CN113092622A - Pretreatment method for detecting content of phenolic impurities in sample - Google Patents
Pretreatment method for detecting content of phenolic impurities in sample Download PDFInfo
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- 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
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- 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
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- G01N30/8679—Target compound analysis, i.e. whereby a limited number of peaks is analysed
Abstract
The invention discloses a pretreatment method for detecting the content of phenolic impurities in a sample, which comprises the following steps: weighing a sample, placing the sample in a volumetric flask, adding an acetonitrile solution of vitamin C, dissolving the sample, shaking up, fixing the volume by using a formic acid solution of the vitamin C, filtering, and using filtrate for instrument analysis; the vitamin C acetonitrile solution and the vitamin C formic acid solution contain 90-110 mu g/ml of vitamin C. By adopting the pretreatment method, the detection limit and the quantification limit can be extremely low in the detection of the phenol impurities, and the repeatability is good; and the stability time of the reference substance solution is prolonged to 30 hours, the stability time of the standard-added recovery solution is prolonged to 11 hours, and meanwhile, the vitamin C and the phenolic target substances can be effectively separated in a corresponding chromatographic system without interfering the determination of the target compounds.
Description
Technical Field
The invention belongs to the field of chemical analysis, and particularly relates to a pretreatment method for detecting the content of phenolic impurities in a sample.
Background
Phenolic genotoxic impurities are a class of impurities that must be controlled in the evaluation of new drugs. Because phenolic substances have strong chemical activity and are easy to oxidize, the existing methods cannot accurately and effectively measure trace phenolic impurities in samples.
The existing measuring method generally uses liquid chromatography to measure the solution in which phenols are directly dissolved and diluted, after the solution is prepared, the phenol impurities can be partially oxidized into quinone products, so that the response value is increased, the measuring result is larger than a normal value, and the result deviation is larger when the solution is placed for a longer time.
Disclosure of Invention
The invention aims to provide a pretreatment method for detecting the content of phenolic impurities in a sample. The invention screens a proper reducing agent vitamin C, and the vitamin C is added in the pretreatment process of detecting the phenolic impurities, so that the oxidation of the phenolic impurities in the sample can be effectively inhibited, and the detection accuracy is improved.
The purpose of the invention is realized by the following technical scheme:
a pretreatment method for detecting the content of phenolic impurities in a sample comprises the following steps:
weighing a sample, placing the sample in a volumetric flask, adding an acetonitrile solution of vitamin C, dissolving the sample, shaking up, fixing the volume by using a formic acid solution of the vitamin C, filtering, and using filtrate for instrument analysis;
the vitamin C acetonitrile solution and the vitamin C formic acid solution, wherein the content of the vitamin C is 90-110 mu g/ml, preferably 100 mu g/ml;
preferably, the solvent of the acetonitrile solution of vitamin C is a 45-55% (volume percent) acetonitrile solution; particularly preferably, the solvent is a 50% (volume percent) acetonitrile solution;
preferably, the solvent of the formic acid solution of vitamin C is 0.05-0.15% (volume percent) formic acid solution; particularly preferably, the solvent is a 0.1% (volume percent) formic acid solution;
as the polarity of the phenolic impurities is larger, the peak is quicker, in order to reduce the solvent effect, the invention firstly adopts 50 percent acetonitrile solution of vitamin C to dissolve a sample, and then 0.1 percent formic acid solution of vitamin C is added to ensure that the diluted sample solvent is close to the initial proportion of a mobile phase, thereby achieving the purpose of reducing the solvent effect.
The phenolic impurities are 1,2, 4-benzenetriol;
the sample can be a raw material drug or a pharmaceutical preparation;
the pharmaceutical preparation can be common pharmaceutical dosage forms such as injection, tablet, suspension, capsule and the like.
