A method for constructing a characteristic chromatogram of a quercus infectoria oral liquid and dynamically monitoring the quality thereof

Abstract

The application discloses a characteristic spectrum construction of shengqi oral liquid, and comprises the following steps: S100, preparation of control sample solution; S200, preparation of test sample solution; S300, high performance liquid chromatography analysis; wherein, the high performance liquid chromatography analysis comprises detecting each control sample solution and the test sample solution by using an evaporative light scattering detector and an ultraviolet PDA detector respectively, and obtaining corresponding fingerprint spectrum and standard characteristic spectrum respectively. The application further discloses a dynamic monitoring method for the quality of shengqi oral liquid. The method can comprehensively monitor various animal and plant components of the shengqi oral liquid, and is simple and fast in operation, realizes real-time monitoring of multiple quality indexes of the shengqi oral liquid, can comprehensively control and evaluate the internal quality of the shengqi oral liquid, and guarantees the stability, effectiveness and safety of the drug quality.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of quality control of traditional Chinese medicine, and particularly relates to a method for constructing a characteristic chromatogram of Yujiqi oral liquid and dynamically monitoring the quality thereof. BACKGROUND

[0002] Yujiqi oral liquid is a kind of traditional Chinese medicine widely used for invigorating deficiency and health care and post-illness conditioning. The current implementation standard is the national drug standard WS-5546 (B-0546)-2014Z. Yujiqi oral liquid is composed of duck (with hair removed, with stomach, intestine and liver left), salvia miltiorrhiza, ligustrum lucidum, astragalus, hawthorn, jujube, medlar and licorice. In the prescription, duck is used as the monarch drug due to its salty and slightly cool properties and its compatibility with blood; astragalus and jujube are used as the ministerial drugs to tonify the spleen and stomach and replenish qi to build the center; medlar and ligustrum lucidum are used as the auxiliary drugs to nourish the kidney and liver and salvia miltiorrhiza is used to treat blood and calm the nerves to enhance the power of the monarch drug; hawthorn pulp is used to help digestion and absorption and licorice is used to detoxify and harmonize the center to enhance the function of digestion and absorption and coordinate the comprehensive effects of nourishing yin and replenishing qi.

[0003] At present, there are few quality detection methods for Yujiqi oral liquid. For example, the Chinese patent ZL2000310110519.7 (publication number CN112834650B) “Yin-yang and qi-replenishing, deficiency-supplementing and health-care oral liquid and its production process” does not involve the quality detection method thereof; the “Research on the synergistic effect and toxicity reduction of Yujiqi oral liquid combined with chemotherapy on Lewis lung cancer mice” published in the Hunan Journal of Chinese Medicine, Vol. 41, No. 5, 2021 and the “Clinical analysis of Yujiqi oral liquid in the treatment of 18 patients with hypertension and diabetes” published in the Electronic Journal of Cardiovascular Disease Integrative Medicine, Vol. 8, No. 36, 2020 also do not involve the quality detection technology of Yujiqi oral liquid.

[0004] In addition, the detection methods of characteristic chromatograms are disclosed in the invention patents “Construction of standard characteristic chromatogram of joint pain-relieving ointment and its quality detection method” (patent number ZL20161058643.5) and “Characteristic chromatogram determination and quality detection method of blood-activating and pain-relieving ointment” (patent number ZL20121108107.4). However, these methods can only determine the content of a single index component, and the efficacy of traditional Chinese medicine is the result of the combined action of multiple active components. The composition of traditional Chinese medicine is complex, and the single quality control index has certain limitations and cannot fully reflect the quality level of traditional Chinese medicine.

[0005] Moreover, in the prior art, there is no comprehensive quality control method for Yujiqi oral liquid to reflect the quality status of multiple animal and plant components in the traditional Chinese medicinal materials-intermediates-finished products, and it is also impossible to effectively and comprehensively control the production process and product quality, which cannot well guarantee the clinical efficacy.

[0006] Therefore, in order to make up for the defects of the quality control method of the existing Chinese patent medicines, it is necessary to construct a standard characteristic spectrum of the product, so as to better control the quality of the product. SUMMARY

[0007] The present application aims to provide a method for constructing a characteristic spectrum of Yuqi oral liquid and dynamically monitoring the quality of Yuqi oral liquid, so as to solve the technical problems of the prior art that the content measurement component is single and the quality of the liquid medicine cannot be monitored in real time during the production process.

[0008] To solve the above technical problems, the present application specifically provides the following technical solutions:

[0009] The present application provides a method for constructing a characteristic spectrum of Yuqi oral liquid, comprising the following steps:

[0010] S100, preparation of reference solution: using methanol to prepare Danshensu sodium, Salvianolic acid B, Rhodioside, Specnuezhenide, Ammonium glycyrrhizinate and Astragaloside A into reference solutions respectively;

[0011] S200, preparation of test solution: taking a certain amount of Yuqi oral liquid sample, passing through a 0.45 micron filter, and then passing through an inorganic membrane needle filter to obtain the test solution;

[0012] S300, high performance liquid chromatography analysis: precisely taking an equal amount of each reference solution and test solution, respectively, for high performance liquid chromatography analysis to obtain a Yuqi oral liquid fingerprint spectrum, and a standard characteristic spectrum is established according to the characteristic spectrum determined by the common peaks of the fingerprint spectrum, so as to establish the peak attribution and quality correlation between each medicinal material of Yuqi oral liquid and the finished product of Yuqi oral liquid;

[0013] In step S300, the high performance liquid chromatography analysis includes using an evaporation light scattering detector and an ultraviolet PDA detector to detect each reference solution and the test solution, respectively, to obtain corresponding fingerprint spectrum and standard characteristic spectrum.

[0014] As a preferred embodiment of the present application, the Yuqi oral liquid comprises the following components by weight: duck liquid 100-150 parts, Danshen 25-35 parts, Nuzhizi 25-35 parts, Huangqi 25-35 parts, Shanzha 15-25 parts, Dazao 10-20 parts, Gouqi 10-20 parts, and Gancao 2-5 parts.

[0015] The effective components in the Yuqi oral liquid at least include Danshensu sodium, Salvianolic acid B, Rhodioside, Specnuezhenide, Ammonium glycyrrhizinate and Astragaloside A.

[0016] As a preferred scheme of the present application, the quercetin oral liquid comprises the following components in parts by weight: 125.04 parts of duck liquid, 31.28 parts of salvia miltiorrhiza, 31.28 parts of ligustrum lucidum, 31.25 parts of astragalus, 18.75 parts of hawthorn, 12.71 parts of jujube, 12.65 parts of medlar, and 3.76 parts of licorice.

[0017] As a preferred scheme of the present application, in step S100, the following steps are specifically included:

[0018] S101, preparing Danshensu sodium into a control solution A containing 0.005-0.015 mg of Danshensu sodium per 1 mL by using methanol with a volume percentage concentration of 0%-50%;

[0019] S102, preparing Salvianolic acid B into a control solution B containing 0.100-0.150 mg of Salvianolic acid B per 1 mL by using methanol with a volume percentage concentration of 20%-60%;

[0020] S103, preparing Rhodioside into a control solution C containing 0.100-0.150 mg of Rhodioside per 1 mL by using methanol with a volume percentage concentration of 0%-50%;

[0021] S104, preparing Specnuezhenide into a control solution D containing 0.150-0.250 mg of Specnuezhenide per 1 mL by using methanol with a volume percentage concentration of 50%-100%;

[0022] S105, preparing Ammonium glycyrrhizinate into a control solution E containing 0.150-0.250 mg of Ammonium glycyrrhizinate per 1 mL by using methanol with a volume percentage concentration of 50%-100%;

[0023] S106, preparing Astragaloside A into a control solution F containing 0.050-0.150 mg of Astragaloside A per 1 mL by using methanol with a volume percentage concentration of 0%-50%.

[0024] As a preferred scheme of the present application, in step S300, the following steps are further included:

[0025] S301, construction of a characteristic spectrum under an evaporative light scattering detector:

[0026] The control solutions and the test solutions are detected by using an evaporative light scattering detector to obtain an evaporative light scattering detector ELSD chromatogram, the evaporative light scattering detector ELSD chromatogram is introduced into a traditional Chinese medicine chromatographic fingerprint similarity evaluation system software, and an evaporative light scattering detector ELSD standard characteristic spectrum of the quercetin oral liquid is obtained, the standard characteristic spectrum has 16 characteristic peaks, the 16 characteristic peaks are sequentially numbered E1-E16 in order of time, and the sum of peak areas of common peaks accounts for more than 90% of the total peak area.

[0027] Among them, there are 3 characteristic peaks belonging to duck liquid; there are 4 characteristic peaks belonging to salvia miltiorrhiza; there are 3 characteristic peaks belonging to ligustrum lucidum; there are 3 characteristic peaks belonging to astragalus; there is 1 characteristic peak belonging to lycium barbarum; there are 2 characteristic peaks belonging to licorice;

[0028] E3 peak is danshensu, as a reference peak, belonging to salvia miltiorrhiza; E6 peak is rhodioside, and E9 peak is specnuzhenoside, both belonging to ligustrum lucidum; E14 peak is salvianolic acid b, belonging to salvia miltiorrhiza; E16 peak is glycyrrhizic acid, belonging to licorice; the common peaks of the characteristic spectrum have 16 characteristic peaks, and the retention time and relative standard deviation of each characteristic peak are:

[0029] E1 peak: retention time 9.262±0.001, relative standard deviation 0.03%

[0030] E2 peak: retention time 13.489±0.003, relative standard deviation 0.02%

[0031] E3 peak: retention time 26.571±0.002, relative standard deviation 0.02%

[0032] E4 peak: retention time 27.274±0.003, relative standard deviation 0.02%

[0033] E5 peak: retention time 30.439±0.004, relative standard deviation 0.03%

[0034] E6 peak: retention time 30.677±0.002, relative standard deviation 0.01%

[0035] E7 peak: retention time 38.947±0.004, relative standard deviation 0.03%

[0036] E8 peak: retention time 42.561±0.003, relative standard deviation 0.02%

[0037] E9 peak: retention time 43.494±0.004, relative standard deviation 0.03%

[0038] E10 peak: retention time 45.375±0.003, relative standard deviation 0.02%

[0039] E11 peak: retention time 48.973±0.004, relative standard deviation 0.03%

[0040] E12 peak: retention time 50.244±0.003, relative standard deviation 0.02%

[0041] E13 peak: retention time 52.58±0.004, relative standard deviation is 0.03%

[0042] E14 peak: retention time 54.981±0.004, relative standard deviation is 0.03%

[0043] E15 peak: retention time 57.149±0.003, relative standard deviation is 0.02%

[0044] E16 peak: retention time 67.14±0.004, relative standard deviation is 0.03%.

[0045] As a preferred scheme of the present application, in step S300, the evaporative light scattering detector detects the following conditions: the chromatographic column is a medium spectrum red C18 reversed-phase chromatographic column, the mobile phase is a gradient eluent composed of organic phase A and aqueous phase B, the organic phase A is acetonitrile, and the aqueous phase B is a 0.1-0.3% glacial acetic acid solution by volume percentage; the drift tube temperature is 90-120℃; the gas flow is 2.0-3.0ml / min; the column temperature is 30-40℃; the flow rate is 0.8mL / min-1.2mL / min; the analysis time is 60-80min; and gradient elution is used;

[0046] The time and mobile phase ratio of gradient elution are as follows: 0-15min, mobile phase A 0%, mobile phase B 100%; 15-30min, mobile phase A 0%→5%, mobile phase B 100%→95%; 30-65min, mobile phase A 5%→50%, mobile phase B 95%→50%; 65-70min, mobile phase A 50→25%, mobile phase B 50%→75%.

[0047] As a preferred scheme of the present application, in step S300, the following steps are further included:

[0048] S302, construction of a characteristic spectrum under a UV detector:

[0049] The control sample solution and the test sample solution are detected using a UV PDA detector to obtain a UV detector PDA chromatogram, the UV detector PDA chromatogram is imported into a traditional Chinese medicine chromatographic fingerprint similarity evaluation system software, and a UV detector PDA standard characteristic spectrum of the Rouqikeli oral liquid is formulated and obtained, the standard characteristic spectrum has 10 characteristic peaks, the 10 characteristic peaks are sequentially numbered P1-P10 in order of time of appearance, and the sum of peak areas of common peaks accounts for more than 95% of the total peak area.

[0050] Among them, 4 characteristic peaks belong to Salvia miltiorrhiza; 2 characteristic peaks belong to Ligustri Lucidi Fructus; 1 characteristic peak belongs to Crataegi Fructus; 1 characteristic peak belongs to Fructus Lycii; 1 characteristic peak belongs to Jujubae Fructus; and 1 characteristic peak belongs to Glycyrrhizae Radix et Rhizoma;

[0051] P3 is Danshensu, as a reference peak, belonging to Salvia miltiorrhiza; P6 is Specnuezhenide, belonging to Ligustri Lucidi Fructus; P9 is Salvianolic acid b, belonging to Salvia miltiorrhiza; the common peaks of the characteristic atlas have 10 characteristic peaks, and the retention time and relative standard deviation of each characteristic peak are:

[0052] P1 peak: retention time 6.464±0.002, relative standard deviation 0.01%

[0053] P2 peak: retention time 14.552±0.001, relative standard deviation 0.01%

[0054] P3 peak: retention time 16.614±0.003, relative standard deviation 0.02%

[0055] P4 peak: retention time 22.287±0.002, relative standard deviation 0.01%

[0056] P5 peak: retention time 23.152±0.001, relative standard deviation 0.01%

[0057] P6 peak: retention time 40.193±0.003, relative standard deviation 0.02%

[0058] P7 peak: retention time 43.087±0.002, relative standard deviation 0.01%

[0059] P8 peak: retention time 45.528±0.004, relative standard deviation 0.03%

[0060] P9 peak: retention time 46.921±0.001, relative standard deviation 0.02%

[0061] P10 peak: retention time 59.625±0.004, relative standard deviation 0.03%.

[0062] As a preferred scheme of the present application, in step S300, the ultraviolet detector detects the following conditions: the chromatographic column is Shim-pack VP-ODS, the mobile phase is a gradient eluent composed of organic phase C and aqueous phase D, the organic phase C is methanol, the aqueous phase D is a 0.5-1.5% (volume fraction) glacial acetic acid solution; the detection wavelength is 280 nm; the column temperature is 30-40 DEG C; the flow rate is 0.9 mL / min-1.1 mL / min; the analysis time is 50-70 min; and gradient elution is adopted;

[0063] The gradient elution time and the mobile phase ratio are as follows: 0-25 min, mobile phase C 0%→21%, mobile phase D 100%→79%; 25-30 min, mobile phase C 21%→40%, mobile phase D 79%→60%; 30-50 min, mobile phase C 40%→40%, mobile phase D 60%→60%; and 50-60 min, mobile phase C 40%→50%, mobile phase D 60%→50%.

[0064] The present application also provides a quality dynamic monitoring method of the Yuqiu oral liquid, which comprises the following steps:

[0065] The effective components in the test solution are selected as the reference solution, and the test solution and the reference solution are subjected to chromatographic analysis to obtain a characteristic spectrum of the Yuqiu oral liquid fingerprint common peaks, and a standard characteristic spectrum is prepared;

[0066] The Yuqiu oral liquid fingerprint is compared with the prepared standard characteristic spectrum, if the same characteristic peaks in the standard characteristic spectrum appear in the chromatogram of the to-be-tested Chinese medicinal material-intermediate-product-finished product, and the peaks corresponding to the reference peaks are E3 or P3 peaks, the relative retention time of each characteristic peak and E3 or P3 peak is calculated, and the relative standard deviation of the relative retention time of each characteristic peak and the relative retention time of each characteristic peak in the standard characteristic spectrum is within ±5%, so that the quality of the Chinese medicinal material-intermediate-product-finished product is qualified, otherwise, the quality is unqualified.

[0067] As a preferred scheme of the present application, the corresponding peak attribution and quality correlation of each medicinal material contained in the Yuqiu oral liquid and the Yuqiu oral liquid finished product are carried out, and the quality of the medicinal product in the production line of the Yuqiu oral liquid Chinese medicinal material-intermediate-product-finished product is dynamically monitored in real time.

[0068] Compared with the prior art, the present application has the following beneficial effects:

[0069] 1. The Yuqiu oral liquid fingerprint and the standard characteristic spectrum are established in the present application, so that each component of the Yuqiu oral liquid is comprehensively monitored, the corresponding peak attribution and quality correlation of each medicinal material of the Yuqiu oral liquid and the Yuqiu oral liquid finished product are effectively carried out, and the quality of the Chinese medicinal material-intermediate-product-finished product is dynamically monitored in real time and traced;

[0070] 2、The present application successfully realizes real-time monitoring of multiple components of the Qianqi oral liquid by using the evaporation light scattering detector (ELSD) in combination with the ultraviolet detector (PDA) technology, and can simultaneously adapt to various animal and plant components in medicinal materials, can comprehensively control and evaluate the internal quality of the Qianqi oral liquid, and ensure the stability, effectiveness and safety of the drug quality. BRIEF DESCRIPTION OF DRAWINGS

[0071] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings needed to be used in the following embodiment or prior art description will be briefly introduced. Obviously, the drawings in the following description are only exemplary, and other implementation drawings can be obtained according to the provided drawings without creative labor for those skilled in the art.

[0072] Figure 1 To provide the Qianqi oral liquid evaporation light scattering detector ELSD fingerprint spectrum shown in Example 1 of the present application;

[0073] Figure 2 To provide the Qianqi oral liquid evaporation light scattering detector ELSD standard characteristic spectrum shown in Example 1 of the present application;

[0074] Figure 3 To provide the Qianqi oral liquid ultraviolet detector PDA full wavelength scan spectrum shown in Example 1 of the present application;

[0075] Figure 4 To provide the Qianqi oral liquid ultraviolet detector PDA fingerprint spectrum shown in Example 1 of the present application;

[0076] Figure 5 To provide the Qianqi oral liquid ultraviolet detector PDA standard characteristic spectrum shown in Example 1 of the present application. DETAILED DESCRIPTION

[0077] The technical solutions in the embodiments of the present application will be described clearly and completely in combination with the drawings in the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

[0078] As shown in Figure 1 The present application provides a construction of the Qianqi oral liquid characteristic spectrum, including the following steps:

[0079] S100, Preparation of control solution: Using methanol to prepare Danshensuan, Salvianolic Acid B, Rhodioside, Specnuezhenide, Ammonium Glycyrrhizinate, Astragaloside into control solution respectively;

[0080] S101, Danshensuan is prepared into control solution A containing 0.005-0.015 mg Danshensuan per 1 mL using methanol with a volume percentage concentration of 0%-50%;

[0081] S102, Salvianolic Acid B is prepared into control solution B containing 0.100-0.150 mg Salvianolic Acid B per 1 mL using methanol with a volume percentage concentration of 20%-60%;

[0082] S103, Rhodioside is prepared into control solution C containing 0.100-0.150 mg Rhodioside per 1 mL using methanol with a volume percentage concentration of 0%-50%;

[0083] S104, Specnuezhenide is prepared into control solution D containing 0.150-0.250 mg Specnuezhenide per 1 mL using methanol with a volume percentage concentration of 50%-100%;

[0084] S105, Ammonium Glycyrrhizinate is prepared into control solution E containing 0.150-0.250 mg Ammonium Glycyrrhizinate per 1 mL using methanol with a volume percentage concentration of 50%-100%;

[0085] S106, Astragaloside is prepared into control solution F containing 0.050-0.150 mg Astragaloside per 1 mL using methanol with a volume percentage concentration of 0%-50%.

[0086] S200, Preparation of test solution: Taking the Astragalus oral liquid sample, measuring 1-5 mL through 0.45 micron organic membrane needle filter, taking the filtrate to obtain;

[0087] The Astragalus oral liquid comprises the following components (existing traditional Chinese medicine formula) by weight: duck liquid 100-150, Danshen 25-35, Nuzhizi 25-35, Huangqi 25-35, Shanzha 15-25, Dazao 10-20, Gouqi 10-20, Gancao 2-5,

[0088] Preferably, duck liquid 125.04, Danshen 31.28, Nuzhizi 31.28, Huangqi 31.25, Shanzha 18.75, Dazao 12.71, Gouqi 12.65, Gancao 3.76.

[0089] S300, High performance liquid chromatography analysis:

[0090] S301. Accurately pipette equal volumes of each reference solution and the test solution, and inject them into the evaporative light scattering detector. The chromatographic column is a Midspect Red C18 reversed-phase column. The mobile phase is a gradient eluent composed of organic phase A and aqueous phase B. Organic phase A is acetonitrile, and aqueous phase B is a 0.1-0.3% (v / v) glacial acetic acid solution. The drift tube temperature is 90-120℃. The gas flow rate is 2.0-3.0 mL / min. The column temperature is 30-40℃. The flow rate is 0.8 mL / min-1.2 mL / min. The analysis time is 60-80 min.

[0091] Gradient elution was employed, as shown in Table 1:

[0092] Table 1:

[0093]

[0094] Based on 18 batches of Wuqi oral liquid, ELSD fingerprint spectra of Wuqi oral liquid were obtained using an evaporative light scattering detector. Figure 1 Based on the characteristic spectrum determined by the common peaks in the evaporative light scattering (ELSD) fingerprint spectrum of Wuqi oral liquid, a standard ELSD characteristic spectrum for Wuqi oral liquid was developed. Figure 2 ).

[0095] S302. Accurately pipette equal volumes of each reference solution and the test solution, and inject them into the UV detector. The chromatographic column is a Shimadzu Shim-pack VP-ODS; the mobile phase is a gradient elution solution composed of organic phase C and aqueous phase D, where organic phase C is methanol and aqueous phase D is a 0.5–1.5% (v / v) glacial acetic acid solution; the detection wavelength is 280 nm; the column temperature is 30–40 °C; the flow rate is 0.9 mL / min–1.1 mL / min; and the analysis time is 50–70 min.

[0096] Gradient elution was employed, see Table 2:

[0097] Table 2:

[0098]

[0099] Based on 18 batches of Wuqi oral liquid, the full wavelength scan image of Wuqi oral liquid using a PDA ultraviolet detector was obtained. Figure 3 ) and the PDA fingerprint spectrum of Wuqi oral liquid using ultraviolet detector ( Figure 4 Based on the characteristic spectrum determined by the common peaks in the fingerprint spectrum of the ultraviolet detector PDA of Wuqi oral liquid, a standard characteristic spectrum of the ultraviolet detector PDA of Wuqi oral liquid was formulated. Figure 5 ).

[0100] Compare the fingerprint spectrum of Wuqi oral liquid with the standard characteristic spectrum obtained above. Samples with a retention time within ±5% are considered qualified.

[0101] The present application mainly formulates the standard characteristic spectrum according to the characteristic spectrum determined by the common peaks of the fingerprint spectrum, thereby establishing the peak attribution and quality correlation corresponding to each medicinal material of the Luqin oral liquid and the finished product of the Luqin oral liquid, realizing the comprehensive monitoring of each component of the Luqin oral liquid by the fingerprint spectrum and the standard characteristic spectrum of the Luqin oral liquid, and more effectively realizing the real-time dynamic monitoring and tracing of the quality of the traditional Chinese medicinal materials-intermediate-finished product.

[0102] Particularly, in step S300, the high performance liquid chromatography analysis comprises detecting each of the control sample solution and the test sample solution by using an evaporative light scattering detector and an ultraviolet PDA detector respectively, obtaining the corresponding fingerprint spectrum and standard characteristic spectrum respectively, and successfully realizing the real-time monitoring of the multiple quality indexes of the Luqin oral liquid by using the combination of the evaporative light scattering detector (ELSD) and the ultraviolet detector (PDA).

[0103] Because the components of traditional Chinese medicines are complex, the components with different properties cannot be monitored by using the same detector or detection method, some components cannot be absorbed under the ultraviolet lamp and need to be monitored by using the evaporative light scattering detector, and some small molecular components have a low response under the evaporative light detector and need to be monitored by using the ultraviolet detector for accurate quantitative monitoring. In particular, in the embodiment, the Luqin oral liquid is composed of duck (with the feathers removed, with the stomach, intestine and liver left), salvia miltiorrhiza, ligustrum lucidum, astragalus, hawthorn, jujube, medlar and licorice, the prescription includes animal medicines and traditional Chinese herbal medicines, the prescription is special, and the two components of the animal medicines and the traditional Chinese herbal medicines have different properties, and the components with different properties cannot be monitored by using the same detector or detection method.

[0104] The scheme combines the evaporative light scattering detector (ELSD) and the ultraviolet PDA detector (PDA) to realize the real-time monitoring of the multiple components of the Luqin oral liquid, can comprehensively control and evaluate the internal quality of the Luqin oral liquid, and guarantees the stability, effectiveness and safety of the drug quality.

[0105] The present application provides a dynamic monitoring method for the quality of the Luqin oral liquid, comprising the following steps:

[0106] The effective components in the test sample solution are selected as the control sample solution, and the control sample solution and the test sample solution are subjected to chromatography analysis, the characteristic spectrum determined by the common peaks of the fingerprint spectrum of the Luqin oral liquid is obtained, and the standard characteristic spectrum is formulated;

[0107] The fingerprint spectrum of the Shengqiu oral liquid is compared with the standard characteristic spectrum, if the characteristic peaks in the chromatogram of the to-be-tested traditional Chinese medicinal material-intermediate-finished product appear in the standard characteristic spectrum, the peaks corresponding to the reference peaks are E3 or P3 peaks, the relative retention time of each characteristic peak is calculated, and the relative standard deviation of the relative retention time of each characteristic peak in the standard characteristic spectrum is within ±5%, so that the quality of the traditional Chinese medicinal material-intermediate-finished product is qualified, otherwise, the quality is unqualified.

[0108] Through the establishment of the fingerprint spectrum and the standard characteristic spectrum of the Shengqiu oral liquid, each component of the Shengqiu oral liquid can be comprehensively monitored, and the peak attribution and quality correlation of each medicinal material in the Shengqiu oral liquid and the finished product of the Shengqiu oral liquid can be effectively realized, and the quality of the traditional Chinese medicinal material-intermediate-finished product can be effectively monitored in real time.

[0109] Through the establishment of the fingerprint spectrum and the standard characteristic spectrum of the Shengqiu oral liquid, different components in the Shengqiu oral liquid can be comprehensively monitored, and most of the existing quality control methods are to follow the registration standard, so that real-time improvement cannot be achieved.

[0110] Preferably, the peak attribution and quality correlation of each medicinal material in the Shengqiu oral liquid and the finished product of the Shengqiu oral liquid are realized, and the quality of the traditional Chinese medicinal material-intermediate-finished product in the production line of the Shengqiu oral liquid is monitored in real time.

[0111] The following provides Example 1:

[0112] (1) Preparation of the reference solution:

[0113] a) Dan Shensu sodium is prepared into a reference solution containing 0.005-0.015 mg of Dan Shensu sodium per 1 mL by using methanol with a volume percentage concentration of 0%-50%;

[0114] b) Danpol B is prepared into a reference solution containing 0.100-0.150 mg of Danpol B per 1 mL by using methanol with a volume percentage concentration of 20%-60%;

[0115] c) Hongjingtian glycoside is prepared into a reference solution containing 0.100-0.150 mg of Hongjingtian glycoside per 1 mL by using methanol with a volume percentage concentration of 0%-50%;

[0116] d) Tenuifolin is prepared into a reference solution containing 0.150-0.250 mg of Tenuifolin per 1 mL by using methanol with a volume percentage concentration of 50%-100%;

[0117] e) Ammonium glycyrrhizinate is prepared into a reference solution containing 0.150-0.250 mg of ammonium glycyrrhizinate per 1 mL by using methanol with a volume percentage concentration of 50%-100%;

[0118] f) Astragaloside is prepared into a control solution containing 0.050-0.150 mg of astragaloside per 1 mL with methanol at a volume percentage concentration of 0%-50%;

[0119] (2) Preparation of the test solution: 5 mL of the Yaqi oral liquid was precisely taken into a 10 mL sample bottle, shaken, filtered with a microporous filter membrane, and the filtered solution was taken, to obtain the test solution;

[0120] The Yaqi oral liquid includes duck liquid 125.04 parts, salvia miltiorrhiza 31.28 parts, ligustrum lucidum 31.28 parts, astragalus 31.25 parts, hawthorn 18.75 parts, Chinese date 12.71 parts, medlar 12.65 parts, and licorice 3.76 parts by weight, which are mainly from Hunan Tianji Pharmaceutical Co., Ltd.;

[0121] (3) High performance liquid chromatography analysis: 10 ul of the control solution and the test solution were precisely taken and injected into a high performance liquid chromatograph, and the Yaqi oral liquid fingerprint spectrum was obtained by determination, and the obtained chromatogram was analyzed to obtain a standard characteristic spectrum.

[0122] High performance liquid chromatography analysis conditions:

[0123] ① Evaporative light scattering detector; the chromatographic column is an RD-C18 chromatographic column with a column length of 250 mm; the mobile phase is a gradient eluent composed of organic phase A and aqueous phase B, the organic phase A is acetonitrile, and the aqueous phase B is a 0.1% glacial acetic acid solution; the drift tube temperature is 90-120°C, the gas flow rate is 2.0-3.0 ml / min; the column temperature is 30-40°C; the flow rate is 1.0 mL / min; the analysis time is 70 min; the gradient elution time and the mobile phase ratio are: 0-15 min, mobile phase A 0%, mobile phase B 100%; 15-30 min, mobile phase A 0%→5%, mobile phase B 100%→95%; 30-65 min, mobile phase A 5→50%, mobile phase B 95%→50%; 65-70 min, mobile phase A 50→25%, mobile phase B 50%→75%.

[0124] ② UV detector: the chromatographic column is Shim-pack VP-ODS (4.6×250 mm, 5 The mobile phase is a gradient eluent composed of organic phase C and water phase D, the organic phase C is methanol, and the water phase D is a 1% (by volume) glacial acetic acid solution; the detection wavelength is 280 nm, the column temperature is 30-40 DEG C; the flow rate is 1.0 mL / min; the analysis time is 60 min; the time and the proportion of the mobile phase in the gradient elution are as follows: 0-25 min, mobile phase C 0% to 21%, mobile phase D 100% to 79%; 25-30 min, mobile phase C 21% to 40%, mobile phase D 79% to 60%; 30-50 min, mobile phase C 40% to 40%, mobile phase D 60% to 60%; 50-60 min, mobile phase C 40% to 50%, mobile phase D 60% to 50%.

[0125] Characterization:

[0126] According to 18 batches of Equisetum arvense oral liquid, the Equisetum arvense oral liquid evaporative light scattering detector ELSD fingerprint spectrum (Fig. 1) is obtained. Figure 1 According to the characteristic spectrum of the Equisetum arvense oral liquid evaporative light scattering detector ELSD fingerprint spectrum common peak, the Equisetum arvense oral liquid evaporative light scattering detector ELSD standard characteristic spectrum (Fig. 2) is formulated. Figure 2

[0127] As can be seen from the figure, the standard characteristic spectrum has 16 characteristic peaks, which are sequentially numbered E1-16 in the order of time of appearance, and the sum of the peak areas of the common peaks accounts for more than 90% of the total peak area.

[0128] Among them, the characteristic peaks belonging to duck liquid are 3; the characteristic peaks belonging to Salvia miltiorrhiza are 4; the characteristic peaks belonging to Ligustri Lucidi Fructus are 3; the characteristic peaks belonging to Astragali Radix are 3; the characteristic peaks belonging to Lycium barbarum are 1; the characteristic peaks belonging to Glycyrrhiza are 2;

[0129] E3 peak is Danshensu, which is a reference peak and belongs to Salvia miltiorrhiza; E6 peak is Rhodioside, and E9 peak is Specnuezhenide, both of which belong to Ligustri Lucidi Fructus; E14 peak is Salvianolic acid b, which belongs to Salvia miltiorrhiza; E16 peak is Glycyrrhizic acid, which belongs to Glycyrrhiza; the common peaks of the characteristic spectrum have 16 characteristic peaks, and the retention time and relative standard deviation of each characteristic peak are as follows:

[0130] E1 peak: retention time 9.262±0.001, relative standard deviation 0.03%

[0131] E2 peak: retention time 13.489±0.003, relative standard deviation 0.02%

[0132] E3 peak: retention time 26.571±0.002, relative standard deviation 0.02%

[0133] ​E4 peak: retention time 27.274 ± 0.003, relative standard deviation 0.02%

[0134] E5 peak: retention time 30.439 ± 0.004, relative standard deviation 0.03%

[0135] E6 peak: retention time 30.677 ± 0.002, relative standard deviation 0.01%

[0136] E7 peak: retention time 38.947 ± 0.004, relative standard deviation 0.03%

[0137] E8 peak: retention time 42.561 ± 0.003, relative standard deviation 0.02%

[0138] E9 peak: retention time 43.494 ± 0.004, relative standard deviation 0.03%

[0139] E10 peak: retention time 45.375 ± 0.003, relative standard deviation 0.02%

[0140] E11 peak: retention time 48.973 ± 0.004, relative standard deviation 0.03%

[0141] E12 peak: retention time 50.244 ± 0.003, relative standard deviation 0.02%

[0142] E13 peak: retention time 52.58 ± 0.004, relative standard deviation 0.03%

[0143] E14 peak: retention time 54.981 ± 0.004, relative standard deviation 0.03%

[0144] E15 peak: retention time 57.149 ± 0.003, relative standard deviation 0.02%

[0145] E16 peak: retention time 67.14 ± 0.004, relative standard deviation 0.03%.

[0146] The fingerprint spectrum of the Equisetum oral liquid is compared with the standard characteristic spectrum obtained above, and the retention time within ± 5% is qualified test product.

[0147] According to the 18 batches of Equisetum oral liquid, the Equisetum oral liquid UV detector PDA full wavelength scanning diagram ( Figure 3 ) and the Equisetum oral liquid UV detector PDA fingerprint spectrum ( Figure 4 ), the characteristic spectrum determined by the common peaks of the Equisetum oral liquid UV detector PDA fingerprint spectrum, the Equisetum oral liquid UV detector PDA standard characteristic spectrum is formulated ( Figure 5).

[0148] The standard characteristic spectrum has 10 characteristic peaks, which are sequentially numbered P1-P10 according to the time of appearance, and the sum of the peak areas accounts for more than 95% of the total peak area;

[0149] Among them, 4 characteristic peaks belong to Salvia miltiorrhiza; 2 characteristic peaks belong to Ligustri Lucidi Fructus; 1 characteristic peak belongs to Fructus Crataegi; 1 characteristic peak belongs to Fructus Lycii; 1 characteristic peak belongs to Fructus Jujubae; and 1 characteristic peak belongs to Glycyrrhizae Radix et Rhizoma;

[0150] P3 peak is Danshensu, which is a reference peak and belongs to Salvia miltiorrhiza; P6 peak is specnuzhenoside, which belongs to Ligustri Lucidi Fructus; P9 peak is salvianolic acid b, which belongs to Salvia miltiorrhiza; the common peaks of the characteristic spectrum have 10 characteristic peaks, and the retention time and relative standard deviation of each characteristic peak are:

[0151] P1 peak: retention time 6.464±0.002, relative standard deviation 0.01%

[0152] P2 peak: retention time 14.552±0.001, relative standard deviation 0.01%

[0153] P3 peak: retention time 16.614±0.003, relative standard deviation 0.02%

[0154] P4 peak: retention time 22.287±0.002, relative standard deviation 0.01%

[0155] P5 peak: retention time 23.152±0.001, relative standard deviation 0.01%

[0156] P6 peak: retention time 40.193±0.003, relative standard deviation 0.02%

[0157] P7 peak: retention time 43.087±0.002, relative standard deviation 0.01%

[0158] P8 peak: retention time 45.528±0.004, relative standard deviation 0.03%

[0159] P9 peak: retention time 46.921±0.001, relative standard deviation 0.02%

[0160] P10 peak: retention time 59.625±0.004, relative standard deviation 0.03%.

[0161] Compare the fingerprint spectrum of the Radix Astragali oral liquid with the standard characteristic spectrum obtained above, and the retention time within ±5% is qualified test product.

[0162] The above embodiments are only exemplary embodiments of the present application, and are not intended to limit the present application. The scope of protection of the present application is defined by the claims. Those skilled in the art can make various modifications or equivalent replacements to the present application within the spirit and protection scope of the present application, and such modifications or equivalent replacements should also be considered to fall within the protection scope of the present application.

Claims

1. A method for constructing a characteristic map of Rougui oral liquid, characterized in that, The method comprises the following steps: S100, preparation of a reference solution: using methanol to prepare sodium danshensu, salvianolic acid B, rhodioside, prunol and ammonium glycyrrhizate into reference solutions, respectively; S200, preparation of a test solution: taking a sample of the Astragalus and Hedysari Oral Liquid, passing it through a 0.45 µm inorganic membrane needle filter, and taking the filtrate to obtain the test solution; S300, high performance liquid chromatography analysis: precisely taking equal amounts of each reference solution and the test solution, and performing high performance liquid chromatography analysis to obtain a fingerprint spectrum of the Astragalus and Hedysari Oral Liquid, and establishing a standard characteristic spectrum according to the characteristic spectrum determined based on the common peaks of the fingerprint spectrum; In step S300, the high performance liquid chromatography analysis comprises detecting each reference solution and the test solution using an evaporative light scattering detector and a UV PDA detector, respectively; In step S300, the evaporative light scattering detector meets the following conditions: the chromatographic column is a medium spectrum red C18 reversed-phase chromatographic column, the mobile phase is a gradient eluent composed of organic phase A and aqueous phase B, the organic phase A is acetonitrile, and the aqueous phase B is a 0.1% to 0.3% glacial acetic acid solution by volume percentage; the drift tube temperature is 90°C to 120°C; the gas flow rate is 2.0 ml / min to 3.0 ml / min; the column temperature is 30°C to 40°C; the flow rate is 0.8 mL / min to 1.2 mL / min; and gradient elution is used; In the gradient elution, the time and the proportion of the mobile phase are as follows: 0 min to 15 min, mobile phase A 0%, mobile phase B 100%; 15 min to 30 min, mobile phase A 0%→5%, mobile phase B 100%→95%; 30 min to 65 min, mobile phase A 5%→50%, mobile phase B 95%→50%; and 65 min to 70 min, mobile phase A 50%→25%, mobile phase B 50%→75%; In step S300, the UV PDA detector meets the following conditions: the chromatographic column is Shim-pack VP-ODS, the mobile phase is a gradient eluent composed of organic phase C and aqueous phase D, the organic phase C is methanol, the aqueous phase D is a 0.5% to 1.5% glacial acetic acid solution by volume percentage, the detection wavelength is 280 nm, the column temperature is 30°C to 40°C, the flow rate is 0.9 mL / min to 1.1 mL / min, and gradient elution is used; In the gradient elution, the time and the proportion of the mobile phase are as follows: 0 min to 25 min, mobile phase C 0%→21%, mobile phase D 100%→79%; 25 min to 30 min, mobile phase C 21%→40%, mobile phase D 79%→60%; 30 min to 50 min, mobile phase C 40%→40%, mobile phase D 60%→60%; and 50 min to 60 min, mobile phase C 40%→50%, mobile phase D 60%→50%.

2. The method according to claim 1, wherein in step S100, the method comprises the following steps: ​ S101, with volume percentage concentration of 0% ~ 50% methanol, prepare Danshensu sodium into 0.005 ~ 0.015 mg Danshensu sodium per 1 mL of control solution A; S102, with volume percentage concentration of 20% ~ 60% methanol, prepare Salvianolic acid B into 0.100 ~ 0.150 mg Salvianolic acid B per 1 mL of control solution B; S103, with volume percentage concentration of 0% ~ 50% methanol, prepare Rhodioloside into 0.100 ~ 0.150 mg Rhodioloside per 1 mL of control solution C; S104, with volume percentage concentration of 50% ~ 100% methanol, prepare Specnuezhenide into 0.150 ~ 0.250 mg Specnuezhenide per 1 mL of control solution D; S105, with volume percentage concentration of 50% ~ 100% methanol, prepare Ammonium glycyrrhizinate into 0.150 ~ 0.250 mg Ammonium glycyrrhizinate per 1 mL of control solution E.

3. The method according to claim 1, wherein the method further comprises the following step in step S300: S301, construction of the characteristic spectrum under the evaporative light scattering detector; The control solution and the test solution are detected by using the evaporative light scattering detector to obtain an evaporative light scattering detector ELSD chromatogram, and the evaporative light scattering detector ELSD chromatogram is introduced into a traditional Chinese medicine chromatographic fingerprint similarity evaluation system software to obtain an evaporative light scattering detector ELSD standard characteristic spectrum of the Yuqi oral liquid, wherein the standard characteristic spectrum has 16 characteristic peaks, and the sum of the peak areas of the 16 characteristic peaks accounts for more than 90% of the total peak area. E3 is Danshensu, which is used as a reference peak and belongs to Danshen; E6 is Rhodioloside, and E9 is Specnuezhenide, both of which belong to Nuzhenzi; E14 is Salvianolic acid B, which belongs to Danshen; and E16 is Glycyrrhizic acid, which belongs to Glycyrrhiza; the common peaks of the characteristic spectrum have 16 characteristic peaks, and the retention time and relative standard deviation of each characteristic peak are as follows: E1: retention time 9.262 ± 0.001, relative standard deviation 0.03% E2: retention time 13.489 ± 0.003, relative standard deviation 0.02% E3: retention time 26.571 ± 0.002, relative standard deviation 0.02% E4: retention time 27.274 ± 0.003, relative standard deviation 0.02% E5: retention time 30.439 ± 0.004, relative standard deviation 0.03% E6: retention time 30.677 ± 0.002, relative standard deviation 0.01% E7: retention time 38.947 ± 0.004, relative standard deviation 0.03% E8: retention time 42.561 ± 0.003, relative standard deviation 0.02% E9: retention time 43.494 ± 0.004, relative standard deviation 0.03% ​ E10 peak: retention time 45.375 ± 0.003, relative standard deviation 0.02% E11 peak: retention time 48.973 ± 0.004, relative standard deviation 0.03% E12 peak: retention time 50.244 ± 0.003, relative standard deviation 0.02% E13 peak: retention time 52.58 ± 0.004, relative standard deviation 0.03% E14 peak: retention time 54.981 ± 0.004, relative standard deviation 0.03% E15 peak: retention time 57.149 ± 0.003, relative standard deviation 0.02% E16 peak: retention time 67.14 ± 0.004, relative standard deviation 0.03%.

4. The method for constructing a characteristic spectrum of Wuqi oral liquid according to claim 1, characterized in that, In step S300, further comprising the following steps: S302, the construction of characteristic spectrum under UV PDA detector: The control solution and the test solution are detected by using the UV PDA detector to obtain the UV detector PDA chromatogram, and the UV detector PDA chromatogram is introduced into the traditional Chinese medicine chromatographic fingerprint similarity evaluation system software to obtain the UV detector PDA standard characteristic spectrum of the liquid, the standard characteristic spectrum has 10 characteristic peaks, which are sequentially numbered P1-10 according to the time order, and the sum of the peak areas of the common peaks accounts for more than 95% of the total peak area; Among them, the characteristic peaks belonging to Salvia miltiorrhiza are 4; the characteristic peaks belonging to Ligustri Lucidi Fructus are 2; the characteristic peaks belonging to Crataegi Fructus are 1; the characteristic peaks belonging to Fructus Lycii are 1; the characteristic peaks belonging to Zizyphi Fructus are 1; the characteristic peaks belonging to Glycyrrhizae Radix et Rhizoma are 1; P3 peak is Danshensu, which is a reference peak and belongs to Salvia miltiorrhiza; P6 peak is spec woman ligustrazine, which belongs to Ligustri Lucidi Fructus; P9 peak is Salvianolic acid b, which belongs to Salvia miltiorrhiza; the common peaks of the characteristic spectrum have 10 characteristic peaks, and the retention time and relative standard deviation of each characteristic peak are: P1 peak: retention time 6.464 ± 0.002, relative standard deviation 0.01% P2 peak: retention time 14.552 ± 0.001, relative standard deviation 0.01% P3 peak: retention time 16.614 ± 0.003, relative standard deviation 0.02% P4 peak: retention time 22.287 ± 0.002, relative standard deviation 0.01% P5 peak: retention time 23.152 ± 0.001, relative standard deviation 0.01% P6 peak: retention time 40.193 ± 0.003, relative standard deviation 0.02% P7 peak: retention time 43.087 ± 0.002, relative standard deviation 0.01% P8 peak: retention time 45.528 ± 0.004, relative standard deviation 0.03% P9 peak: retention time 46.921 ± 0.001, relative standard deviation 0.02% P10 peak: retention time 59.625 ± 0.004, relative standard deviation 0.03%.

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