CN114062544B - Feature map of four-material capsule extract volatile oil or inclusion compound thereof, and establishment method and application thereof - Google Patents
Feature map of four-material capsule extract volatile oil or inclusion compound thereof, and establishment method and application thereof Download PDFInfo
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Abstract
The application relates to a method for detecting four-material capsule extract volatile oil or inclusion compound thereof by using a characteristic spectrum of the four-material capsule extract volatile oil or inclusion compound thereof. The method adopts a Gas Chromatography (GC) method, and uses 3-n-butenyl phthalide as a reference substance to carry out GC detection through a gas chromatographic column. According to the application, by analyzing the characteristic spectrum of the volatile oil or the inclusion compound, the four-material capsule extract can be detected, the quality control and evaluation of the volatile oil and the inclusion compound of the four-material capsule extract can be comprehensively carried out, and the quality stability, uniformity, safety and effectiveness of the medicine are ensured.
Description
Technical Field
The application relates to the technical field of medicine analysis, in particular to a characteristic spectrum of four-material capsule extract volatile oil or inclusion compound and application thereof in detection and quality control of the four-material capsule extract volatile oil or the inclusion compound.
Background
Volatile oils are a generic term for oily liquids which are present in plants and which have a volatile, aromatic odor and which distill off with water vapor. Volatile oil is used as a main effective component in Chinese herbal medicines and widely exists in traditional Chinese medicines, and the quality of medicinal material treatment is judged by the content of the volatile oil. In the development of modern medicines, the application of volatile oil is also increasingly involved, and the fact that the volatile oil has volatility and instability, how to add the extracted volatile oil into a traditional Chinese medicine preparation ensures the therapeutic effect of the preparation and ensures the maximum exertion of the curative effect of the medicine becomes a key problem to be solved.
The traditional Chinese medicine volatile oil treatment method comprises extracting volatile oil, decocting or pulverizing the medicinal materials to obtain extract powder or medicinal powder, spraying the volatile oil directly on the medicinal powder or adsorbing the volatile oil in starch and other adjuvants, and making into various dosage forms. The method is simple and easy to operate, does not need excessive cost and equipment, but has poor stability of the medicine, can not be stored for a long time, and meanwhile, volatile oil can be gradually dissipated, and meanwhile, mildew is easy to generate.
The process of clathrating volatile oil with beta-cyclodextrin is one of the new preparation methods of modern preparations. The cyclodextrin is prepared by enzymatic cyclization of starch, has a special structure of a ring-shaped hollow cylinder, and is a good inclusion material. The volatile oil is embedded into the cylindrical structure to form an ultrafine particle dispersion, so that the dispersion effect is good, the absorption is easy, the medicine is slowly released, the bioavailability of the medicine is higher, and the adverse reaction is low. The inclusion of beta-cyclodextrin with volatile oil is a physical process, and the volatile oil still maintains the original properties and functions without chemical changes. The inclusion of the volatile oil with beta-cyclodextrin can increase the solubility and dissolution rate of the volatile oil and improve the stability of the volatile oil; the volatile oil is powdered, the volatility of the volatile oil is reduced, and the utilization rate of the volatile oil is improved; improve bad smell of volatile oil, reduce irritation of volatile oil, reduce toxic side effect, etc.
Volatile oil has volatility and inevitably causes the loss of the content in the preparation process. In order to ensure the curative effect of the preparation, the quality control of volatile oil or inclusion compound in the preparation is required. The property, relative density, content and the like of the volatile oil can be measured to be used as a quality control method of the volatile oil or the inclusion compound, however, the method cannot reflect the whole condition of the volatile oil or the inclusion compound, and the volatile oil component is a key factor for determining the curative effect of the preparation. How to control the quality of the volatile oil more comprehensively is important.
The four-material capsule is composed of 4 medicines of ligusticum wallichii, angelica, white paeony root and prepared rehmannia root, has the functions of regulating menstruation and nourishing blood, and is used for treating deficiency of nutrient and blood and irregular menstruation. In the preparation method of the four-material capsule, volatile oil of the ligusticum wallichii and the angelica is extracted to prepare volatile oil beta-cyclodextrin inclusion compound which participates in the preparation process. Therefore, the quality control before and after the inclusion of the volatile oil is one of the key links in the preparation process.
