；27.1~30 min，10 mmol/L；温度为30℃，流速为1 mL/min；然后分离后的液体经过第一抑制器9和第一检测器10进行检测，检测后经过第三六通阀排出；参见图5，a表示尿液和b表示尿液加标，通过抑制电导检测；得到的峰中：1表示亚硝酸根；2表示草铵膦；3表示草甘膦，说明可以实现目标离子的检测；Step 2. Referring to Figure 2, the injection end of the automatic sampler 16 is disconnected, the first pump 1 sends the eluent into the quantitative loop 6, and then the mixed liquid to be measured enters the first guard column 7 and the first analysis Column 8 separates target ions and impurity ions; the eluent is a gradient elution of 10~40 mol/L KOH solution, specifically: 0~5 min, 10 mmol/L; 5~25 min, 10~40mmol /L; 25~27 min, 40 mmol/L; 27.1~30 min, 10 mmol/L; temperature is 30°C, flow rate is 1 mL/min; then the separated liquid passes through the first suppressor 9 and the first detection The detector 10 is used for detection, and is discharged through the third six-way valve after detection; see Figure 5, a represents urine and b represents urine spiked, and is detected by inhibiting conductivity; among the peaks obtained: 1 represents nitrite; 2 represents grass Ammonium phosphorus; 3 represents glyphosate, indicating that the detection of target ions can be achieved;
步骤三、当待测物浓度较低，即小于10 mg/L时，一维检测单元无法准确检测时，参见图3，经过一维检测单元预处理的待测尿液在第三六通阀的切换下可以进入二维检测单元的富集柱11，其中根据各目标物质的分离时间，控制草铵膦、亚硝酸盐、草铵膦的阀切换时间分别为：9.7~11.4 min、15~15.9 min、26.4~29 min；在富集柱11中完成对低浓度的目标物质的富集；Step 3. When the concentration of the substance to be tested is low, that is, less than 10 mg/L, and the one-dimensional detection unit cannot accurately detect it, see Figure 3. The urine to be tested that has been preprocessed by the one-dimensional detection unit is in the third and six-way valve. The enrichment column 11 of the two-dimensional detection unit can be entered by switching. According to the separation time of each target substance, the valve switching times for controlling glufosinate, nitrite, and glufosinate are: 9.7~11.4 min, 15~ 15.9 min, 26.4~29 min; the enrichment of low-concentration target substances is completed in the enrichment column 11;
步骤四、参见图4，此时断开富集柱11的进料端，同时第二泵2将洗脱液送入富集柱11中，随后混合的待测尿液进入第二保护柱12和第二分析柱13,进行洗脱分离，淋洗液为23mmol的KOH溶液等度淋洗，柱温为30℃，流速为1mL/min，然后分离后的液体经过第二抑制器14和第二检测器15进行检测，参见图6，a表示尿液和b表示尿液加标，通过抑制电导检测；得到的峰中：1表示草铵膦；2表示亚硝酸根；3表示草甘膦，说明可以实现低浓度的目标离子的检测；Step 4. Refer to Figure 4. At this time, the feed end of the enrichment column 11 is disconnected. At the same time, the second pump 2 sends the eluent into the enrichment column 11, and then the mixed urine to be tested enters the second guard column 12. and the second analytical column 13 for elution and separation. The eluent is 23 mmol of KOH solution for isocratic elution. The column temperature is 30°C and the flow rate is 1 mL/min. Then the separated liquid passes through the second suppressor 14 and the third Two detectors 15 are used for detection, see Figure 6, a represents urine and b represents urine spiked, detected by inhibiting conductivity; among the peaks obtained: 1 represents glufosinate; 2 represents nitrite; 3 represents glyphosate , indicating that the detection of low-concentration target ions can be achieved;
