Some standard content:
Chemical Industry Standard of the People's Republic of China
HG2613—94
Paclobutrazol technical
Published on April 4, 1994
Ministry of Chemical Industry of the People's Republic of China
Implementation on October 1, 1994
Chemical Industry Standard of the People's Republic of China
Paclobutrazol technical
Other names, structural formulas and basic physicochemical parameters of paclobutrazol are as follows: ISO common name: Paclobutrazol
CIPAC digital code: 445
HG2613—94
Chemical name: (2RS, 3RS)-1-(4-chlorophenyl)-4,4-dimethyl-2-(1H-1,2,4-triazol-1-yl)-pentanol-3-ol Structural formula:
Empirical formula: C16H20CIN0
Relative molecular mass: 293.5 (according to the 1989 international relative atomic mass) Biological properties: plant growth regulator and fungicide Melting point: 165~166℃
Vapor pressure (20℃): 1×10-6Pa
Solubility (20℃): water 35mg/L, methanol 150g/L, acetone 110g/L, cyclohexanone 180g/L, propylene glycol 50g/L, dichloromethane 100g/L, hexane 10g/L, xylene 60g/L. Stability: It can be stored for more than 2 years at 20℃; it is stable at 50℃ for 6 months. It is stable to acid and alkali. Subject content and scope of application
This standard specifies the technical requirements, test methods, inspection rules, and marking, packaging, transportation and storage of paclobutrazol original drug. This standard applies to paclobutrazol original drug composed of paclobutrazol and impurities generated in its production, and should not contain added modifiers. 2 Reference standards
GB/T601 Preparation of standard solution for titration analysis GB/T603 Preparation of preparations and products used in test methods GB/T1604 Acceptance rules for pesticides
GB/T1605 Sampling methods for commercial pesticides
GB3796 General rules for pesticide packaging
3 Technical requirements
3.1 Appearance: white to light yellow solid.
3.2 Paclobutrazol original drug should meet the following index requirements. Approved by the Ministry of Chemical Industry of the People's Republic of China on April 4, 1994, and implemented on October 1, 1994
Paclobutrazol content,
Loss on drying,
Acetone insoluble matter\)
Acidity (in HSO.)
Note: 1) Test at least once every three months.
4 Test method
4.1 Determination of paclobutrazol technical
4.1.1 Identification test
HG2613-94
Superior product
%(m/m)
Qualified product
This identification test can be carried out simultaneously with the determination of paclobutrazol content. The retention time of the main peak of the sample solution and the retention time of paclobutrazol of the standard solution under the same conditions should have a deviation within 1.5%. 4.1.2 Determination of paclobutrazol content
4.1.2.1 High performance liquid chromatography internal standard method (arbitration method) 4.1.2.1.1 Method summary
The sample was dissolved in anhydrous ethanol, diethyl fumarate (diethyl maleate) was used as the internal standard, a mixture of methanol, acetonitrile and water was used as the mobile phase, and paclobutrazol was separated and determined by high performance liquid chromatography using a Nova-pakC1 column and a variable wavelength ultraviolet detector. 4.1.2.1.2 Reagents and solutions
Paclobutrazol standard: known content, ≥99.0% (m/m); internal standard: diethyl fumarate, must not contain impurities that interfere with the analysis; methanol (GB/T683): analytical grade, redistilled; acetonitrile (GB/T3329): analytical grade, redistilled or HPLC grade; anhydrous ethanol (GB/T678): analytical grade;
water, double distilled water;
internal standard solution: weigh 0.200g (accurate to 0.0002g) of diethyl fumarate in a 500mL volumetric flask, dilute to the scale with anhydrous ethanol, and shake well.
4.1.2.1.3 Instruments
High performance liquid chromatograph: with variable wavelength UV detector; Chromatographic column: stainless steel column, filled with Nova-pak C18 4μm, 150×4.6mm (ID); Data processor;
Ultrasonic degassing device;
Micro syringe: 25uL.
4.1.2.1.4 Chromatographic conditions
Mobile phase: methanol + ethyl + water = 46+18+36 (V/V); Flow rate: 1.0mL/min;
Column temperature: room temperature;
Measurement wavelength: 225nm
Injection volume: 20uL;
Retention time: internal standard, about 3.4min; paclobutrazol 1, about 4.2min;
paclobutrazol, about 5min.
HG2613—94
The above operating conditions are typical operating parameters. According to different instruments and chromatographic columns, the given parameters can be appropriately adjusted to obtain the best separation effect.
4.1.2.1.5 Operation steps
a. Preparation of standard solution
Weigh about 0.100g (accurate to 0.0002g) of paclobutrazol standard into a 100mL volumetric flask, dissolve it with anhydrous ethanol, dilute to the scale, and shake well. Then use a pipette to transfer 5mL of standard solution and 5mL of internal standard solution into the same 25mL volumetric flask, dilute to the scale with anhydrous ethanol, and shake well.
b. Preparation of sample solution
Weigh a sample containing about 0.100g (accurate to 0.0002g) of paclobutrazol into a 100mL volumetric flask, dissolve it with anhydrous ethanol, dilute to the mark, and shake well. Use a pipette to accurately transfer 5mL of sample solution and 5mL of internal standard solution into the same 25mL volumetric flask, dilute to the mark with anhydrous ethanol, and shake well.
c. Determination
Under the above operating conditions, after the instrument is stable, continuously inject several needles of standard solution until the relative response value of two adjacent needles changes by less than 1.5%, and then analyze in the following order: standard solution;
b. sample solution;
c. sample solution;
standard solution.
Figure 1 HPLC chromatogram of paclobutrazol technical (internal standard method) 1-Solvent; 24H-paclobutrazol; 3-Internal standard; 4-paclobutrazol 1; 5-paclobutrazol; 6-1H-chlorazolone
4.1.2.1.6 Calculation
Calculate the average value of the peak area ratio of paclobutrazol to internal standard in the two sample solutions and the two standard solutions before and after the sample, respectively. The mass percentage of paclobutrazol in the sample is X, according to the formula (1) Calculation: 3
HG2613-94
X,=2miP
Wherein: --- the average value of the peak area ratio of paclobutrazol to the internal standard in the standard solution; ↑2 -- the average value of the peak area ratio of paclobutrazol to the internal standard in the sample solution; m1 -- the mass of the standard,;
m2 -- the mass of the sample, 8;
P -- the percentage content of the standard, (m/m). 4.1.2.1.7 Allowable difference
The difference between the results of two parallel determinations should not exceed 1.5%. 4.1.2.2 High performance liquid chromatography external standard method
4.1.2.2.1 Method summary
The sample is dissolved in methanol, and a mixture of methanol, acetonitrile and water is used as the mobile phase. The Nova-pak C18 column and a variable wavelength ultraviolet detector are used to separate and determine paclobutrazol by high performance liquid chromatography. 4.1.2.2.2 Reagents
Paclobutrazol standard sample: known content, >99.0% (m/m); methanol (GB/T623): analytical grade, redistilled; acetonitrile (GB/T3329): analytical grade;
Water: double distilled water.
4.1.2.2.3 Instruments
High performance liquid chromatograph: with variable wavelength UV detector; chromatographic column: stainless steel column, filled with Nova-pak C18, 4um, 150×4.6mm (ID); data processor;
ultrasonic degasser;
micro syringe: 25uL.
4.1.2.2.4 Liquid chromatography operating conditions
Mobile phase: methanol + acetonitrile + water = 46 + 18 + 36 (V/V); flow rate: 1.0mL/min;
Column temperature: room temperature;
Measurement wavelength: 230nm;
Injection volume: 20uL;
Retention time, 4H-paclobutrazol, about 2.8min; paclobutrazol I, about 4.2min; paclobutrazol, about 5min; 1H-chlorazine, about 5.8min. The above operating conditions are typical operating parameters. According to different instruments and chromatographic columns, the given parameters can be appropriately adjusted to obtain the best separation effect.
4.1.2.2.5 Operation stepswww.bzxz.net
a. Preparation of standard solution
Weigh about 0.100g (accurate to 0.0002g) of paclobutrazol standard sample into a 100mL volumetric flask, dissolve it with methanol and dilute it to the mark, and shake it well. Use a 5mL pipette to accurately transfer 5mL of the solution into another 25mL volumetric flask, dilute it with the mobile phase to the mark, and shake it well. b. Preparation of sample solution
Weigh about 0.100g (accurate to 0.0002g) of paclobutrazol sample into a 100mL volumetric flask, dissolve it with methanol, dilute it to the mark, and shake it well. Use a 5mL pipette to accurately transfer 5mL of the sample solution into another 25mL volumetric flask, dilute it with the mobile phase to the mark, and shake it well.
Under the above operating conditions, after the instrument is stable, continuously inject several needles of standard solution. After the response value change between two adjacent needles is less than 1.5%4
, perform the sample analysis in the following order:
a. Standard solution;
Test solution;
c. Test solution,
Standard solution.
HG2613-94
Figure 2 High Performance Liquid Chromatography of Paclobutrazol Technical
1—Solvent; 2—4H-paclobutrazol; 3—paclobutrazol 1; 4—paclobutrazol; 6—1H-chlorazolone 4.1.2.2.6 Calculation
Calculate the average value of the paclobutrazol peak area in the two standard solution and the two sample solutions, and the mass percentage of paclobutrazol in the sample X2, according to formula (2):
In the formula: A1, A2
ms, m4
are the average values of the paclobutrazol peak area in the two standard solution and the sample solution; a is the mass of the standard and the sample, g; p
a is the percentage of the standard, (m/m).
4.1.2.2.7 Allowable Difference
The difference between the results of two parallel determinations should not be greater than 1.5%. 4.2 Determination of loss on drying
Use a weighing bottle with known weight to weigh 5 grams (accurate to 0.0002g) of the sample, flatten it to a thickness of no more than 5mm, place it in an oven at 105±2℃ to dry for 2h, then take it out and put it in a desiccator, cool it to room temperature, and weigh it. Loss on drying X expressed as mass percentage: Calculate according to formula (3): Xa-
Where: a——mass of weighing bottle and sample, g; gb——mass of weighing bottle and sample after drying, g; gb
m——mass of sample, g.
4.3 Determination of acetone insoluble matter
4.3.1 Reagents
Acetone (GB/T686): Dry it over anhydrous sodium sulfate. 4.3.2 Apparatus
Erlenmeyer flask: with ground glass joint, 250mL; Reflux condenser: matched with the flask;
Gooch crucible or glass sand core crucible: No. 3;
4.3.3 Determination steps
HG2613-94
Weigh 5g of sample (accurate to 0.01g), put it into an Erlenmeyer flask, add 150mL of acetone and heat to reflux until all soluble substances are dissolved. Filter out the solution with a crucible that has reached a constant weight, then wash the Erlenmeyer flask with 60mL of acetone three times, and filter it by suction. Dry it in an oven at 110℃ for 30min, take it out and cool it to room temperature and weigh it. The mass percentage X of acetone insoluble matter is calculated according to formula (4): m1-mo×100
Wherein: m1——the mass of the crucible and the insoluble matter after constant weight, g; mo—the mass, g;
m—the mass of the sample.
4.4 Determination of acidity
4.4.1 Reagents and solutions
Acetone (GB/T686): analytical grade;
Sodium hydroxide (GB/T629): standard titration solution c(NaOH)=0.02mol/L, prepared and calibrated according to GB/T601 and then diluted.
Methyl red (HG/T3-958): 2g/L methyl red ethanol solution. 4.4.2 Operation steps
Weigh 2~5g (accurate to 0.01g) of sample, place in a 250mL conical flask, add 25mL acetone. After the sample is dissolved, add 75mL distilled water, cover with a stopper, shake vigorously, add 3~4 drops of indicator, and titrate with sodium hydroxide standard solution until the color changes from red to yellow. At the same time, perform a blank determination. The acidity X of the sample is expressed in mass percentage. Calculate according to formula (5): Xs=c(V1-V)×0. 049)
Where: c——actual concentration of sodium hydroxide standard solution, mol/L; V. ——Volume of sodium hydroxide standard solution consumed in blank test, mL; V1——Volume of sodium hydroxide standard solution consumed in test, mL; m
-Mass of sample, g;
(5)
-Mass of sulfuric acid in grams equivalent to 1.00mL sodium hydroxide standard titration solution Cc(NaOH)=1.000mol/LJ.
5 Inspection rules
5.1 Sampling method
Sampling shall be carried out in accordance with the "technical drug sampling" method in GB/T1605. The sampling package shall be determined by random method, and the final sampling volume shall be not less than 250g.
5.2 Acceptance rules
Acceptance shall be carried out in accordance with GB/T1604 standard.
Marking, packaging, transportation and storage
HG2613—-94
6.1 The marking and packaging of paclobutrazol technical shall comply with the relevant provisions of GB3796 and shall be marked with trademarks. 6.2 Paclobutrazol technical shall be packed in double-layer plastic bags and iron drums, with a net weight of 50kg per piece. If the user requires packaging of other specifications, it shall be agreed upon by both parties, but it shall comply with the relevant requirements of GB3796. 6.3 During transportation and storage, it must be protected from moisture and sunlight, maintained in good ventilation, and must not be mixed with food, feed, and seeds. Avoid contact with skin and eyes, and prevent inhalation through the mouth and nose.
6.4 Warranty period: Under the specified storage and transportation conditions, the warranty period of paclobutrazol technical is two years from the date of production. 7
A1 Method Summary
HG2613—94
Appendix A
Gas chromatography determination of paclobutrazol content
(Supplement)
The sample was dissolved in chloroform, dicyclohexyl phthalate was used as the internal standard, and paclobutrazol was separated and determined by gas chromatography using a stainless steel column filled with ChromosorbWAW-DMCS treated with FFAP composite cross-linking and a hydrogen flame ionization detector.
Reagents and solutions
Paclobutrazol standard: known content, >99.0% (m/m); internal standard: dicyclohexyl phthalate, must not contain impurities that interfere with the analysis; stationary phase: ChromosotbWAW-DMCS treated with FFAP composite cross-linking bond, particle size 150180um Chloroform (GB/T682): analytical grade;
Hydrogen: deoxygenated;
Internal standard solution: weigh 11g of dicyclohexyl phthalate in a 1000mL volumetric flask, dissolve it with chloroform, dilute to the scale, and shake well.
A3 Instruments
Gas chromatograph: with hydrogen flame ionization detector; Chromatographic column: 1000X2mm(1.0) stainless steel column, filled with ChromosorbWAW-DM-CS stationary phase treated with FFAP composite cross-linking bonding;
Data processor;
Micro-injector: 10uL.
A4 Preparation of Chromatographic Column
a. Filling of Chromatographic Column
Connect a glass funnel to the inlet of the clean and dry chromatographic column, and a joint with a screen to the outlet, and connect it to the vacuum pump with a rubber tube. Turn on the vacuum pump, slowly and evenly add the prepared filler from the funnel end, and keep tapping the column wall up and down with rubber to make it tightly and evenly filled (about 1g can be loaded). Remove the chromatographic column, plug a small ball of silanized glass wool at both ends of the column, and bend it into a disc shape. b.
Aging of the chromatographic column
Connect the inlet of the chromatographic column to the vaporization chamber, and do not connect the outlet to the detector for the time being. Replace the air in the column with 80mL/min of nitrogen. After 1 hour, increase the temperature to 210℃ at 20mL/min of nitrogen and 3℃/min, and maintain it for 24 hours. After cooling, connect the outlet of the column to the detector. Gas chromatography operating conditions
Temperature: 200℃ for column chamber, vaporization chamber, and detection chamber; Gas flow: carrier gas (high-purity H2), 42mL/min; Tail gas: nitrogen, 28mL/min;
Air, 500mL/min;
Injection volume: about 0.6uL;
Retention time: paclobutrazol, about 10min;
paclobutrazol I, about 12.3min;
Dicyclohexyl phthalate, about 14.5min. HG2613—-94
The above operating conditions can be adjusted appropriately according to different instruments and chromatographic columns to obtain the best results. A6 Determination steps
a. Preparation of standard solution
Weigh 0.090g (accurate to 0.0002g) of paclobutrazol standard sample into a stoppered glass bottle, accurately add 10mL of internal standard solution with a pipette, and shake well.
Preparation of sample solution
Weigh a sample containing about 0.090g (accurate to 0.0002g) of paclobutrazol into a stoppered glass bottle, accurately add 10mL of internal standard solution with a pipette, and shake well.
Under the above operating conditions, after the instrument is stable, inject several needles of standard solution continuously. After the relative response value of two adjacent needles changes less than 1.5%, inject and analyze in the following order: standard solution;
b. sample solution;
c. sample solution;
standard solution.
Figure 3 Gas chromatogram of paclobutrazol original drug
1 solvent; 2-chlorazine; 3-paclobutrazol; 4-paclobutrazol 1; 5-dicyclohexyl phthalate A7 Calculation
Calculate the average value of the peak area ratio of paclobutrazol to internal standard in the two needles of sample solution and the two needles of standard solution before and after the sample, and the mass percentage content X of paclobutrazol in the sample. Calculate according to formula (A1): 9
HG2613—94
Xg--T2mip
Wherein: r1——the average value of the peak area ratio of paclobutrazol to the internal standard in the standard solution; T2——the average value of the peak area ratio of paclobutrazol to the internal standard in the test solution; m1——the mass of the standard, g,
m2——the mass of the test sample, g;
p——the percentage content of the standard (m/m).
Allowable difference
The difference between the results of two parallel determinations should not exceed 1.5%. Additional remarks:
This standard was proposed by the Technical Supervision Department of the Ministry of Chemical Industry. This standard is under the technical jurisdiction of the Shenyang Chemical Industry Research Institute of the Ministry of Chemical Industry. This standard was drafted by the Jiangsu Pesticide Research Institute. The main drafters of this standard are Yu Youfen, Yin Shangzheng, Liang Qinying, Xu Xiangsheng, and Qiao Zhengfu. 10
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