In the prior art, there is still a need for an improved quality control method for the volatile oil and the clathrate thereof, which can comprehensively and accurately evaluate the quality of the volatile oil and the clathrate of the four-ingredient capsule extract, and ensure the uniformity and stability of the quality of the volatile oil and the clathrate.
Disclosure of Invention
Aiming at the technical problems, the application provides a detection method of four-material capsule extract volatile oil or inclusion compound thereof. The method adopts GC method to establish characteristic spectrum of the volatile oil or the inclusion compound thereof, can detect the volatile oil and the inclusion compound thereof, comprehensively evaluate the quality of the four-material capsule extract, and ensure uniformity and stability of the quality. In addition, the application also provides a GC characteristic spectrum of the four-material capsule extract volatile oil or the inclusion compound thereof.
The characteristic spectrum of the traditional Chinese medicine refers to a spectrum which can mark common peaks in the traditional Chinese medicine obtained by adopting a certain analysis means after the traditional Chinese medicine is properly treated, and is a novel quality control mode of the traditional Chinese medicine.
Specifically, the application provides a detection method of four-material capsule extract volatile oil or inclusion compound thereof, which comprises the following steps:
1) Determining a Gas Chromatography (GC) characteristic spectrum of the pharmaceutical preparation to be detected by using a GC method;
2) Determining the GC spectrum of the 3-n-butenyl phthalide by using a GC method, and comparing the GC spectrum with the characteristic spectrum in the step 1);
wherein the GC characteristic spectrum of the four-material capsule extract volatile oil or the inclusion compound comprises 9 characteristic peaks, and the 3-n-butenyl phthalide peak obtained in the step 2) is taken as a reference peak (S peak);
3) And when the relative retention time and the relative deviation of each characteristic peak meet the following conditions, judging that the to-be-detected pharmaceutical preparation contains four capsule extract volatile oil or inclusion compound:
peak No. 1: the relative retention time is 0.121+/-0.012, and the relative deviation is 0-10%;
peak No. 2: the relative retention time is 0.226+/-0.023, and the relative deviation is 0-10%;
peak No. 3: the relative retention time is 0.389+/-0.039, and the relative deviation is 0-10%;
peak No. 4: the relative retention time is 0.464 plus or minus 0.046, and the relative deviation is 0-10%;
peak No. 5: the relative retention time is 0.872 plus or minus 0.087, and the relative deviation is 0-10%;
peak No. 6: the relative retention time is 0.970 plus or minus 0.097, and the relative deviation is 0-10%;
peak No. 7: the relative retention time is 1.000, and the relative deviation is 0-10%;
peak No. 8: the relative retention time is 1.082 plus or minus 0.108, and the relative deviation is 0-10%;
peak No. 9: the relative retention time is 1.111 plus or minus 0.111, and the relative deviation is 0-10%;
wherein the conditions of the gas chromatography include the following: a capillary chromatographic column taking (5% -phenyl) -methyl polysiloxane as a stationary phase; a FID detector; the program is run at elevated temperature conditions.
The term "four-substance capsule extract volatile oil" refers to volatile oil extracted from angelica and/or ligusticum wallichii, for example, according to the preparation process of four-substance capsules.
The term "clathrate" refers to, for example, a product obtained by clathrating a volatile oil of a four-substance capsule extract with a clathrating material, and for example, beta-cyclodextrin is used as the clathrating material.
The term "drug to be tested" refers to, for example, including but not limited to, four-substance capsules and the like, volatile oils and the like from which the four-substance capsules are made, or clathrates and the like of volatile oils of the four-substance capsules.
In a preferred embodiment, in the detection method of the present application, the chromatographic column has a length of 30m or 60m, preferably 30m; an inner diameter of 0.32mm or 0.53mm, preferably 0.32mm; the film thickness is 0.25 μm, 0.5 μm or 1 μm, preferably 0.25 μm.
In a preferred embodiment, in the detection method of the present application, the carrier gas used is nitrogen or helium, preferably nitrogen.
In a preferred embodiment, in the detection method of the present application, a carrier gas flow rate of 0.6 to 0.8ml/min, preferably 0.7ml/min is used.
In a preferred embodiment, in the detection method of the present application, the sample inlet temperature is 250 ℃ to 260 ℃, preferably 250 ℃; the detector temperature is 260℃to 270℃and preferably 260 ℃.
In a preferred embodiment, in the detection method of the application, the split ratio is 10 to 20:1, preferably 20:1.
In a preferred embodiment, in the detection method of the present application, the amount of sample introduced is 1 to 2. Mu.l, preferably 2. Mu.l.
In a preferred embodiment, in the detection method of the present application, the temperature-raising condition is an initial temperature of 100 ℃, and is maintained for 15 minutes; raising the temperature to 110 ℃ at a rate of 5 ℃ per minute for 25 minutes; raising the temperature to 150 ℃ at a rate of 4 ℃ per minute and keeping for 30 minutes; the temperature was raised to 250℃at a rate of 10℃per minute and maintained for 10 minutes.
In a preferred embodiment, the detection method of the present application is applied to a four-substance capsule formulation process.
In one embodiment, the application provides a four-material capsule extract volatile oil and inclusion compound characteristic spectrum, the method comprises the following steps and conditions:
(1) Reference solution preparation: taking about 4mg of 3-n-butenyl phthalide, placing in a 100mL volumetric flask, dissolving with absolute ethyl alcohol, diluting to scale, and shaking uniformly to obtain the final product.
(2) Preparing volatile oil test sample solution: and (3) taking 10 μl of the volatile oil, placing in a 20mL volumetric flask, dissolving with absolute ethanol, diluting to a scale, shaking uniformly, and filtering.
(3) Preparing a clathrate compound test sample solution: about 122mg of clathrate powder is precisely weighed, placed in a conical flask with a plug, precisely added with 20ml of absolute ethyl alcohol, sealed, weighed, treated by ultrasound (the temperature should not be higher than 35 ℃ and the power is 100W and the frequency is 40 kHz) for 40 minutes, cooled, weighed again, complemented with the lost weight by absolute ethyl alcohol, shaken uniformly, filtered, and the subsequent filtrate is taken, thus obtaining the clathrate.
(4) Analysis by gas chromatography
The chromatographic conditions are as follows: capillary chromatographic column with (5% -phenyl) -methyl polysiloxane as stationary phase; a FID detector; the carrier gas is nitrogen, and the flow rate of the carrier gas is 0.7ml/min; the temperature of the sample inlet is 250 ℃; the detector temperature was 260 ℃; the split ratio is 20:1; sample injection amount: 2 μl; and (5) programming temperature conditions.
(5) And (3) measuring: precisely sucking 2 μl of each of the reference solution, volatile oil test solution and clathrate test solution, respectively, and injecting into gas chromatograph for measurement, and recording chromatogram.
The GC-MS method is combined with comparison of reference substance information to determine that two main active components in volatile oil of radix Angelicae sinensis and rhizoma Ligustici Chuanxiong are 3-n-butenyl phthalide (peak 7) and ligustilide (peak 9), respectively. When ligustilide is used as reference, the number 9 peak is used as reference peak (S peak), and the relative retention time of other 8 peaks is less than 1.000; because of the larger peak area of peak 9, the relative peak areas of peak 1, peak 2, peak 3, peak 4, peak 5, peak 6, peak 7, peak 8 are smaller, all being less than 0.100; the peak of the compound is positioned in the middle of a chromatogram, the separation degree between the compound and other chromatographic peaks is good, the peak corresponding to the 3-n-butenyl phthalide is taken as an S peak, and the calculated relative retention time and relative peak area of each characteristic peak are moderate. Obviously, compared with ligustilide (peak 9), 3-n-butenyl phthalide (peak 7) is more suitable as a reference peak to determine GC characteristic patterns of the volatile oil and the inclusion compound of the four-material capsule extract.
Thus, the 3-n-butenyl phthalide peak was selected as a reference peak to determine the four capsule extract volatile oil or clathrate.
The GC characteristic spectrum of the four-material capsule extract volatile oil or inclusion compound comprises 9 common main components, 3-n-butenyl phthalide is taken as a reference substance, a No. 7 peak is taken as a reference substance peak (S peak), 9 common characteristic peaks are arranged in the characteristic spectrum, the corresponding peak of the reference substance peak is taken as the S peak, and the relative retention time of each characteristic peak and the S peak is calculated and is within +/-10% of a specified value. The prescribed value is peak 1:0.121, peak 2:0.226, peak 3:0.389, peak 4:0.464, peak 5:0.872, peak 6:0.970, peak 7:1.000 (S peak), peak 8:1.082, peak 9:1.111.
in a preferred embodiment, the present application provides a device for detecting a four-substance capsule extract volatile oil or its inclusion compound, the device comprising at least one processor, and
a memory storing instructions that, when executed by at least one processor, perform the comparing steps of the method according to any of the embodiments of the present application.
The application has at least the following beneficial effects:
1) The application establishes the characteristic patterns of the volatile oil and the inclusion compound of the four-material capsule extract by a Gas Chromatography (GC) method, and determines the precision, the repeatability and the stability of the characteristic patterns by experiments;
2) The characteristic spectrum of the obtained four-material capsule extract volatile oil and the clathrate compound is established, and the characteristic spectrum can be used for detecting the four-material capsule extract volatile oil and the clathrate compound;
3) By analyzing the characteristic patterns of the volatile oil and the inclusion compound of the four-material capsule extract, the quality of the volatile oil and the inclusion compound in the four-material capsule preparation process can be comprehensively controlled and evaluated, and the stability and consistency of the quality of the medicine are ensured, so that the method is used for the quality standard and quality control of the medicine of the four-material capsule extract.
Description of the drawings
FIG. 1 gas chromatography of ligustilide
FIG. 2 gas chromatography of blank adjuvant
FIG. 3.3-n-butenyl phthalide (reference) gas chromatography
FIG. 4 characteristic spectrum of volatile oil
FIG. 5 clathrate characteristic spectrum
Detailed Description
Example 1
The method for constructing the characteristic spectrum of the volatile oil and the inclusion compound of the four-material capsule extract comprises the following steps and conditions:
(1) Reference solution preparation: taking 2 μl of ligustilide, placing into a 10ml volumetric flask, dissolving with absolute ethanol, diluting to scale, and shaking.
(2) Preparing volatile oil test sample solution: placing 10 μl of volatile oil in 20ml volumetric flask, dissolving with absolute ethanol, diluting to scale, shaking, and filtering.
(3) Preparing a clathrate compound test sample solution: about 122mg of clathrate powder is precisely weighed, placed in a conical flask with a plug, precisely added with 20ml of absolute ethyl alcohol, sealed, weighed, treated by ultrasound (the temperature should not be higher than 35 ℃ and the power is 100W and the frequency is 40 kHz) for 40 minutes, cooled, weighed again, complemented with the lost weight by absolute ethyl alcohol, shaken uniformly, filtered, and the subsequent filtrate is taken, thus obtaining the clathrate.
(4) Analysis by gas chromatography
The chromatographic conditions are as follows: capillary chromatographic column with (5% -phenyl) -methyl polysiloxane as stationary phase; a FID detector; the carrier gas is nitrogen, and the flow rate of the carrier gas is 0.7ml/min; the temperature of the sample inlet is 250 ℃; the detector temperature was 260 ℃; the split ratio is 20:1; sample injection amount: 2 μl; the temperature programming condition is that the initial temperature is 100 ℃, and the temperature is kept for 15 minutes; raising the temperature to 110 ℃ at a rate of 5 ℃ per minute for 25 minutes; raising the temperature to 150 ℃ at a rate of 4 ℃ per minute and keeping for 30 minutes; the temperature was raised to 250℃at a rate of 10℃per minute and maintained for 10 minutes.
(5) And (3) measuring: precisely sucking 2 μl of each of the reference solution, volatile oil test solution and clathrate test solution, respectively, and injecting into gas chromatograph for measurement, and recording chromatogram.
The GC characteristic spectrum of the four-material capsule extract volatile oil and the clathrate compound comprises 9 common main components, ligustilide is taken as a reference substance, a No. 9 peak is taken as a reference substance peak (S peak), 9 common characteristic peaks are arranged in the characteristic spectrum, the corresponding peak of the reference substance is taken as the S peak, the relative retention time and the relative peak area of each characteristic peak and the S peak are calculated, and the relative retention time of the other 8 peaks is less than 1.000; and the relative peak area values of the peak 1, the peak 2, the peak 3, the peak 4, the peak 5, the peak 6, the peak 7 and the peak 8 are smaller and smaller than 0.100. The results are shown in tables 1 to 2.
TABLE 1 relative retention time results for volatile oils and clathrates
TABLE 2 results of relative peak area of volatile oil and clathrate
Example 2
The method for constructing the characteristic spectrum of the volatile oil and the inclusion compound of the four-material capsule extract comprises the following steps and conditions:
(1) Reference solution preparation: taking about 4mg of 3-n-butenyl phthalide, placing in a 100ml volumetric flask, dissolving with absolute ethyl alcohol, diluting to scale, and shaking uniformly to obtain the final product.
(2) Preparing volatile oil test sample solution: placing 10 μl of volatile oil in 20ml volumetric flask, dissolving with absolute ethanol, diluting to scale, shaking, and filtering.
(3) Preparing a clathrate compound test sample solution: about 122mg of clathrate powder is precisely weighed, placed in a conical flask with a plug, precisely added with 20ml of absolute ethyl alcohol, sealed, weighed, treated by ultrasound (the temperature should not be higher than 35 ℃ and the power is 100W and the frequency is 40 kHz) for 40 minutes, cooled, weighed again, complemented with the lost weight by absolute ethyl alcohol, shaken uniformly, filtered, and the subsequent filtrate is taken, thus obtaining the clathrate.
(4) Analysis by gas chromatography
The chromatographic conditions are as follows: capillary column with (5% -phenyl) -methyl polysiloxane as stationary phase; a FID detector; the carrier gas is nitrogen, and the flow rate of the carrier gas is 0.7ml/min; the temperature of the sample inlet is 250 ℃; the detector temperature was 260 ℃; the split ratio is 20:1; sample injection amount: 2 μl; the temperature programming condition is that the initial temperature is 100 ℃, and the temperature is kept for 15 minutes; raising the temperature to 110 ℃ at a rate of 5 ℃ per minute for 25 minutes; raising the temperature to 150 ℃ at a rate of 4 ℃ per minute and keeping for 30 minutes; the temperature was raised to 250℃at a rate of 10℃per minute and maintained for 10 minutes.
(5) And (3) measuring: precisely sucking 2 μl of each of the reference solution, volatile oil test solution and clathrate test solution, respectively, and injecting into gas chromatograph for measurement, and recording chromatogram. The GC characteristic spectrum of the four-material capsule extract volatile oil and the inclusion compound comprises 9 common main components, 3-n-butenyl phthalide is taken as a reference object, a No. 7 peak is taken as a reference object peak (S peak), 9 common characteristic peaks are arranged in the characteristic spectrum, the corresponding peak of the reference object peak is taken as the S peak, and the relative retention time of each characteristic peak and the S peak is calculated, wherein the relative retention time is within +/-10% of a specified value. The prescribed value is peak 1:0.121, peak 2:0.226, peak 3:0.389, peak 4:0.464, peak 5:0.872, peak 6:0.970, peak 7:1.000 (S peak), peak 8:1.082, peak 9:1.111.
the peak of the compound is positioned in the middle of a chromatogram, the separation degree between the compound and other chromatographic peaks is good, the peak corresponding to the 3-n-butenyl phthalide is taken as an S peak, and the calculated relative retention time and relative peak area of each characteristic peak are moderate. The results are shown in tables 3 to 4.
Obviously, compared with ligustilide (peak 9), 3-n-butenyl phthalide (peak 7) is more suitable as a reference peak to determine GC characteristic patterns of the volatile oil and the inclusion compound of the four-material capsule extract.
Thus, the 3-n-butenyl phthalide peak was selected as a reference peak to determine the four capsule extract volatile oil or clathrate.
TABLE 3 relative retention time results for volatile oil and clathrate
TABLE 4 results of relative peak area of volatile oil and clathrate
Example 3
The method for constructing the four-material capsule characteristic map comprises the following steps and conditions:
(1) Reference solution preparation: taking about 4mg of 3-n-butenyl phthalide, placing in a 100ml volumetric flask, dissolving with absolute ethyl alcohol, diluting to scale, and shaking uniformly to obtain the final product.
(2) Preparation of four-substance capsule test sample solution: taking 15g of the four-material capsule, precisely weighing, placing into a conical bottle with a plug, precisely adding 400ml of absolute ethyl alcohol, sealing, performing ultrasonic treatment (the temperature is not higher than 35 ℃, the power is 100W, the frequency is 40 kHz) for 40 minutes, cooling, filtering, concentrating to 15ml, transferring the concentrated solution into a 20ml volumetric flask, adding absolute ethyl alcohol to a scale, shaking uniformly, filtering, and taking the subsequent filtrate.
(3) Analysis by gas chromatography
The chromatographic conditions are as follows: capillary column with (5% -phenyl) -methyl polysiloxane as stationary phase; a FID detector; the carrier gas is nitrogen, and the flow rate of the carrier gas is 0.7ml/min; the temperature of the sample inlet is 250 ℃; the detector temperature was 260 ℃; the split ratio is 20:1; sample injection amount: 2 μl; the temperature programming condition is that the initial temperature is 100 ℃, and the temperature is kept for 15 minutes; raising the temperature to 110 ℃ at a rate of 5 ℃ per minute for 25 minutes; raising the temperature to 150 ℃ at a rate of 4 ℃ per minute and keeping for 30 minutes; the temperature was raised to 250℃at a rate of 10℃per minute and maintained for 10 minutes.
(4) And (3) measuring: 2 mu l of each of the reference substance solution and the four-substance capsule sample solution is precisely sucked, injected into a gas chromatograph, measured and recorded. The GC characteristic spectrum of the four-material capsule constructed by the application comprises 9 common main components, 3-n-butenyl phthalide is taken as a reference object, a No. 7 peak is taken as a reference object peak (S peak), 9 common characteristic peaks are arranged in the characteristic spectrum, the corresponding peak of the reference object peak is taken as the S peak, and the relative retention time of each characteristic peak and the S peak is calculated, wherein the relative retention time is within +/-10% of a specified value. The prescribed value is peak 1:0.121, peak 2:0.226, peak 3:0.389, peak 4:0.464, peak 5:0.872, peak 6:0.970, peak 7:1.000 (S peak), peak 8:1.082, peak 9:1.111.
similar to the results in example 2, for the detection of four-substance capsules, the 3-n-butenyl phthalide is taken as a reference substance, the peak of the compound is positioned in the middle of a chromatogram, the separation degree between the compound and other chromatographic peaks is good, the peak corresponding to the 3-n-butenyl phthalide is taken as an S peak, and the calculated relative retention time and relative peak area of each characteristic peak are moderate. The results are shown in tables 5 to 6.
The GC characteristic spectrum of the four-material capsule comprises 9 common main components, ligustilide is taken as a reference object, a No. 9 peak is taken as a reference object peak (S peak), 9 common characteristic peaks are arranged in the characteristic spectrum, the corresponding peak of the reference object is taken as the S peak, the relative retention time and the relative peak area of each characteristic peak and the S peak are calculated, and the relative retention time of the other 8 peaks is less than 1.000; and the relative peak area values of the peak 1, the peak 2, the peak 3, the peak 4, the peak 5, the peak 6, the peak 7 and the peak 8 are smaller and smaller than 0.100.
Obviously, 3-n-butenyl phthalide (peak 7) is more suitable as a reference peak to determine the GC characteristic of the four-substance capsule than ligustilide (peak 9).
Thus the 3-n-butenyl phthalide peak was chosen as the reference peak to determine the four-material capsules.
Table 5 relative retention time results for four-substance capsules
TABLE 6 relative peak area results for four capsules
Example 4
The embodiment provides methodology investigation of the construction of the characteristic spectrum of the volatile oil and the inclusion compound of the four-material capsule extract.
(1) Specificity investigation: taking a blank auxiliary material sample, and preparing a blank auxiliary material sample solution according to a preparation method of a sample solution in the characteristic spectrum construction method in the embodiment 2. Meanwhile, a reference solution and a test solution are prepared. The above solution was taken and analyzed according to the chromatographic conditions in the characteristic spectrum construction method in example 2.
Specificity conclusion: the blank auxiliary materials have no interference on the detection of the volatile oil and the inclusion compound characteristic spectrum of the four-material capsule extract. The characteristic patterns of the volatile oil and the clathrate compound to be tested show 9 characteristic peaks, wherein the peak corresponding to the 3-n-butenyl phthalide peak is a reference peak (S peak). The results are shown in figures 2-5 of the drawings.
(2) Repeatability: taking the same volatile oil and clathrate compound sample solution, continuously sampling for 6 times according to the chromatographic conditions in the characteristic spectrum construction method described in the above embodiment 2, detecting the characteristic spectrum, recording the retention time and the peak area of each common peak, and calculating the relative retention time and the relative peak area of each common peak by taking the retention time and the peak area of the 3-n-butenyl phthalide peak as references.
Repeatability conclusion: each common peak has a relative retention time RSD value of less than 5.0%; the RSD value of each common peak relative peak area is less than 10.0%. The method has good repeatability.
TABLE 7 repeatability test results-volatile oil test solution (1)
TABLE 8 precision test results-volatile oil test solution (2)
TABLE 9 repeatability test results-clathrate test sample solution (1)
TABLE 10 repeatability test results-clathrate test sample solution (2)
(3) Precision: different experimenters use different instruments on different dates to take the same part of volatile oil and clathrate compound sample solution, sample the characteristic spectrum continuously for 6 times according to the chromatographic conditions in the characteristic spectrum construction method described in the above embodiment 2, record the retention time and the peak area of each common peak, and calculate the relative retention time and the relative peak area of each common peak by taking the retention time and the peak area of the 3-n-butenyl phthalide peak as references.
Conclusion of precision: each common peak has a relative retention time RSD value of less than 5.0%; the RSD value of each common peak relative peak area is less than 10.0%. The method has good precision.
TABLE 11 precision test results-volatile oil test solution (1)
TABLE 12 precision test results-volatile oil test sample solution (2)
TABLE 13 precision test results-clathrate test sample solution (1)
TABLE 14 precision test results-clathrate test sample solution (2)
(4) Solution stability: taking the same volatile oil and clathrate compound sample solution, respectively carrying out sample injection at 0h, 6h, 12h, 24h, 48h and 60h according to chromatographic conditions in the characteristic spectrum construction method described in the above embodiment 2, detecting the characteristic spectrum, recording the retention time and the peak area of each common peak, and calculating the relative retention time and the relative peak area of each common peak by taking the retention time and the peak area of the 3-n-butenyl phthalide peak as references.
Solution stability conclusion: each common peak has a relative retention time RSD value of less than 5.0%; the RSD value of each common peak relative peak area is less than 10.0%. The stability of the test sample solution is indicated to be good.
TABLE 15 stability test results-volatile oil test solution (1)
TABLE 16 stability test results-volatile oil test solution (2)
TABLE 17 stability test results-clathrate test sample solution (1)
TABLE 18 stability test results-clathrate test sample solution (2)
The detection method of the feature spectrum of the volatile oil and the inclusion compound of the four-material capsule extract provided by the application is simple and easy to operate, the corresponding chromatographic peak of the feature components is kept complete, and the solution stability is good, so that the detection method can be used for detecting the volatile oil and the inclusion compound of the four-material capsule extract. Meanwhile, the method has high precision, good reproducibility and certain specificity, and the separation effect among various chromatographic peaks in the obtained characteristic spectrum is good. The four-material capsule extract volatile oil or the inclusion compound characteristic spectrum constructed by the construction method of the four-material capsule extract volatile oil and the inclusion compound GC characteristic spectrum provided by the application can effectively characterize the quality of the four-material capsule extract volatile oil or the inclusion compound characteristic spectrum, ensure the stability and consistency of the quality of medicines, and is beneficial to comprehensively controlling the quality of four-material capsule preparations.