步骤五、通过在空白尿液中添加0.1~100 mg/L的目标离子对建立起的二维离子交换柱切换法进行了检出限、定量限、线性和回收率的评价。通过分别对草甘膦、草铵膦和亚硝酸盐峰面积和溶液浓度建立线性回归方程，每个浓度峰面积分别取六次计算平均值，计算得到一维检测单元中草甘膦、草铵膦和亚硝酸盐的分别在10~100 mg/L、10~100 mg/L、10~100 mg/L有良好的线性关系，线性相关系数（R2）分别为0.9999、0.9992、0.9992，检出限分别为0.40、5.88、0.97 mg/L（S/N=3），定量限分别为1.32、19.6、3.25 mg/L（S/N=10）。二维检测单元中草甘膦、草铵膦和亚硝酸盐的分别在0.1~10 mg/L、0.1~10 mg/L、0.1~10 mg/L有良好的线性关系，线性相关系数（R2）分别为0.9996、0.9993、0.9993，检出限分别为0.05、0.02、0.03 mg/L（S/N=3），定量限分别为0.19、0.06、0.11 mg/L（S/N=10）。本发明的检测方法中，需要确定一维检测单元和二维检测单元的检测浓度分界点，由于确定柱切换时间的范围时使用2 mg/L 的标准样品，当柱切换时间确定后发现当样品浓度增加到一定值时，从第一维到第二维的回收效率反而降低，即当浓度大于10 mg/L时，回收效率降低，故选择10mg/L 为二维检测浓度的最高值，在这个条件下测定第二维的标准曲线，当目标离子浓度为0.1~10 mg/L时线性相关性R2均＞0.999；测定第一维的线性曲线，当浓度为10~100 mg/L时，线性相关性R2均＞0.999，故第一维的测定范围是10~100 mg/L。Step 5: The detection limit, quantitation limit, linearity and recovery rate of the established two-dimensional ion exchange column switching method were evaluated by adding 0.1~100 mg/L target ions to blank urine. By establishing linear regression equations for the peak areas and solution concentrations of glyphosate, glufosinate and nitrite respectively, and taking the average of six calculations for each concentration peak area, the glyphosate, glufosinate and nitrite in the one-dimensional detection unit were calculated. Phosphine and nitrite have good linear relationships at 10~100 mg/L, 10~100 mg/L, and 10~100 mg/L respectively. The linear correlation coefficients (R 2 ) are 0.9999, 0.9992, and 0.9992 respectively. The limits of detection were 0.40, 5.88, and 0.97 mg/L (S/N=3), and the limits of quantitation were 1.32, 19.6, and 3.25 mg/L (S/N=10). In the two-dimensional detection unit, glyphosate, glufosinate and nitrite have good linear relationships at 0.1~10 mg/L, 0.1~10 mg/L, and 0.1~10 mg/L respectively. The linear correlation coefficient (R 2 ) are 0.9996, 0.9993, and 0.9993 respectively, the detection limits are 0.05, 0.02, and 0.03 mg/L (S/N=3) respectively, and the quantification limits are 0.19, 0.06, and 0.11 mg/L (S/N=10) respectively. . In the detection method of the present invention, it is necessary to determine the detection concentration dividing point of the one-dimensional detection unit and the two-dimensional detection unit. Since a 2 mg/L standard sample is used to determine the range of column switching time, when the column switching time is determined, it is found that when the sample When the concentration increases to a certain value, the recovery efficiency from the first dimension to the second dimension decreases. That is, when the concentration is greater than 10 mg/L, the recovery efficiency decreases. Therefore, 10 mg/L is selected as the highest value for the two-dimensional detection concentration. Under this condition, when the second-dimensional standard curve is measured, when the target ion concentration is 0.1~10 mg/L, the linear correlation R2 is >0.999; when the first-dimensional linear curve is measured, when the concentration is 10~100 mg/L , linear correlation R2 are all >0.999, so the measurement range of the first dimension is 10~100 mg/L.
