A Zhengqi Tablet, its preparation process and a method for quantitatively and qualitatively determining multiple components therein

Abstract

The present invention provides a Zhengqi tablet, which comprises the following components by mass: 0.5-1.5 parts of Pogostemon oil; 0.1-1 part of Perilla leaf oil; 150-250 parts of Aucklandia lappa; 100-150 parts of Atractylodes lancea; 10-100 parts of Glycyrrhiza uralensis; 150-250 parts of Poria cocos; 100-150 parts of Citrus reticulata Blanco; 100-150 parts of Rhizoma Pinelliae preparatum; 100-150 parts of Magnolia officinalis var. biloba; 100-150 parts of Zingiber officinale. The present invention also provides a preparation process of the Zhengqi tablet. The present invention further provides a method for the quantitative and qualitative determination of multiple components in the Zhengqi tablet. The Zhengqi tablet, the preparation process thereof and the method for the quantitative and qualitative determination of multiple components provided by the present invention can quickly identify 10 components in the Zhengqi tablet, quickly quantify 7 of them, with simple operation, shortened detection time, good linear relationship, good repeatability, good precision, high accuracy and good stability.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine preparation and detection of traditional Chinese medicine components, and relates to a Zhengqi tablet and a preparation process thereof and a quantitative and qualitative determination method of multiple components therein. Background Art

[0002] Zhengqi Tablets are traditional Chinese patent medicines, mainly used to treat colds, headaches, chest tightness, vomiting, diarrhea, abdominal distension, etc. It can relieve heat and dampness, and is mostly used for fever, chest tightness, abdominal distension, vomiting and diarrhea caused by exogenous heat and dampness. It can also soothe the stomach and stop vomiting, and is mostly used for nausea and vomiting caused by excessive dampness and turbidity. At the same time, it can aromaticize turbidity, and is often used for loss of appetite, thick and greasy tongue coating, diarrhea and other symptoms caused by spleen dampness and stomach turbidity. It belongs to the Huoxiang Zhengqi series of products, but there are significant differences in the prescription, prescription amount and process between Zhengqi Tablets and Huoxiang Zhengqi Tablets (pills, drops). The material basis of the two is quite different, and they belong to different Chinese patent medicines. Therefore, the composition, preparation and quantitative and qualitative detection of the components of Zhengqi Tablets cannot be referenced from Huoxiang Zhengqi Tablets.

[0003] At present, various studies are being carried out on Zhengqi tablets. Traditional Chinese medicine fingerprints have the characteristics of overall, macroscopic, and fuzzy analysis. They can achieve the purpose of overall quality control by describing the overall characteristics of traditional Chinese medicine and using appropriate fuzzy processing methods. Therefore, they have become an effective means of quality control of traditional Chinese medicine. Among them, chromatographic fingerprint analysis can visualize the overall characteristics of various chemical components contained in traditional Chinese medicine, thereby revealing quality problems that are difficult to find in conventional inspections. However, traditional Chinese medicine fingerprints cannot quantitatively and qualitatively determine many components in traditional Chinese medicine. Chinese patent 2018109246518 established a high-performance liquid fingerprint of Zhengqi tablets, but did not quantitatively and qualitatively determine the components contained in Zhengqi tablets. Chinese patent 2020113871985 established a gas phase fingerprint of Zhengqi tablets, only pointing out 3 index components, but also did not quantitatively and qualitatively determine the components contained in Zhengqi tablets. At the same time, Chinese patent 2022110331062 only quantitatively determined 4 components in the Zhengqi tablet extract, which is an intermediate of Zhengqi tablets. It can be seen that the research on Zhengqi Tablets is not comprehensive, and there is a lack of rapid quantitative and qualitative determination of the many components in Zhengqi Tablets. Summary of the invention

[0004] In view of the shortcomings of the prior art mentioned above, the purpose of the present invention is to provide a Zhengqi tablet and its preparation process and a method for quantitative and qualitative determination of multiple components therein, which can provide Zhengqi tablets with stable quality and low unqualified rate, and through a reliable preparation method and quantitative and qualitative determination of 10 components in Zhengqi tablets, the lack of the prior art is solved. By conducting quantitative and qualitative determination of multiple components in Zhengqi tablets, the quality of Zhengqi tablets can be objectively, comprehensively and accurately evaluated, which is of great significance for controlling the quality of Zhengqi tablets and ensuring clinical efficacy.

[0005] To achieve the above and other related objectives, the first aspect of the present invention provides a Zhengqi tablet, which, by mass fraction, comprises the following components:

[0006] Patchouli essential oil 0.5 - 1.5 parts; specifically, such as 0.5 - 0.8 parts, 0.8 - 1.2 parts, 1.2 - 1.5 parts; Perilla leaf oil 0.1 - 1 part; specifically, such as 0.1 - 0.4 parts, 0.4 - 0.8 parts, 0.8 - 1 part;

[0007] Costus root 150 - 250 parts; specifically, such as 150 - 180 parts, 180 - 220 parts, 220 - 250 parts; Atractylodes lancea 100 - 150 parts; specifically, such as 100 - 130 parts, 130 - 140 parts, 140 - 150 parts; Licorice root 10 - 100 parts; specifically, such as 10 - 60 parts, 60 - 70 parts, 70 - 100 parts;

[0008] Poria cocos 150 - 250 parts; specifically, such as 150 - 180 parts, 180 - 220 parts, 220 - 250 parts; Dried tangerine peel 100 - 150 parts; specifically, such as 100 - 130 parts, 130 - 140 parts, 140 - 150 parts; Prepared Pinellia ternata 100 - 150 parts; specifically, such as 100 - 130 parts, 130 - 140 parts, 140 - 150 parts; Ginger-processed Magnolia officinalis 100 - 150 parts; specifically, such as 100 - 130 parts, 130 - 140 parts, 140 - 150 parts; Fresh ginger 100 - 150 parts; specifically, such as 100 - 130 parts, 130 - 140 parts, 140 - 150 parts. Preferably, the Zhengqi tablet, by mass fraction, comprises the following components:

[0009] Patchouli essential oil 0.8 - 1.2 parts;

[0010] Perilla leaf oil 0.4 - 0.8 parts;

[0011] Costus root 180 - 220 parts;

[0012] Atractylodes lancea 130 - 140 parts;

[0013] Licorice root 60 - 70 parts;

[0014] Poria cocos 180 - 220 parts;

[0015] Dried tangerine peel 130 - 140 parts;

[0016] Prepared Pinellia ternata 130 - 140 parts;

[0017] Ginger-processed Magnolia officinalis 130 - 140 parts;

[0018] Fresh ginger 130 - 140 parts.