The instrumental analysis is preferably liquid chromatography;
the liquid chromatographic analysis adopts an ultraviolet detector to directly sample;
the liquid chromatography analysis, some of the analysis conditions are shown in table 1:
the liquid chromatographic analysis also comprises the preparation of a reference solution, and the specific steps are as follows: dissolving the reference substance according to the same steps as the sample dissolution and constant volume, and diluting (using formic acid solution of vitamin C) into reference substance solutions with different concentrations according to the requirements of different drug determination limits.
Compared with the prior art, the invention has the following advantages and effects:
1. the pretreatment method for detecting the phenolic impurities can effectively solve the stability problem of the phenolic impurities in the determination process, and the vitamin C as a reducing agent can effectively inhibit the oxidation of the phenolic target compounds, so that the stability time of a reference substance solution is prolonged to 30 hours, the stability time of a standard-added recovery solution is prolonged to 11 hours, and meanwhile, the vitamin C and the phenolic target compounds can be effectively separated in a corresponding chromatographic system without interfering the determination of the target compounds. The method has the advantages of more accurate, stable and reliable detection result and good method durability.
2. By adopting the pretreatment method, the detection limit and the quantification limit can be extremely low in detection of the phenol impurities, and the repeatability is good.
Drawings
FIG. 1 is a liquid chromatography spectrum of an empty solvent (100. mu.g/ml vitamin C solution) in example 1.
FIG. 2 is a liquid chromatography chromatogram of phenolic impurities in the test sample of example 1 (vitamin C solution as diluent).
FIG. 3 is a liquid chromatogram of phenolic impurities (vitamin C content 90. mu.g/ml) in a control solution of example 1 (vitamin C solution as diluent).
FIG. 4 is a liquid chromatogram of phenolic impurities (vitamin C content 100. mu.g/ml) in a control solution of example 1 (vitamin C solution as diluent).
FIG. 5 is a liquid chromatogram of phenolic impurities (vitamin C content 110. mu.g/ml) in a control solution of example 1 (vitamin C solution as diluent).
FIG. 6 is a liquid chromatography chromatogram of phenolic impurities in the test sample solution of example 1 (vitamin C solution as diluent).
FIG. 7 is a liquid chromatography chromatogram of phenolic impurities in a control solution of example 1 (vitamin C in 45% acetonitrile as diluent).
FIG. 8 is a liquid chromatography chromatogram of phenolic impurities in a control solution of example 1 (vitamin C in 50% acetonitrile as diluent).
FIG. 9 is a liquid chromatography chromatogram of phenolic impurities in a control solution of example 1 (vitamin C in 55% acetonitrile as diluent).
FIG. 10 is a liquid chromatography chromatogram of 0.02mg/L phenolic impurities of the control solution of example 1 (vitamin C solution as diluent).
FIG. 11 is a liquid chromatography chromatogram of 0.05mg/L phenolic impurities of the control solution of example 1 (vitamin C solution as diluent).
FIG. 12 is a liquid chromatography chromatogram of 0.1mg/L phenolic impurities of the control solution of example 1 (vitamin C solution as diluent).
FIG. 13 is a liquid chromatography chromatogram of 0.2mg/L phenolic impurities of the control solution of example 1 (vitamin C solution as diluent).
FIG. 14 is a liquid chromatography chromatogram of 0.3mg/L phenolic impurities of the control solution of example 1 (vitamin C solution as diluent).
FIG. 15 is a liquid chromatography chromatogram of 0.4mg/L phenolic impurities of the control solution of example 1 (vitamin C solution as diluent).
FIG. 16 is a line graph of the control solution of example 1 (vitamin C solution as diluent).
FIG. 17 is a liquid chromatography chromatogram of a phenolic impurity (0.01 mg/L) in a detection limiting solution of example 1 (vitamin C solution is used as a diluent).
FIG. 18 is a liquid chromatography chromatogram of a limiting solution for the quantification of phenolic impurities (0.02 mg/L) in example 1 (vitamin C solution as diluent).
FIG. 19 is a liquid chromatography chromatogram of phenolic impurities in sample solution batch 1 of example 1 (vitamin C solution as diluent).
FIG. 20 is a liquid chromatography chromatogram of phenolic impurities in sample solution of batch 2 of example 1 (vitamin C solution as diluent).
FIG. 21 is a liquid chromatography chromatogram of phenolic impurities in sample solution batch 3 of example 1 (vitamin C solution as diluent).
FIG. 22 is a liquid chromatography chromatogram of phenolic impurities in sample solution of example 1 (vitamin C solution as diluent) batch 4.
FIG. 23 is a liquid chromatography chromatogram of phenolic impurities in sample solution of example 1 (vitamin C solution as diluent) batch 5.
FIG. 24 is a liquid chromatogram of a control solution of example 2 (vitamin C solution as diluent) set for 0h phenolic impurities.
FIG. 25 is a liquid chromatogram of a control solution of example 2 (vitamin C solution as diluent) after standing for 2h of phenolic impurities.
FIG. 26 is a liquid chromatogram of a control solution of example 2 (vitamin C solution as diluent) set aside for 4h of phenolic impurities.
FIG. 27 is a liquid chromatography chromatogram of a control solution of example 2 (vitamin C solution as diluent) after standing for 8h of phenolic impurities.
FIG. 28 is a liquid chromatogram of a control solution of example 2 (vitamin C solution as diluent) after 12h standing for phenolic impurities.
FIG. 29 is a liquid chromatography chromatogram of a control solution of example 2 (vitamin C solution as diluent) after 17h standing of phenolic impurities.
FIG. 30 is a liquid chromatogram of a control solution of example 2 (vitamin C solution as diluent) set for 30h of phenolic impurities.
FIG. 31 is a liquid chromatography chromatogram of a control solution of example 2 (vitamin C solution as diluent) after standing for 40h of phenolic impurities.
FIG. 32 is a liquid chromatography chromatogram of example 2 (vitamin C solution as diluent) spiked with the recovered solution for 0h of phenolic impurities.
FIG. 33 is a liquid chromatography chromatogram of example 2 (vitamin C solution as diluent) spiked with the recovered solution for 2 h.
FIG. 34 is a liquid chromatography chromatogram of example 2 (vitamin C solution as diluent) spiked with the recovered solution for 4 h.
FIG. 35 is a liquid chromatography chromatogram of example 2 (vitamin C solution as diluent) spiked with the recovered solution for 8 h.
FIG. 36 is a liquid chromatography chromatogram of example 2 (vitamin C solution as diluent) spiked with the recovered solution for 11 h.
FIG. 37 is a liquid chromatography chromatogram of example 2 (vitamin C solution as diluent) spiked with the recovered solution for 16 h.
FIG. 38 is a liquid chromatography chromatogram of example 2 (vitamin C solution as diluent) spiked with the recovered solution for 30 h.
FIG. 39 is a liquid chromatography chromatogram of example 2 (vitamin C solution as diluent) spiked with the recovered solution for 40 h.
FIG. 40 is a liquid chromatography spectrum of phenolic impurities in a control solution of comparative example 1 (10% sodium hydroxide solution as diluent).
FIG. 41 is a liquid chromatography chromatogram of phenolic impurities in comparative example 1 (10% sodium hydroxide solution as diluent) plus standard test sample solution.
FIG. 42 is a liquid chromatography spectrum of phenolic impurities in a blank solvent of comparative example 2 (0.3% hydrogen peroxide solution as diluent).
FIG. 43 is a liquid chromatographic chart of phenolic impurities in the test sample of comparative example 2 (0.3% hydrogen peroxide solution as diluent).
FIG. 44 is a liquid chromatography spectrum of phenolic impurities in a control solution of comparative example 2 (0.3% hydrogen peroxide solution as diluent).
FIG. 45 is a liquid chromatography chromatogram of comparative example 2 (0.3% hydrogen peroxide solution as diluent) spiked with phenolic impurities in the recovered solution.
FIG. 46 is a liquid chromatography spectrum of phenolic impurities in 0.06% hydrogen peroxide solution of comparative example 3.
FIG. 47 is a liquid chromatography spectrum of phenolic impurities in 0.15% hydrogen peroxide solution of comparative example 3.
FIG. 48 is a liquid chromatography spectrum of phenolic impurities in 0.3% hydrogen peroxide solution of comparative example 3.
FIG. 49 is a liquid chromatography chromatogram of phenolic impurities in a blank solvent of comparative example 4 (0.3% hydrogen peroxide solution as diluent with manganese dioxide catalyst added).
FIG. 50 is a liquid chromatography chromatogram of phenolic impurities in a control solution of comparative example 4 (0.3% hydrogen peroxide solution as diluent with manganese dioxide catalyst added).
FIG. 51 is a liquid chromatography chromatogram of comparative example 4 (0.3% hydrogen peroxide solution as diluent with manganese dioxide catalyst added) spiked to recover phenolic impurities in solution.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.
Example 1
The method for measuring the content of phenols impurities in 5 batches of paroxetine hydrochloride raw materials sold in the market comprises the following steps:
(1) preparation of control solutions
Precisely weighing 10mg of a 1,2, 4-benzenetriol reference substance, placing the reference substance in a 10mL volumetric flask, firstly adding 0.5mL of a 50% acetonitrile solution of vitamin C (the content is 100 mu g/mL) to dissolve the reference substance, then adding a 0.1% formic acid solution of the vitamin C (the content is 100 mu g/mL) to dilute the reference substance, fixing the volume to a scale to obtain a phenolic impurity reference substance stock solution I with the concentration of 1 mg/mL, and then diluting the reference substance I with the 0.1% formic acid solution of the vitamin C (the content is 100 mu g/mL) to prepare reference substance solutions with different concentrations according to the requirements of different medicament determination limits;
(2) sample pretreatment
And (3) testing the sample: precisely weighing 0.1g of raw material medicine powder, placing the raw material medicine powder into a 10mL volumetric flask, firstly adding 0.5mL of 50% acetonitrile solution of vitamin C (with the content of 100 mu g/mL), dissolving a sample, shaking up, fixing the volume to a scale by using 0.1% formic acid solution of the vitamin C (with the content of 100 mu g/mL), shaking up, and filtering;
adding a standard and recovering a sample: precisely weighing 0.1g of raw material medicine powder, adding a proper amount (200 mu L, 400 mu L or 600 mu L) of 1,2, 4-benzenetriol reference substance solution (the concentration is about 5 mu g/mL), putting the solution into a 10mL volumetric flask, dissolving the solution according to the same steps as the test sample, fixing the volume and filtering;
(3) conditions for liquid chromatography
The results of the liquid chromatography analysis of the phenolic impurities are shown in FIGS. 1 to 9.
The blank solvent (100. mu.g/ml of vitamin C solution) was not interfering with the detection of impurities (FIG. 1). The impurity 1,2, 4-benzenetriol is not detected in the test solution (fig. 2).
The content of vitamin C is 90 μ g/ml, and the control solution of phenol impurities with 200ng/ml has good peak shape and no interference peak at peak position at 1.492min, and the peak area is 0.043mAU min (figure 3).
The content of vitamin C is 100 μ g/ml, and the control solution of phenol impurities with 200ng/ml has good peak shape and no interference peak at peak position at 1.492min (FIG. 4).
The content of vitamin C is 110 μ g/ml, and the control solution of phenol impurities with 200ng/ml has good peak shape and no interference peak at peak position at 1.492min, and the peak area is 0.043mAU min (FIG. 5).
In the solution with standard recovery rate, the impurity peak is not affected, and is completely separated from the bulk drug, the peak area is 0.046mAU min (figure 6), the recovery rate reaches 104.5%, and the recovery rate is good.