Claims (6)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111320527.9A CN114113373B (en) | 2021-11-09 | 2021-11-09 | System and method for measuring glyphosate, glufosinate and nitrite in urine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111320527.9A CN114113373B (en) | 2021-11-09 | 2021-11-09 | System and method for measuring glyphosate, glufosinate and nitrite in urine |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114113373A CN114113373A (en) | 2022-03-01 |
| CN114113373B true CN114113373B (en) | 2023-12-26 |
Family
ID=80377798
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111320527.9A Active CN114113373B (en) | 2021-11-09 | 2021-11-09 | System and method for measuring glyphosate, glufosinate and nitrite in urine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114113373B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN118584019B (en) * | 2024-08-02 | 2024-10-25 | 则正(济南)生物科技有限公司 | A method for detecting nitrite content in pharmaceutical excipients by pre-column derivatization liquid chromatography |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH07181173A (en) * | 1993-12-24 | 1995-07-21 | Kawatetsu Techno Res Corp | Method and apparatus for gradient analysis of liquid chromatography |
| CN102967681A (en) * | 2012-11-26 | 2013-03-13 | 中国科学院青岛生物能源与过程研究所 | Multi-dimensional ion chromatographic analysis system |
| CN104678041A (en) * | 2015-02-26 | 2015-06-03 | 华东理工大学 | Method for simultaneously testing inorganic anions and aniline compounds by two-dimensional system |
| CN104713973A (en) * | 2013-12-13 | 2015-06-17 | 中国科学院大连化学物理研究所 | Two-dimensional preparative chromatography instrument system having online enrichment function, and applications thereof |
| CN110568092A (en) * | 2019-08-21 | 2019-12-13 | 兰州市食品药品检验所 | method for simultaneously detecting phosphine, ethephon, glufosinate-ammonium and glyphosate in water |
| CN113219112A (en) * | 2021-04-24 | 2021-08-06 | 康龙化成(北京)新药技术股份有限公司 | Two-dimensional high-pressure preparation liquid chromatography system and method for separating and purifying low-content target components in medicine |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040019430A1 (en) * | 2001-11-21 | 2004-01-29 | Patrick Hurban | Methods and systems for analyzing complex biological systems |
| US8101422B2 (en) * | 2005-09-16 | 2012-01-24 | Dionex Corporation | Multidimensional chromatography apparatus and method |
| CN102128885B (en) * | 2010-12-08 | 2013-01-30 | 广东中烟工业有限责任公司 | Method for quickly and simultaneously detecting 16 inorganic anions and organic acids in tobacco |
| JP6232067B2 (en) * | 2012-08-31 | 2017-11-15 | 西安奥▲嵐▼科技▲開▼▲発▼有限▲責▼任公司 | Separation system and separation method for multidimensional liquid chromatography for protein separation |
-
2021
- 2021-11-09 CN CN202111320527.9A patent/CN114113373B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH07181173A (en) * | 1993-12-24 | 1995-07-21 | Kawatetsu Techno Res Corp | Method and apparatus for gradient analysis of liquid chromatography |
| CN102967681A (en) * | 2012-11-26 | 2013-03-13 | 中国科学院青岛生物能源与过程研究所 | Multi-dimensional ion chromatographic analysis system |
| CN104713973A (en) * | 2013-12-13 | 2015-06-17 | 中国科学院大连化学物理研究所 | Two-dimensional preparative chromatography instrument system having online enrichment function, and applications thereof |
| CN104678041A (en) * | 2015-02-26 | 2015-06-03 | 华东理工大学 | Method for simultaneously testing inorganic anions and aniline compounds by two-dimensional system |
| CN110568092A (en) * | 2019-08-21 | 2019-12-13 | 兰州市食品药品检验所 | method for simultaneously detecting phosphine, ethephon, glufosinate-ammonium and glyphosate in water |
| CN113219112A (en) * | 2021-04-24 | 2021-08-06 | 康龙化成(北京)新药技术股份有限公司 | Two-dimensional high-pressure preparation liquid chromatography system and method for separating and purifying low-content target components in medicine |
Non-Patent Citations (1)
| Title |
|---|
