CN112834650B - Quality detection method of bone-strengthening and blood-generating oral liquid - Google Patents
Quality detection method of bone-strengthening and blood-generating oral liquid Download PDFInfo
- Publication number
- CN112834650B CN112834650B CN202110035108.4A CN202110035108A CN112834650B CN 112834650 B CN112834650 B CN 112834650B CN 202110035108 A CN202110035108 A CN 202110035108A CN 112834650 B CN112834650 B CN 112834650B
- Authority
- CN
- China
- Prior art keywords
- bone
- blood
- strengthening
- oral liquid
- generating
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The application discloses a quality detection method of bone-strengthening and blood-generating oral liquid, compared with the prior art, the quality detection method comprises the following steps: preparing a first reference substance solution; preparing a first test solution; establishing a first fingerprint of the bone-strengthening and blood-generating oral liquid; formulating a standard fingerprint of the bone strengthening and blood generating oral liquid; preparing a second reference solution; preparing a second test solution; establishing a second fingerprint map of the bone-strengthening and blood-generating oral liquid; and comparing the second fingerprint of the bone-strengthening and blood-generating oral liquid with the standard fingerprint, and keeping the second test sample solution within +/-5% of the retention time to be qualified in detection. The application provides a quality detection method of bone-strengthening and blood-generating oral liquid, can carry out comprehensive detection to the active ingredient of bone-strengthening and blood-generating oral liquid, does benefit to the effective control to bone-strengthening and blood-generating oral liquid finished product quality to ensure the clinical curative effect of bone-strengthening and blood-generating oral liquid.
Description
Technical Field
The application relates to the field of quality detection methods, in particular to a quality detection method of an oral liquid for strengthening bone and generating blood.
Background
The oral liquid for strengthening bone and generating blood is prepared from bone liquid, codonopsis pilosula, astragalus, lucid ganoderma, chinese date, black fungus and other medicinal materials, has the functions of benefiting marrow, strengthening bones, tonifying qi and generating blood, is mainly the bone liquid as a monarch, has the functions of tonifying kidney and marrow, ensures that marrow and foot have strong bone and essence and can be converted into blood, and the bone liquid is rich in nutrient components such as calcium, so that the oral liquid can not only supplement deficiency of the bone and essence but also prevent deficiency of the bone and essence for the old, children and women who are pregnant and lack of calcium and have anemia, can treat symptoms such as soreness of waist, weakness of knees, weak feet, shaking teeth and falling teeth and developmental retardation caused by the deficiency of calcium and the anemia, can supplement qi of spleen and stomach by the codonopsis pilosula and the astragalus and enrich the source of life and transformation, ensures that the acquired origin exists, and qi and blood can be generated by oneself, and can treat shortness of spirit, fatigue and weakness and can also promote the recovery of the anemia. The combination of Shen and Qin with bone fluid can nourish both spleen and kidney, so as to mutually transform bone marrow and qi and blood to nourish yin and blood. The lucid ganoderma and the Chinese date are used as ministerial drugs to strengthen the qi-tonifying effect to generate blood. Black fungus is sweet, neutral and nontoxic, and moist but not greasy, and is used as a guide to coordinate various medicines. Therefore, all the medicines are mutually matched, have mild property and taste, and tonify but not dry, have the effects of benefiting marrow, strengthening bones, tonifying qi and generating blood, and are a good prescription for treating the symptoms of marrow deficiency and bone weakness caused by calcium deficiency and anemia and deficiency of qi and blood. The bone-strengthening and blood-generating oral liquid has the following clinical indications: it can be used for treating anemia and calcium deficiency of the elderly, children, and women before and after pregnancy, with the symptoms of listlessness, asthenia, pale complexion, soreness of waist and knees, muscle spasm, marasmus, odontoptosis, and infantile delayed fontanel closure, delayed teeth, delayed erection, and delayed movement.
The existing quality standard of the existing bone-strengthening and blood-generating oral liquid only measures the content of a single index component, other effective components are not monitored, certain limitation exists, and the quality level of the bone-strengthening and blood-generating oral liquid cannot be comprehensively reflected. The quality detection method for the bone-strengthening and blood-generating oral liquid is rarely disclosed, and for example, chinese patents 201210012628.4 (publication number CN 201210012628.4), 202030219270.8 (publication number CN 202030219270.8) and 202030219275.0 (publication number CN 202030219275.0) do not relate to the quality detection method; the clinical effect observation of the oral liquid for strengthening bone and generating blood in combination with the litseus sodium phosphate tablets for treating the osteoporosis of postmenopausal women in the 27 th volume and the 8 th volume of 2019 in the journal of China family planning science, the treatment effect and mechanism of the oral liquid for strengthening bone and generating blood on ovaries-removing osteoporosis rats in the 48 th volume and the 23 th volume of 2017 in the Chinese herbal medicine, the protection effect and mechanism research of the oral liquid for strengthening bone and generating blood on the bone marrow inhibition of mice after chemotherapy in the 26 th volume and the 22 th volume in the 2017 th volume in the journal of China Chinese herbal medicine, and the quality detection technology of the oral liquid for strengthening bone and generating blood in the 1 st volume in the 2004 in the Guangdong pharmacy are not related.
In the prior art, no comprehensive quality control method reflects the quality conditions of the traditional Chinese medicinal materials, intermediates and finished products, can not effectively control the production process and the product quality, and can not better ensure the clinical curative effect.
Therefore, it is an urgent need for the skilled in the art to provide a quality detection method for a bone-strengthening and blood-generating oral liquid, which can comprehensively detect the effective components of the bone-strengthening and blood-generating oral liquid.
Disclosure of Invention
In order to solve the technical problems, the application provides a quality detection method of the bone-strengthening and blood-generating oral liquid, which can comprehensively detect the effective components of the bone-strengthening and blood-generating oral liquid.
The technical scheme provided by the application is as follows:
the application provides a quality detection method of an oral liquid for strengthening bone and generating blood, which comprises the following steps: preparation of a first control solution: preparing the calycosin glucoside into a first reference substance solution containing 0.001-0.004 mg of the calycosin glucoside per 1mL by using 50-100% methanol by volume percentage; preparing a first test solution: weighing 5mL of bone-strengthening and blood-generating oral liquid, placing the oral liquid in a 50mL volumetric flask, adding 0-50% methanol or 0-50% ethanol to 50mL scale positions, shaking up, filtering with a microporous membrane, and taking subsequent filtrate to prepare a first test solution; preparing a first fingerprint of the bone strengthening and blood generating oral liquid: precisely absorbing the first reference substance solution and the first test solution with the same amount respectively, injecting the first reference substance solution and the first test solution into a liquid chromatograph for high performance liquid chromatography, and measuring to obtain a first fingerprint of the bone-strengthening and blood-generating oral liquid, wherein the specific conditions of the high performance liquid chromatography are as follows: the chromatographic column is a C18 reversed phase chromatographic column; the mobile phase is gradient eluent composed of an organic phase A and a water phase B, the organic phase A is methanol or acetonitrile, and the water phase B is formic acid solution with the volume percentage concentration of 0.1-0.4%; the detection wavelengths are 254nm, 270nm and 285nm, and the switching time of the detection wavelengths is 0min, 8min and 25min; the column temperature is 20-40 ℃; the flow rate is 0.8mL/min to 1.2mL/min; the analysis time is 50-65 min; gradient elution was used:
preparing a standard fingerprint spectrum of the bone-strengthening and blood-generating oral liquid: preparing a standard fingerprint of the bone-strengthening and blood-generating oral liquid according to the first fingerprint of the bone-strengthening and blood-generating oral liquid; preparing a second reference solution; preparing a second test solution; and (3) establishing a second fingerprint map of the bone strengthening and blood generating oral liquid: accurately sucking the second reference solution and the second test solution in equal amount respectively, injecting into a liquid chromatograph for high performance liquid chromatography, and measuring to obtain a second fingerprint of the bone-strengthening and blood-generating oral liquid; and comparing the second fingerprint of the bone-strengthening and blood-generating oral liquid with the standard fingerprint, and keeping the second test sample solution which is qualified in detection within +/-5 percent of the retention time.
Preferably, in the step of establishing the first fingerprint of the bone-strengthening and blood-generating oral liquid, 10 μ l of each of the first control solution and the first test solution is sucked.
Preferably, the column length of the C18 reverse phase chromatography column is 250mm.
Preferably, the formic acid solution has a concentration of 0.2% by volume.
Preferably, the column temperature is 30 to 40 ℃.
Preferably, the flow rate is 1.0mL/min.
Preferably, the analysis time is 55min.
Further, in a preferred mode of the present invention, the oral liquid for strengthening bone and generating blood comprises the following components in parts by weight: 600 to 700 parts of bone fluid, 40 to 50 parts of codonopsis pilosula, 70 to 80 parts of astragalus root, 10 to 20 parts of lucid ganoderma, 30 to 40 parts of Chinese date and 10 to 20 parts of black fungus.
Further, in a preferred embodiment of the present invention, the step of "preparing the standard fingerprint of the bone-strengthening and blood-generating oral liquid" specifically comprises: and establishing the standard fingerprint of the bone-strengthening and blood-generating oral liquid according to the first fingerprint of the 15 batches of bone-strengthening and blood-generating oral liquid.
Further, in a preferred embodiment of the present invention, the standard fingerprint spectrum is detected by switching wavelengths at 254nm, 270nm, and 285nm, and has 12 common peaks, the sum of the areas of the common peaks accounts for more than 90% of the total peak area, the fingerprint spectrum attributed to synovial fluid has 3 common peaks, the fingerprint spectrum attributed to codonopsis pilosula has 2 common peaks, the fingerprint spectrum attributed to astragalus membranaceus has 2 common peaks, the fingerprint spectrum attributed to ganoderma lucidum has 2 common peaks, the fingerprint spectrum attributed to jujube has 2 common peaks, and the fingerprint spectrum attributed to black fungus has 1 common peak.
Further, in a preferred mode of the present invention, the standard fingerprint spectrum switches wavelengths at 254nm, 270nm and 285nm, and the retention time of the detected 12 common peaks is: (1) 5.373 to 5.406, (2) 5.721 to 5.741, (3) 6.826 to 6.855, (4) 8.011 to 8.045, (5) 9.140 to 9.174, (6) 10.372 to 10.397, (7) 11.220 to 11.259, (8) 14.127 to 14.159, (9) 16.213 to 16.244, (10) 18.475 to 18.519, (11) 19.767 to 19.812, and (12) 44.546 to 44.617.
Further, in a preferred embodiment of the present invention, the similarity between the first fingerprint of the bone-strengthening and blood-generating oral liquid and the standard fingerprint of the bone-strengthening and blood-generating oral liquid is 0.944 to 0.998.
Further, in a preferred mode of the present invention, the common peak of the first fingerprint spectrum has 11 characteristic peaks, and the retention time and the relative standard deviation of each characteristic peak are as follows:
peak No. 1: retention time 5.721 ± 0.020 with relative standard deviation of 0.35%;
peak No. 2: retention time 6.826 ± 0.029, relative standard deviation 0.42%;
peak No. 3: the retention time is 8.011 +/-0.034, and the relative standard deviation is 0.43 percent;
peak No. 4: retention time 9.140 ± 0.034, relative standard deviation 0.37%;
peak No. 5: retention time 10.372 ± 0.025, relative standard deviation 0.24%;
peak No. 6: retention time 11.220 ± 0.039, relative standard deviation 0.35%;
peak No. 7: retention time 14.127 ± 0.032, relative standard deviation 0.23%;
peak No. 8: retention time 16.213 ± 0.031, relative standard deviation 0.19%;
peak No. 9: retention time 18.475 ± 0.044, relative standard deviation 0.24%;
peak No. 10: retention time 19.767 ± 0.045, relative standard deviation 0.23%;
peak No. 11: retention time 44.546 ± 0.071 with a relative standard deviation of 0.16%.
Further, in a preferred embodiment of the present invention, the step "preparing the second control solution" is the same as the step "preparing the first control solution".
Further, in a preferred mode of the present invention, the step "preparing the second sample solution" is the same as the step "preparing the first sample solution".
Further, in a preferred mode of the present invention, the specific conditions of the hplc analysis in the step of "preparing the second fingerprint of the bone-strengthening and blood-generating oral liquid" are the same as the specific conditions of the hplc analysis in the step of "preparing the first fingerprint of the bone-strengthening and blood-generating oral liquid".
Compared with the prior art, the technical scheme provided by the invention is that the quality detection method of the bone-strengthening and blood-generating oral liquid comprises the following steps: preparation of a first control solution: preparing the calycosin glucoside into a first reference substance solution containing 0.001-0.004 mg of the calycosin glucoside per 1mL by using 50-100% methanol by volume percentage; preparing a first test solution: weighing 5mL of bone-strengthening and blood-generating oral liquid, placing the oral liquid in a 50mL volumetric flask, adding 0-50% methanol or 0-50% ethanol to 50mL scale positions, shaking up, filtering with a microporous membrane, and taking subsequent filtrate to prepare a first test solution; a first fingerprint spectrum of the bone strengthening and blood generating oral liquid is formulated: precisely absorbing the first reference substance solution and the first test solution with the same amount respectively, injecting the first reference substance solution and the first test solution into a liquid chromatograph for high performance liquid chromatography, and measuring to obtain a first fingerprint of the bone-strengthening and blood-generating oral liquid, wherein the specific conditions of the high performance liquid chromatography are as follows: the chromatographic column is a C18 reversed phase chromatographic column; the mobile phase is gradient eluent composed of an organic phase A and a water phase B, the organic phase A is methanol or acetonitrile, and the water phase B is formic acid solution with the volume percentage concentration of 0.1-0.4%; the detection wavelengths are 254nm, 270nm and 285nm, and the switching time of the detection wavelengths is 0min, 8min and 25min; the column temperature is 20-40 ℃; the flow rate is 0.8mL/min to 1.2mL/min; the analysis time is 50-65 min; gradient elution is adopted:
preparing a standard fingerprint spectrum of the bone-strengthening and blood-generating oral liquid: preparing a standard fingerprint of the bone-strengthening and blood-generating oral liquid according to the first fingerprint of the bone-strengthening and blood-generating oral liquid; preparing a second reference solution; preparing a second test solution; and (3) establishing a second fingerprint of the bone-strengthening and blood-generating oral liquid: accurately sucking the second reference solution and the second test solution in equal amount respectively, injecting into a liquid chromatograph for high performance liquid chromatography, and measuring to obtain a second fingerprint of the bone-strengthening and blood-generating oral liquid; the second fingerprint of the bone-strengthening and blood-generating oral liquid is compared with the standard fingerprint, the qualified second test solution is reserved within +/-5% of the time, so that the components of the bone-strengthening and blood-generating oral liquid are comprehensively monitored by establishing the standard fingerprint of the bone-strengthening and blood-generating oral liquid, peak attribution and quality association of various medicinal materials of the bone-strengthening and blood-generating oral liquid and a finished product of the bone-strengthening and blood-generating oral liquid can be effectively carried out, and dynamic monitoring and tracing on the quality of a Chinese medicinal material-intermediate (sampling in the preparation process of the finished product of the bone-strengthening and blood-generating oral liquid) -finished product can be effectively carried out.
Drawings
In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present application, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
Fig. 1 is a standard fingerprint of a bone-strengthening and blood-generating oral liquid according to a quality detection method of the bone-strengthening and blood-generating oral liquid provided by an embodiment of the present invention;
fig. 2 is a first control solution chromatogram of a quality detection method of a bone-strengthening and blood-generating oral liquid according to an embodiment of the present invention;
fig. 3 is a full-wavelength scanning spectrum of the bone-strengthening and blood-generating oral liquid according to the quality detection method of the bone-strengthening and blood-generating oral liquid provided by the embodiment of the present invention;
fig. 4 is a first fingerprint of 15 batches of bone-strengthening and blood-generating oral liquid according to the quality detection method of the bone-strengthening and blood-generating oral liquid provided by the embodiment of the invention.
Detailed Description
In order to make those skilled in the art better understand the technical solutions in the present application, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, and it is obvious that the described embodiments are only a part of the embodiments of the present application, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.
It will be understood that when an element is referred to as being "fixed" or "disposed" on another element, it can be directly on the other element or be indirectly disposed on the other element; when an element is referred to as being "connected to" another element, it can be directly connected to the other element or be indirectly connected to the other element.
It will be understood that the terms "length," "width," "upper," "lower," "front," "rear," "first," "second," "vertical," "horizontal," "top," "bottom," "inner," "outer," and the like, as used herein, refer to an orientation or positional relationship indicated in the drawings that is solely for the purpose of facilitating the description and simplifying the description, and do not indicate or imply that the referenced device or element must have a particular orientation, be constructed and operated in a particular orientation, and thus should not be considered as limiting the present application.
Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present application, "plurality" or "a plurality" means two or more unless specifically limited otherwise.
It should be understood that the structures, ratios, sizes, etc. shown in the drawings are only used for understanding and reading the disclosure, and are not used for limiting the practical limitations of the present application, so that the present invention has no technical significance, and any modifications of the structures, changes of the ratio relationships, or adjustments of the sizes, should still fall within the scope of the technical contents disclosed in the present application without affecting the efficacy and the achievable purpose of the present application.
As shown in fig. 1-4, the present application provides a quality detection method of an oral liquid for strengthening bone and generating blood, comprising the following steps: preparation of a first control solution: preparing the calycosin glucoside into a first reference substance solution containing 0.001-0.004 mg of the calycosin glucoside per 1mL by using 50-100% methanol by volume percentage; preparing a first test solution: weighing 5mL of bone-strengthening and blood-generating oral liquid, placing the oral liquid in a 50mL volumetric flask, adding 0-50% methanol or 0-50% ethanol to 50mL scale positions, shaking up, filtering with a microporous membrane, and taking subsequent filtrate to prepare a first test solution; preparing a first fingerprint of the bone strengthening and blood generating oral liquid: accurately sucking equivalent first reference substance solution and first test solution respectively, injecting into a liquid chromatograph for high performance liquid chromatography, and determining to obtain a first fingerprint of the bone-strengthening and blood-generating oral liquid, wherein the specific conditions of the high performance liquid chromatography are as follows: the chromatographic column is a C18 reversed phase chromatographic column; the mobile phase is gradient eluent composed of an organic phase A and a water phase B, the organic phase A is methanol or acetonitrile, and the water phase B is formic acid solution with the volume percentage concentration of 0.1-0.4%; the detection wavelengths are 254nm, 270nm and 285nm, and the switching time of the detection wavelengths is 0min, 8min and 25min; the column temperature is 20-40 ℃; the flow rate is 0.8mL/min to 1.2mL/min; the analysis time is 50-65 min; gradient elution was used:
preparing a standard fingerprint of the bone strengthening and blood generating oral liquid: preparing a standard fingerprint of the bone-strengthening and blood-generating oral liquid according to the first fingerprint of the bone-strengthening and blood-generating oral liquid; preparing a second reference solution; preparing a second test solution; and (3) establishing a second fingerprint of the bone-strengthening and blood-generating oral liquid: accurately sucking the second reference solution and the second test solution in equal amount respectively, injecting into a liquid chromatograph for high performance liquid chromatography, and measuring to obtain a second fingerprint of the bone-strengthening and blood-generating oral liquid; and comparing the second fingerprint of the bone-strengthening and blood-generating oral liquid with the standard fingerprint, and keeping the second test sample solution which is qualified in detection within +/-5 percent of the retention time.
Compared with the prior art, the quality detection method of the bone-strengthening and blood-generating oral liquid comprises the following steps: preparation of a first control solution: preparing the calycosin glucoside into a first reference substance solution containing 0.001-0.004 mg of the calycosin glucoside per 1mL by using 50-100% methanol by volume percentage; preparing a first test solution: measuring 5mL of bone-strengthening and blood-generating oral liquid, placing the oral liquid in a 50mL volumetric flask, adding 0-50% methanol or 0-50% ethanol to the 50mL scale, shaking up, filtering by using a microporous filter membrane, and taking subsequent filtrate to prepare a first test sample solution; a first fingerprint spectrum of the bone strengthening and blood generating oral liquid is formulated: precisely absorbing the first reference substance solution and the first test solution with the same amount respectively, injecting the first reference substance solution and the first test solution into a liquid chromatograph for high performance liquid chromatography, and measuring to obtain a first fingerprint of the bone-strengthening and blood-generating oral liquid, wherein the specific conditions of the high performance liquid chromatography are as follows: the chromatographic column is a C18 reversed phase chromatographic column; the mobile phase is gradient eluent composed of an organic phase A and a water phase B, the organic phase A is methanol or acetonitrile, and the water phase B is formic acid solution with the volume percentage concentration of 0.1-0.4%; the detection wavelengths are 254nm, 270nm and 285nm, and the switching time of the detection wavelengths is 0min, 8min and 25min; the column temperature is 20-40 ℃; the flow rate is 0.8mL/min to 1.2mL/min; the analysis time is 50-65 min; gradient elution is adopted:
preparing a standard fingerprint spectrum of the bone-strengthening and blood-generating oral liquid: preparing a standard fingerprint of the bone-strengthening and blood-generating oral liquid according to the first fingerprint of the bone-strengthening and blood-generating oral liquid; preparing a second reference solution; preparing a second test solution; and (3) establishing a second fingerprint map of the bone strengthening and blood generating oral liquid: accurately sucking the second reference substance solution and the second test solution in equal amount, respectively, injecting into a liquid chromatograph for high performance liquid chromatography, and measuring to obtain a second fingerprint of the bone-strengthening and blood-generating oral liquid; the second fingerprint of the bone-strengthening and blood-generating oral liquid is compared with the standard fingerprint, the qualified second test solution is reserved within +/-5% of the time, so that the components of the bone-strengthening and blood-generating oral liquid are comprehensively monitored by establishing the standard fingerprint of the bone-strengthening and blood-generating oral liquid, the peak attribution and quality association of various medicinal materials of the bone-strengthening and blood-generating oral liquid and a finished product of the bone-strengthening and blood-generating oral liquid can be effectively carried out, and the dynamic monitoring and source tracing of the quality of a Chinese medicinal material-intermediate (sampling in the preparation process of the finished product of the bone-strengthening and blood-generating oral liquid) -finished product can be more effectively carried out.
It should be added that, in the embodiment of the present invention, the hplc analysis employs three wavelengths with detection wavelengths of 254nm, 270nm, and 285nm, determines the maximum absorption wavelength of each component of the bone-strengthening and blood-generating oral liquid by full-wavelength scanning, and then determines the detection wavelengths in different intervals according to the spectrum absorption of the specific component, so that the quality monitoring of each component in the bone-strengthening and blood-generating oral liquid can be more accurately performed, the purpose of increasing the quality coverage can be achieved by using the interval segmented wavelength detection method, the detection quality and the detection sensitivity can be effectively improved, the quality of the bone-strengthening and blood-generating oral liquid can be comprehensively, objectively, and scientifically evaluated, and the method can be used as an important basis for identifying the authenticity of the bone-strengthening and blood-generating oral liquid.
Specifically, in the embodiment of the invention, the oral liquid for strengthening bone and generating blood comprises the following components in parts by weight: 600 to 700 parts of bone fluid, 40 to 50 parts of codonopsis pilosula, 70 to 80 parts of astragalus root, 10 to 20 parts of lucid ganoderma, 30 to 40 parts of Chinese date and 10 to 20 parts of black fungus.
Specifically, in the embodiment of the present invention, the step of "preparing the standard fingerprint of the bone-strengthening and blood-generating oral liquid" specifically comprises: and formulating the standard fingerprint of the bone-strengthening and blood-generating oral liquid according to the first fingerprint of the 15 batches of bone-strengthening and blood-generating oral liquid.
Specifically, in the embodiment of the invention, the standard fingerprint spectrum is detected by switching wavelengths under 254nm, 270nm and 285nm, and has 12 common peaks, the sum of the peak areas of the common peaks accounts for more than 90% of the total peak area, the fingerprint spectrum attributed to bone fluid has 3 common peaks, the fingerprint spectrum attributed to codonopsis pilosula has 2 common peaks, the fingerprint spectrum attributed to astragalus membranaceus has 2 common peaks, the fingerprint spectrum attributed to ganoderma lucidum has 2 common peaks, the fingerprint spectrum attributed to jujube has 2 common peaks, the fingerprint spectrum attributed to black fungus has 1 common peak, the established standard fingerprint spectrum has 12 common peaks, the information content is large, and chemical components in the test solution can be completely reserved.
Specifically, in the embodiment of the present invention, the standard fingerprint spectrum switches wavelengths at 254nm, 270nm, and 285nm, and retention times of the detected 12 common peaks are: (1) 5.373-5.406, (2) 5.721-5.741, (3) 6.826-6.855, (4) 8.011-8.045, (5) 9.140-9.174, (6) 10.372-10.397, (7) 11.220-11.259, (8) 14.127-14.159, (9) 16.213-16.244, (10) 18.475-18.519, (11) 19.767-19.812, and (12) 44.546-44.617.
It should be noted that, in the present embodiment, the retention time of the common peak is calculated by using the bone-strengthening and blood-generating oral liquid with lot number of 2006021 as the reference map S1.
Specifically, in the embodiment of the invention, the similarity between the first fingerprint of the bone-strengthening and blood-generating oral liquid and the standard fingerprint of the bone-strengthening and blood-generating oral liquid is 0.944-0.998.
It should be added that, in the embodiment of the present invention, a standard feature spectrum is prepared by using traditional Chinese medicine fingerprint similarity evaluation software according to a first feature spectrum determined by a common peak of a first fingerprint of the oral liquid for bone-strengthening and blood-nourishing in 15 batches, the standard feature spectrum has 11 feature peaks, the oral liquid for bone-strengthening and blood-nourishing in 2006021 is used as a reference spectrum S1, and retention time and relative standard deviation of each feature peak are as follows:
peak No. 1: retention time 5.721 ± 0.020 with relative standard deviation of 0.35%;
peak No. 2: retention time 6.826 ± 0.029, relative standard deviation 0.42%;
peak No. 3: the retention time is 8.011 +/-0.034, and the relative standard deviation is 0.43 percent;
peak No. 4: retention time 9.140 ± 0.034, relative standard deviation 0.37%;
peak No. 5: retention time 10.372 ± 0.025, relative standard deviation 0.24%;
peak No. 6: retention time 11.220 ± 0.039, relative standard deviation 0.35%;
peak No. 7: retention time 14.127 ± 0.032, relative standard deviation 0.23%;
peak No. 8: retention time 16.213 ± 0.031, relative standard deviation 0.19%;
peak No. 9: retention time 18.475 ± 0.044, relative standard deviation 0.24%;
peak No. 10: retention time 19.767 ± 0.045, relative standard deviation 0.23%;
peak No. 11: retention time 44.546 ± 0.071 with a relative standard deviation of 0.16%.
More specifically, the preferred retention time of each peak in the standard fingerprint is as follows: (1) 5.340 to 5.406, (2) 5.701 to 5.741, (3) 6.797 to 6.855, (4) 7.977 to 8.045, (5) 9.106 to 9.174, (6) 10.347 to 10.397, (7) 11.181 to 11.259, (8) 14.095 to 14.159, (9) 16.182 to 16.244, (10) 18.431 to 18.519, (11) 19.722 to 19.812, and (12) 44.475 to 44.617.
Specifically, in the present example, the step "preparing the second control solution" is the same as the step "preparing the first control solution".
More specifically, the "preparing the second control solution" is specifically: the calycosin glucoside is prepared into a second reference solution containing 0.001-0.004 mg of calycosin glucoside per 1mL by using 50-100% methanol by volume percentage.
Specifically, in the present embodiment, the step "preparing the second sample solution" is the same as the step "preparing the first sample solution".
More specifically, the "preparing the second sample solution" includes: taking 5mL of bone-strengthening and blood-generating oral liquid, placing the oral liquid in a 50mL volumetric flask, adding 0-50% methanol or 0-50% ethanol to 50mL scale positions, shaking up, filtering with a microporous membrane, taking the subsequent filtrate, and preparing a second test solution.
Specifically, in the embodiment of the present invention, the specific conditions of the high performance liquid chromatography analysis for "preparing the second fingerprint of the bone-tonifying and blood-generating oral liquid" in the step are the same as the specific conditions of the high performance liquid chromatography analysis for "preparing the first fingerprint of the bone-tonifying and blood-generating oral liquid".
It should be noted that, in the embodiment of the present invention, through traditional Chinese medicine fingerprint similarity evaluation software, a second feature spectrum is determined according to 15 batches of common peaks of a second fingerprint of the bone-strengthening and blood-generating oral liquid, and the second feature spectrum is compared with the standard fingerprint, and the second test solution that is qualified in detection is determined if the retention time is within ± 5%.
The quality detection method of the bone-strengthening and blood-generating oral liquid provided by the invention comprises the following specific operation steps or working principle:
preparing the calycosin glucoside into a first reference solution containing 0.001-0.004 mg of the calycosin glucoside per 1mL by using 50-100% of methanol, measuring 5mL of bone-tonifying oral liquid, putting the bone-tonifying oral liquid into a 50mL volumetric flask, adding 0-50% of methanol or 0-50% of ethanol to 50mL scale positions, shaking uniformly, filtering by using a microporous membrane, taking subsequent filtrate to prepare a first sample solution, respectively and precisely absorbing the first reference solution and the first sample solution in equal amount, injecting the first sample solution and the first sample solution into a liquid chromatograph for high performance liquid chromatography analysis, determining to obtain a first fingerprint of the bone-tonifying oral liquid, formulating a standard fingerprint of the bone-tonifying oral liquid according to the first fingerprint of the bone-tonifying oral liquid, preparing a second reference solution containing 0.001-0.004 mg of the calycosin glucoside per 1mL by using 50-100% of methanol, measuring 5mL of the bone-strengthening and blood-generating oral liquid, placing the oral liquid in a 50mL volumetric flask, adding 0-50% methanol or 0-50% ethanol to 50mL scale positions, shaking up, filtering with a microporous membrane, taking a subsequent filtrate to prepare a second test solution, respectively and precisely absorbing the second reference solution and the second test solution in equal amount, injecting the solution into a liquid chromatograph for high performance liquid chromatography analysis, determining to obtain a second fingerprint of the bone-strengthening and blood-generating oral liquid, comparing the second fingerprint of the bone-strengthening and blood-generating oral liquid with a standard fingerprint, retaining the qualified second test solution within +/-5% of the time, determining a second characteristic map according to the common peak of the second fingerprint of the bone-strengthening and blood-generating oral liquid in 15 batches by traditional Chinese medicine fingerprint similarity evaluation software, comparing the second characteristic map with the standard fingerprint, the second test solution qualified for detection is reserved within +/-5%, the high performance liquid chromatography fingerprint detection method provided by the invention has the advantages of higher precision, better stability, shorter analysis time and certain specificity, the separation degree of each characteristic peak in the detected fingerprint is better, the fingerprint of the bone-strengthening and blood-generating oral liquid and the fingerprint of the intermediate of the bone-strengthening and blood-generating oral liquid have better correlation, the application of the intermediate of the bone-strengthening and blood-generating oral liquid and the finished product fingerprint thereof can enhance the quality controllability in the production process, the application of the fingerprint of the bone-strengthening and blood-generating oral liquid can comprehensively reflect whether the production process is uniform and stable, and the quality stability and the clinical curative effect of the bone-strengthening and blood-generating oral liquid can be ensured.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (5)
1. A quality detection method of an oral liquid for strengthening bone and generating blood is characterized by comprising the following steps:
preparation of a first control solution: preparing the calycosin glucoside into a first reference solution containing 0.001-0.004 mg of calycosin glucoside per 1mL by using 50-100% methanol by volume percentage concentration;
preparing a first test solution: weighing 5mL of bone-strengthening and blood-generating oral liquid, placing the oral liquid in a 50mL volumetric flask, adding 0-50% methanol or 0-50% ethanol to 50mL scale positions, shaking up, filtering with a microporous membrane, and taking subsequent filtrate to prepare a first test solution;
a first fingerprint spectrum of the bone strengthening and blood generating oral liquid is formulated: accurately sucking equivalent first reference substance solution and first test solution respectively, injecting into a liquid chromatograph for high performance liquid chromatography, and determining to obtain a first fingerprint of the bone-strengthening and blood-generating oral liquid, wherein the specific conditions of the high performance liquid chromatography are as follows: the chromatographic column is a C18 reversed phase chromatographic column; the mobile phase is gradient eluent composed of an organic phase A and a water phase B, the organic phase A is methanol or acetonitrile, and the water phase B is formic acid solution with the volume percentage concentration of 0.2%; the detection wavelengths are 254nm, 270nm and 285nm, and the switching time of the detection wavelengths is 0min, 8min and 25min; the column temperature is 20-40 ℃; the flow rate is 0.8mL/min to 1.2mL/min; the analysis time is 55min; gradient elution was used:
The first control solution and the first test solution are respectively sucked to be 10 mu l;
preparing a standard fingerprint spectrum of the bone-strengthening and blood-generating oral liquid: preparing a standard fingerprint of the bone-strengthening and blood-generating oral liquid according to the first fingerprint of the bone-strengthening and blood-generating oral liquid;
preparation of a second control solution: preparing the calycosin glucoside into a second reference solution containing 0.001-0.004 mg of the calycosin glucoside per 1mL by using 50-100% methanol by volume percentage;
preparing a second test solution: measuring 5mL of bone-strengthening and blood-generating oral liquid, placing the oral liquid in a 50mL volumetric flask, adding 0-50% methanol or 0-50% ethanol to the 50mL scale, shaking up, filtering by using a microporous filter membrane, and taking subsequent filtrate to prepare a second test solution;
and (3) establishing a second fingerprint of the bone-strengthening and blood-generating oral liquid: precisely absorbing the second reference substance solution and the second test solution with the same amount respectively, injecting the second reference substance solution and the second test solution into a liquid chromatograph for high performance liquid chromatography analysis, and determining to obtain a second fingerprint of the bone-strengthening and blood-generating oral liquid;
comparing the second fingerprint of the bone-strengthening and blood-generating oral liquid with the standard fingerprint, and keeping the second sample solution qualified in detection within +/-5 percent of the retention time;
the bone-strengthening and blood-generating oral liquid comprises the following components in parts by weight: 600 to 700 parts of bone fluid, 40 to 50 parts of codonopsis pilosula, 70 to 80 parts of astragalus root, 10 to 20 parts of lucid ganoderma, 30 to 40 parts of Chinese date and 10 to 20 parts of black fungus.