Vitamin C45% acetonitrile solution, 200ng/ml phenol impurity control solution showed good peak shape at 1.492min, and no interference peak at peak position, peak area was 0.043mAU min (FIG. 7).
Vitamin C50% acetonitrile solution, 200ng/ml phenol impurity control solution showed good peak shape at 1.492min, and no interference peak at peak position, peak area was 0.044mAU min (FIG. 8).
Vitamin C55% acetonitrile solution, and 200ng/ml phenol impurity control solution showed good peak shape at 1.492min, and no interference peak at peak position, and the peak area was 0.042mAU min (FIG. 9).
Therefore, for the detection of the phenolic impurities, the vitamin C solution can be used as a diluent, the recovery rate is qualified, and the blank solvent has no interference.
(4) Calculation of results
(5) Linear relationship and quantitative limits of the method
Plotting the peak areas as ordinate and the concentrations of the phenolic impurities as abscissa according to the chromatograms of the control solutions with different concentrations in fig. 10-15 (fig. 16), to obtain a linear equation of the phenolic impurities of y 0.0002x +0.0009 with a correlation coefficient of 0.9996;
the detection limit of the method was estimated with a signal-to-noise ratio of 3, the detection limit of the phenolic impurities was 0.01mg/L (FIG. 17), and the quantitation limit was 0.02mg/L (FIG. 18), with a signal-to-noise ratio of 10.
(6) Actual sample detection
By applying the method and equation established in the above steps, the content of the phenolic impurities (1, 2, 4-benzenetriol) in 5 batches of commercially available raw material samples except the sample to be tested is determined, and the result shows that the content of the 1,2, 4-benzenetriol in the 5 batches of samples is between 0.01mg/L and 30mg/L (fig. 19 to fig. 23). The method has good repeatability, 6 parts of samples with the same concentration are prepared in parallel, the RSD of the determination result is 2.4 percent, different operators prepare 6 parts of the same samples in addition, and the RSD of the determination result of 12 parts of samples is 2.6 percent.
Example 2
Investigating the stability time of the phenolic impurities in the determination process, comprising the following steps:
(1) preparing a reference substance solution in the same way as in example 1;
(2) sample pretreatment, same as example 1;
(3) liquid chromatography conditions were the same as in example 1;
(4) analysis of results
After standing for different periods of time, the concentrations of phenolic impurities in the control solution and the spiked recovered solution were measured, and the results of liquid chromatography analysis are shown in fig. 24-fig. 39, table 4, and table 5:
the impurity control solution had a concentration of 197.10ng/ml measured at 0h, a concentration of 177.73ng/ml measured at 30h, an absolute value of the concentration change rate of 9.8%, and a concentration of 157.80ng/ml measured at 40h, an absolute value of the concentration change rate of 19.9%, exceeding 15%, and it was found that the stabilization time of the control solution could be extended to 30 hours by the addition of the vitamin C solution.
The impurity spiked recovery solution had a concentration of 199.10ng/ml measured at 0h, a concentration of 227.48ng/ml measured at 11h, an absolute value of the concentration change rate of 14.3%, and a concentration of 240.91ng/ml measured at 16h, an absolute value of the concentration change rate of 21.0%, exceeding 15%, thus showing that the stabilization time of the spiked recovery solution reached 11 hours by adding the vitamin C solution.
The vitamin C solution is added, so that the oxidation of the phenolic target compounds can be effectively inhibited, the stability time of the reference solution is prolonged to 30 hours, the stability time of the standard-added recovery solution is prolonged to 11 hours, and meanwhile, the vitamin C and the phenolic target compounds can be effectively separated in a corresponding chromatographic system without interfering the determination of the target compounds. The method has the advantages of more accurate, stable and reliable detection result and good method durability.
Comparative example 1 (10% sodium hydroxide solution was used as the sample diluting solvent)
A method for detecting the content of phenolic impurities in a sample comprises the following steps:
(1) preparation of control solutions
Precisely weighing 10mg of 1,2, 4-benzenetriol reference substance, placing the reference substance in a 10mL volumetric flask, dissolving the reference substance by 10 percent (mass-volume ratio; the same below) of sodium hydroxide solution, fixing the volume to scale to obtain a phenolic impurity reference substance stock solution I with the concentration of about 1 mg/mL, and preparing reference substance solutions with different concentrations by using 10 percent of sodium hydroxide solution according to the requirements of different drug determination limits.
(2) Sample pretreatment
Adding a standard and recovering a solution: precisely weighing 0.1g of medicine raw material sample powder (weighing amount of a preparation sample is calculated by the contained main component), adding 400 mu L of impurity (1, 2, 4-benzenetriol) reference substance solution (the concentration is about 5 mu g/mL), placing the solution into a 10mL volumetric flask, adding a proper amount of 10% sodium hydroxide solution, shaking up, fixing the volume to scale by using the 10% sodium hydroxide solution, and filtering;
(3) conditions for liquid chromatography
Liquid chromatogram of phenolic impurities 40-41.
It can be seen from the figure that the peak height and peak area of the phenolic impurities in the spiked recovered solution are much smaller than those of the control solution, the peak area in the control solution is 0.021mAU min (FIG. 40), the peak area in the spiked recovered solution is 0.009mAU min (FIG. 41), the recovery rate is 42.8%, the recovery rate is not good, the peak shape is not good, the response is too poor, and therefore, the 10% sodium hydroxide solution is not suitable for use as a diluent.
Comparative example 2 (0.3% hydrogen peroxide solution as sample dilution solvent)
A method for detecting the content of phenolic impurities in a sample comprises the following steps:
(1) preparation of control solutions
Precisely weighing about 10mg of 1,2, 4-benzenetriol reference substance, placing the reference substance in a 10mL volumetric flask, dissolving the reference substance by 0.3% (volume percentage) of hydrogen peroxide solution, fixing the volume to scale to obtain a phenolic impurity reference substance stock solution I with the concentration of about 1 mg/mL, and preparing reference substance solutions with different concentrations by 0.3% of hydrogen peroxide according to the requirements of different drug determination limits.
(2) Sample pretreatment
And (3) testing the sample: precisely weighing about 0.1g of medicine raw material sample powder (weighing amount of preparation sample is calculated by main components contained), placing the medicine raw material sample powder into a 10mL volumetric flask, adding a proper amount of 0.3% (volume percentage; the same below) hydrogen peroxide solution, shaking up, fixing the volume to scale by 0.3% hydrogen peroxide, and filtering;
adding a standard and recovering a solution: precisely weighing about 0.1g of medicine raw material sample powder (weighing amount of preparation sample is calculated by main components contained in the preparation sample), placing the medicine raw material sample powder into a 10mL volumetric flask, adding 400 μ L of impurity (1, 2, 4-benzenetriol) reference solution (the concentration is about 5 μ g/mL), adding a proper amount of 0.3% hydrogen peroxide solution, shaking up, fixing the volume to scale by 0.3% hydrogen peroxide, and filtering;
(3) conditions for liquid chromatography were the same as in comparative example 1
Liquid chromatogram 42-45 of phenolic impurities is shown.
The blank solvent and the sample both have impurity detection signals (figure 42 and figure 43), the impurity responses in the reference solution and the standard-added recovery solution are equivalent, the peak area in the reference solution is 1.057mAU min (figure 44), the peak area in the standard-added recovery solution is 1.005mAU min (figure 45), the recovery rate is 95.1%, and the recovery rate is good. However, 0.3% hydrogen peroxide solution is not suitable because of its interference.
Comparative example 3 (different concentrations of hydrogen peroxide solution for the blank solvent)
A method for detecting the content of phenolic impurities in a sample comprises the following steps:
(1) preparation of blank solvent
Solutions of 0.06%, 0.15%, 0.3% (volume percent) hydrogen peroxide were formulated to examine the interfering effects of low concentrations of hydrogen peroxide blank solvent.
(2) Conditions for liquid chromatography were the same as in example
Liquid chromatography of the blank solvent is shown in FIGS. 46-48.
0.06%, 0.15% and 0.3% hydrogen peroxide solutions all have impurity detection signals, and the detection of an ultraviolet spectrophotometer shows that the hydrogen peroxide has signals under the wavelength of 200 nm-400 nm, so that the hydrogen peroxide solution is not applicable.
Comparative example 4 (0.3% hydrogen peroxide solution as diluent with manganese dioxide catalyst)
A method for detecting the content of phenolic impurities in a sample comprises the following steps:
(1) preparation of control solutions
Precisely weighing 10mg of 1,2, 4-benzenetriol reference substance, placing the reference substance in a 10mL volumetric flask, dissolving the reference substance by 0.3 percent (volume percentage) of hydrogen peroxide solution, fixing the volume to a scale to obtain a phenolic impurity reference substance stock solution I with the concentration of 1 mg/mL, adding 50mg of manganese dioxide (5 g/L), and preparing reference substance solutions with different concentrations by 0.3 percent of hydrogen peroxide according to the requirements of different drug determination limits.
(2) Sample pretreatment
Precisely weighing 0.1g of medicine raw material sample powder (the weighing amount of a preparation sample is calculated by the contained main component), placing the medicine raw material sample powder into a 10mL volumetric flask, adding a proper amount of 0.3% hydrogen peroxide solution, shaking up, adding 50mg of manganese dioxide, fixing the volume to the scale by 0.3% hydrogen peroxide, and filtering;
(3) conditions for liquid chromatography
Liquid chromatogram 49-FIG. 51.
As can be seen, the blank solvent still had interference after the addition of manganese dioxide (FIG. 49), and the impurity responses in the control solution and the spiked recovery solution were decreased (FIGS. 50 and 51), and it was difficult to achieve the analytical sensitivity, so that it was not suitable.
And (4) conclusion: the 0.3% hydrogen peroxide solution is used as a diluent, and manganese dioxide is added, so that the sensitivity is greatly reduced, and the method is not suitable for use.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. A pretreatment method for detecting the content of phenolic impurities in a sample is characterized by comprising the following steps:
weighing a sample, placing the sample in a volumetric flask, adding an acetonitrile solution of vitamin C, dissolving the sample, shaking up, fixing the volume by using a formic acid solution of the vitamin C, filtering, and using filtrate for instrument analysis;
the vitamin C acetonitrile solution and the vitamin C formic acid solution contain 90-110 mu g/ml of vitamin C.
2. The pretreatment method according to claim 1, wherein: the vitamin C acetonitrile solution and the vitamin C formic acid solution contain 100 mu g/ml of vitamin C.
3. The pretreatment method according to claim 1, wherein:
the solvent of the acetonitrile solution of the vitamin C is 45-55 percent (volume percentage) of acetonitrile solution;
the solvent of the formic acid solution of the vitamin C is 0.05-0.15% (volume percentage) formic acid solution.
4. The pretreatment method according to claim 1, wherein: the solvent is 50% (volume percentage) acetonitrile solution of vitamin C.
5. The pretreatment method according to claim 1, wherein: the solvent of the formic acid solution of the vitamin C is 0.1 percent (volume percentage) of formic acid solution.
6. The pretreatment method according to claim 1, wherein: the phenolic impurity is 1,2, 4-benzenetriol.
7. The pretreatment method according to claim 1, wherein: the sample is a raw material drug or a pharmaceutical preparation.
8. The pretreatment method according to claim 1, wherein: the instrumental analysis adopts liquid chromatography.
9. The pretreatment method according to claim 8, wherein: the liquid chromatographic analysis adopts an ultraviolet detector and directly samples.
10. The pretreatment method according to claim 8, wherein: the analysis conditions of the liquid chromatographic analysis are shown in the table 1:
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