| Determination of glyphosate and phosphate in water by ion chromatography—inductively coupled plasma mass spectrometry detection;Zhong-Xian Guo 等;Journal of Chromatography A;第1100卷 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114113373A (en) | 2022-03-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Thomas et al. | Determination of trace amounts of arsenic species in natural waters by high-performance liquid chromatography–inductively coupled plasma mass spectrometry | |
| Emteborg et al. | Speciation of organic and inorganic selenium in a biological certified reference material based on microbore ion-exchange chromatography coupled to inductively coupled plasma atomic emission spectrometry via a direct injection nebulizer or coupled to electrothermal atomic absorption spectrometry | |
| Hama et al. | Pyrrolizidine alkaloids quantified in soil and water using UPLC-MS/MS | |
| Shan et al. | A fully automated system with on-line micro solid-phase extraction combined with capillary liquid chromatography–tandem mass spectrometry for high throughput analysis of microcystins and nodularin-R in tap water and lake water | |
| Gregoire | Determination of boron in fresh and saline waters by inductively coupled plasma mass spectrometry | |
| CN112461960B (en) | Method for simultaneously measuring various heterocyclic pesticides, degradation products and intermediates in water | |
| CN114113373B (en) | System and method for measuring glyphosate, glufosinate and nitrite in urine | |
| Zheng et al. | Reversed-phase liquid chromatography with mixed ion-pair reagents coupled with ICP-MS for the direct speciation analysis of selenium compounds in human urine | |
| CN109342638B (en) | Method for detecting content of nitrile, quaternary ammonium salt and impurities ammonium, potassium and sodium ions in nitrile and quaternary ammonium salt by using cation exchange inhibition conductivity method | |
| Le Bot et al. | Optimization and performance evaluation of the analysis of glyphosate and AMPA in water by HPLC with fluorescence detection | |
| Prados et al. | A fully automated HPLC method for the determination of catecholamines in biological samples utilizing ethylenediamine condensation and peroxyoxalate chemiluminescence detection | |
| Chen et al. | Development of a high-performance liquid chromatography method for the simultaneous quantification of four organoarsenic compounds in the feeds of swine and chicken | |
| Jiang et al. | Simultaneous determination of pentachlorophenol, niclosamide and fenpropathrin in fishpond water using an LC-MS/MS method for forensic investigation | |
| Sitanurak et al. | Simultaneous determination of iodide and creatinine in human urine by flow analysis with an on-line sample treatment column | |
| Hogendoorn et al. | Automated sample clean‐up and fractionation of organochlorine pesticides and polychlorinated biphenyls in human milk using NP‐HPLC with column‐switching | |
| Hils et al. | Speciation of trace amounts of aluminium in percolating water of forest soil by online coupling HPLC-ICP-MS | |
| Khoschsorur et al. | Simple and rapid HPLC method for simultaneous determination of multiple antiepileptic drugs in human serum | |
| CN210775360U (en) | Ion chromatographic valve switching and separating system for analyzing ammonia release amount in flue gas | |
| Gummow et al. | Paired-ion extraction and high-performance liquid chromatographic determination of diminazene in cattle plasma: a modified method | |
| CN115236259B (en) | High performance liquid chromatography determination method for residual citric acid in Fmoc-amino acid | |
| CN106525802A (en) | Measure method and device for lead form in cigarette smoke | |
| CN114324635B (en) | Method for measuring bromate in drinking water by utilizing ion chromatography on-line matrix elimination system | |
| Wahlen | Fast and accurate determination of arsenobetaine in fish tissues using accelerated solvent extraction and HPLC-ICP-MS determination | |
| CN106770777A (en) | HPLC detects method of the desmopressin acetate about material | |
| Piatina et al. | Direct quantification of metal‐organic interactions by size‐exclusion chromatography (SEC) and inductively coupled plasma mass spectrometry (ICP‐MS) |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |