CN118362665B - Method for measuring multi-component content of infant throat-clearing particles - Google Patents
Method for measuring multi-component content of infant throat-clearing particles Download PDFInfo
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- arctiin
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- 238000000034 method Methods 0.000 title claims abstract description 37
- 239000002245 particle Substances 0.000 title claims description 15
- WBYWAXJHAXSJNI-VOTSOKGWSA-M .beta-Phenylacrylic acid Natural products [O-]C(=O)\C=C\C1=CC=CC=C1 WBYWAXJHAXSJNI-VOTSOKGWSA-M 0.000 claims abstract description 36
- WBYWAXJHAXSJNI-SREVYHEPSA-N Cinnamic acid Chemical compound OC(=O)\C=C/C1=CC=CC=C1 WBYWAXJHAXSJNI-SREVYHEPSA-N 0.000 claims abstract description 36
- 229930016911 cinnamic acid Natural products 0.000 claims abstract description 36
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- 239000013558 reference substance Substances 0.000 claims abstract description 24
- 238000001514 detection method Methods 0.000 claims abstract description 23
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- 238000004458 analytical method Methods 0.000 description 5
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- 238000012360 testing method Methods 0.000 description 3
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- 206010068319 Oropharyngeal pain Diseases 0.000 description 2
- UILPJVPSNHJFIK-UHFFFAOYSA-N Paeonol Chemical compound COC1=CC=C(C(C)=O)C(O)=C1 UILPJVPSNHJFIK-UHFFFAOYSA-N 0.000 description 2
- 201000007100 Pharyngitis Diseases 0.000 description 2
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- QQILFGKZUJYXGS-UHFFFAOYSA-N Indigo dye Chemical compound C1=CC=C2C(=O)C(C3=C(C4=CC=CC=C4N3)O)=NC2=C1 QQILFGKZUJYXGS-UHFFFAOYSA-N 0.000 description 1
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- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention provides a multi-component content determination method of infant throat-clearing granules, and relates to the technical field of traditional Chinese medicine detection. The determination method adopts an HPLC method to determine, and chromatographic conditions are as follows: ZORBAX SB-C18 column, mobile phase: the A phase is acetonitrile-B phase 0.1% phosphoric acid, gradient elution is carried out, the flow rate is 1.0mL/min, the column temperature is 35 ℃, and the detection wavelength is 260-290nm; the sample injection amount is 5-15 mu L; precisely sucking the stock solution of the mixed reference substance and the solution of the sample, respectively measuring the areas of the forsythin, the arctiin and the cinnamic acid under the chromatographic conditions, and calculating to obtain the product. The method is simple and convenient to operate, can rapidly and stably separate and detect the content of the forsythin, the arctiin and the cinnamic acid within 35min, has good separation degree, has a linear relation r of more than or equal to 0.9998, has average sample recovery rate of more than 95%, improves the current standard, and can be used for quality control of the infant throat clearing granules.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicine detection, and particularly relates to a multi-component content determination method of infant throat-clearing granules.
Background
The granule for clearing heat from throat of children comprises radix scrophulariae, herba Taraxaci, fructus Arctii (parched), herba Menthae, periostracum Cicadae, radix Isatidis, fructus forsythiae, cortex moutan, and indigo naturalis, and adjuvant such as sucrose and dextrin, has effects of clearing heat from exterior, removing toxic substance and relieving sore throat, and can be used for treating fever headache, cough hoarseness, and sore throat caused by infantile exogenous wind-heat. However, the infant throat-clearing granule is not yet recorded in pharmacopoeia at present, the quality standard is the ministerial standard (WS 3-B-1092-92), only the thin-layer chromatography identification of the burdock fruit and the natural indigo 2 medicinal materials is contained, the content measurement of characteristic components is lacked, and the quality control is greatly improved.
