GB 29382-2012Mesotrione technical material (English PDF)

4.2 Sampling Carried out in

GB/T 1605-2001 of "original drug product sampling" approach. Determining Sample packages with a random number table method; final sample size Not less than 100g.

4.3 Identification Test Infrared spectroscopy

--- mesotrione sample and standard ~ 400cm-1 in the infrared range of 1 4000cm-absorption spectrum should not expressly Significant difference. Mesotrione standard infrared spectra of Figure 1. FIG 1 mesotrione standard infrared spectrum of FIG. The present liquid chromatography

--- identification test can be conducted simultaneously with the determination of one mass fraction sulcotrione nitrate. Chromatography under the same operating conditions, Main peaks of the sample solution and the retention time of the solution of the mesotrione sample peak retention time scale, the relative difference of 1.5% should be Or less. Determination of mass sulcotrione nitro ketone score 4.4

4.4.1 Method summary After drying in vacuo a sample was dissolved in mobile phase, acetonitrile and water (phosphate adjusted to pH 3) as the mobile phase, used as a filler C

18 Stainless steel column and a UV detector at a wavelength of 270nm, the sample of mesotrione reverse phase high performance liquid chromatography and determination, External standard.

4.4.2 Reagents and solutions Acetonitrile. HPLC grade; Water. The new secondary steam distilled water; Phosphoric acid; Mesotrione Sample. a known mass fraction, w>=99.0%.

4.4.3 Instruments HPLC. a variable wavelength UV detector; Chromatographic data processor or chromatography workstation; Column. 250mm × 4.6mm (id) stainless steel column, C18,5µm filler contents (or column with equivalent effect); Filter. membrane pore size of about

0.45 m; Microsyringe. 50µL; Quantitative sampling tube. 5µL; Ultrasonic cleaning.

4.4.4 HPLC operating conditions Mobile phase. [Psi] [acetonitrile. water (adjusted to pH 3 phosphate to)] = 40.60, by membrane filtration, and degassing; Flow rate. 1.0mL/min; Column temperature. room temperature (temperature change should be less than 2 deg.] C); Detection wavelength. 270nm; Injection Volume. 5µL; Retention time. mesotrione about 10.1min. Are typical operating parameters of the above, according to the characteristics of different instruments, for a given operating parameters appropriately adjusted to obtain the best result. Typical mesotrione HPLC TC shown in Figure 2. 1

--- mesotrione. Figure 2 HPLC mesotrione original drug

4.4.5.1 Preparation of standard solution

0.1 g of weighed (accurate to 0.0002g) mesotrione standard in 50mL volumetric flask, dilute to the mark with acetonitrile, ultrasonic oscillation 5min to dissolve the sample, cooled to room temperature, shaking. Pipette with a pipette 5mL above solution in 50mL volumetric flask, dilute with the mobile phase Release to the mark.

4.4.5.2 Preparation of sample solution Weigh mesotrione containing

0.1 g of (accurate to 0.0002g) sample had been dried (drying method of section 4.5.2) in a 50mL volumetric flask , Washed with acetonitrile to volume, to dissolve the sample ultrasonic vibration 5min, cooled to room temperature, shaking. Pipette with a pipette to the solution 5mL, 50mL volumetric flask, diluted in phase with the flow to the mark.

4.4.5.3 Determination Under the above operating conditions, after the instrument is stable, the number of consecutive doses of the standard solution is injected until the two adjacent needles mesotrione peak area becomes relatively After of less than 1.5%, measured according to the standard solution, sample solution, the sample solution, the order of standard solution.

4.4.5.4 computing The two needle and the sample solution was measured before and after the sample solution two needle mesotrione standard peak area are averaged. Mesotrione sample Oxadiazon mass fraction according to formula (1). w1 = A2 · M1 · w A1 · m2 (1) Where. W1

--- mesotrione sample mass fraction in%; A2

--- sample solution, nitro sulcotrione one average peak area; M1

--- mesotrione standard mass in grams (G); W

--- the mass fraction mesotrione standard sample, expressed in%; A1

--- the standard solution, mesotrione average peak area;

--- M2 sample mass, in grams (g).

4.4.6 allows poor Ketone mass difference mesotrione Results replicates grass two fractions should be not more than 1.5%, and the arithmetic mean value as a measurement result.

4.5 Determination of loss on drying

4.5.1 Instruments Vacuum pump; Vacuum oven. 60 °C ± 2 °C, exemplar vacuum pressure. -90kPa ± 5kPa.

4.5.2 Determination Step Weigh 10g weighing bottle with the sample constant weight (accurate to 0.01g), paving, placed in a vacuum oven. Vacuum pump was turned on 60 °C ± 2 °C, exemplar vacuum dried to constant weight (about 1h) -90kPa ± 5kPa depressed condition in a desiccator, cooled to room temperature, Weighed.

4.5.3 computing Loss on drying the sample by the formula (2). w2 = m0-m1m × 100 (2) Where.

--- w2 of the sample mass fraction of drying loss, expressed in%; Mass M0

--- weighing bottle and the sample in grams (G); Mass M1

--- dried weighing bottle and the sample in grams (G);

--- m sample mass, in grams (g). Determination of the mass fraction of

4.6 Nitro thioxanthone xanthone

4.6.1 Method summary Sample was dissolved in acetonitrile, using a liquid chromatography tandem quadrupole mass spectrometer, using MRM mode, the sample nitro xanthones Tandem mass spectrometry for detection, based on the extracted ion chromatogram obtained external standard.

4.6.2 Reagents and solutions Acetonitrile. HPLC grade; Water. The new secondary steam distilled water; Ammonium acetate; Ammonium acetate solution [c (CH3COONH4) = 5mmol/L]. 0.375g weighed (accurate to 0.0002g) ammonium acetate in a 1L capacity Volumetric flask, dilute to the mark with water, shake, filter; Mesotrione sample. no nitro xanthone; Nitro xanthone Sample. a known mass fraction, w>=90.0%.

4.6.3 Instruments Liquid chromatography - tandem quadrupole mass spectrometer; Column. 150mm × 4.6mm (id) stainless steel column, C18,5µm filler contents (or column with equivalent effect); Filter. membrane pore size of about

0.22 m; Autosampler. 5µL; Ultrasonic cleaning.

4.6.4 HPLC operating conditions Mobile phase. [Psi] (acetonitrile. ammonium acetate solution) = 50.50, by membrane filtration, and degassing; Flow rate. 0.5mL/min, 5.5min ~ 10min into the mass; Column temperature. 40 °C; Injection Volume. 5µL; Retention time. nitro xanthone about 7.2min.

4.6.5 Mass operating conditions Ion source. APCI (-); Source temperature. 130 °C; Desolvation temperature. 550 °C; Cone voltage. 20V; Collision gas. argon; Collision energy. 30V; Scan mode. MRM, parent ion 344; 207 ions; The above are typical operating parameters, depending on the instrument type may, for a given operating parameters appropriately adjusted to obtain the best result. Typical mesotrione TC extracted ion chromatograms shown in Figure 3. 1

--- nitro xanthone. LC extracted ion chromatograms of FIG. 3 mesotrione TC nitro xanthone

4.6.6 Determination Step

4.6.6.1 Preparation of working standard Preparation of standard solution. Weigh 0.01g (accurate to 0.0002g) xanthone-nitro-Standard in 100mL volumetric flask to volume with acetonitrile To the mark to dissolve the sample ultrasonic vibration 5min, cooled to room temperature, shake (solution 1). Pipette solution 1mL to 10mL pipetted Volumetric flask, dilute to the mark with acetonitrile, shake (solution 2). Then pipette 5mL aliquot solution in 50mL volumetric flask with acetonitrile Dilute to volume, mix (solution 3). Weigh 0.5g (accurate to 0.0002g) containing no nitro xanthone mesotrione sample in 10mL stoppered vial, added with a pipette 3 into 5mL solution, to dissolve the sample ultrasonic vibration 5min, cooled to room temperature, shaken and filtered to give standard solution.

4.6.6.2 Preparation of sample solution Weigh 0.5g (accurate to 0.0002g) mesotrione sample in 10mL stoppered vials, with a pipette 5mL of acetonitrile was added, over To dissolve the sample acoustic shock 5min, cooled to room temperature, shaken and filtered.

4.6.6.3 Determination Under the above operating conditions, after the instrument is stable, the number of consecutive doses of the standard solution is injected until the extracted ion chromatogram two adjacent needles nitro xanthene Thioxanthone peak area relative changes of less than 30%, measured according to the standard solution, sample solution, the sample solution, the order of standard solution.

4.6.6.4 computing The two needle and the sample solution was measured before and after the two doses of the standard sample solution nitro xanthone peak area are averaged. Sample Xanthone content nitro peaks by formula (3) is calculated. w3 = A2 · M1 · w 2000A1 · m2 × 106 (3) Where.

--- the mass fraction of the sample W3 nitro xanthone peaks to in mg/kg; A2

--- sample solution, nitro xanthone average peak area; M1

--- nitro xanthone standard mass in grams (G); W

--- the mass fraction nitro xanthone standard sample, expressed in%;

--- 2000 dilution; A1

--- the standard solution, nitro xanthone average peak area;

--- M2 sample mass, in grams (g).

4.6.7 allows poor Nitro ketone mass xanthene parallel relative deviation measurement result two fractions should be not more than 30%, and the arithmetic mean value as a measurement result.

4.7 pH value measured Carried out in GB/T 1601.

4.8 Determination of acetone insolubles 5g weighed (accurate to 0.01g) sample, according to