CN104897807B - A kind of method applying triacontanol content in gas chromatographic detection foliar fertilizer - Google Patents
A kind of method applying triacontanol content in gas chromatographic detection foliar fertilizer Download PDFInfo
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- CN104897807B CN104897807B CN201510255107.5A CN201510255107A CN104897807B CN 104897807 B CN104897807 B CN 104897807B CN 201510255107 A CN201510255107 A CN 201510255107A CN 104897807 B CN104897807 B CN 104897807B
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Abstract
The present invention relates to a kind of detection method of triacontanol, particularly a kind of method applying triacontanol content in gas chromatographic detection foliar fertilizer.The method of triacontanol content in this application gas chromatographic detection foliar fertilizer, comprise and take triacontanol standard items and be dissolved in methenyl choloride, configure the triacontanol solution of a series of concentration known, this triacontanol solution is carried out gas chromatographic analysis, obtain the triacontanol peak area of series concentration, with triacontanol concentration for horizontal ordinate, peak area is that ordinate obtains its typical curve and equation of linear regression; By laggard for testing sample process promoting the circulation of qi analysis of hplc, obtain the peak area of triacontanol in sample, after bringing peak area into above-mentioned equation of linear regression, obtain corresponding triacontanol concentration.The inventive method processing procedure is quick, and content detection is comparatively accurate, can tackle the quality monitoring of violated interpolation or excessive this material of interpolation on market.
Description
(1) technical field
The present invention relates to a kind of detection method of triacontanol, particularly a kind of method applying triacontanol content in gas chromatographic detection foliar fertilizer.
(2) background technology
The long-chain fatty alcohol that triacontanol (1-Triacontanol) is made up of 30 carbon atoms, also known as melissyl alcohol.The research such as Rics in 1977 proves that triacontanol is a kind of new plant growth stimulator, there is hestening rooting, germinate, bloom, cauline leaf grows and precocity acts on, and improve the different physiological roles such as chlorophyll content, enhancing photosynthesis, in addition, can also promote that rice seedling and cucumber seedling are to the absorption of nitrogen, phosphatic manure.Triacontanol always coexists with other long-chain alcohol at nature, experiment proves wherein only have triacontanol just to have physiologically active, and other long chain alkanes not only do not have facilitation to plant growth, there is inhibiting effect on the contrary, so the purity of triacontanol and the correlativity of its physiologically active are particularly evident, in addition, the impact of its working concentration on regulating action is fairly obvious.
Triacontanol is present in the wax of various plants and insect, finds no toxic action to people and animals and beneficial organism, is the plant growth regulator that a kind of Environmental compatibility is good, is widely used in agricultural production, and numerous commercial preparation is developed.But, if the excessive use of triacontanol, can cause seedling stage bud scale bend, root deformity, strain then causes young leaflet tablet curling, and in the new-type fertilizer based on foliar fertilizer on market, the situation of violated interpolation or excessive interpolation plant growth regulator happens occasionally, therefore, very important to the monitoring of triacontanol formulation products quality.
(3) summary of the invention
The present invention is in order to make up the deficiencies in the prior art, and invented a kind of processing procedure fast, content detection is more a kind of method applying triacontanol content in gas chromatographic detection foliar fertilizer.
The present invention is achieved through the following technical solutions:
A kind of method applying triacontanol content in gas chromatographic detection foliar fertilizer, it is characterized in that: take triacontanol standard items and be dissolved in organic solvent, configure the triacontanol solution of a series of concentration known, this triacontanol solution is carried out gas chromatographic analysis, obtain the chromatographic peak area of series concentration triacontanol, with triacontanol concentration for horizontal ordinate, peak area is that ordinate obtains its typical curve and equation of linear regression; By laggard for testing sample process promoting the circulation of qi analysis of hplc, obtain the peak area of triacontanol in sample, after bringing peak area into above-mentioned equation of linear regression, obtain corresponding triacontanol concentration; Wherein, the condition of gas chromatographic analysis comprises HP-5 capillary chromatographic column, 30mm × 320 μm × 0.25 μm; Injector temperature: 280-300 DEG C, does not shunt; Detector temperature: 320-330 DEG C; Column temperature: adopt temperature programme, initial temperature 160-200 DEG C, keeps 1min, is raised to 320 DEG C with 15 DEG C/min, keeps 3min; Carrier gas is nitrogen: flow velocity is 1.0-2.0mL/min; Sample size: 1 μ L.