The literature' content determination of 8 components in the infant throat-clearing granule and main component analysis [ J ] "(Zhang Bo, zhixue branch, tian Lan, etc.. In south China pharmacy, 2023, 21 (11): 3008-3013.) establishes a high performance liquid chromatography method for simultaneously determining the content of harpagoside, (R, S) -epigoitrin, forsythoside A, chicoric acid, forsythin, arctiin, harpagoside and paeonol in the infant throat-clearing granule, but the analysis time of the method is longer, the whole course analysis is more than 80 minutes, and the separation degree between peaks needs to be further improved; the HPLC method is established in the literature to simultaneously measure 6 components [ J ] "(He Yan, hu Xiaoxiang, liu Yangga, and the like; chinese patent medicines, 2023, 45 (12): 3906-3909.) in the infant throat-clearing granules, and simultaneously measure the content of monocaffeoyltartaric acid, forsythoside A, chicoric acid, forsythin, arctiin and harpagoside in the infant throat-clearing granules, but the analysis time of the method is still longer, and the whole analysis time is more than 50 minutes.
Therefore, there is a need to develop a method for rapidly and stably measuring the content of multiple components in the infant throat-clearing granules with good separation degree of each component.
Disclosure of Invention
Aiming at the problems existing in the prior art, the invention provides a multi-component content determination method for the infant throat-clearing particles, which is simple and convenient to operate, can rapidly and stably separate and detect the content of the forsythin, the arctiin and the cinnamic acid within 35min, has good separation degree, has a linear relation r of more than or equal to 0.9998, and has average sample-adding recovery rate of more than 95 percent.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
the first aspect of the invention provides a multi-component content determination method of the pediatric throat-clearing particles, wherein the determination method adopts an HPLC method to determine the chromatographic conditions: ZORBAX SB-C18 column, mobile phase: acetonitrile (A) -0.1% phosphoric acid (B), gradient eluting, wherein the flow rate is 1.0mL/min, the column temperature is 35 ℃, and the detection wavelength is 260-290nm; the sample injection amount is 5-15 mu L;
Preparation of mixed control solution stock: precisely weighing reference substances of 300-600 mug of the forsythin, 1500-2500 mug of the arctiin and 30-60 mug of the cinnamic acid in each 1mL of mixed reference substance solution stock solution prepared by dissolving the reference substances of the forsythin, the arctiin and the cinnamic acid in methanol;
Preparation of test solution: taking 1.5-2.5g of infant throat-clearing particles, precisely weighing, placing in a20 mL volumetric flask, precisely adding methanol to scale, sealing, performing ultrasonic treatment for 30-45min at a power of 200-300W and a frequency of 40-60 kHz, standing at room temperature, adding methanol to constant volume, shaking uniformly, filtering, and taking filtrate to obtain a sample solution;
The measuring method comprises the following steps: precisely sucking the stock solution of the mixed reference substance and the solution of the sample, respectively measuring the areas of the forsythin, the arctiin and the cinnamic acid under the chromatographic conditions, and calculating to obtain the product.
Preferably, the gradient elution is: 0-10min,20% A;10-38min,20% A to 29% A;38-44min,29% A to 90% A;44-45min,90% A;45-55min,20% A.
Preferably, the chromatographic column specification in the chromatographic conditions is 250mm×4.6mm,5 μm.
Preferably, the detection wavelength is 278nm.
Preferably, the sample injection amount is 10 μl.
Preferably, the preparation of the mixed reference solution stock solution specifically comprises: precisely weighing reference substances of the forsythin, the arctiin and the cinnamic acid, respectively placing the reference substances in a volumetric flask, adding methanol for dissolution and dilution to a scale, and shaking uniformly to prepare mixed reference substance solution stock solution containing 511.4762 mug of the forsythin, 2144.538 mug of the arctiin and 54.47084 mug of the cinnamic acid in each 1 mL.
Preferably, the preparation of the sample solution specifically comprises: taking 2.0 g of infant throat-clearing granule, precisely weighing, placing into a volumetric flask with a plug 20 mL, precisely adding methanol, sealing, performing ultrasonic treatment 40 min with ultrasonic treatment power of 250W and frequency of 50 kHz, cooling to room temperature, adding methanol to constant volume of 20 mL, shaking, filtering, and collecting filtrate to obtain sample solution.
Preferably, the standard curve is drawn by using the mixed reference solution stock solution under the chromatographic condition, specifically: precisely measuring 0.1, 0.2, 0.5, 1,2 and 5mL of mixed reference substance stock solution respectively, fixing the volume of methanol to 10mL to obtain mixed reference substance solutions with serial concentrations, respectively carrying out sample injection measurement, taking the mass concentration as an abscissa and the peak area as an ordinate, and calculating a regression equation to obtain the product.
Further preferably, the detection limit and the quantification limit are determined according to the regression equation with a signal to noise ratio S/n=3 and S/n=10.
Preferably, in the determination method, theoretical plate numbers of 3 components to be detected, namely, the forsythin, the arctiin and the cinnamic acid are not lower than 21500,3, and the separation degree of chromatographic peaks and adjacent peaks of the components to be detected is larger than 2.1.
Compared with the prior art, the invention has the following beneficial effects: (1) The detection method has the separation degree of 3 components of the forsythin, the arctiin and the cinnamic acid from adjacent peaks of more than 2.1, and particularly has the average separation degrees of 3.645, 3.159 and 2.782 respectively; (2) In the detection method, the forsythin is in good linear relation with the peak area in the range of 5.115-511.5 mug/mL, the arctiin is in the range of 21.45-2145 mug/mL, the cinnamic acid is in the range of 0.5447-54.47 mug/mL, the correlation coefficient (r) is not lower than 0.9998, the average sample recovery rate is 99.14%, 99.90% and 95.04%, and the RSD is 1.08%, 0.97% and 1.43% respectively; (3) the detection method of the invention is stable and has good repeatability.
Drawings
FIG. 1 HPLC diagram of the mixed control of forsythin, arctiin and cinnamic acid;
FIG. 2 HPLC diagram of a sample of pediatric throat-clearing granules;
FIG. 3 HPLC diagram of a fructus forsythiae negative sample of the pediatric throat-clearing granule;
FIG. 4 HPLC diagram of burdock fruit-deficient negative sample of the pediatric throat-clearing granule;
FIG. 5 HPLC diagram of radix scrophulariae-deficient negative sample of the pediatric throat-clearing granule;
FIG. 6 is an HPLC plot of different batches (9 batches) of pediatric throat-clearing pellet samples;
FIG. 7 is an HPLC chart of comparative example 1 (replacement of chromatographic column);
FIG. 8 is an HPLC chart of comparative example 2 (mobile phase acetonitrile to methanol);
FIG. 9 is an HPLC chart of comparative example 3 (exchange elution gradient program);
FIG. 10 is an HPLC chart of comparative example 4 (column temperature is changed to 30 ℃);
FIG. 11 is an HPLC chart of comparative example 5 (flow rate 1.5 mL/min);
FIG. 12 is an HPLC chart of comparative example 6 (phosphoric acid solution concentration is changed to 0.3%).
Detailed Description
It is to be noted that the raw materials used in the present invention are all common commercial products, and the sources thereof are not particularly limited.
Example 1
1. Instrument and reagent
1.1 Instruments
Agilent 1260 Infinity ii high performance liquid chromatograph (diode array detector, agilent, usa).
1.2 Reagent
Pediatric throat-clearing granules 9 batches, from 3 companies: manufacturer A (lot number: SB20003, SB20013, SB20024, SB20031, SB20039, UB20006, UB 20007); manufacturer B (lot number: ZJB 2202); manufacturer C (batch number: 221212). Forsythin (lot number: 110821-202318, content: 95.8%), arctiin (lot number: 110819-202213, content: 99.0%), cinnamic acid (lot number: 110786-202305, content: 99.8%) are from the national food and drug verification institute. Acetonitrile and phosphoric acid were both chromatographically pure (Merck).
2. Method and results
2.1 Chromatographic conditions
Chromatographic column: ZORBAX SB-C18X 250X 4.6 mm SN:USCL125194
Mobile phase: acetonitrile-B0.1% phosphoric acid
Elution gradient: 0-10min,20% A;10-38min,20% A to 29% A;38-44min,29% A to 90% A;44-45min,90% A;45-55min,20% A.
Flow rate: 1.0 mL/min; column temperature: 35. the temperature is lower than the temperature; sample injection amount: 10. 2, L; detection wavelength: 278nm.
2.2 Preparation of the solution
2.2.1 Mixing reference substance solution
Accurately weighing a proper amount of each reference substance, and adding methanol to prepare mixed reference substance stock solution containing 511.4762 mug, 2144.538 mug and 54.47084 mug of the forsythin, the arctiin and the cinnamic acid in each 1 mL. Sample injection measurement is carried out according to the chromatographic condition under the condition of 2.1, and the chromatogram is shown in figure 1.
2.2.2 Test solution
Mixing 10 bags of the product (batch number: UB 20007), grinding, precisely weighing 2.0 g, placing into a volumetric flask with a plug of 20 mL, precisely adding methanol, sealing, performing ultrasonic treatment (250W, 50 kHz) of 40 min, cooling to room temperature, adding methanol to volume of 20 mL, shaking, filtering, and collecting the filtrate. Sample injection measurement is carried out according to the chromatographic condition under the condition of 2.1, and the chromatogram is shown in figure 2.
2.2.3 Negative sample solution
Negative samples lacking fructus forsythiae, fructus Arctii (parched) and radix scrophulariae are prepared according to the prescription process of the granule for clearing heat from throat of children, negative sample solution is prepared according to the method under item 2.2.2, and sample injection measurement is carried out according to the chromatographic condition under item 2.1, and the result is shown in figures 3-5. The negative sample can not interfere with the measurement of the forsythin, the arctiin and the cinnamic acid; the forsythin is from fructus forsythiae, the arctiin is from fructus arctii, and the cinnamic acid is from radix scrophulariae.
2.3 Linear relationship, detection limit and quantification limit
Precisely measuring the stock solution of the mixed reference substance, fixing the volume by methanol, carrying out gradient dilution to 2, 5, 10, 20, 50 and 100 times to obtain a series of mixed reference substance solutions, respectively carrying out sample injection measurement, taking the mass concentration as an abscissa and the peak area as an ordinate, and calculating a regression equation. And the detection limit and the quantitative limit were measured according to the signal-to-noise ratios S/n=3 and S/n=10, respectively, and the results are shown in table 1.
TABLE 1 regression equation, correlation coefficient, linear range, detection limit and quantitative limit for forsythin, arctiin, cinnamic acid
| Composition of the components | Regression equation | r | Range/(mug mL -1) | Detection limit/(mug mL -1) | Quantitative limit/(mug mL -1) |
| Forsythin glycoside | y = 6666.9x + 3916.2 | 0.9999 | 5.115~511.5 | 0.098 | 0.382 |
| Arctiin | y = 5361.4x + 29075 | 0.9999 | 21.45~2145 | 0.140 | 0.412 |
| Cinnamic acid | y = 90208x - 4869.7 | 0.9998 | 0.5447~54.47 | 0.010 | 0.041 |
2.4 Precision test
The same mixed reference substance solution is precisely measured, the sample is continuously injected for 6 times, the peak areas of 3 components are recorded, and the RSD of the peak areas of the forsythin, the arctiin and the cinnamic acid are respectively 0.15%, 0.08% and 0.14%, which indicates that the instrument precision is good.
2.5 Stability test
Taking the same batch of infant throat-clearing particles (batch number: UB 20007), preparing according to the method under the item of '2.2.2', and respectively carrying out sample injection measurement on 0, 4, 8, 12, 16, 24 and 48 h to obtain the sample solution with RSD of peak areas of the forsythin, the arctiin and the cinnamic acid of 0.76%, 0.58 and 0.55%, wherein the sample solution is stable in 48 h.
2.6 Repeatability test
Taking the same batch of infant throat-clearing particles (batch number: UB 20007), preparing 6 parts of sample solution according to the method under the item of '2.2.2', and carrying out sample injection measurement, wherein the average values of the contents of the forsythin, the arctiin and the cinnamic acid are 2.517, 13.360 and 0.309 and mg/bag respectively, the contents of the RSD are 0.45%, 0.42% and 0.39% respectively, and the relative retention time RSD is 0.04%, 0.03% and 0.01% respectively, so that the method is good in repeatability.
2.7 Sample addition recovery test
6 Parts of the sample (batch No. UB 20007) with known content of each component are precisely weighed, 1.0g of each part is placed in a 20mL volumetric flask, a reference substance solution is added according to the ratio of 1:1, methanol is added to 20mL, a sample solution is prepared according to the method under the item "2.2.2", and the recovery rate is calculated by sampling respectively. As a result, the average recovery rates of the forsythin, the arctiin and the cinnamic acid were 99.14%, 99.90% and 95.04% respectively, and the RSD was 1.08%, 0.97% and 1.43% respectively.
2.8 Sample content determination
9 Batches of samples from 3 manufacturers are respectively taken and prepared according to the method under the item "2.2.2", and the sample is sampled and measured according to the chromatographic condition under the item "2.1", and the chromatogram is shown in figure 2. The degree of separation of each component and the results of measurement of the content are shown in Table 2.
From the above measurement results, the separation degree of the chromatographic peaks and the adjacent peaks of the 3 components to be measured is more than 2.1, specifically, the average separation degrees of the forsythin, the arctiin and the cinnamic acid are 3.645, 3.159 and 2.782 respectively. The determination method can be used for rapidly and stably separating and detecting the content of the forsythin, the arctiin and the cinnamic acid within 35min, has good separation degree, and can be used for quality control of the children throat clearing granules.
Comparative example 1
Chromatographic conditions
Chromatographic column: diamonsil C18.5u250.times.4.6: 4.6 mm
Mobile phase: acetonitrile-B0.1% phosphoric acid
Elution gradient: 0-10min,20% A;10-38min,20% A to 29% A;38-44min,29% A to 90% A;44-45min,90% A;45-55min,20% A.
Flow rate: 1.0 mL/min; column temperature: 35. the temperature is lower than the temperature; sample injection amount: 10. 2, L; detection wavelength: 278nm.
The chromatographic conditions differ from those of example 1 only in the chromatographic column.
Mixing 10 bags of the product (batch number: UB 20007), grinding, precisely weighing 2.0 g, placing into a volumetric flask with a plug of 20 mL, precisely adding methanol, sealing, performing ultrasonic treatment (250W, 50 kHz) of 40 min, cooling to room temperature, adding methanol to volume of 20 mL, shaking, filtering, and collecting the filtrate. The chromatogram is shown in FIG. 7.
From this figure, it can be seen that the chromatographic column in example 1 was changed to Diamonsil C18.5u.250×4.6: 4.6 mm, and the separation degree of cinnamic acid was lowered, and it was not completely separated from other components (the separation degree of cinnamic acid was 1.263, < 1.50).
Comparative example 2
Chromatographic conditions
Chromatographic column: ZORBAX SB-C18X 250X 4.6 mm SN:USCL125194
Mobile phase: a is methanol-B0.1% phosphoric acid
Elution gradient: 0-10min,20% A;10-38min,20% A to 29% A;38-44min,29% A to 90% A;44-45min,90% A;45-55min,20% A.
Flow rate: 1.0 mL/min; column temperature: 35. the temperature is lower than the temperature; sample injection amount: 10. 2, L; detection wavelength: 278nm.
The chromatographic conditions differ from those of example 1 only in the mobile phase composition.
Mixing 10 bags of the product (batch number: UB 20007), grinding, precisely weighing 2.0 g, placing into a volumetric flask with a plug of 20 mL, precisely adding methanol, sealing, performing ultrasonic treatment (250W, 50 kHz) of 40 min, cooling to room temperature, adding methanol to volume of 20 mL, shaking, filtering, and collecting the filtrate. The chromatogram is shown in FIG. 8.
From this figure, it can be seen that, when the mobile phase in example 1 was changed to A: methanol-B: 0.1% phosphoric acid, no separation of the other components was possible from the forsythin, arctiin and cinnamic acid.
Comparative example 3
Chromatographic conditions
Chromatographic column: ZORBAX SB-C18X 250X 4.6 mm SN:USCL125194
Mobile phase: acetonitrile-B0.1% phosphoric acid
Elution gradient: 0-10min,18% A;10-40min,18% A-30% A;40-45min,30% A to 90% A;45-46min,90% A;46-56min,18% A.
Flow rate: 1.0 mL/min; column temperature: 35. the temperature is lower than the temperature; sample injection amount: 10. 2, L; detection wavelength: 278nm.
The chromatographic conditions differ from those of example 1 only in the elution gradient.