Claims (15)
1. A method for detecting volatile oil or inclusion compound thereof from a four-material capsule extract, the method comprising the following steps:
1) Determining a Gas Chromatography (GC) characteristic spectrum of the drug to be detected by using a GC method;
2) Determining the GC profile of 3-n-butenyl phthalide using GC method and comparing it with the GC profile in step 1);
3) Taking the 3-n-butenyl phthalide peak obtained in the step 2) as a reference peak (S peak), and judging that the medicine to be detected contains four capsule extract volatile oil or inclusion compound thereof when the GC characteristic spectrum comprises 9 characteristic peaks and the relative retention time and the relative deviation of the characteristic peaks meet the following conditions:
peak No. 1: the relative retention time is 0.121+/-0.012, and the relative deviation is 0-10%;
peak No. 2: the relative retention time is 0.226+/-0.023, and the relative deviation is 0-10%;
peak No. 3: the relative retention time is 0.389+/-0.039, and the relative deviation is 0-10%;
peak No. 4: the relative retention time is 0.464+/-0.046, and the relative deviation is 0-10%;
peak No. 5: the relative retention time is 0.872+/-0.087, and the relative deviation is 0-10%;
peak No. 6: the relative retention time is 0.970+/-0.097, and the relative deviation is 0-10%;
peak No. 7: the relative retention time is 1.000, and the relative deviation is 0-10%;
peak No. 8: the relative retention time is 1.082+/-0.108, and the relative deviation is 0-10%;
peak No. 9: the relative retention time is 1.111+/-0.111, and the relative deviation is 0-10%;
wherein the conditions of the gas chromatography include the following: a capillary chromatographic column taking (5% -phenyl) -methyl polysiloxane as a stationary phase; a FID detector; the program is run at elevated temperature conditions.
2. The method of claim 1, wherein the chromatographic column is 30m or 60m long; an inner diameter of 0.32mm or 0.53mm; the film thickness was 0.25 μm, 0.5 μm or 1 μm.
3. The method of claim 1, wherein the carrier gas used is nitrogen or helium.
4. The method of claim 1, wherein a carrier gas flow rate of 0.6-0.8 ml/min is used.
5. The method of claim 4, wherein a carrier gas flow of 0.7ml/min is used.
6. The method of claim 1, wherein the sample inlet temperature is 250 ℃ to 260 ℃; the temperature of the detector is 260-270 ℃.
7. The method of claim 6, wherein the sample inlet temperature is 250 ℃.
8. The method of claim 6, wherein the detector temperature is 260 ℃.
9. The method of claim 1, wherein the split ratio is 10-20:1.
10. The method of claim 9, wherein the split ratio is 20:1.
11. The method of claim 1, wherein the sample amount is 1-2 μl.
12. The method of claim 11, wherein the sample is introduced in an amount of 2 μl.
13. The method of claim 1, wherein the elevated temperature condition is an initial temperature of 100 ℃ for 15 minutes; raising the temperature to 110 ℃ at a rate of 5 ℃ per minute for 25 minutes; raising the temperature to 150 ℃ at a rate of 4 ℃ per minute and keeping for 30 minutes; the temperature was raised to 250℃at a rate of 10℃per minute and maintained for 10 minutes.
14. A quality control method for four-material capsule preparation process, comprising the detection method of any one of claims 1-4.
15. A device for detecting volatile oil or its clathrate of four-substance capsule extract comprises at least one processor, and
memory storing instructions which, when executed by at least one processor, perform the comparing steps of the method according to any one of claims 1-13.
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| WO2020239054A1 (en) * | 2019-05-30 | 2020-12-03 | 特丰制药有限公司 | Method for determining chloralkane content in chloral hydrate or preparation thereof |
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| CN101183091A (en) * | 2007-12-18 | 2008-05-21 | 天津中新药业集团股份有限公司达仁堂制药厂 | Measuring method of agastache for dispelling turbidity soft capsules fingerprint |
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