CN102253034A - Method for on-line detection of gibberellin with flow-injection chemiluminescence method - Google Patents
Method for on-line detection of gibberellin with flow-injection chemiluminescence method Download PDFInfo
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- CN102253034A CN102253034A CN2011100943153A CN201110094315A CN102253034A CN 102253034 A CN102253034 A CN 102253034A CN 2011100943153 A CN2011100943153 A CN 2011100943153A CN 201110094315 A CN201110094315 A CN 201110094315A CN 102253034 A CN102253034 A CN 102253034A
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Abstract
The invention discloses a method for on-line detection of gibberellin with a flow-injection chemiluminescence method. The method comprises the steps of: preparing a cerium ammonium sulfate solution (Ce(IV)), a Na2SO3 solution, a gibberellin standard solution and a sample gibberellin solution; performing three-way mixing on the Na2SO3 solution and the gibberellin standard solution respectively by an active pump through corresponding pipes at a flow rate of 30-60 r/min, and inputting into a flow-injection chemiluminescence analyzer; after a baseline is stabilized, injecting the cerium ammonium sulfate solution through a sixteen-way injection valve at a flow rate of 30-60 r/min by an auxiliary pump for 10-20 seconds, mixing all solutions, leading the mixed solution to flow into a flowing cell, detecting chemiluminescence intensity of the mixed solution, recording data, and establishing a standard curve equation; and repeating the operation to detect the chemiluminescence intensity of the sample gibberellin, and substituting a chemiluminescence intensity value into the standard curve equation to compute the content of the sample gibberellin. The detection method is simple and is low in cost, can be applied to on-line regular detection and is suitable for application to industrial or agricultural production.
Description
Technical field
The invention belongs to the physiology field of phytochemistry, plant physiology, Plant Pathology, plant protection, natural product chemistry, Food Chemistry, analytical chemistry, organic chemistry, biological chemistry, Secondary Metabolism of Plant material, the method for the online detection gibberellin of particularly a kind of portable injection chemiluminescence method.
Background technology
Gibberellin is a class plant hormone that extensively exists, and is one of five big class plant hormones.Its chemical constitution belongs to Diterpenes acid, derive and get by the Fourth Ring skeleton, can break the Seed Dormancy phase, promote the plant growth, usually produce gibberellin with the gibberella submerged fermentation now, have tens of kinds more than in the product, wherein topmost active component is a gibberellin A 3, claim gibberellic acid (gibberellic acid) again, gibberellin A
3Molecular structure has redox characteristics.
Gibberellin not only has multiple effect to industrial crops such as paddy rice, cottons, and can be medicinal; Medical gibberellin can be used as externally applied drug, and the effect of antipruritic, convergence, myogenic, skin is arranged, and skin injury, erosion, ulcer focus and some common diseases of internal medicine are all had certain curative effect.
The height of the fungi of plant Nei Neisheng-gibberella content is very important to the growth of plant, if the mould of interior life of plant or external source breeding in a large number can induce excessive gibberellin, excessive gibberellin can cause wheat, rice crop etc. that damping-off, dead leaf disease, withertip disease and root rot etc. take place; Endogenetic fungus-gibberella in the plant too shortage can cause plant inner gibberellin content to cross low and induce of short stem or disease such as dwarfing, and the method for the online detection gibberellin of portable injection chemiluminescence method of the present invention can be used for monitoring breeding, cultivation and the plant protection etc. of plant.This method is that the chemiluminescence of physical flow injection and chemistry combines, utilize chemical substance that fluorescence or phosphorescence are arranged in the redox reaction process, this part optical signals is transformed into electric signal, and further signal amplifies again, reaches the purpose of quantitative test according to luminous intensity.
Analyzing detecting method has at present: 1. (Zhang Chaoying etc. 1993 for high performance liquid chromatography; Li Jianfei, Wang De good 2010), 2. thin-layered chromatography (Cheng Guohua etc. 1991), 3. fluorescence spectrophotometry (Luo Guoan etc. 1990; Suo Landi etc. 1996), 4. ultraviolet-visible light spectrophotometric method (Wang Baoqin, Niu Jishan 2007), 5. infra-red sepectrometry (He Wei flat 1991) etc.