2. The method for detecting the quality of the oral liquid for strengthening bone and generating blood according to claim 1, wherein the step of establishing the standard fingerprint of the oral liquid for strengthening bone and generating blood specifically comprises the following steps: and establishing the standard fingerprint of the bone-strengthening and blood-generating oral liquid according to the first fingerprint of the 15 batches of bone-strengthening and blood-generating oral liquid.
3. The method for detecting the quality of oral liquid for strengthening bone and generating blood as claimed in claim 2, wherein the standard fingerprint spectrum switches wavelengths at 254nm, 270nm and 285nm, and the retention time of 11 detected common peaks are respectively as follows:
peak No. 1: the retention time is 5.721-5.741 min;
peak No. 2: the retention time is 6.826-6.855 min;
peak No. 3: the retention time is 8.011-8.045 min;
peak No. 4: the retention time is 9.140-9.174 min;
peak No. 5: the retention time is 10.372-10.397 min;
peak No. 6: the retention time is 11.220-11.259 min;
peak No. 7: the retention time is 14.127-14.159 min;
peak No. 8: the retention time is 16.213-16.244 min;
peak No. 9: the retention time is 18.475 to 18.519 min;
peak No. 10: the retention time is 19.767-19.812 min;
peak No. 11: the retention time is 44.546-44.617 min.
4. The method for detecting the quality of the oral liquid for strengthening bone and generating blood according to claim 3, wherein the similarity between the first fingerprint of the oral liquid for strengthening bone and generating blood and the standard fingerprint of the oral liquid for strengthening bone and generating blood is 0.944-0.998.
5. The method for detecting the quality of oral liquid for strengthening bone and generating blood as claimed in claim 4, wherein the common peaks of the first fingerprint have 11 characteristic peaks, and the retention time and relative standard deviation of each characteristic peak are as follows:
peak No. 1: retention time 5.721 + -0.020 min, relative standard deviation 0.35%;
peak No. 2: retention time 6.826 + -0.029 min, relative standard deviation 0.42%;
peak No. 3: the retention time is 8.011 +/-0.034 min, and the relative standard deviation is 0.43 percent;
peak No. 4: retention time 9.140 plus or minus 0.034 min, relative standard deviation 0.37%;
peak No. 5: the retention time is 10.372 +/-0.025 min, and the relative standard deviation is 0.24 percent;
peak No. 6: retention time 11.220 + -0.039 min, relative standard deviation 0.35%;
peak No. 7: the retention time is 14.127 +/-0.032 min, and the relative standard deviation is 0.23%;
peak No. 8: retention time 16.213 ± 0.031 min, relative standard deviation 0.19%;
peak No. 9: the retention time is 18.475 plus or minus 0.044 min, and the relative standard deviation is 0.24 percent;
peak No. 10: retention time 19.767 ± 0.045 min, relative standard deviation 0.23%;
peak No. 11: retention time 44.546 ± 0.071 min with relative standard deviation of 0.16%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110035108.4A CN112834650B (en) | 2021-01-12 | 2021-01-12 | Quality detection method of bone-strengthening and blood-generating oral liquid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110035108.4A CN112834650B (en) | 2021-01-12 | 2021-01-12 | Quality detection method of bone-strengthening and blood-generating oral liquid |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112834650A CN112834650A (en) | 2021-05-25 |
| CN112834650B true CN112834650B (en) | 2022-10-21 |
Family
ID=75929708
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110035108.4A Active CN112834650B (en) | 2021-01-12 | 2021-01-12 | Quality detection method of bone-strengthening and blood-generating oral liquid |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112834650B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000007607A1 (en) * | 1998-08-04 | 2000-02-17 | Kosbab, John, V. | Nutrient and therapeutic compositions for the treatment of cancer |
| CN105675750A (en) * | 2016-01-18 | 2016-06-15 | 吉林修正药业新药开发有限公司 | Method for constructing HPLC characteristic chromatogram of Chinese patent medicine 'Yishen Bugu liquid' |
| CN107576739A (en) * | 2017-09-12 | 2018-01-12 | 健民药业集团股份有限公司 | A kind of HPLC fingerprint atlas detection methods of LONGMU ZHUANGGU KELI |
| CN109718304A (en) * | 2019-03-20 | 2019-05-07 | 江西天元药业有限公司 | Improve the Chinese medicine composition of the microcirculation of human body cardio and vascular function and pre- preventing thrombosis |
Family Cites Families (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2004286279A1 (en) * | 2003-10-29 | 2005-05-12 | The Samuel Roberts Noble Foundation, Inc. | Plant isoflavonoid hydroxylases and methods of use thereof |
| TWI285262B (en) * | 2005-06-20 | 2007-08-11 | Dev Center Biotechnology | Characteristic mass spectrum fingerprinting setting method and rapid identification method of Chinese herb medicine and prescription |
| CN1733100A (en) * | 2005-09-02 | 2006-02-15 | 北京润德康医药科技有限公司 | Chinese medicinal preparation for strengthening bone and generating blood and process for preparing the same |
| EP2294402A2 (en) * | 2008-06-05 | 2011-03-16 | Bionovo Inc. | Method of quantification of multiple bioactives from botanical compositons |
| CN101822765B (en) * | 2010-01-28 | 2012-02-29 | 江西新赣江药业有限公司 | Quality inspection method of traditional Chinese medicine compound preparation with functions of tonifying kidney, strengthening bones, enriching blood and being beneficial to sperm |
| CN103202858A (en) * | 2012-01-16 | 2013-07-17 | 湖南天劲制药有限责任公司 | Pig bone nutrient and bone-strengthening and blood-generating oral liquid prepared by same |