The condition of above-mentioned gas chromatographic analysis comprises HP-5 capillary chromatographic column, 30mm × 320 μm × 0.25 μm; Injector temperature: 300 DEG C, do not shunt; Detector temperature: 320 DEG C; Column temperature: adopt temperature programme, initial temperature 180 DEG C, keeps 1min, is raised to 320 DEG C with 15 DEG C/min, keeps 3min; Carrier gas is nitrogen: flow velocity is 1.5mL/min; Sample size: 1 μ L.
Described extraction agent is that organic solvent and phosphoric acid mix, and extractant and phosphoric acid ratio are 5-10:1.
Described organic solvent is methenyl choloride, the mixing of one or more in phenixin and ether.
Described extraction agent is the mixed liquor of methenyl choloride and phosphoric acid, methenyl choloride: phosphoric acid=6:1.
Described testing sample is foliage fertilizer, and described foliage fertilizer comprises solid foliage fertilizer, liquid foliage fertilizer or pulvis foliage fertilizer.
Testing sample disposal route is:
Fluid sample: accurately take foliage fertilizer described in 1.0g, with in 30mL extraction agent, ultrasonic 30min, after vibration 30min, static 30min, gets organic phase after separatory, revolves and steams near dry, be settled to 25mL, get 1mL cross 0.22 μm of filter membrane after for subsequent use.
Powder samples: accurately take 1.0g pulvis foliage fertilizer, dissolves with 30mL methenyl choloride, and adds 5mL phosphoric acid wherein, ultrasonic 30min, after vibration 30min, static 30min, the centrifugal 5min of 8000rpm, removing precipitation, adds in 20mL ultrapure water by supernatant, move in separating funnel, interrupted oscillation 5min, leave standstill 30min, treat methenyl choloride phase and aqueous phase layering, pipette methenyl choloride phase, revolve and steam near dry, be settled to 25mL, get 1mL cross 0.22 μm of filter membrane after for subsequent use.Take after solid foliage fertilizer grind into powder.When configuring the triacontanol solution of a series of concentration known, triacontanol solution concentration is 10 ~ 100 μ g/mL.
Described detecting device is flame ionization ditector.
Because triacontanol is low pole compound, water-soluble hardly, cold ethanol, is insoluble in benzene, dissolves in methenyl choloride, tetrachloromethane, in ether.Therefore tried out methenyl choloride, phenixin two kinds of reagent extract the triacontanol in sample, and result shows that the extraction effect of triacontanol in methenyl choloride is best, therefore selects methenyl choloride extraction agent the most.When extracting triacontanol, the simple carbon trichloride that uses is difficult to be proposed, and wherein adds appropriate phosphoric acid, can accelerate demulsification, effectively improve extraction ratio, therefore in test use methenyl choloride: the ratio of phosphoric acid=6:1 is extracted if think.
To during testing sample process need purify, by the method for high speed refrigerated centrifuge, some insoluble matters in foliar fertilizer are separated removing, again because containing numerous inorganic ions and large polar material in foliar fertilizer, these inorganic ions and large polar material are water miscible, so sample is carried out liquid-liquid extraction, triacontanol is low pole, and be assigned in organic phase, inorganic ion is then removed with aqueous phase.
Wherein, triacontanol standard items are purchased from German Dr.ehrenstorfer company, and methenyl choloride and phenixin are chromatographically pure.The instrument adopted is: Agilent7890A gas chromatograph is furnished with fid detector (flame ionization ditector); Sigma3-18K high speed freezing centrifuge (German Sigma company).
Under the determined chromatographic condition of this method, measure by concentration sample introduction analysis from low to high the mark liquid of triacontanol in a series of mass concentration of 10 ~ 100 μ g/mL, its range of linearity, equation of linear regression, related coefficient and quantitative limit are in table 1.
The range of linearity of table 1 triacontanol, linear equation, related coefficient and quantitative limit
Beneficial effect of the present invention is: the inventive method processing procedure is quick, and content detection is comparatively accurate, can tackle the quality monitoring of violated interpolation or excessive this material of interpolation on market.
(4) embodiment
Embodiment 1
Accurately pipette the liquid foliage fertilizer of 1.0g, dissolve with 30mL methenyl choloride, and add 5mL phosphoric acid wherein, ultrasonic 30min, after vibration 30min, static 30min, takes out lower floor's organic phase, upper strata emulsion continues to use 30mL chloroform extraction twice, merge methenyl choloride phase, revolve and steam near dry, be settled to 25mL, get 1mL and cross 0.22 μm of filter membrane, to be measured;
Analyzed by gas chromatograph by above-mentioned liquid to be measured, gas chromatograph condition is: HP-5 capillary chromatographic column, 30mm × 320 μm × 0.25 μm; Injector temperature: 280 DEG C, do not shunt; Detector temperature: 320 DEG C; Column temperature: adopt temperature programme, initial temperature 160 DEG C, keeps 1min, is raised to 320 DEG C with 15 DEG C/min, keeps 3min; Carrier gas is nitrogen: flow velocity is 1.0mL/min; Sample size: 1 μ L; Adopt external standard method to obtain the concentration of triacontanol in liquid to be measured, and then obtain its content in foliage fertilizer.
Embodiment 2
Accurately pipette 1.0g pulvis foliage fertilizer, dissolve with 30mL methenyl choloride, and add 5mL phosphoric acid wherein, ultrasonic 30min, after vibration 30min, static 30min, the centrifugal 5min of 8000rpm, removing precipitation, adds in 20mL ultrapure water by supernatant, move in separating funnel, interrupted oscillation 5min, leave standstill 30min, treat methenyl choloride phase and aqueous phase layering, pipette methenyl choloride phase, revolve and steam near dry, be settled to 25mL, get 1mL and cross 0.22 μm of filter membrane, to be measured;
Analyzed by gas chromatograph by above-mentioned liquid to be measured, gas chromatograph condition is: HP-5 capillary chromatographic column, 30mm × 320 μm × 0.25 μm; Injector temperature: 300 DEG C, do not shunt; Detector temperature: 330 DEG C; Column temperature: adopt temperature programme, initial temperature 200 DEG C, keeps 1min, is raised to 320 DEG C with 15 DEG C/min, keeps 3min; Carrier gas is nitrogen: flow velocity is 2.0mL/min; Sample size: 1 μ L; Adopt external standard method to obtain the concentration of triacontanol in liquid to be measured, and then obtain its content in foliage fertilizer.
Embodiment 3
Accurately pipette the solid foliage fertilizer after 1.0g pulverizing, dissolve with 30mL methenyl choloride, and add 5mL phosphoric acid wherein, ultrasonic 30min, after vibration 30min, static 30min, the centrifugal 5min of 8000rpm, removing precipitation, adds in 20mL ultrapure water by supernatant, move in separating funnel, interrupted oscillation 5min, leave standstill 30min, treat methenyl choloride phase and aqueous phase layering, pipette methenyl choloride phase, revolve and steam near dry, be settled to 25mL, get 1mL and cross 0.22 μm of filter membrane, to be measured;
Analyzed by gas chromatograph by above-mentioned liquid to be measured, gas chromatograph condition is: the condition of gas chromatographic analysis comprises HP-5 capillary chromatographic column, 30mm × 320 μm × 0.25 μm; Injector temperature: 300 DEG C, do not shunt; Detector temperature: 320 DEG C; Column temperature: adopt temperature programme, initial temperature 180 DEG C, keeps 1min, is raised to 320 DEG C with 15 DEG C/min, keeps 3min; Carrier gas is nitrogen: flow velocity is 1.5mL/min; Sample size: 1 μ L; Adopt external standard method to obtain the concentration of triacontanol in liquid to be measured, and then obtain its content in foliage fertilizer.
Embodiment 4
Accurately pipette the liquid foliage fertilizer of 1.0g, dissolve with 30mL methenyl choloride, and add 5mL phosphoric acid wherein, ultrasonic 30min, after vibration 30min, static 30min, takes out lower floor's organic phase, upper strata emulsion continues to use 30mL chloroform extraction twice, merge methenyl choloride phase, revolve and steam near dry, be settled to 25mL, get 1mL and cross 0.22 μm of filter membrane, to be measured;
Analyzed by gas chromatograph by above-mentioned liquid to be measured, gas chromatograph condition is: HP-5 capillary chromatographic column, 30mm × 320 μm × 0.25 μm; Injector temperature: 290 DEG C, do not shunt; Detector temperature: 330 DEG C; Column temperature: adopt temperature programme, initial temperature 190 DEG C, keeps 1min, is raised to 320 DEG C with 15 DEG C/min, keeps 3min; Carrier gas is nitrogen: flow velocity is 1.5mL/min; Sample size: 1 μ L; Adopt external standard method to obtain the concentration of triacontanol in liquid to be measured, and then obtain its content in foliage fertilizer.
Embodiment 5
Accurately pipette the liquid foliage fertilizer of 1.0g, dissolve with 30mL methenyl choloride, and add 5mL phosphoric acid wherein, ultrasonic 30min, after vibration 30min, static 30min, takes out lower floor's organic phase, upper strata emulsion continues to use 30mL chloroform extraction twice, merge methenyl choloride phase, revolve and steam near dry, be settled to 25mL, get 1mL and cross 0.22 μm of filter membrane, to be measured;
Analyzed by gas chromatograph by above-mentioned liquid to be measured, gas chromatograph condition is: HP-5 capillary chromatographic column, 30mm × 320 μm × 0.25 μm; Injector temperature: 280 DEG C, do not shunt; Detector temperature: 330 DEG C; Column temperature: adopt temperature programme, initial temperature 170 DEG C, keeps 1min, is raised to 320 DEG C with 15 DEG C/min, keeps 3min; Carrier gas is nitrogen: flow velocity is 1.6mL/min; Sample size: 1 μ L; Adopt external standard method to obtain the concentration of triacontanol in liquid to be measured, and then obtain its content in foliage fertilizer.
Embodiment 6
Accurately take same batch sample 6 parts, carry out sample pre-treatments, adopt same chromatographic condition by method shown in embodiment 3, to six increment product continuous sample introductions, identify the repeatability of detection method, test findings is in table 2.As can be seen from the table, the mean concentration that detection obtains is 85.69 μ g/mLRSD values is 2.17%, illustrates that the repeatability of detection method is good.
Table 2 triacontanol gas-chromatography detection method replica test
Embodiment 7
Precision takes 1 part, sample, sample pre-treatments is carried out by method shown in embodiment 3, adopt same chromatographic condition, same sample continuous sample introduction six times, the accuracy of detection method is identified, test findings is in table 3, as can be seen from the table, detecting the mean concentration obtained is 36.44 μ g/mL, and RSD value is 0.68%, illustrates that the accuracy of detection method is good.
Table 3 triacontanol gas-chromatography detection method accuracy is tested
Embodiment 8
In blank sample, add triacontanol standard solution, establish high, medium and low three Pitch-based sphere respectively, each level five repetition.Same chromatographic condition is adopted to detect sample successively.
By aforementioned chromatographic condition and detecting step, carry out TIANZHU XINGNAO Capsul test, test findings is in table 4.The average TIANZHU XINGNAO Capsul scope of different Pitch-based sphere is 80.9% ~ 118.3%, and coefficient of variation scope is 5.6 ~ 8.9%, all meets analysis of agricultural drugs standard-required.
The recovery of table 4 triacontanol in foliar fertilizer sample
Embodiment 9
Application the inventive method has carried out analyzing detecting to the foliar fertilizer sample of embodiment 1-5 successively.
The testing result of table 5 triacontanol in foliar fertilizer sample
Claims (7)
1. apply the method for triacontanol content in gas chromatographic detection foliar fertilizer for one kind, it is characterized in that: take triacontanol standard items and be dissolved in organic solvent, configure the triacontanol solution of a series of concentration known, this triacontanol solution is carried out gas chromatographic analysis, obtain the peak area of series concentration triacontanol, with triacontanol concentration for horizontal ordinate, peak area is that ordinate obtains its typical curve and equation of linear regression; By laggard for testing sample process promoting the circulation of qi analysis of hplc, obtain the peak area of triacontanol in sample, after bringing peak area into above-mentioned equation of linear regression, obtain corresponding triacontanol concentration; Wherein, the condition of gas chromatographic analysis comprises HP-5 capillary chromatographic column, 30mm × 320 μm × 0.25 μm; Injector temperature: 280-300 DEG C, does not shunt; Detector temperature: 320-330 DEG C; Column temperature: adopt temperature programme, initial temperature 160-200 DEG C, keeps 1min, is raised to 320 DEG C with 15 DEG C/min, keeps 3min; Carrier gas is nitrogen: flow velocity is 1.0-2.0mL/min; Sample size: 1 μ L; Testing sample process: (1) accurately takes the liquid foliage fertilizer of 1.0g, dissolves, ultrasonic 30min with extraction agent, after vibration 30min, static 30min, gets organic phase after separatory, revolves and steams near dry, be settled to 25mL, get 1mL cross 0.22 μm of filter membrane after for subsequent use; (2) accurately take 1.0g pulvis foliage fertilizer, dissolve with 30mL methenyl choloride, and add 5mL phosphoric acid wherein, ultrasonic 30min, after vibration 30min, static 30min, the centrifugal 5min of 8000rpm, removing precipitation, adds in 20mL ultrapure water by supernatant, move in separating funnel, interrupted oscillation 5min, leave standstill 30min, treat methenyl choloride phase and aqueous phase layering, pipette methenyl choloride phase, revolve and steam near dry, be settled to 25mL, get 1mL cross 0.22 μm of filter membrane after for subsequent use; (3) accurately take the solid foliage fertilizer after 1.0g pulverizing, dissolve with 30mL methenyl choloride, and add 5mL phosphoric acid wherein, ultrasonic 30min, after vibration 30min, static 30min, the centrifugal 5min of 8000rpm, removing precipitation, adds in 20mL ultrapure water by supernatant, move in separating funnel, interrupted oscillation 5min, leave standstill 30min, treat methenyl choloride phase and aqueous phase layering, pipette methenyl choloride phase, revolve and steam near dry, be settled to 25mL, get 1mL cross 0.22 μm of filter membrane after for subsequent use; Described detecting device is flame ionization ditector.
2. the method for triacontanol content in application gas chromatographic detection foliar fertilizer according to claim 1, is characterized in that: the condition of described gas chromatographic analysis comprises HP-5 capillary chromatographic column, 30mm × 320 μm × 0.25 μm; Injector temperature: 300 DEG C, do not shunt; Detector temperature: 320 DEG C; Column temperature: adopt temperature programme, initial temperature 180 DEG C, keeps 1min, is raised to 320 DEG C with 15 DEG C/min, keeps 3min; Carrier gas is nitrogen: flow velocity is 1.5mL/min; Sample size: 1 μ L.
3. the method for triacontanol content in application gas chromatographic detection foliar fertilizer according to claim 1, is characterized in that: described extraction agent is that organic solvent and phosphoric acid mix, and organic solvent and phosphoric acid ratio are 5-10:1.
4. the method for triacontanol content in application gas chromatographic detection foliar fertilizer according to claim 3, is characterized in that: described organic solvent is methenyl choloride, the mixing of one or more in phenixin and ether.
5. the method for triacontanol content in the application gas chromatographic detection foliar fertilizer according to claim 1 or 3 or 4, is characterized in that: described extraction agent is the mixed liquor of methenyl choloride and phosphoric acid, methenyl choloride: phosphoric acid=6:1.
6. the method for triacontanol content in application gas chromatographic detection foliar fertilizer according to claim 1, is characterized in that: described testing sample is foliage fertilizer, and described foliage fertilizer comprises solid foliage fertilizer, liquid foliage fertilizer or pulvis foliage fertilizer.
7. the method for triacontanol content in application gas chromatographic detection foliar fertilizer according to claim 1, is characterized in that: when configuring the triacontanol solution of a series of concentration known, triacontanol solution concentration is 10 ~ 100 μ g/mL.
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| CN101701948A (en) * | 2009-11-23 | 2010-05-05 | 北京联合大学生物化学工程学院 | Methods for establishing fingerprints of cistanche extracts and identifying quality of drugs |
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| CN101701948A (en) * | 2009-11-23 | 2010-05-05 | 北京联合大学生物化学工程学院 | Methods for establishing fingerprints of cistanche extracts and identifying quality of drugs |
Non-Patent Citations (3)
| Title |
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| Wang, Mei-Fei;Lian, Hong-Zhen;Mao, Li.Comparison of various extraction methods for policosanol from rice bran wax and establishment of chromatographic fingerprint of policosano.《JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY》.2007,第55卷(第14期),5552-5558. * |
| 陈芳 等.气相色谱法测定糠蜡提取物中的二十八烷醇和三十烷醇.《食品科学》.2003,第24卷(第4期),实验部分第1.2-1.3节. * |
| 黄明立.毛细管气相色谱法测定多廿醇片中5种主要成分的含量.《药物分析杂志》.2007,第27卷(第8期),1168-1171. * |
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