Mixing 10 bags of the product (batch number: UB 20007), grinding, precisely weighing 2.0 g, placing into a volumetric flask with a plug of 20 mL, precisely adding methanol, sealing, performing ultrasonic treatment (250W, 50 kHz) of 40 min, cooling to room temperature, adding methanol to volume of 20 mL, shaking, filtering, and collecting the filtrate. The chromatogram is shown in FIG. 9.
From this figure, it can be seen that the elution gradient in example 1 is replaced by: 0-10min,18% A;10-40min,18% A-30% A;40-45min,30% A to 90% A;45-46min,90% A;46-56min,18% A. Cinnamic acid is not separated from other ingredients.
Comparative example 4
Chromatographic conditions
Chromatographic column: ZORBAX SB-C18X 250X 4.6 mm SN:USCL125194
Mobile phase: acetonitrile-B0.1% phosphoric acid
Elution gradient: 0-10min,20% A;10-38min,20% A to 29% A;38-44min,29% A to 90% A;44-45min,90% A;45-55min,20% A.
Flow rate: 1.0 mL/min; column temperature: 30. the temperature is lower than the temperature; sample injection amount: 10. 2, L; detection wavelength: 278nm.
The chromatographic conditions differ from those of example 1 only in the column temperature.
Mixing 10 bags of the product (batch number: UB 20007), grinding, precisely weighing 2.0 g, placing into a volumetric flask with a plug of 20 mL, precisely adding methanol, sealing, performing ultrasonic treatment (250W, 50 kHz) of 40min, cooling to room temperature, adding methanol to volume of 20 mL, shaking, filtering, and collecting the filtrate. The chromatogram is shown in FIG. 10.
From this figure, it can be seen that the column temperature in example 1 was changed to 30 ℃, and there was no significant effect on the peak time of the forsythin, arctiin, and cinnamic acid, but the separation degree of cinnamic acid was decreased (the separation degrees of cinnamic acid at 35 ℃, 30 ℃ in fig. 2 and 10 were 3.094 and 2.215, respectively).
Comparative example 5
Chromatographic conditions
Chromatographic column: ZORBAX SB-C18X 250X 4.6 mm SN:USCL125194
Mobile phase: acetonitrile-B0.1% phosphoric acid
Gradient: 0-10min,20% A;10-38min,20% A to 29% A;38-44min,29% A to 90% A;44-45min,90% A;45-55min,20% A.
Flow rate: 1.5 mL/min; column temperature: 35. the temperature is lower than the temperature; sample injection amount: 10. 2, L; detection wavelength: 278nm.
The chromatographic conditions differ from those of example 1 only in the flow rate.
Mixing 10 bags of the product (batch number: UB 20007), grinding, precisely weighing 2.0 g, placing into a volumetric flask with a plug of 20 mL, precisely adding methanol, sealing, performing ultrasonic treatment (250W, 50 kHz) of 40min, cooling to room temperature, adding methanol to volume of 20 mL, shaking, filtering, and collecting the filtrate. The chromatogram is shown in FIG. 11.
From this figure, it can be seen that cinnamic acid cannot be completely separated from other components by changing the flow rate in example 1 to 1.5 mL/min.
Comparative example 6
Chromatographic conditions
Chromatographic column: ZORBAX SB-C18X 250X 4.6 mm SN:USCL125194
Mobile phase: acetonitrile-B0.3% phosphoric acid
Gradient: 0-10min,20% A;10-38min,20% A to 29% A;38-44min,29% A to 90% A;44-45min,90% A;45-55min,20% A.
Flow rate: 1.0 mL/min; column temperature: 35. the temperature is lower than the temperature; sample injection amount: 10. 2, L; detection wavelength: 278nm.
The chromatographic conditions differ from those of example 1 only in the concentration of the mobile phase phosphoric acid solution.
Mixing 10 bags of the product (batch number: UB 20007), grinding, precisely weighing 2.0 g, placing into a volumetric flask with a plug of 20 mL, precisely adding methanol, sealing, performing ultrasonic treatment (250W, 50 kHz) of 40 min, cooling to room temperature, adding methanol to volume of 20 mL, shaking, filtering, and collecting the filtrate. The chromatogram is shown in FIG. 12.
From this figure, it can be seen that cinnamic acid cannot be completely separated from other components (the degree of separation is 1.219, < 1.50) by changing the phosphoric acid solution in example 1 to 0.3%.
Finally, it should be noted that the above description is only for illustrating the technical solution of the present invention, and not for limiting the scope of the present invention, and that the simple modification and equivalent substitution of the technical solution of the present invention can be made by those skilled in the art without departing from the spirit and scope of the technical solution of the present invention.
Claims (7)
1. A multi-component content determination method of infant throat-clearing particles adopts an HPLC method to determine the content of the components, and the chromatographic conditions are as follows: ZORBAX SB-C18 column, mobile phase: the A phase is acetonitrile-B phase 0.1% phosphoric acid, gradient elution is carried out, the flow rate is 1.0mL/min, the column temperature is 35 ℃, and the detection wavelength is 260-290nm; the sample injection amount is 5-15 mu L;
The preparation of the mixed reference solution stock solution is specifically as follows: precisely weighing reference substances of the forsythin, the arctiin and the cinnamic acid, respectively placing the reference substances in a volumetric flask, adding methanol for dissolution, diluting to a scale, and shaking uniformly to prepare mixed reference substance solution stock solution containing 511.4762 mug of the forsythin, 2144.538 mug of the arctiin and 54.47084 mug of the cinnamic acid in each 1 mL;
The preparation of the sample solution comprises the following steps: taking 2.0 g of infant throat-clearing particles, precisely weighing, placing into a volumetric flask with a plug 20mL, precisely adding methanol, sealing, performing ultrasonic treatment with power of 250W and frequency of 50 kHz at 40 min, cooling to room temperature, adding methanol to volume of 20mL, shaking, filtering, and collecting filtrate to obtain sample solution;
drawing a standard curve under the chromatographic condition by using the mixed reference substance solution stock solution, wherein the standard curve specifically comprises the following steps: precisely measuring 0.1mL, 0.2mL, 0.5mL, 1mL, 2mL and 5mL of mixed reference substance stock solution respectively, and fixing the volume of methanol to 10mL to obtain mixed reference substance solutions with serial concentrations, respectively carrying out sample injection measurement, taking the mass concentration as an abscissa and the peak area as an ordinate, and calculating a regression equation to obtain the compound reference substance;
The measuring method comprises the following steps: precisely sucking the sample solution, measuring the areas of the forsythin, the arctiin and the cinnamic acid under the chromatographic conditions, and calculating to obtain the final product.
2. The method for determining the multicomponent content of the pediatric throat-clearing particles according to claim 1, wherein the gradient elution is as follows: 0-10min,20% A;10-38min,20% A to 29% A;38-44min,29% A to 90% A;44-45min,90% A;45-55min,20% A.
3. The method for measuring the multicomponent content of the pediatric throat-clearing particles according to claim 1, wherein the chromatographic conditions are characterized by a chromatographic column specification of 250mm x 4.6mm,5 μm.
4. The method for measuring the multicomponent content of the pediatric throat-clearing particles according to claim 1, wherein the detection wavelength is 278nm.
5. The method for measuring the multicomponent content of the pediatric throat-clearing particles according to claim 1, wherein the sample injection amount is 10 μl.
6. The method for measuring the multicomponent content of the pediatric throat-clearing particles according to claim 1, wherein the detection limit and the quantitative limit are determined according to the regression equation according to the signal-to-noise ratio S/n=3 and S/n=10.
7. The method for measuring the multicomponent content of the pediatric throat-clearing particles according to claim 1, wherein the separation degree of 3 components to be measured from adjacent peaks in the measuring method is more than 2.1.
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| CN105726623A (en) * | 2016-02-28 | 2016-07-06 | 甘肃复兴厚生物医药科技有限公司 | Astragalus membranaceus ultramicro decoction pieces as well as preparation method and quality testing method thereof |
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| CN101574486A (en) * | 2008-05-09 | 2009-11-11 | 北京亚东生物制药有限公司 | Quality control method for Chinese medicinal capsule for treating anemofrigid cold |
| CN105726623A (en) * | 2016-02-28 | 2016-07-06 | 甘肃复兴厚生物医药科技有限公司 | Astragalus membranaceus ultramicro decoction pieces as well as preparation method and quality testing method thereof |
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