The shortcoming of various detection methods: 1. the high performance liquid chromatography instrument is somewhat expensive, simultaneously this professional and technical personnel to be arranged, general gibberellin processing factory is difficult for realizing that special-purpose in addition chromatographically pure moving phase and chromatographic column are somewhat expensive, and instrument such as high pressure lipuid chromatography (HPLC) is inconvenient to carry; 2. the used chemical reagent of thin-layered chromatography is many, and the experimentation complexity is suitable for the purifying of small sample; 3. the detectability of fluorescence spectrophotometry is than higher; 4. the ultraviolet-visible light spectrophotometric method reaction time long, test sample speed is slow; 5. the range of linearity of infra-red sepectrometry is narrow, disturbed by other impurity.This several method all is suitable for the research to gibberellin of university and research institute; By contrast, the portable injection chemiluminescence method is highly sensitive, and the range of linearity is wide, and instrument and equipment is simple, favored by people.Because in the H2SO4 acid medium, gibberellin can enhanced sensitivity Ce (IV)-Na2SO3 system produce this phenomenon of chemiluminescence, in view of the above, the present invention proposes the chemical luminous system of this mensuration gibberellin, and, set up a kind of new method of simple, fast measuring gibberellin in conjunction with the flow injection technology.This method not only has enough sensitivity and stability, and good selectivity is arranged.
Summary of the invention
The method that the purpose of this invention is to provide the online detection gibberellin of a kind of portable injection chemiluminescence method.
Concrete steps are:
With 0.08～0.12mol/L H
2SO
4As solvent preparation 5.0 * 10
-4The Cericammoniumsulfate of mol/L (Ce (IV)) prepares 1.0 * 10 with redistilled water as solvent
-2The Na of mol/L
2SO
3Solution, with pH be sodium hydrogen phosphate-sodium dihydrogen phosphate phosphate buffer of 5～7 (0.1～0.3mol/L) for the solvent compound concentration be 2.5mg/mL gibberellin standard solution and sample Gibberellins solution.Active efflux is respectively with the current-carrying Na of above-mentioned concentration
2SO
3Solution and gibberellin standard solution carry out the threeway mixing with the flow velocity of 30～60r/min by corresponding pipeline, the mobile injected chemical luminescence analyzer of input.After treating that baseline is steadily, secondary moving pump with Cericammoniumsulfate solution with the flow velocity of 30～60r/min by 16 logical introduction valves, annotate 10～20 seconds sample time, each liquid mixes, and flows into flow cell and detects its chemiluminescence intensity, record data are made the typical curve equation; Repeat the aforesaid operations process then, the chemiluminescence intensity of specimen gibberellin, substitution typical curve equation, the content of calculation sample gibberellin.Above-mentioned test process and various chemical reagent (sodium sulphite, Cericammoniumsulfate and treat test sample etc.) all remain under 35 ℃ of constant temperatures, mentioned reagent be analyze pure.
Under optimized conditions, the concentration of gibberellin standard solution is 5.0 * 10
-3～1.0 * 10
-1Linear with luminous intensity Δ I in the mg/mL scope, this shows and can utilize this method that gibberellin is done quantitative test.
Portable injection chemiluminescence method of the present invention is highly sensitive owing to it, good reproducibility, and the range of linearity is wide, and instrument and equipment is simple, and is simple to operate, and it is low to detect cost, can onlinely measure routinely, is easy to be applied in industry or the agricultural production.
Description of drawings:
Accompanying drawing is gibberellin A of the present invention
3Molecular structure.
Gibberellin A
3Molecular structure has redox characteristics, there is fluorescence to discharge when oxidized, this light can be had the instrument detecting of distinct electrical chemiluminescence detection function, and oxidized signal intensity is directly proportional with hydroxyl quantity on its molecular structure, analyzes gibberellin thus.
Embodiment
Embodiment:
Use 0.1mol/L H
2SO
4As solvent preparation 5.0 * 10
-4The Cericammoniumsulfate of mol/L (Ce (IV)) prepares 1.0 * 10 with redistilled water as solvent
-2The Na of mol/L
2SO
3Solution is that sodium hydrogen phosphate-sodium dihydrogen phosphate phosphate buffer (0.2mol/L) of 6.2 is 2.5mg/mL gibberellin standard solution and sample Gibberellins solution for the solvent compound concentration with pH.Active efflux is respectively with the current-carrying Na of above-mentioned concentration
2SO
3Solution and gibberellin standard solution carry out the threeway mixing with the flow velocity of 45r/min by corresponding pipeline, the mobile injected chemical luminescence analyzer of input.After treating that baseline is steadily, secondary moving pump with Cericammoniumsulfate solution with the flow velocity of 45r/min by 16 logical introduction valves, annotate 15 seconds sample time, each liquid mixes, and flows into flow cell and detects its chemiluminescence intensity, record data are made the typical curve equation; Repeat the aforesaid operations process then, the chemiluminescence intensity of specimen gibberellin, substitution typical curve equation, the content of calculation sample gibberellin.Above-mentioned test process and various chemical reagent (sodium sulphite, Cericammoniumsulfate and treat test sample etc.) all remain under 35 ℃ of constant temperatures, mentioned reagent be analyze pure.
Claims (1)
1. the method for the online detection gibberellin of portable injection chemiluminescence method is characterized in that concrete steps are: with 0.08～0.12mol/L H
2SO
4As solvent preparation 5.0 * 10
-4The Cericammoniumsulfate Ce (IV) of mol/L prepares 1.0 * 10 with redistilled water as solvent
-2The Na of mol/L
2SO
3Solution is 5～7 with pH, and concentration is that sodium hydrogen phosphate-sodium dihydrogen phosphate phosphate buffer of 0.1～0.3mol/L is 2.5mg/mL gibberellin standard solution and sample Gibberellins solution for the solvent compound concentration; Active efflux is respectively with the current-carrying Na of above-mentioned concentration
2SO
3Solution and gibberellin standard solution carry out the threeway mixing with the flow velocity of 30～60r/min by corresponding pipeline, the mobile injected chemical luminescence analyzer of input; After treating that baseline is steadily, secondary moving pump with Cericammoniumsulfate solution with the flow velocity of 30～60r/min by 16 logical introduction valves, annotate 10～20 seconds sample time, each liquid mixes, and flows into flow cell and detects its chemiluminescence intensity, record data are made the typical curve equation; Repeat the aforesaid operations process then, the chemiluminescence intensity of specimen gibberellin, substitution typical curve equation, the content of calculation sample gibberellin; Described test process and various chemical reagent all remain under 35 ℃ of constant temperatures, and it is pure that described reagent is analysis.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102253033A (en) * | 2011-04-13 | 2011-11-23 | 桂林理工大学 | On-line rapid detection method of stevioside |
| CN106018386A (en) * | 2016-04-20 | 2016-10-12 | 杭州大茗堂生物科技有限公司 | Analytical method of antioxidant capacity in tea or tea products |
| CN106404507A (en) * | 2016-08-31 | 2017-02-15 | 广东省农业科学院农产品公共监测中心 | Leaf vegetable gibberellins residue detection sample pretreatment method and detection method |
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| JPH03107749A (en) * | 1989-09-21 | 1991-05-08 | Fuaamu Tec:Kk | Method for measuring sulfur dioxide by chemiluminescence reaction and method for measuring carbon dioxide by chemiluminescence reaction |
| CN1869663A (en) * | 2006-05-30 | 2006-11-29 | 黑龙江双河松嫩大豆生物工程有限责任公司 | Portable injection chemiluminescence analysis method of isoflavone compound |
| CN101493419A (en) * | 2009-03-06 | 2009-07-29 | 桂林工学院 | Chemiluminescence method for detecting chlorophyll alpha |
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2011
- 2011-04-13 CN CN2011100943153A patent/CN102253034A/en active Pending
Patent Citations (3)
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| JPH03107749A (en) * | 1989-09-21 | 1991-05-08 | Fuaamu Tec:Kk | Method for measuring sulfur dioxide by chemiluminescence reaction and method for measuring carbon dioxide by chemiluminescence reaction |
| CN1869663A (en) * | 2006-05-30 | 2006-11-29 | 黑龙江双河松嫩大豆生物工程有限责任公司 | Portable injection chemiluminescence analysis method of isoflavone compound |
| CN101493419A (en) * | 2009-03-06 | 2009-07-29 | 桂林工学院 | Chemiluminescence method for detecting chlorophyll alpha |
Non-Patent Citations (4)
| Title |
|---|
| JINGKAI ZHANG ET AL.: "Flow injecion determination of benzhexol based on its sensitizing effect on the chemiluminescent reaction of Ce(IV)-sulfite", 《LUMINESCENCE》 * |
| 孙述文等: "流动注射后化学发光法测定舒必利", 《陕西师范大学学报( 自然科学版)》 * |
| 慕苗等: "硝酸铈铵-Na2SO3化学发光体系测定萘普生", 《分析试验室》 * |
| 罗国安等: "荧光分光光度法测定赤霉素的含量", 《中国药科大学学报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102253033A (en) * | 2011-04-13 | 2011-11-23 | 桂林理工大学 | On-line rapid detection method of stevioside |
| CN106018386A (en) * | 2016-04-20 | 2016-10-12 | 杭州大茗堂生物科技有限公司 | Analytical method of antioxidant capacity in tea or tea products |
| CN106018386B (en) * | 2016-04-20 | 2019-04-16 | 浙江大茗堂生物科技有限公司 | The analysis method of oxidation resistance in a kind of tealeaves or tea product |
| CN106404507A (en) * | 2016-08-31 | 2017-02-15 | 广东省农业科学院农产品公共监测中心 | Leaf vegetable gibberellins residue detection sample pretreatment method and detection method |
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Application publication date: 20111123 |