[0019] Preferably, the Zhengqi tablet, by mass fraction, comprises the following components:

[0020] 1.0 part of patchouli oil;

[0021] 0.6 part of perilla leaf oil;

[0022] 200 parts of costus root;

[0023] 133 parts of atractylodes rhizome;

[0024] 67 parts of licorice root;

[0025] 200 parts of poria cocos;

[0026] 133 parts of tangerine peel;

[0027] 133 parts of processed pinellia tuber;

[0028] 133 parts of ginger-processed magnolia bark;

[0029] 133 parts of fresh ginger.

[0030] Preferably, the patchouli oil and perilla leaf oil should be accurately weighed.

[0031] Preferably, the costus root, atractylodes rhizome, licorice root, poria cocos, tangerine peel, pinellia tuber, magnolia bark, and fresh ginger should be accurately weighed and then ground into powder. The pinellia tuber is the processed pinellia tuber, and the magnolia bark is the ginger-processed magnolia bark. The fresh ginger is the fresh rhizome of Zingiber officinale.

[0032] The patchouli oil is the extracted oil of the plant of the genus Pogostemon in the family Lamiaceae. The perilla leaf oil is the extracted oil of the perilla leaf of the annual upright herb of the dicotyledonous class in the tribe Saturejeae of the family Lamiaceae. The costus root is the dried root of the plant Aucklandia lappa in the family Asteraceae. The atractylodes rhizome is the dried rhizome of the plant Atractylodes lancea or Atractylodes chinensis in the family Asteraceae. The licorice root is the dried root and rhizome of the plant Glycyrrhiza glabra, Glycyrrhiza inflata, or Glycyrrhiza glabra var. glabra in the family Fabaceae. The poria cocos is the dried sclerotium of the fungus Poria cocos in the family Polyporaceae. The tangerine peel is the dried ripe pericarp of the plant Citrus reticulata Blanco and its cultivated varieties in the family Rutaceae. The pinellia tuber is the dried tuber of the plant Pinellia ternata in the family Araceae. The magnolia bark is the dried stem bark, root bark, and branch bark of the plant Magnolia officinalis or Magnolia officinalis var. biloba in the family Magnoliaceae. The fresh ginger is the fresh rhizome of the perennial herb Zingiber officinale in the family Zingiberaceae.

[0033] The second aspect of the present invention provides a preparation process of Zhengqi tablets, comprising the following steps:

[0034] 1) Mix the first part of Aucklandia lappa, Atractylodes lancea, and Glycyrrhiza glabra according to the ratio, crush them into powder, and sieve to obtain the powdered material;

[0035] 2) Decoct the second part of Poria cocos, Citrus reticulata Blanco, Pinellia ternata (Thunb.) Breit., Magnolia officinalis Rehd. et Wils. var. biloba Rehd. et Wils., Zingiber officinale Rosc., and Glycyrrhiza glabra with water according to the ratio, take the decoction, filter, and concentrate to obtain the pasty material;

[0036] 3) Mix the pasty material with the powdered material, granulate, and then add Pogostemon cablin oil and Perilla frutescens oil according to the ratio, and press into tablets.

[0037] Preferably, in step 1) or 2), the mass ratio of the first part of Glycyrrhiza glabra to the second part of Glycyrrhiza glabra is 55 - 65:35 - 45, specifically such as 55 - 60:40 - 45, 60 - 65:35 - 40, and preferably 60:40.

[0038] Preferably, in step 1), sucrose is also added during the mixing.

[0039] Preferably, the mass ratio of the sucrose to the added Aucklandia lappa is 25 - 30:150 - 250, specifically such as 25 - 30:150 - 180, 25 - 30:180 - 220, 25 - 30:220 - 250, preferably 27:180 - 220, and more preferably 27:200.

[0040] Preferably, in step 1), the mesh number of the sieve used for sieving is 80 - 120 meshes, and preferably 100 meshes.

[0041] Preferably, in step 2), decoct until boiling.

[0042] Preferably, in step 2), the number of decoction times is at least 2 times, and preferably 2 times.

[0043] Preferably, when the number of decoction times is 2 times, the first decoction time is 2 - 4 hours, and preferably 3 hours; the second decoction time is 1 - 3 hours, and preferably 2 hours.

[0044] Preferably, when the number of decoction times is 2 times, the water addition amount for the first decoction is 7.5 - 8.5 times the amount, and preferably 8 times the amount; the water addition amount for the second decoction is 5.5 - 6.5 times the amount, and preferably 6 times the amount.

[0045] The above-mentioned decoction is obtained by combining at least 2 times of decoctions.

[0046] Preferably, in step 2), the filtration is carried out using a basket filter. The basket filter is a commonly used basket filter.

[0047] Preferably, in step 2), the concentration temperature is 50 - 70 °C.

[0048] Preferably, in step 2), it is concentrated to a relative density of 1.03 - 1.05, thereby obtaining a dense paste.

[0049] Preferably, in step 3), when the paste is mixed with the powder, an appropriate amount of water is added to the paste. The appropriate amount of water addition means adding water until the paste can flow.

[0050] Preferably, in step 3), the granulation is a commonly used granulation process. Specifically, according to the divided pot batches of the granulation ingredients, the powder is used as the bottom material and put into a fluidized bed granulator, and the paste is sprayed in for granulation.

[0051] Preferably, in step 3), when adding patchouli essential oil and perilla leaf oil, a lubricant is also added.

[0052] Preferably, the lubricant is talc powder.

[0053] Preferably, the addition amount of the lubricant is 1.5 - 2.5% of the weight of the granules, preferably 2%.

[0054] Preferably, in step 3), the tabletting is a commonly used tabletting process for traditional Chinese medicine tablets.

[0055] The third aspect of the present invention provides a Zhengqi tablet prepared by the above process.

[0056] The fourth aspect of the present invention provides a method for quantitative and qualitative determination of multiple components in Zhengqi tablets, including: adding Zhengqi tablets to methanol for dissolution, ultrasonic extraction, cooling, taking the supernatant for filtration, and detecting the obtained test solution by ultra - high performance liquid chromatography (UPLC) to qualitatively determine 10 components in the test solution: liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylodin, perillaketone, nobiletin, and tangeretin, and quantitatively determining the contents of 7 components in the test solution: liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, and atractylodin.

[0057] Preferably, the CAS number of liquiritin is 551 - 15 - 5, the CAS number of hesperidin is 520 - 26 - 3, the CAS number of honokiol is 35354 - 74 - 6, the CAS number of costunolide is 553 - 21 - 9, the CAS number of dehydrocostus lactone is 74299 - 48 - 2, the CAS number of magnolol is 528 - 43 - 8, the CAS number of atractylodin is 55290 - 63 - 6, the CAS number of perillaketone is 142546 - 15 - 4, the CAS number of nobiletin is 478 - 01 - 3, and the CAS number of tangeretin is 481 - 53 - 8.

[0058] Preferably, the ratio of the mass g of the Zhengqi tablets added to the volume mL of methanol added is 0.3:15 - 25, preferably 0.3:20.

[0059] Preferably, the time for ultrasonic extraction is 30 - 60 minutes, preferably 40 - 50 minutes, and more preferably 45 minutes.

[0060] Preferably, the power of ultrasonic extraction is 200 - 350 W, preferably 250 W; the frequency of ultrasonic extraction is 40 - 60 kHz, preferably 40 kHz.

[0061] Preferably, it is cooled to room temperature, and the room temperature is 20 - 30 °C.

[0062] Preferably, the filtration is to take the supernatant and filter it through a membrane.

[0063] Preferably, the membrane is a 0.22 μm membrane.

[0064] Preferably, the detection is carried out by ultra - performance liquid chromatography (UPLC), including the following steps:

[0065] A1) Preparation of reference solution: Add reference substances of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylodin, perillaketone, nobiletin, and tangeretin to methanol, dissolve and make up the volume to prepare a reference solution.

[0066] A2) Sample detection: Use ultra - performance liquid chromatography (UPLC) to detect the test solution and the reference solution in step A1) respectively. Qualitative analysis is carried out by comparing the retention times, and quantitative analysis is carried out by the external standard method. Qualitatively determine 10 components in the test solution: liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylodin, perillaketone, nobiletin, and tangeretin. Quantitatively determine the contents of 7 components in the test solution: liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, and atractylodin.

[0067] The above ultra - performance liquid chromatography (UPLC) is different from the traditional high - performance liquid chromatography (HPLC). It covers new technologies such as small - particle fillers, very low system volume, and rapid detection methods, increasing the analysis throughput, sensitivity, and chromatographic peak capacity. Compared with the problems of long detection time, complex chromatographic peaks, and affecting resolution in the HPLC method, the detection speed, sensitivity, and resolution of UPLC are incomparable to those of HPLC. The speed, sensitivity, and resolution of UPLC are 9 times, 3 times, and 1.7 times that of HPLC respectively. It shortens the analysis time, reduces the solvent consumption, and lowers the analysis cost.

[0068] Preferably, in step A1), the reference solution should be shaken well, filtered, and the subsequent filtrate should be taken.

[0069] Preferably, in step A1), the reference solution is prepared by successive dilution. Specifically, a stock solution is first prepared and then diluted with methanol to obtain the reference solution. The stock solution is stored in a refrigerator at 4°C in the dark.

[0070] Preferably, in step A1), the content range of liquiritin in the reference solution is 4.52 - 45.22 μg / ml, preferably 10 μg / ml; the content range of hesperidin is 9.50 - 237.61 μg / ml, preferably 50 μg / ml; the content range of honokiol is 1.98 - 49.50 μg / ml, preferably 10 μg / ml; the content range of costunolide is 10.44 - 261.14 μg / ml, preferably 50 μg / ml; the content range of dehydrocostus lactone is 21.49 - 531.30 μg / ml, preferably 100 μg / ml; the content range of magnolol is 1.79 - 44.86 μg / ml, preferably 10 μg / ml; the content range of atractylodin is 1.92 - 47.96 μg / ml, preferably 10 μg / ml; the content range of perillaketone is 15 - 25 μg / ml, preferably 20 μg / ml; the content range of nobiletin is 40 - 60 μg / ml, preferably 50 μg / ml; the content range of tangeretin is 15 - 25 μg / ml, preferably 20 μg / ml.

[0071] Preferably, in step A2), in the ultra - performance liquid chromatography (UPLC), the detector is a diode array detector (DAD).

[0072] Preferably, in step A2), the chromatographic column in the ultra - performance liquid chromatography is a T3 chromatographic column.

[0073] More preferably, the chromatographic column in the ultra - performance liquid chromatography is a Waters Acquity UPLC HSS T3 chromatographic column (column length: 100 mm, inner diameter: 2.1 mm, packing particle size: 1.8 μm).

[0074] Preferably, in step A2), the detection wavelength in the ultra - performance liquid chromatography is 224 - 226 nm. More preferably, the detection wavelength is 225 nm.

[0075] Preferably, in step A2), the column temperature in the ultra - performance liquid chromatography is 30 - 45°C, specifically such as 30 - 35°C, 35 - 45°C, 32 - 38°C, preferably 35°C.

[0076] Preferably, in step A2), the flow rate in the ultra-high performance liquid chromatography is 0.1 - 0.3 mL / min. More preferably, the flow rate in the ultra-high performance liquid chromatography is 0.2 mL / min.

[0077] Preferably, in step A2), the injection volume in the ultra-high performance liquid chromatography is 0.5 - 5 μL. More preferably, the injection volume in the ultra-high performance liquid chromatography is 1 μL.

[0078] Preferably, in step A2), in the ultra-high performance liquid chromatography, the mobile phase is acetonitrile - 0.05 - 0.15% formic acid aqueous solution, where phase A is acetonitrile and phase B is 0.05 - 0.15% formic acid aqueous solution; the analysis time is 19 min; gradient elution is performed.

[0079] More preferably, in the ultra-high performance liquid chromatography, the mobile phase is acetonitrile - 0.10% formic acid aqueous solution, where phase A is acetonitrile and phase B is 0.10% formic acid aqueous solution; the analysis time is 19 min; gradient elution is performed.

[0080] The above 0.05 - 0.15% formic acid aqueous solution is a formic acid aqueous solution with a volume percentage of 0.05 - 0.15%. The above 0.10% formic acid aqueous solution is a formic acid aqueous solution with a volume percentage of 0.10.

[0081] More preferably, the specific program of the gradient elution is shown in Table 1 and is as follows:

[0082] 0 - 4 min, the volume ratio of phase A to phase B is 20:80 - 35:65;

[0083] 4 - 5 min, the volume ratio of phase A to phase B is 35:65 - 40:60;

[0084] 5 - 6 min, the volume ratio of phase A to phase B is 40:60 - 55:45;

[0085] 6 - 15 min, the volume ratio of phase A to phase B is 55:45 - 60:40;

[0086] 15 - 16 min, the volume ratio of phase A to phase B is 60:40 - 90:10;

[0087] 16 - 19 min, the volume ratio of phase A to phase B is 90:10 - 90:10.

[0088] The analysis time of this determination method is only 19 min, which can quickly determine 10 components in Zhengqi tablets in a short time, not only saving reagents but also greatly reducing the daily QC detection work.

[0089] Table 1

[0090]

[0091] Preferably, in step A2), the external standard method includes the following steps:

[0092] A) Prepare a series of reference substance solutions with different concentrations according to step A1), perform UPLC detection respectively, obtain the linear relationships between the chromatographic peak areas of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, and atractylodin and the corresponding contents of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, and atractylodin, plot the corresponding standard working curves, and calculate the regression equations of the standard working curves of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, and atractylodin respectively;

[0093] B) Perform UPLC detection on the test solution, substitute the obtained chromatographic peak areas of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, and atractylodin into the regression equations of the corresponding standard working curves of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, and atractylodin in step A), and calculate the contents of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, and atractylodin in the test solution.

[0094] More preferably, in the standard working curve, the chromatographic peak areas of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, and atractylodin are used as the ordinate (Y-axis), and the corresponding contents (i.e., concentrations) of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, and atractylodin are used as the abscissa (X-axis).

[0095] In the ultra-high performance liquid chromatography (UPLC) detection of the above perillaketone, nobiletin, and tangeretin, due to the too small peak areas and too low contents of perillaketone and tangeretin, which do not meet the general content requirements (≥0.02%), they are not suitable as quantitative indicators. At the same time, according to the provisions of the Chinese Pharmacopoeia, when quantifying, the resolution between the chromatographic peaks of the substance to be measured and the adjacent chromatographic peaks should be greater than 1.5, and perillaketone, nobiletin, and tangeretin do not meet this requirement, so they can only be qualitatively determined.

[0096] As described above, the present invention provides a Zhengqi tablet, its preparation process, and a method for quantitative and qualitative determination of multiple components therein. The composition and preparation of the Zhengqi tablet are determined. By using the pretreatment under optimized conditions and the UPLC instrument detection method, 10 components in the Zhengqi tablet can be quickly qualitatively analyzed within an extremely short time, such as 19 minutes, and 7 components (liquiritin, hesperidin, honokiol, magnolol, costunolide, dehydrocostus lactone, and atractylodin) can be quickly quantitatively analyzed. This method is simple to operate, has strong repeatability, and is easy to control. The pretreatment of this method is simple, which improves the problems of complicated pretreatment and poor parallelism of hesperidin in the past. This method can detect multiple components simultaneously, and in addition, it greatly shortens the detection time. This method is beneficial to the control of front-end medicinal materials. Liquiritin is the main component of licorice, hesperidin is the main component of tangerine peel, honokiol and magnolol are the main components of ginger-processed magnolia bark, costunolide and dehydrocostus lactone are the main components of Aucklandia lappa Decne., and atractylodin is the main component of Atractylodes lancea (Thunb.) DC. Therefore, when controlling the quality of the compound prescription, 5 medicinal materials, including licorice, tangerine peel, ginger-processed magnolia bark, Aucklandia lappa Decne., and Atractylodes lancea (Thunb.) DC., can be controlled, ensuring stable production and clinical efficacy. The standard curves of the 7 components determined by this method have good linear relationships within their respective ranges, with good repeatability, precision, accuracy, and stability, and can be used for data accumulation of multiple batches of samples, formulating reasonable content limits, and ensuring stable production and clinical efficacy.

[0097] Compared with the prior art, the present invention has obvious detection advantages. For example, Chinese Patent No. 2018109246518 only provides the HPLC fingerprint of the Zhengqi tablet obtained by a gradient elution program with an analysis time of 115 minutes, and does not perform quantitative and qualitative analysis on any components in the Zhengqi tablet. Another example is that Chinese Patent No. 2020113871985 only provides the GC fingerprint of the Zhengqi tablet obtained by a temperature programming procedure, and does not perform quantitative and qualitative analysis on any components in the Zhengqi tablet. More specifically, although Chinese Patent No. 2022110331062 performs quantitative and qualitative analysis of components, its analysis object is the extract of the Zhengqi tablet, and only the quantitative and qualitative analysis of 4 components in the extract of the Zhengqi tablet obtained by a gradient elution program with an analysis time of 34 minutes is carried out.

[0098] Compared with Chinese Patent No. 2018109246518, the detection instrument UPLC adopted in the present invention is superior to the HPLC adopted therein; compared with Chinese Patent No. 2020113871985, the detection instrument UPLC adopted in the present invention is completely different from the GC adopted therein; compared with Chinese Patent No. 2022110331062, the analysis object of the present invention is Zhengqi Tablets, which is different from the Zhengqi Tablet extract as the intermediate of Zhengqi Tablets. It should be particularly noted that the present invention only provides a gradient elution program with an analysis time of 19 minutes. Its detection time is much shorter than the above-mentioned prior art, and it can simultaneously identify 10 components and quantify 7 components, greatly saving the solvent amount and reducing the cost. Description of the Drawings

[0099] Figure 1 It shows the chromatogram for the specificity study of the components in the present invention. Among them, S1 is the Zhengqi Tablet sample without Aucklandia lappa and Atractylodes lancea, S2 is the Zhengqi Tablet sample without Atractylodes lancea, S3 is the Zhengqi Tablet sample without Aucklandia lappa, S4 is the Zhengqi Tablet sample without Magnolia officinalis var. biloba, S5 is the Zhengqi Tablet sample without Citrus reticulata Blanco, S6 is the Zhengqi Tablet sample without Glycyrrhiza uralensis Fisch., S7 is the Zhengqi Tablet sample, and S8 is the reference substance; 1 is liquiritin, 2 is hesperidin, 3 is honokiol, 4 is costunolide, 5 is dehydrocostus lactone, 6 is magnolol, and 7 is atractylodin.

[0100] Figure 2 It shows the UPLC chromatograms of the test sample and the reference substance in the present invention. Among them, S1 is the UPLC chromatogram of the test sample, and S2 is the UPLC chromatogram of the reference substance; 1 is liquiritin, 2 is hesperidin, 3 is perillaketone, 4 is nobiletin, 5 is tangeretin, 6 is honokiol, 7 is costunolide, 8 is dehydrocostus lactone, 9 is magnolol, and 10 is atractylodin. Detailed Embodiments

[0101] The present invention will be further elaborated below in combination with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the protection scope of the present invention.

[0102] The following specific examples illustrate the implementation manners of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific implementation manners. Various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention.

[0103] The reagents and instruments used in the following examples are as follows:

[0104] 1. Reagents

[0105] 10 batches of Zhengqi Tablets (batch numbers 200401, 200402, 210101, 210102, 21010601, 21010701, 210201, 210204, 210901, 210902, prepared by Shanghai Hutchison Pharmaceutical Co., Ltd.); negative samples (lacking licorice Zhengqi Tablets, lacking tangerine peel Zhengqi Tablets, lacking magnolia bark Zhengqi Tablets, and lacking atractylodes rhizome Zhengqi Tablets) were all provided by Shanghai Hutchison Pharmaceutical Co., Ltd.

[0106] Reference substances: liquiritin (batch number 111610 - 201908, mass fraction 95%), hesperidin (batch number 110721 - 201818, mass fraction 96.2%), honokiol (batch number 110730 - 201915, mass fraction 99.8%), magnolol (batch number 110729 - 201513, mass fraction 98.8%), costunolide (batch number 111524 - 201911, mass fraction 99.9%), dehydrocostus lactone (batch number 111525 - 201912, mass fraction 99.5%), atractylodin (batch number 111924 - 201806, mass fraction 99.5%), all of the above reference substances were purchased from the National Institutes for Food and Drug Control. Perillaketone (batch number 1681 - 025A1, mass fraction 98.7%) was purchased from TLC Pharmaceutical Standards Ltd., Canada. Nobiletin (batch number 6082, mass fraction 98.7%) and tangeretin (batch number 6083, mass fraction 99.5%) were purchased from Shanghai Standard Technology Service Co., Ltd.

[0107] Reagents: anhydrous methanol (analytical pure AR, Sinopharm Chemical Reagent Co., Ltd.); acetonitrile (chromatographic pure, TEDIA Company, USA); formic acid (chromatographic pure, TEDIA Company, USA); ultrapure water was prepared by Milli - Q ultrapure water treatment system.

[0108] 2. Instruments

[0109] Waters Acquity UPLC H - Class ultra - high performance liquid chromatography instrument (equipped with Empower 3 chromatographic workstation, quaternary ultra - high pressure solvent manager, automatic injection sample manager, column oven, PDAeλ detector, Waters Company, USA); SB - 5200DTD ultrasonic cleaner (Ningbo Xinzhi Biotechnology Co., Ltd.); AL204 and XS205 analytical electronic balances (METTLER TOLEDO Instrument Shanghai Co., Ltd.); Milli - Q Advantage A10 ultrapure water system (Merck Millipore, Germany).

[0110] Example 1

[0111] Take the first part of 200 g of Aucklandia lappa, 133 g of Atractylodes lancea, and 40.2 g of Glycyrrhiza uralensis, add 27 g of sucrose, and crush them together into fine powder. Pass through a 100-mesh sieve to obtain powder 1#. Take the second part of 200 g of Poria cocos, 133 g of Citrus reticulata Blanco, 133 g of Pinellia ternata (Thunb.) Breit., 133 g of Magnolia officinalis Rehd. et Wils. var. biloba Rehd. et Wils., 133 g of Zingiber officinale Roscoe, and 26.8 g of Glycyrrhiza uralensis, decoct with water twice. The first water addition is 8 times the amount, decoct until boiling for 3 hours, the second water addition is 6 times the amount, decoct until boiling for 2 hours. Combine the decoction liquids, filter through a basket filter, and concentrate at 60 °C to a relative density of 1.04 to obtain extract paste 1#. Mix extract paste 1# with powder 1# to granulate. When mixing the extract paste and the powder, add an appropriate amount of water to the extract paste until it can flow, then add 1.0 g of Pogostemon cablin oil, 0.6 g of Perilla frutescens oil, and 2% of the granule weight of talc powder, and press into 1000 tablets to obtain Zhengqi Tablet sample 1#.

[0112] Example 2

[0113] Take the first part of 190 g of Aucklandia lappa, 130 g of Atractylodes lancea, and 36 g of Glycyrrhiza uralensis, add 27 g of sucrose, and crush them together into fine powder. Pass through a 110-mesh sieve to obtain powder 2#. Take the second part of 190 g of Poria cocos, 130 g of Citrus reticulata Blanco, 130 g of Pinellia ternata (Thunb.) Breit., 130 g of Magnolia officinalis Rehd. et Wils. var. biloba Rehd. et Wils., 130 g of Zingiber officinale Roscoe, and 24 g of Glycyrrhiza uralensis, decoct with water twice. The first water addition is 8.5 times the amount, decoct until boiling for 3.5 hours, the second water addition is 6.5 times the amount, decoct until boiling for 2.5 hours. Combine the decoction liquids, filter through a basket filter, and concentrate at 65 °C to a relative density of 1.05 to obtain extract paste 2#. Mix extract paste 2# with powder 2# to granulate. When mixing the extract paste and the powder, add an appropriate amount of water to the extract paste until it can flow, then add 0.9 g of Pogostemon cablin oil, 0.5 g of Perilla frutescens oil, and 2.5% of the granule weight of talc powder, and press into 1000 tablets to obtain Zhengqi Tablet sample 2#.

[0114] Example 3

[0115] 1. Preparation of test solution

[0116] Take 0.3 g of Zhengqi Tablet sample 1# prepared in Example 1, accurately weigh it, place it in a 50 mL centrifuge tube, accurately add 20 mL of methanol, ultrasonically extract (250 w, 40 kHz) for 45 minutes, cool to room temperature, take the supernatant and filter through a 0.22 μm filter membrane, and take the subsequent filtrate to obtain test solution 1#.

[0117] 2. Preparation of reference solution

[0118] Take the reference substances of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylodin, perillaketone, nobiletin, and tangeretin, accurately weigh them, dissolve them in methanol and make up to the mark in a 100 mL volumetric flask to prepare a reference stock solution. Store the reference stock solution in a refrigerator at 4 °C in the dark.

[0119] The reference substance stock solution was then diluted and made up to volume with methanol to prepare a series of reference substance solutions with different concentrations. In the series of reference substance solutions with different concentrations, the content range of liquiritin was 4.52 - 45.22 μg / ml; the content range of hesperidin was 9.50 - 237.61 μg / ml; the content range of honokiol was 1.98 - 49.50 μg / ml; the content range of costunolide was 10.44 - 261.14 μg / ml; the content range of dehydrocostus lactone was 21.49 - 531.30 μg / ml; the content range of magnolol was 1.79 - 44.86 μg / ml; the content range of atractylenolide was 1.92 - 47.96 μg / ml; the content range of perillaketone was 20 μg / ml; the content range of nobiletin was 50 μg / ml; the content range of tangeretin was 20 μg / ml. The reference substance solution was also shaken well, filtered, and the subsequent filtrate was taken.

[0120] 3. Determination

[0121] The test solution 1# and a series of reference substance solutions with different concentrations were respectively detected by ultra - high performance liquid chromatography (UPLC). Qualitative analysis was carried out by comparing the retention times, and quantitative analysis was carried out by the external standard method. That is, the retention times were compared to determine 10 components in the test solution: liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylenolide, perillaketone, nobiletin and tangeretin. Then, a series of reference substance solutions with different concentrations were respectively detected by UPLC to obtain the linear relationships between the chromatographic peak areas of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylenolide and their corresponding contents, and the corresponding standard working curves were plotted. The regression equations of the standard working curves of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylenolide were respectively calculated. Then, the test solution 1# was detected by UPLC, and the chromatographic peak areas of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylenolide obtained were substituted into the regression equations of the corresponding standard working curves of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylenolide to calculate the contents of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylenolide in the test solution.

[0122] Among them, the high - performance liquid chromatography method includes the following detection conditions:

[0123] The detector is a diode array detector (DAD); the chromatographic column is a Waters Acquity UPLC HSS T3 chromatographic column (column length is 100 mm, inner diameter is 2.1 mm, packing particle size is 1.8 μm); the detection wavelength is 225 nm; the column temperature is 35 °C; the flow rate is 0.2 mL / min; the injection volume is 1 μl; the mobile phase is acetonitrile - 0.10% formic acid aqueous solution, where phase A is acetonitrile and phase B is 0.10% formic acid aqueous solution; the analysis time is 19 min; gradient elution is performed.

[0124] The specific program for gradient elution is as follows:

[0125] From 0 to 4 min, the volume ratio of phase A to phase B is 20:80 - 35:65;

[0126] From 4 to 5 min, the volume ratio of phase A to phase B is 35:65 - 40:60;

[0127] From 5 to 6 min, the volume ratio of phase A to phase B is 40:60 - 55:45;

[0128] From 6 to 15 min, the volume ratio of phase A to phase B is 55:45 - 60:40;

[0129] From 15 to 16 min, the volume ratio of phase A to phase B is 60:40 - 90:10;

[0130] From 16 to 19 min, the volume ratio of phase A to phase B is 90:10 - 90:10.

[0131] Example 4

[0132] 1. Preparation of the test solution

[0133] Take 0.3 g of the Zhengqi Tablet sample 1# prepared in Example 1, accurately weigh it, place it in a 50 mL centrifuge tube, accurately add 15 mL of methanol, ultrasonically extract (240 w, 35 kHz) for 40 minutes, cool to room temperature, take the supernatant and filter it through a 0.22 μm filter membrane, and take the subsequent filtrate to obtain the test solution 2#.

[0134] 2. Preparation of the reference solution

[0135] Take the reference substances of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylodin, perillaketone, nobiletin, and tangeretin, accurately weigh them, dissolve them in methanol and make up to the mark in a 100 mL volumetric flask to prepare a reference stock solution. The reference stock solution is stored in a refrigerator at 4 °C in the dark. Then, dilute and make up the reference stock solution with methanol to prepare a series of reference solutions with different concentrations.

[0136] In a series of reference substance solutions with different concentrations, the concentration ranges of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylodin, perillaketone, nobiletin and tangeretin are the same as those in step 2 of Example 3.

[0137] 3. Determination

[0138] The test solution 2# and a series of reference substance solutions with different concentrations were respectively detected by ultra-high performance liquid chromatography (UPLC). Qualitative analysis was carried out by comparing the retention times, and quantitative analysis was carried out by the external standard method. The retention times were compared to determine 10 components in the test solution: liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylodin, perillaketone, nobiletin and tangeretin. The concentrations of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol and atractylodin in the test solution 2# were determined. The specific quantitative process was the same as that in step 3 of Example 3.

[0139] Among them, the high performance liquid chromatography includes the following detection conditions:

[0140] The detector is a diode array detector (DAD); the chromatographic column is a Waters Acquity UPLC HSS T3 chromatographic column (column length: 100 mm, inner diameter: 2.1 mm, packing particle size: 1.8 μm); the detection wavelength is 224 nm; the column temperature is 30 °C; the flow rate is 0.1 mL / min; the injection volume is 0.5 μL; the mobile phase is acetonitrile - 0.15% formic acid aqueous solution, where phase A is acetonitrile and phase B is 0.15% formic acid aqueous solution; the analysis time is 34 min; gradient elution is carried out.

[0141] The specific program of gradient elution is the same as that in step 3 of Example 3.

[0142] Example 5

[0143] 1. Preparation of the test solution

[0144] Take 0.3 g of the Zhengqi tablets sample 1# prepared in Example 1, accurately weigh it, place it in a 50 mL centrifuge tube, accurately add 25 mL of methanol, extract by ultrasound (260 w, 45 kHz) for 50 minutes, cool to room temperature, take the supernatant and filter it through a 0.22 μm filter membrane, and take the subsequent filtrate to obtain the test solution 3#.

[0145] 2. Preparation of the reference substance solution

[0146] Take the reference substances of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylodin, perillaketone, nobiletin and tangeretin, accurately weigh them, dissolve them in methanol and make up to the mark in a 100 mL volumetric flask to prepare a reference substance stock solution. The reference substance stock solution is stored in a refrigerator at 4 °C in the dark. Then, dilute and make up the reference substance stock solution with methanol to prepare a series of reference substance solutions with different concentrations.

[0147] In a series of reference substance solutions with different concentrations, the concentration ranges of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylodin, perillaketone, nobiletin and tangeretin are the same as those in step 2 of Example 3.

[0148] 3. Determination

[0149] Use ultra-high performance liquid chromatography (UPLC) to detect the test solution 3# and a series of reference substance solutions with different concentrations respectively. Compare the retention times for qualitative analysis, and use the external standard method for quantitative analysis. Compare the retention times to determine 10 components in the test solution: liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylodin, perillaketone, nobiletin and tangeretin, and determine the concentrations of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol and atractylodin in the test solution 3#. The specific quantitative process is the same as that in step 3 of Example 3.

[0150] Among them, the high performance liquid chromatography includes the following detection conditions:

[0151] The detector is a diode array detector (DAD); the chromatographic column is a Waters Acquity UPLC HSS T3 chromatographic column (column length is 100 mm, inner diameter is 2.1 mm, packing particle size is 1.8 μm); the detection wavelength is 226 nm; the column temperature is 45 °C; the flow rate is 0.3 mL / min; the injection volume is 2 μL; the mobile phase is acetonitrile - 0.05% formic acid aqueous solution, where phase A is acetonitrile and phase B is 0.05% formic acid aqueous solution; the analysis time is 19 min; gradient elution.

[0152] The specific program of gradient elution is the same as that in step 3 of Example 3.

[0153] Example 6

[0154] Take the Zhengqi tablets sample 1# prepared in Example 1 and prepare the test solution according to step 1 in Example 3. At the same time, respectively take the Zhengqi tablets negative samples lacking flavors of licorice, tangerine peel, ginger-processed magnolia bark, Aucklandia lappa and Atractylodes lancea, and prepare the negative test solutions according to step 1 in Example 3. In addition, prepare the reference substance solution according to step 2 in Example 3.

[0155] The above test solution, negative test solution, and reference solution were respectively determined according to Step 3 in Example 3, and the retention times were compared for qualitative analysis. The specific test results are shown in Figure 1 . It can be seen from Figure 1 that there was no negative interference in the 7 chromatographic peaks, indicating good method specificity. Among them, Peak 1 was liquiritin, derived from Glycyrrhiza uralensis Fisch. herbs; Peak 2 was hesperidin, derived from Citrus reticulata Blanco herbs; Peak 3 was honokiol, and Peak 6 was magnolol, both derived from Magnoliae Officinalis Cortex herbs; Peak 4 was costunolide, and Peak 5 was dehydrocostus lactone, both derived from Aucklandia lappa Decne. herbs; Peak 7 was atractylodin, derived from Atractylodes lancea (Thunb.) DC. and Aucklandia lappa Decne. herbs.

[0156] Example 7

[0157] The methodology verification was carried out for the method for the quantitative and qualitative determination of multiple components in the Zhengqi tablets sample of the present invention, and the results of its performance indicators are as follows.

[0158] 1. Linear relationship

[0159] An appropriate amount of reference substances of liquiritin, hesperidin, honokiol, magnolol, costunolide, dehydrocostus lactone, and atractylodin was accurately weighed, and methanol was added according to Step 2 in Example 3 to prepare a series of reference solutions with different concentrations. According to the chromatographic conditions in Step 3 of Example 3, 1 μL of the reference solution was accurately pipetted and injected into the ultra-high performance liquid chromatograph. Using the reference substance concentration (μg / mL) as the abscissa (x-axis) and the peak area of each target component as the ordinate (y-axis), a standard curve was plotted, and the signal-to-noise ratio of the quantitative limit S / N≥10 and the detection limit S / N≥3 were ensured. The standard regression equations, correlation coefficients, linear ranges, quantitative limits, and detection limits of multiple components were calculated, and the specific results are shown in Table 2.

[0160] It can be seen from Table 2 that the 7 components had good linear relationships within their respective injection mass concentration ranges, indicating that the method of the present invention had a wide linear range and high accuracy.

[0161] Table 2 Linear regression equations and correlation coefficients

[0162]

[0163] 2. Stability

[0164] The Zhengqi tablets sample with batch number 200402 was taken, and the test solution was prepared according to Step 1 in Example 3. According to the chromatographic conditions in Step 3 of Example 3, the sample was injected for analysis at 0 h, 1 h, 3 h, 8 h, 12 h, and 24 h respectively, and the chromatographic peak area data of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, and atractylodin were recorded. The results showed that the RSDs of their chromatographic peak areas were all less than 0.92% within 24 h, indicating that the detection of the test solution within 24 h had no influence on the results and the stability was good.

[0165] 3. Precision

[0166] Take any one of the reference substance solutions prepared in Step 2 of Example 3, and according to the chromatographic conditions in Step 3 of Example 3, inject samples continuously for analysis 6 times respectively, and record the peak area data of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol and atractylodin. The results show that the RSDs of the peak areas of the 7 components for 6 consecutive injections are all less than 0.32%, indicating good precision of the instrument.

[0167] 4. Repeatability

[0168] Take 6 samples of Zhengqi Tablets with batch number 200402, accurately weigh them, prepare 6 samples of test solutions in parallel according to Step 1 of Example 3, determine them according to the chromatographic conditions in Step 3 of Example 3, record the peak area data of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol and atractylodin, and calculate the contents. The results show that the RSDs of the contents of the 7 components are all less than 1.19%, indicating good repeatability of the method.

[0169] 5. Standard addition recovery

[0170] Weigh 9 samples of Zhengqi Tablets (batch number 200402) with known concentrations, accurately weigh them, and add any one of the reference substance solutions prepared in Step 2 of Example 1 with low, medium and high 3 mass concentrations (equivalent to 50%, 100% and 150% of the original mass fraction) respectively. Take 3 samples for each mass concentration, prepare test solutions according to Step 1 of Example 3, and inject samples for analysis according to the chromatographic conditions in Step 3 of Example 3. Calculate the standard addition recoveries and RSDs of each component according to the measured amount and the added amount. The results are shown in Table 3. It can be seen from Table 3 that the average recoveries of the 7 components are 94.44% - 104.24%, and the RSDs are 1.08% - 2.80%, indicating good accuracy of the method.

[0171] Table 3 Results of standard addition recovery test (n = 3)

[0172]

[0173]

[0174] Example 8

[0175] Take 10 batches of Zhengqi Tablets samples with different batch numbers, prepare test solutions according to Step 1 of Example 3, and inject samples for analysis according to the chromatographic conditions in Step 3 of Example 3. Calculate the contents of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol and atractylodin respectively. The results are shown in Table 4.

[0176] As can be seen from Table 4, using the established method for determining the content of Zhengqi Tablets, a total of 10 batches of Zhengqi Tablet samples were determined. According to the current quality standard, each tablet contains not less than 0.8 mg of tangerine peel calculated as hesperidin (C 28 H 34 O 15 ). As can be seen from the results in Table 4, all 10 batches of samples meet the current standard requirements.

[0177] Table 4 Results of the content determination of samples

[0178]

[0179] Example 9

[0180] Take the Zhengqi Tablet sample 1# prepared in Example 1 and 10 reference substances, and prepare the test solution respectively according to the steps in Step 1 of Example 3. According to the chromatographic conditions in Step 3 of Example 3, the results show that 10 components can be qualitatively determined in Zhengqi Tablets, so as to better control the quality of Zhengqi Tablets. The specific results are shown in Figure 2 . As Figure 2 shown, the 1st peak is liquiritin, the 2nd peak is hesperidin, the 3rd peak is perillaketone, the 4th peak is nobiletin, the 5th peak is tangeretin, the 6th peak is honokiol, the 7th peak is costunolide, the 8th peak is dehydrocostus lactone, the 9th peak is magnolol, and the 10th peak is atractylodin. Among them, perillaketone and tangeretin (the 3rd peak and the 5th peak) are not suitable as quantitative indicators because their peak areas are too small and their contents are too low to meet the general content requirements (≥0.02%). At the same time, according to the provisions of the Chinese Pharmacopoeia, when quantifying, the resolution between the chromatographic peak of the substance to be measured and the adjacent chromatographic peak should be greater than 1.5. Perillaketone, nobiletin and tangeretin (the 3rd peak, the 4th peak, the 5th peak) do not meet this requirement, so they can only be qualitatively determined.

[0181] In summary, a kind of Zhengqi Tablet provided by the present invention, its preparation process and the method for quantitative and qualitative determination of multiple components therein can quickly qualitatively determine 10 components in Zhengqi Tablets, and quickly quantify 7 of them. The operation is simple, the detection time is shortened, the linear relationship is good, the repeatability is good, the precision is good, the accuracy is high, and the stability is good. Therefore, the present invention effectively overcomes various disadvantages in the prior art and has high industrial utilization value.

[0182] The above embodiments are only illustrative of the principles and effects of the present invention, and are not used to limit the present invention. Any person familiar with this technology can modify or change the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or changes completed by those with ordinary knowledge in the technical field without departing from the spirit and technical idea disclosed by the present invention should still be covered by the claims of the present invention.

Claims

1. A method for quantitatively and qualitatively determining multiple components in Zhengqi tablets, comprising: Dissolve Zhengqi tablets in methanol, extract by ultrasonic wave, let it cool, take the supernatant and filter it. The obtained test solution is detected by ultra-high performance liquid chromatography to qualitatively determine 10 components in the test solution: liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylodin, perillaketone, nobiletin and tangeretin, and quantitatively determine the contents of 7 components in the test solution: liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylodin; The Zhengqi tablets are composed of the following components by mass parts: Patchouli essential oil 0.5 - 1.5 parts; Perilla leaf oil 0.1 - 1 part; Aucklandia lappa 150 - 250 parts; Atractylodes lancea 100 - 150 parts; Glycyrrhiza uralensis 10 - 100 parts; Poria cocos 150 - 250 parts; Tangerine peel 100 - 150 parts; Pinellia ternata processed with ginger 100 - 150 parts; Magnolia officinalis processed with ginger 100 - 150 parts; Fresh ginger 100 - 150 parts; In the chromatographic conditions of the ultra-high performance liquid chromatography, the detector is a diode array detector; the chromatographic column is a T3 chromatographic column; the detection wavelength is 224 - 226 nm; the mobile phase is acetonitrile - 0.05 - 0.15% formic acid aqueous solution, where phase A is acetonitrile and phase B is 0.05 - 0.15% formic acid aqueous solution; the analysis time is 19 min; gradient elution; The specific procedure of the gradient elution is as follows: 0 - 4 min, the volume ratio of phase A to phase B is 20:80 - 35:65; 4 - 5 min, the volume ratio of phase A to phase B is 35:65 - 40:60; 5 - 6 min, the volume ratio of phase A to phase B is 40:60 - 55:45; 6 - 15 min, the volume ratio of phase A to phase B is 55:45 - 60:40; 15 - 16 min, the volume ratio of phase A to phase B is 60:40 - 90:10; 16 - 19 min, the volume ratio of phase A to phase B is 90:10 - 90:

10.

2. The method for quantitative and qualitative determination of multiple components in Zhengqi tablets according to claim 1, characterized in that, The Zhengqi tablets are composed of the following components by mass parts: Patchouli essential oil 0.8 - 1.2 parts; Perilla leaf oil 0.4 - 0.8 parts; Aucklandia lappa 180 - 220 parts; Atractylodes lancea 130 - 140 parts; Glycyrrhiza uralensis 60 - 70 parts; Poria cocos 180 - 220 parts; Tangerine peel 130 - 140 parts; Pinellia ternata processed with ginger 130 - 140 parts; Magnolia officinalis processed with ginger 130 - 140 parts; Fresh ginger 130 - 140 parts.

3. The method for quantitatively and qualitatively determining multiple components in Zhengqi tablets according to claim 1, characterized in that, Including any one or more of the following conditions: B1) The mass ratio of the added Zhengqi tablets to the volume of added methanol is 0.3:15 - 25, g / mL; B2) The ultrasonic extraction time is 30 - 60 minutes; B3) The power of the ultrasonic extraction is 200 - 350 W; the frequency of the ultrasonic extraction is 40 - 60 kHz; B4) Cooling to room temperature during the cooling process; B5) The filtration is to take the supernatant and filter it through a filter membrane, and the filter membrane is a 0.22 μm filter membrane.

4. The method for quantitative and qualitative determination of multiple components in Zhengqi tablets according to claim 1, characterized in that, The detection by ultra-high performance liquid chromatography includes the following steps: A) Prepare a reference solution: Dissolve the reference substances of liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylodin, perillaketone, nobiletin and tangeretin in methanol and make up the volume to prepare a reference solution; B) Sample detection: The test solution and the reference solution in step A) are respectively detected by ultra-high performance liquid chromatography. Qualitative analysis is carried out by comparing the retention times, and quantitative analysis is carried out by the external standard method. Ten components in the test solution are qualitatively determined: liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylodin, perillaketone, nobiletin and tangeretin. The contents of seven components in the test solution are quantitatively determined: liquiritin, hesperidin, honokiol, costunolide, dehydrocostus lactone, magnolol, atractylodin.

5. The method for quantitative and qualitative determination of multiple components in Zhengqi tablets according to claim 4, characterized in that, The chromatographic conditions of the ultra-high performance liquid chromatography described in step B) include any one or more of the following conditions: C1) The chromatographic column is Waters Acquity UPLC HSS T3 chromatographic column; C2) The column temperature is 30~45°C; C3) The flow rate is 0.1~0.3 mL / min; C4) The injection volume is 0.5~5 μL.

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