| CN103808840B (en) * | 2012-11-02 | 2015-09-09 | 丽珠集团利民制药厂 | A kind of method for building up of ginseng and astragalus injection for strengthening body finger-print |
| US20190219550A1 (en) * | 2012-11-02 | 2019-07-18 | Li Min Pharmaceutical Factory Of Livzon Pharmaceutical Group | System and Method for Identifying Shenqi Fuzheng Injection |
| CN103926351B (en) * | 2014-05-06 | 2015-05-27 | 湖南康寿制药有限公司 | Blood-production preparation quality test method and construction method of standard fingerprint spectrum thereof |
| CN104459003B (en) * | 2014-12-16 | 2016-01-20 | 湖南汉森制药股份有限公司 | The construction method of contracting spring standard preparation finger-print and characteristic spectrum and quality determining method |
| CN104569199B (en) * | 2014-12-30 | 2016-08-24 | 上海现代中医药股份有限公司 | A kind of assay method of Radix Astragali finger printing |
| CN106501397B (en) * | 2016-10-19 | 2019-04-09 | 辽宁康辰药业有限公司 | The method for building up and purposes for the treatment of osteoporosis preparation HPLC fingerprint pattern |
| CN107505418B (en) * | 2017-06-29 | 2020-03-06 | 中山大学 | Construction method of HPLC fingerprint of lung-tonifying and blood-activating capsule and standard fingerprint |
| CN107367567B (en) * | 2017-07-17 | 2019-02-26 | 中山市中智药业集团有限公司 | The fingerprint map construction and its quality determining method of stilbene jujube oral liquid |
| CN107929420B (en) * | 2017-11-23 | 2021-01-19 | 江西天元药业有限公司 | Traditional Chinese medicine composition for improving qi and blood functions and immunity of human body and detection method |
| CN112014481B (en) * | 2019-05-29 | 2022-10-25 | 湖北梦阳药业股份有限公司 | Detection method of raw white oral liquid fingerprint spectrum |
-
2021
- 2021-01-12 CN CN202110035108.4A patent/CN112834650B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000007607A1 (en) * | 1998-08-04 | 2000-02-17 | Kosbab, John, V. | Nutrient and therapeutic compositions for the treatment of cancer |
| CN105675750A (en) * | 2016-01-18 | 2016-06-15 | 吉林修正药业新药开发有限公司 | Method for constructing HPLC characteristic chromatogram of Chinese patent medicine 'Yishen Bugu liquid' |
| CN107576739A (en) * | 2017-09-12 | 2018-01-12 | 健民药业集团股份有限公司 | A kind of HPLC fingerprint atlas detection methods of LONGMU ZHUANGGU KELI |
| CN109718304A (en) * | 2019-03-20 | 2019-05-07 | 江西天元药业有限公司 | Improve the Chinese medicine composition of the microcirculation of human body cardio and vascular function and pre- preventing thrombosis |
Non-Patent Citations (2)
| Title |
|---|
| Immuno-enhancement effects of Shenqi Fuzheng Injection on cyclophosphamide-induced immunosuppression in Balb/c mice;Jinxu Wang et al;《Journal of Ethnopharmacology》;20121231;第788-795页 * |
| 益肾补骨液高效液相色谱法特征图谱研究;徐云 等;《长春中医药大学学报》;20180228;第34卷(第1期);第31-33页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112834650A (en) | 2021-05-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101912440B (en) | Quality control method of complex salvia tablet and application thereof | |
| CN103364506A (en) | Quality control method for effective parts of pomelo flavedo and pomelo flavedo preparation | |
| CN100518768C (en) | Cassia twig tuckahoe effervescence tablet and testing method | |
| CN108459090B (en) | Quality control method of Jichuan decoction composition | |
| CN102507834B (en) | Quality control method for eight-flavor agilawood preparations | |
| CN104965033B (en) | Determination method for HPLC (High Performance Liquid Chromatography) fingerprint of rhizoma atractylodis and cortex phellodendri pain removing preparation | |
| CN112834650B (en) | Quality detection method of bone-strengthening and blood-generating oral liquid | |
| CN104007198B (en) | A kind of glossy ganoderma emperor's preparation HPLC standard finger-print and construction method thereof and application | |
| CN109507312A (en) | A kind of identification method of Cortex Phellodendri and its application | |
| CN105158352B (en) | A kind of method for building up of finger-print of perhexiline piece and its application | |
| CN110274962B (en) | Method for measuring content of salvianolic acid component in astragalus-ginseng qi-tonifying dropping pills | |
| CN113341007B (en) | HPLC (high Performance liquid chromatography) characteristic spectrum-based method for measuring content of all ingredients of jujube kernel nerve-soothing capsules | |
| CN109270181A (en) | The method for building up and its standard diagram of first luxuriant growth Tongbian capsule HPLC finger-print | |
| CN114324667B (en) | HPLC (high Performance liquid chromatography) characteristic spectrum construction method and content determination method of DACHAIHU decoction | |
| CN103575823B (en) | The detection method of 8 kinds of chemical compositions in a kind of Tangminling preparation | |
| CN113759056B (en) | Characteristic spectrum of Chinese lobelia and preparation thereof and construction method thereof | |
| CN111896637B (en) | Detection method of Jinqing intermediate and fingerprint spectrum construction method thereof | |
| CN109298116A (en) | A kind of detection method of nauclea officinalis medicinal extract piece | |
| CN113189225A (en) | Method for simultaneously determining content of 3 alkaloids in strong loquat syrup | |
| CN101920001B (en) | Finger-print quality determination method on wuji powder preparation and intermediate thereof | |
| CN107941927B (en) | Method for determining lobetyolin content by UPLC/Q-TOF-MS | |
| CN105738499A (en) | Method for constructing refined coronary tablet specific chromatogram and measuring six components | |
| CN102038856A (en) | Content measuring method for four components in Chinese medicinal composition | |
| CN104198634A (en) | Method for establishing Sishen tablet fingerprint spectrum | |
| CN104458954A (en) | Semen cuscutae formula particle fingerprint spectrum and building method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |