CN113640398B - Construction method of gypsum standard decoction feature map - Google Patents

Construction method of gypsum standard decoction feature map Download PDF

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CN113640398B
CN113640398B CN202110706336.XA CN202110706336A CN113640398B CN 113640398 B CN113640398 B CN 113640398B CN 202110706336 A CN202110706336 A CN 202110706336A CN 113640398 B CN113640398 B CN 113640398B
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peak
gypsum
solution
standard decoction
decoction
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CN113640398A (en
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李国卫
黄跃前
吴文平
钟春琳
刘晓琳
杨晓东
程学仁
陈向东
邱韵静
黄上书
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Guangdong Yifang Pharmaceutical Co Ltd
Shandong Yifang Pharmaceutical Co Ltd
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Guangdong Yifang Pharmaceutical Co Ltd
Shandong Yifang Pharmaceutical Co Ltd
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    • 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
    • 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
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • 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
    • G01N30/86—Signal analysis
    • G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
    • G01N30/8679—Target compound analysis, i.e. whereby a limited number of peaks is analysed
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention relates to a construction method of a characteristic spectrum of gypsum standard decoction, which comprises the following steps: providing a reference solution, preparing a sample solution, detecting the sample solution and the reference solution by adopting an ion chromatography, introducing a chromatogram obtained by detecting the reference solution and a chromatogram obtained by detecting the sample solution into a traditional Chinese medicine chromatographic fingerprint similarity evaluation system, and making a characteristic map of a gypsum standard decoction; the reference substance solution is a solution obtained by dissolving a reference substance in a solvent, wherein the reference substance contains sodium ions, potassium ions and magnesium ions; the preparation of the test solution comprises the following steps: extracting the gypsum standard decoction by using an extraction solvent, and collecting an extracting solution to obtain a sample solution, wherein the extraction solvent is methane sulfonic acid solution. The invention constructs the characteristic spectrum of the gypsum standard decoction for the first time. In the characteristic spectrum, the relative retention time of each characteristic peak is stable, and the relative peak area is obviously different from the integral difference of other mineral medicines, so that the specificity is presented.

Description

Construction method of gypsum standard decoction feature map
Technical Field
The invention relates to the technical field of traditional Chinese medicine detection, in particular to a construction method of a gypsum standard decoction feature map.
Background
Gypsum, which is aqueous calcium sulfate (CaSO) 4 ·2H 2 The ore of O) is used for the treatment of, the aliases are fine stone, fine marble (miscellaneous records), soft gypsum (materia medica derived from materia medica) and the like Cold water stone (outline of the project) and white tiger (medicine change of the definition). Gypsum is a monoclinic mineral, is usually produced in sedimentary rock formed in salt lakes and inland lakes in gulf, is a fibrous aggregate, is in the form of long blocks, plate blocks or irregular blocks, is white, off-white or light yellow, is semitransparent, heavy, soft, has spun silk-like luster in longitudinal section, is slightly gas and has light taste.
Gypsum has the following pharmacological actions: (1) antipyretic effect: recent researches show that the gypsum and white tiger soup have certain antipyretic effect on experimental heat-producing rabbits. The natural gypsum 1:1 decoction is administered to rabbit rectum by 4mL, and has antipyretic effect on cow milk and vaccine-heated rabbits, but pure gypsum has no antipyretic effect. It has been shown that gypsum can inhibit the central thermoregulation of excessive excitation during fever, has strong and rapid antipyretic effect, but is not durable. (2) thirst quenching effect: it has thirst quenching effect on animals caused by inhibiting drinking water, heating caused by endotoxin, dehydration caused by diuretic or dehydration caused by hypertonic saline, and radiation heat. (3) enhancing phagocytosis: the gypsum Hanks liquid with the ratio of 1:1 can enhance the capability of the rabbit alveolar macrophages cultured in vitro to engulf the white staphylococcus and the colloidal gold, and can promote the maturation of phagocytes. The above-mentioned effects of gypsum may be closely related to the calcium ions contained therein. The white tiger decoction can enhance the phagocytic rate and phagocytic index of macrophages in the abdominal cavity of mice, has the effect of promoting the function of phagocytizing staphylococcus aureus by white cells in vitro, can enhance the lymphocyte conversion rate, and promotes the generation of mouse antibodies and the content of serum lysozyme. The phagocytosis promotion of Gypsum Fibrosum and BAIHU decoction may be one of the pharmacological bases for treating infectious diseases. (4) sedative and antispasmodic effects: gypsum Fibrosum is taken orally, and part of Gypsum Fibrosum is changed into soluble calcium salt to be absorbed, so that the blood calcium concentration is increased, and the excitability of muscle is inhibited, thereby playing a certain role in tranquilizing and antispasmodic. (5) antiviral action: gypsum Fibrosum has antiviral effect, and metal ion may be the effective component for resisting viruses. (6) other actions: studies show that gypsum has synergistic effect on the hypoglycemic effect of ginseng and rhizoma anemarrhenae. The 2.5% gypsum supernatant is used to replace water for free drinking for 1 month, the calcium content in the organs such as pituitary gland, adrenal gland, submaxillary gland, prostate gland, pancreas, testis is reduced, and the calcium content in spleen and thymus is increased. The gypsum extract can be used for exciting isolated Bufo siccus and rabbit heart at small dosage, and inhibiting at large dosage. For the isolated small intestine and uterus of rabbits, the amplitude is increased when a small amount of gypsum supernatant is used, and the tension is reduced when a large amount of gypsum supernatant is used, so that the amplitude is reduced. The gypsum supernatant can inhibit the delivery of the mouse intestinal contents. Gypsum also can reduce vascular permeability, shorten clotting time, and promote urination.
Conventionally, gypsum fingerprint is mainly constructed by adopting X-ray diffraction analysis, but the price of an X-ray diffractometer is higher, and the X-ray diffraction analysis mainly aims at unit cells forming a mineral drug and is not an element in the unit cells, so that the established fingerprint can only aim at the most main part, and other parts with other characteristics (such as inorganic elements) can be ignored.
Currently, there is still an urgent need for developing an accurate and reliable detection means for gypsum decoction.
Disclosure of Invention
Based on the background technology, one of the purposes of the invention is to provide a construction method of a gypsum standard decoction feature map. By the method, the trace element characteristic spectrum of the gypsum standard decoction containing the sodium ion characteristic peak, the potassium ion characteristic peak and the magnesium ion characteristic peak is successfully constructed for the first time.
The above object of the present invention can be achieved by the following technical solutions:
a construction method of a characteristic map of gypsum standard decoction comprises the following steps:
providing a reference solution, preparing a sample solution, detecting the reference solution by adopting an ion chromatography, detecting the sample solution by adopting an ion chromatography, introducing a chromatogram obtained by detecting the reference solution and a chromatogram obtained by detecting the sample solution into a traditional Chinese medicine chromatographic fingerprint similarity evaluation system, and making a characteristic map of a gypsum standard decoction;
The reference object solution is a solution obtained by dissolving a reference object in a solvent, wherein the reference object comprises sodium ions, potassium ions and magnesium ions;
the preparation of the test solution comprises the following steps: extracting the gypsum standard decoction by adopting an extraction solvent, and collecting an extracting solution to obtain a sample solution; the extraction solvent is methane sulfonic acid solution.
In one embodiment, the mass percentage of the methanesulfonic acid in the extraction solvent is 0.5% -0.8%.
In one embodiment, the mass percentage of the methanesulfonic acid in the extraction solvent is 0.5% -0.65%.
In one embodiment, the eluent used for detection is a methane sulfonic acid solution, and the concentration of methane sulfonic acid in the eluent is 19-21 mmol/L.
In one embodiment, the flow rate of the eluent used for detection is 0.75mL/min to 0.9mL/min.
In one embodiment, the column temperature used for the detection is 28℃to 32 ℃.
In one embodiment, the detected condition comprises: the guard column is IonPac CG12A, the inner diameter of the guard column is 4mm, the column length is 50mm, and the particle size of the filler is 8 mu m; the analytical column is IonPac CS12A, the inner diameter of the analytical column is 4mm, the column length is 250mm, and the particle size of the filler is 8 mu m; the suppressor is CSRS 4mm; the temperature of the detection tank is 32-36 ℃; the inhibiting current is 45 mA-49 mA; the sensitivity of the detector is 4.5 mu S/cm-5.5 mu S/cm; the sample injection amount is 4.5-5.5. Mu.l.
In one embodiment, the amount of the extraction solvent used is 15 mL-25 mL for each 0.1g of gypsum standard decoction.
In one embodiment, the solvent is water.
In one embodiment, the concentration of sodium ions is 0.8 μg/mL to 1.2 μg/mL.
In one embodiment, the concentration of magnesium ions is 0.8 μg/mL to 1.2 μg/mL.
In one embodiment, the concentration of potassium ions is 0.8 μg/mL to 1.2 μg/mL.
In one embodiment, the extraction is performed by shaking.
In one embodiment, the extraction solution is collected by filtration.
In one embodiment, the characteristic spectrum of the gypsum standard decoction comprises 3 characteristic peaks, wherein peak 1 is Na + Peak, peak 2 is K + Peak, peak 3 is Mg 2+ Peak to peak 3 (Mg 2+ ) The corresponding peak of the reference object is a reference peak S, the relative retention time of each other characteristic peak is within + -10% of the corresponding specified value, and the specified values of the relative retention time of the characteristic peaks are respectively: peak 1 was 0.48 and peak 2 was 0.66.
The characteristic spectrum of the gypsum standard decoction is obtained by the construction method. In one embodiment, the mineral drug decoction is a mirabilite standard decoction, an oyster standard decoction or a talcum standard decoction.
The mineral medicine decoction generally contains sodium ions, potassium ions and magnesium ions, the contents of the metal ions are obviously different from one another in all decoction of different kinds of mineral medicines, and the decoction identification of the mineral medicines is realized by detecting the contents of the sodium ions, the potassium ions and the magnesium ions in the decoction of the mineral medicines and comparing the contents with the contents of the sodium ions, the potassium ions and the magnesium ions reflected by the characteristic patterns.
Compared with the prior art, the invention has the following beneficial effects:
the characteristic spectrum of the gypsum standard decoction is constructed by adopting an ion chromatography for the first time, and the characteristic spectrum is a trace element characteristic spectrum containing sodium ion characteristic peaks, potassium ion characteristic peaks and magnesium ion characteristic peaks.
According to the invention, the effective extraction of sodium ions, potassium ions and magnesium ions in the gypsum standard decoction is realized by adopting a proper type of extraction solvent, and the characteristic spectrum of metal ions in the gypsum standard decoction is constructed by further matching with proper ion chromatography detection conditions.
In the characteristic spectrum, the relative retention time of each characteristic peak is stable, and the relative peak area is obviously different from the decoction of other mineral drug varieties (such as mirabilite and the like), so that the specificity is presented. Therefore, the characteristic spectrum constructed by the invention not only can realize accurate and reliable detection and effective identification of sodium ions, potassium ions and magnesium ions in the gypsum standard decoction, and is convenient for ensuring uniform and stable medicament quality, but also can be used for decoction of other kinds of mineral medicaments, such as: the mirabilite standard decoction, the oyster standard decoction and the talcum standard decoction are distinguished, so that the purpose of qualitative identification is achieved.
Drawings
FIG. 1 is a characteristic chromatogram of a standard decoction of gypsum (eluent flow rate 0.8 mL/min);
FIG. 2 is a characteristic chromatogram of a standard decoction of gypsum (eluent flow rate 1.0 mL/min);
FIG. 3 is a characteristic chromatogram of a standard decoction of gypsum (eluent flow rate 1.2 mL/min);
FIG. 4 is a characteristic chromatogram of a standard decoction of gypsum (18 mmol/L methanesulfonic acid solution);
FIG. 5 is a characteristic chromatogram of a standard decoction of gypsum (20 mmol/L methanesulfonic acid solution);
FIG. 6 is a characteristic chromatogram of a standard decoction of gypsum (22 mmol/L methanesulfonic acid solution);
FIG. 7 is a characteristic chromatogram of a standard decoction of gypsum (28 ℃, 30 ℃, 32 ℃);
FIG. 8 is a graph showing the concentration of the extracted acid species (0.1% hydrochloric acid solution, 0.1% sulfuric acid solution, 0.1% methanesulfonic acid solution), in mass%;
fig. 9 shows the inspection results of the characteristic spectrum extraction acid types (0.1% hydrochloric acid solution, 0.1% sulfuric acid solution and 0.1% methanesulfonic acid solution) of the standard gypsum decoction, wherein "%" is the mass percent;
FIG. 10 shows the concentration of extracted acid (0.26%, 0.34%, 0.44%, 0.54%, 0.64%, 0.80%, 1.00%), "%" is mass%;
fig. 11 shows the inspection results of the concentration (0.26%, 0.34%, 0.44%, 0.54%, 0.64%, 0.80%, 1.00%) of acid extracted from the characteristic spectrum of the standard decoction of gypsum, wherein "%" is mass percent;
FIG. 12 is an examination of the extracted acid volumes (10 mL, 20mL, 30 mL);
FIG. 13 shows the results of examining the acid usage amount (10 mL, 20mL, 30 mL) extracted from the gypsum standard decoction feature map;
FIG. 14 is a superposition of characteristic maps of 23 batches of gypsum standard decoction;
FIG. 15 is a graph of peak assignments for gypsum standard decoction features;
FIG. 16 is a specific investigation of gypsum standard decoction feature patterns;
FIG. 17 is a chart of different column temperature investigation chromatograms of the gypsum standard decoction feature spectrum durability;
FIG. 18 is a chart of different flow rates for examining the durability of a gypsum standard decoction feature;
FIG. 19 is a chromatogram of a gypsum standard decoction with different feature spectrum durability for inspection by chromatographic columns;
FIG. 20 is a chromatogram of a gypsum standard decoction feature spectrum for investigation of different leacheate concentrations;
FIG. 21 is a characteristic map of a standard decoction of mirabilite;
FIG. 22 is a characteristic spectrum of oyster decoction;
FIG. 23 is a characteristic spectrum of a standard decoction of talc.
Detailed Description
The present invention will be described in more detail below in order to facilitate understanding of the present invention. It should be understood, however, that the invention may be embodied in many different forms and is not limited to the implementations or embodiments described herein. Rather, these embodiments or examples are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments or examples only and is not intended to be limiting of the invention. As used herein, the optional scope of the term "and/or" includes any one of the two or more related listed items, as well as any and all combinations of related listed items, including any two or more of the related listed items, or all combinations of related listed items.
The ion chromatography is widely applied to the determination of inorganic element content, and can be applied to the detection of heavy metals in the environment and the detection of inorganic elements in industry. For example: the patent literature discloses a method for detecting trace metal ion content in high-purity thiourea by ion chromatography, which uses ion chromatography to detect sodium, ammonium, potassium, magnesium and calcium cations, and belongs to quantitative detection. Ion chromatography is currently also applied to gypsum to a relatively small extent and is mainly applied to the detection of industrial gypsum, for example: li Tao in light (Li Tao in light, song Wenji, zhang Jianjun, feng Ziping. Determination of calcium sulfate content in desulfurized gypsum powder by ion chromatography [ J ]. Physical and chemical examination (chemical handbook), 2014,50 (01): 89-92) the desulfurized gypsum is studied by ion chromatography, and the calcium sulfate content in desulfurized gypsum powder is determined; xu Danhua and Mei Yi fly (Xu Danhua, mei Yi Fu, liu Shihua ion chromatography for simultaneous determination of fluorine and chlorine in gypsum and gypsum products [ J ]. Chinese building materials science and technology, 2014,23 (01))
1-2) ion chromatography is used to simultaneously determine fluorine and chlorine in gypsum and gypsum products. To date, ion chromatography technology has not been cited in research on mineral medicinal gypsum, and in particular, ion chromatography has been applied to research on establishing gypsum or gypsum standard decoction feature patterns, so that establishing a gypsum standard decoction feature pattern by using ion chromatography belongs to a technical blank in the research at the present stage. However, ion chromatography is not applied to gypsum standard decoction feature map construction at present.
Based on the method, the invention adopts proper extraction solvents to realize the effective extraction of sodium ions, potassium ions and magnesium ions in the gypsum standard decoction, and further combines proper ion chromatography detection conditions to construct the characteristic spectrum of metal ions in the gypsum standard decoction.
The invention provides a construction method of a characteristic map of gypsum standard decoction, which comprises the following steps:
providing a reference solution, preparing a sample solution, detecting the reference solution by adopting an ion chromatography, detecting the sample solution by adopting an ion chromatography, introducing a chromatogram obtained by detecting the reference solution and a chromatogram obtained by detecting the sample solution into a traditional Chinese medicine chromatographic fingerprint similarity evaluation system, and making a characteristic map of a gypsum standard decoction;
The reference object solution is a solution obtained by dissolving a reference object in a solvent, wherein the reference object comprises sodium ions, potassium ions and magnesium ions;
the preparation of the test solution comprises the following steps: extracting the gypsum standard decoction by adopting an extraction solvent, and collecting an extracting solution to obtain a sample solution; the extraction solvent is methane sulfonic acid solution.
The invention discloses a traditional Chinese medicine chromatographic fingerprint similarity evaluation system which is software published by the national formulary committee and capable of comparing the similarity of fingerprints.
The extraction solvent disclosed by the invention is the extraction acid in the specific embodiment, and the concentration of the extraction acid is expressed in mass percent under the condition of no special description.
It should be noted that, in the construction method of the present invention, the steps are not limited in time sequence.
The mass percentage of methane sulfonic acid in the extraction solvent may be selected from, including but not limited to, any one of the following concentrations or a concentration range between any two of the following concentrations: 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%. In one example, the mass percentage of methane sulfonic acid in the extraction solvent is 0.5% -0.8%. Preferably, the mass percentage of methane sulfonic acid in the extraction solvent is 0.5% -0.65%. More preferably, the mass percentage of methane sulfonic acid in the extraction solvent is 0.5% -0.55%, for example 0.54%.
In one example, the detection uses a solution of methane sulfonic acid with a concentration of methane sulfonic acid of 19mmol/L to 21mmol/L, and the molar concentration of methane sulfonic acid in the solution can be selected from any one of the following concentrations or a concentration range between any two of the following concentrations: 19mmol/L, 19.5mmol/L, 20mmol/L, 20.5mmol/L, 21mmol/L.
In one example, the flow rate of the eluent used for detection is from 0.75mL/min to 0.9mL/min, for example, the flow rate of the eluent may be selected from, including but not limited to, any one or a range of flow rates between any two of the following: 0.75mL/min, 0.8mL/min, 0.85mL/min, 0.9mL/min.
In one example, the column temperature used for the detection is 28℃to 32 ℃. The column temperature used for detection may be selected from, but is not limited to, any one of the following temperatures, a temperature range of + -0.5deg.C for any one of the following temperatures, or a temperature range between any two temperatures: 28 ℃, 29 ℃, 30 ℃, 31 ℃, 32 ℃.
In one example, the detected condition includes: the guard column is IonPac CG12A, the inner diameter of the guard column is 4mm, the column length is 50mm, and the particle size of the filler is 8 mu m; the analytical column is IonPac CS12A, the inner diameter of the analytical column is 4mm, the column length is 250mm, and the particle size of the filler is 8 mu m; the suppressor is CSRS 4mm; the temperature of the detection tank is 32-36 ℃; the inhibiting current is 45 mA-49 mA; the sensitivity of the detector is 4.5 mu S/cm-5.5 mu S/cm; the sample injection amount is 4.5-5.5. Mu.l. The temperature of the detection cell can be 32 ℃, 33 ℃, 34 ℃, 35 ℃, 36 ℃, the inhibiting current can be 45mA, 46mA, 47mA, 48mA and 49mA, the sensitivity of the detector can be 4.5 mu S/cm, 5 mu S/cm and 5.5 mu S/cm, and the sample injection amount can be 4.5 mu l, 5 mu l and 5.5 mu l. It will be appreciated that the detection conditions employed in the present invention may be recommended conditions for the instrument used, for example, a recommended suppression current of 47mA.
In one example, the amount of the extraction solvent used is 15 mL-25 mL for each 0.1g of gypsum standard decoction. The amount of the extraction solvent corresponding to each 0.1g of gypsum standard decoction may be selected from, but is not limited to, any one or a range of amounts between any two of the following: 15mL, 17mL, 19mL, 21mL, 23mL, 25mL.
In one example, the solvent is water. For example ultrapure water.
In one example, the concentration of sodium ions is 0.8. Mu.g/mL to 1.2. Mu.g/mL. The concentration of sodium ions may be selected from the group including, but not limited to, any one of the following or a range of concentrations between any two of the following: 0.8. Mu.g/mL, 0.9. Mu.g/mL, 1.0. Mu.g/mL, 1.1. Mu.g/mL, 1.2. Mu.g/mL.
In one example, the concentration of magnesium ions is 0.8 μg/mL to 1.2 μg/mL. The concentration of magnesium ions may be selected from the group including, but not limited to, any one of the following or a range of concentrations between any two of the following: 0.8. Mu.g/mL, 0.9. Mu.g/mL, 1.0. Mu.g/mL, 1.1. Mu.g/mL, 1.2. Mu.g/mL.
In one example, the concentration of potassium ions is 0.8. Mu.g/mL to 1.2. Mu.g/mL. The concentration of potassium ions may be selected from the group including, but not limited to, any one of the following concentrations or a range of concentrations between any two of the following: 0.8. Mu.g/mL, 0.9. Mu.g/mL, 1.0. Mu.g/mL, 1.1. Mu.g/mL, 1.2. Mu.g/mL.
In one example, the extraction is performed using shock extraction.
In one example, the extraction fluid is collected by filtration.
In one example, the construction method includes the steps of:
providing a reference solution, preparing a sample solution, detecting the reference solution by adopting an ion chromatography, detecting the sample solution by adopting an ion chromatography, introducing a chromatogram obtained by detecting the reference solution and a chromatogram obtained by detecting the sample solution into a traditional Chinese medicine chromatographic fingerprint similarity evaluation system, and making a characteristic map of a gypsum standard decoction;
the reference object solution is a solution obtained by dissolving a reference object in a solvent, wherein the reference object comprises sodium ions, potassium ions and magnesium ions;
the preparation of the test solution comprises the following steps: extracting the gypsum standard decoction by adopting an extraction solvent, and collecting an extracting solution to obtain a sample solution; the extraction solvent is methane sulfonic acid solution;
the mass percentage of the methane sulfonic acid in the extraction solvent is 0.5-0.65%;
the eluent used for detection is a methanesulfonic acid solution, and the concentration of methanesulfonic acid in the eluent is 19 mmol/L-21 mmol/L;
The flow rate of the eluent used for detection is 0.75-0.9 mL/min;
the column temperature adopted for detection is 28-32 ℃.
In one example, the characteristic spectrum of the gypsum standard decoction comprises 3 characteristic peaks, and peak 1 is Na + Peak, peak 2 is K + Peak, peak 3 is Mg 2+ Peak to peak 3 (Mg 2+ ) The corresponding peak of the reference object is a reference peak S, the relative retention time of each other characteristic peak is within + -10% of the corresponding specified value, and the specified values of the characteristic peaks are respectively: peak 1 was 0.48 and peak 2 was 0.66. Because of Na + 、K + 、Mg 2+ Controls are readily available and it is recommended that each characteristic peak be matched consistent with its reference retention time.
The invention also provides application of the characteristic spectrum of the gypsum standard decoction in detecting the mineral medicine decoction, wherein the characteristic spectrum of the gypsum standard decoction is obtained by the construction method.
In the characteristic spectrum provided by the invention, the relative retention time of each peak is stable, and although the relative peak areas of each characteristic peak in different batches of gypsum standard decoction are different, the difference is limited in the gypsum standard decoction, and the difference is very remarkable relative to the decoction of other mineral medicine varieties (such as mirabilite, oyster and talcum), and the difference of the relative retention areas in the decoction of different varieties of mineral medicines is enough to distinguish the gypsum standard decoction from the decoction of other mineral medicine varieties.
The term "mineral drug" as used herein refers to a pharmaceutically acceptable mineral or processed product of minerals or minerals of some naturally occurring minerals such as rock, including also fossil of some archaea, for example, cinnabar, realgar, gypsum, calamine, etc. In particular, for the purposes of the present invention, a "mineral drug" may be selected from one of the following: mirabilite, oyster shell and talcum.
The preparation method of the gypsum standard decoction provided by the embodiment of the invention comprises the following steps: decocting Gypsum Fibrosum decoction pieces in water, and drying the decoction.
The number of times of the decoction according to the embodiment of the present invention is preferably a plurality of times (e.g., 2 times, 3 times, 4 times, etc.), and if the number of times of the decoction is a plurality of times, the decoctions obtained by the plurality of times of the decoction may be combined and then dried. Preferably, the number of times of decoction is two.
Further, in the first decoction process, the water amount is 7-9 times of that of the gypsum decoction pieces, the gypsum decoction pieces are soaked for 25-35 min before decoction, and the decoction is heated and boiled by strong fire (the power is 450-550W) and then kept slightly boiled by slow fire (the power is 180-220W) for 40-50 min. After the decoction is finished, filtering the decoction while the hot by adopting a screen (namely decoction) to obtain filtrate, wherein the screen can be a 200-mesh screen, and the obtained filtrate can be cooled by cold water. In the step, the water adding amount can be 7 times, 7.5 times, 8 times, 8.5 times and 9 times of that of the gypsum decoction pieces, the time for soaking the gypsum decoction pieces before decoction can be 25min, 30min and 35min, the corresponding power of strong fire can be 450W, 470W, 490W, 500W, 520W and 550W, the corresponding power of slow fire can be 180W, 200W, 210W and 220W, and the time for keeping micro boiling can be controlled to be 40min, 45min and 50min.
Further, in the second decoction process, the water addition amount is 5-6.5 times of that of the gypsum decoction pieces, and the decoction is carried out by boiling with strong fire (the power is 450-550W) and then keeping micro-boiling with slow fire (the power is 180-220W) for 25-35 min. After the decoction is finished, filtering the decoction while the hot by adopting a screen (namely decoction) to obtain filtrate, wherein the screen can be a 200-mesh screen, and the obtained filtrate can be cooled by cold water. In the step, the water adding amount can be 5 times, 5.5 times, 6 times and 6.5 times of that of the gypsum decoction pieces, the corresponding power of the strong fire can be 450W, 470W, 490W, 500W, 520W and 550W, the corresponding power of the slow fire can be 180W, 200W, 210W and 220W, and the duration of keeping micro boiling can be controlled to be 25min, 30min and 35min.
It is understood that the resulting decoction may be concentrated (including but not limited to concentrated under reduced pressure) to a fluid extract prior to drying, including but not limited to vacuum freeze drying.
Example 1 creation of a feature map analysis method
1. Instrument, reagent and reagent
1.1 instrument: ion chromatograph (Dionex ICS-6000, siemens technologies Co., ltd.); cation suppressor (CSRS 300 4mm, siemens technologies Co., ltd.); ionPac CG12A guard column (4X 50mm,8 μm); ionPac CS12A analytical column (4X 250mm,8 μm); one ten-thousandth analytical balance (ME 204E, mertrel-tolidox); ultrapure water systems (Milli-Q Direct, merck Co., ltd.).
1.2 reagent: hydrochloric acid (guangzhou chemical reagent plant, top grade purity); sulfuric acid (Guangzhou chemical reagent plant, top grade purity); methanesulfonic acid (MSA, ala Ding Shiji (Shanghai), 99.5%); the water was ultrapure water (laboratory homemade).
1.3 reagents: the sodium component analysis standard substance in water (national defense science and technology industry application chemical first-class metering station, concentration: 1000 mug/mL, lot number: GBW (E) 080526); potassium unit element solution standard substance (Beijing North great bureau of metering technology, concentration: 1000. Mu.g/mL, lot number: GBW (E) 083296); magnesium unit solution standard substance (Beijing northern Weijimetric technical institute, concentration: 1000 μg/mL, lot number: GBW (E) 083298); the lot number information for 23 standard lots of gypsum decoction is shown in Table 1 below.
Table 1, 23 Gypsum herbs and Standard decoction information Table
Sequence number Lot number of medicinal materials Standard decoction lot number Production area
1 G161022 GT161022 Ore Lin Zhen of Lanling county in Linyi city in Shandong province
2 G161023 GT161023 Linyi city Lanling county Lanlin town No. 3 mine in Shandong province
3 G161024 GT161024 Zaozhuang City, Shandong Province
4 G1901008 GT1901008 City and county Yang Lingzhen of filial sense of Hubei province
5 G1901009 GT1901009 City and county Yang Lingzhen of filial sense of Hubei province
6 G1901010 GT1901010 New river industrial area in city and county of Xiaozhong city of Hubei province
7 G1903178 GT1903178 Yao Fengshen town in Shanxi province, fortune city, summer county
8 G1903179 GT1903179 Sizhen in summer county in city of fortune of Shanxi province
9 G1903180 GT1903180 Shanxi province fortune city, summer county, temple front town
10 G1906106 GT1906106 Sanlong town in Lingshan county in Guangxi Qinzhou
11 G2004040 GT2004040 Yongzhou City, Hunan Province
12 G2005079 GT2005079 Guangdong province
13 G2005080 GT2005080 Guangdong province
14 G2005083 GT2005083 Xindu District, Chengdu City, Sichuan Province
15 G2005084 GT2005084 Guangxi Yulin Yuzhou district
16 G2005085 GT2005085 Lotus pool in city of si chuan province
17 G2005086 GT2005086 Lotus pool in city of si chuan province
18 G2005087 GT2005087 Guangxi Yulin City
19 G2005094 GT2005094 Urban area of Bozhou city of Anhui province
20 G2005095 GT2005095 Urban area of Bozhou city of Anhui province
21 G2005096 GT2005096 Urban area of Bozhou city of Anhui province
22 G2005097 GT2005097 Urban area of Bozhou city of Anhui province
23 G2006002 GT2006002 Urban area of Bozhou city of Anhui province
The 23 batches of gypsum standard decoction are prepared by taking gypsum decoction pieces in batches shown in Table 1 and processing the gypsum decoction pieces by the following process:
(1) Taking 100g of gypsum decoction pieces, adding water and decocting twice:
adding 8 times of water for the first time, soaking for 30 minutes, heating with strong fire (power 500W), boiling with slow fire (power 200W), keeping slight boiling for 45 minutes, filtering with 200 mesh screen, and cooling the filtrate with cold water;
adding 6 times of water for the second decoction, boiling with strong fire (power 500W), keeping micro-boiling with slow fire (power 200W) for 30 min, filtering with 200 mesh screen, and cooling with cold water;
(2) Mixing the filtrates, concentrating under reduced pressure to obtain clear paste with volume of about 100ml, packaging into 10ml penicillin bottles, packaging into 3ml bottles, vacuum freeze drying, taking out, and rolling aluminum cap.
2. Preparation of reference solutions
The sodium component analysis standard substance, potassium unit element solution standard substance and magnesium unit element solution standard substance in water are precisely weighed, placed in a 30mL plastic vial, and added with ultrapure water to prepare each 1mL of Na-containing solution + 、K + 、Mg 2+ 1 μg of each solution.
3. Determination of chromatographic conditions
The chromatographic method of the gypsum standard decoction characteristic spectrum is inspected, and the influence of different flow rates and different leaching solution concentrations on the ion chromatogram is inspected.
(1) Determination of optimal flow rate
The experiment selects three flow rates of 0.8ml/min, 1.0ml/min and 1.2ml/min for comparison, and determines the optimal flow rate.
Sample solution preparation: taking a proper amount of gypsum standard decoction (GT 1903178), grinding, taking about 0.1g, precisely weighing, placing into a 30ml plastic bottle, precisely adding 20ml of 0.54% methane sulfonic acid solution, fully shaking to dissolve, shaking uniformly, filtering, and collecting subsequent filtrate.
Chromatographic conditions: selecting an IonPac CG12A (4X 50 mm) guard column, an IonPac CS12A (4X 250 mm) analysis column, and a CSRS 4mm inhibitor; taking 20mmol/L methane sulfonic acid solution as eluent; the flow rates are respectively 0.8ml per minute, 1.0ml per minute and 1.2ml per minute; the column temperature is 30 ℃; the temperature of the detection tank is 35 ℃; the suppression current was 47mA; the detector sensitivity was 5 mus per cm; the sample injection amount is 5 μl; the results are shown in FIG. 1, FIG. 2 and FIG. 3.
By comparing the chromatograms of 3 different flow rates, when the flow rate is selected to be 1.0ml/min and 1.2ml/min, the peak information is complete, the peak separation degree is good, but the inverted peak appears; when 0.8ml/min is selected as the flow rate, the peak information is complete, the peak separation degree is good, and no inverse peak appears, so that 0.8ml/min is selected as the flow rate of the eluent.
(2) Examination of the leacheate
In the experiment, leacheate with 3 concentrations of 18mmol/L methane sulfonic acid solution, 20mmol/L methane sulfonic acid solution and 22mmol/L methane sulfonic acid solution are selected for comparison, and leacheate with proper concentration is determined.
Preparation of the sample solution is described in the section "determination of optimal flow rate" above with reference to the above (1).
Chromatographic conditions: ionPac CG12A (4X 50mm,8 μm) guard column, ionPac CS12A (4X 250mm,8 μm) analytical column, CSRS 4mm inhibitor was selected; respectively taking 18mmol/L methane sulfonic acid solution, 20mmol/L methane sulfonic acid solution and 22mmol/L methane sulfonic acid solution as leacheate; the flow rate is 0.8ml per minute; the column temperature is 30 ℃; the temperature of the detection tank is 35 ℃; the suppression current was 47mA; the detector sensitivity was 5 mus per cm; the sample injection amount is 5 μl; the results are shown in FIG. 4, FIG. 5 and FIG. 6.
By comparing chromatograms of the 3 leaches with different concentrations, when 18mmol/L, 20mmol/L and 22mmol/L methane sulfonic acid solutions are selected as the leaches, the separation degree effect of each characteristic peak is consistent and the baseline is stable, but when the concentration of the leacheate is 18mmol/L, the elution time is longer; when the concentration of the eluent is 22mmol/L, the peak outlet time of each characteristic peak is relatively early, and each characteristic peak is relatively compact, so that the concentration of the eluent is 20mmol/L for saving analysis time and analysis results.
(3) Investigation of column temperature
The experiment selects 3 different column temperatures at 28 ℃,30 ℃ and 32 ℃ for comparison, and determines the proper chromatographic column temperature.
Preparation of the sample solution is described in the section "determination of optimal flow rate" above with reference to the above (1).
Chromatographic conditions: ionPac CG12A (4X 50mm,8 μm) guard column, ionPac CS12A (4X 250mm,8 μm) analytical column, CSRS 4mm inhibitor was selected; taking 20mmol/L methane sulfonic acid solution as eluent; the flow rate is 0.8ml per minute; the column temperatures were 28 ℃,30 ℃ and 32 ℃ respectively; the temperature of the detection tank is 35 ℃; the suppression current was 47mA; the detector sensitivity was 5 mus per cm; the sample injection amount is 5 μl; the results are shown in FIG. 7.
By comparing the chromatograms of 3 different column temperatures, the baseline of the chromatograms of 3 column temperatures is stable, the separation degree is good, and the response of each characteristic peak is higher at 30 ℃ so that 30 ℃ is selected as the optimal column temperature.
(4) Determination of chromatographic conditions
IonPac CG12A (4X 50mm,8 μm) guard column, ionPac CS12A (4X 250mm,8 μm) analytical column, CSRS 4mm inhibitor was selected; taking 20mmol/L methane sulfonic acid solution as eluent; the flow rate is 0.8ml per minute; the column temperature is 30 ℃; the temperature of the detection tank is 35 ℃; the suppression current was 47mA; the detector sensitivity was 5 mus per cm; the sample loading was 5. Mu.l.
4. Examination of pretreatment method of sample solution
The sample pretreatment method of the gypsum standard decoction feature map is examined, and the extracted acid type, the extracted acid concentration and the extracted acid dosage are mainly examined.
(1) Investigation of the extracted acid species
The experiment examines the influence of different extracted acid types on the characteristic spectrum of the gypsum standard decoction, selects 0.1% (w/w) hydrochloric acid solution, 0.1% (w/w) sulfuric acid solution and 0.1% (w/w) methane sulfonic acid solution as extraction solvents, and observes 3 trace ion peaks Na + 、K + 、Mg 2+ The peak pattern and the separation effect of the peaks are calculated, and the total peak area/sample weighing of 3 characteristic peaks is calculated to compare the influence of different acid types on the characteristic spectrum of the gypsum standard decoction, so that the large total peak area/sample weighing is obtained, and the optimal extracted acid type is selected as a standard.
Taking a proper amount of gypsum standard decoction (GT 1903180), grinding, taking about 0.1g, precisely weighing, parallel 3 groups, 2 parts of each group, placing into a 30ml plastic bottle, precisely adding 0.1% (w/w) hydrochloric acid solution, 0.1% (w/w) sulfuric acid solution and 0.1% (w/w) methanesulfonic acid solution respectively, fully shaking to dissolve, shaking uniformly, filtering, taking a subsequent filtrate, injecting into an ion chromatograph, analyzing by sample injection according to chromatographic conditions determined under the term of 'determination of chromatographic conditions', recording peak areas of all characteristic peaks, and calculating 'total peak areas/sample amounts', wherein the results are shown in tables 2, 8 and 9.
TABLE 2 extraction of characteristic patterns of Standard decoction of Gypsum Fibrosum acid investigation result table
The results show that the characteristic patterns are obviously different by adopting different acid types as extraction solvents, and the total peak area/sample amount is larger than other two acids by adopting 0.1% (w/w) methane sulfonic acid as the extraction solvent, so that the methane sulfonic acid is selected as the extraction acid type.
(2) Investigation of the concentration of the extracted acid
The experiment examines the influence of different extraction acid concentrations on the characteristic spectrum of the gypsum standard decoction, selects 0.26% (w/w) methane sulfonic acid solution, 0.34% (w/w) methane sulfonic acid solution, 0.44% (w/w) methane sulfonic acid solution, 0.54% (w/w) methane sulfonic acid solution, 0.64% (w/w) methane sulfonic acid solution, 0.80% (w/w) methane sulfonic acid solution and 1.00% (w/w) methane sulfonic acid solution as extraction solvents, and observes 3 trace ion peaks Na + 、K + 、Mg 2+ The peak pattern and the separation effect of the peaks, and the total peak area/sample weighing of 3 characteristic peaks are calculated to compare the influence of different acid concentrations on the characteristic spectrum of the gypsum standard decoction, and the optimal extraction acid concentration is selected.
Taking a proper amount of gypsum standard decoction (GT 1906106), grinding, taking about 0.1g, precisely weighing, parallel 7 groups, 2 parts of each group, placing into a 30ml plastic bottle, precisely adding 0.26% (w/w) methane sulfonic acid solution, 0.34% (w/w) methane sulfonic acid solution, 0.44% (w/w) methane sulfonic acid solution, 0.54% (w/w) methane sulfonic acid solution, 0.64% (w/w) methane sulfonic acid solution, 0.80% (w/w) methane sulfonic acid solution and 1.00% (w/w) methane sulfonic acid solution respectively, weighing, fully shaking to dissolve, shaking uniformly, filtering, taking a subsequent filtrate, recording peak areas of each characteristic peak according to chromatographic conditions determined under the term of '3. Chromatographic conditions', carrying out sample injection analysis, and calculating 'total peak areas/sample amounts', wherein the results are shown in tables 3, 10 and 11.
TABLE 3 extraction of characteristic patterns of Standard decoction of Gypsum Fibrosum acid concentration investigation result table
The results show that the characteristic patterns are obviously different by adopting the methanesulfonic acid with different concentrations as the extraction solvent, and the 0.54% (w/w) methanesulfonic acid is used as the extraction solvent, so that the total peak area/sample weight is maximum, and the 0.54% (w/w) methanesulfonic acid solution is selected as the optimal concentration of the extraction acid.
(3) Investigation of the amount of acid extracted
The experiment examines the influence of different extraction acid consumption on the characteristic spectrum of the gypsum standard decoction, selects 10ml, 20ml and 30ml of 0.54% (w/w) methane sulfonic acid solution as an extraction solvent, and observes 3 trace ion peaks Na + 、K + 、Mg 2 + The peak pattern and separation effect of the peaks, and the total peak area/sample weighing of 3 characteristic peaks are calculated to compare the influence of different acid dosage on the characteristic spectrum of the gypsum standard decoction, and the optimal acid extraction dosage is selected.
Taking a proper amount of gypsum standard decoction (GT 161022), grinding, taking about 0.1g, precisely weighing, parallel 3 groups, 2 parts of each group, placing into a 30ml plastic bottle, precisely adding 10ml, 20ml and 30ml of 0.54% methane sulfonic acid solution respectively, weighing, fully shaking to dissolve, shaking uniformly, filtering, taking a subsequent filtrate, determining chromatographic conditions according to the term of ' 3 ' chromatographic conditions ', carrying out sample injection analysis, recording the peak areas of all characteristic peaks, and calculating ' total peak areas/sample amounts ', wherein the results are shown in tables 4, 12 and 13.
TABLE 4 extraction of acid quantity from Standard decoction of Gypsum Fibrosum and examination result table
The results show that the characteristic patterns are obviously different by adopting different amounts of methane sulfonic acid as an extraction solvent, and 20ml of 0.54% (w/w) methane sulfonic acid is used as the extraction solvent, so that the total peak area/sample weight is maximum, and 20ml of the extraction solvent is selected as the optimal amount of the extraction acid.
(4) Determination of the method for preparing a sample solution
According to the experimental results, the pretreatment method of the gypsum standard decoction characteristic spectrum sample is determined as follows:
taking a proper amount of gypsum standard decoction, grinding, taking about 0.1g, precisely weighing, placing into a 30ml plastic bottle, precisely adding 20ml of 0.54% methane sulfonic acid solution, weighing, fully shaking to dissolve, shaking uniformly, filtering, and taking subsequent filtrate to obtain the plaster.
5. Determination of characteristic peaks
Taking 23 batches of gypsum standard decoction samples, examining the preparation method of the sample solution determined under the item of '4. Pretreatment method of the sample solution', preparing the sample solution, respectively precisely sucking the sample solution, determining chromatographic conditions determined under the item of '3. Determination of chromatographic conditions', sample injection measurement, common peak identification is carried out on 23 batches of gypsum standard decoction feature patterns by using Chinese medicine chromatographic fingerprint similarity evaluation software, wherein the components are known, the peak type and the separation degree are better, and trace ion peak 1 (Na) with stable content and higher content is selected + ) Peak 2 (K) + ) Peak 3 (Mg) 2+ ) As shown in FIG. 14, the peak is used as the characteristic peak of gypsum standard decoction, wherein the peak area is larger, and the reference substance is easily available Mg 2+ The chromatographic peak is used as a reference peak S, the relative retention time of each characteristic peak and the S peak is calculated, and the characteristic peak is positioned by utilizing the relative retention time. See table 5 below.
Table 5, 23 sets of gypsum standard decoction characteristic patterns relative retention time
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Table 6, 23 batches of gypsumStandard decoction feature map relative peak area
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In peak 3 (Mg) 2+ ) For reference peak S, the relative retention time RSD value of each characteristic peak and S peak of 23 batches of gypsum standard decoction is in the range of 0.58-0.89%, which shows that the relative retention time of each characteristic peak is relatively stable. The relative peak area RSD value of each characteristic peak and the S peak is in the range of 40.62-59.42%, and the relative peak areas of samples of different batches are greatly different, which indicates that the proportion of chemical components represented by each characteristic peak of the gypsum standard decoction of different batches is greatly different.
6. Identification of characteristic peaks
As can be seen from FIG. 15, na can be found in the characteristic spectrum of the standard decoction of Gypsum Fibrosum + 、K + 、Mg 2+ Characteristic peaks with consistent retention time, peak 1 was determined to be Na + Peak 2 is K + Peak 3 is Mg 2+ 。
Example 2, characterization atlas methodology validation
(1) Investigation of specificity
With reference to the detection conditions and the method for preparing a sample solution determined in example 1, a sample solution, a reference solution and a blank solvent (0.54% methanesulfonic acid) were prepared by precisely sucking a gypsum standard decoction (GT 161022) in 5 μl each, and were measured by ion chromatography. The results are shown in FIG. 16.
TABLE 7 chromatographic peak separation parameter Table
The result shows that the sample solution has the same chromatographic peak and blank at the retention time corresponding to the chromatographic reference substanceThe solvent is Na + 、K + The chromatographic peaks, but the peak areas are only 2.54% and 4.42% of the sample respectively, have little influence on the measurement of the sample, and show that the method has good specificity.
(2) Precision investigation
Taking appropriate amount of Gypsum standard decoction (GT 161022), grinding, taking about 0.1g, precisely weighing, preparing sample solution according to sample solution preparation method determined in example 1, repeatedly sampling for 6 times according to chromatographic conditions determined in example 1, and collecting sample solution with Mg 2+ Peak (peak 3) is a reference peak S, the relative retention time and relative peak area of each characteristic peak and S peak are calculated, RSD value is calculated, and experimental results are shown in tables 8 and 9.
Table 8, gypsum Standard decoction characteristics precision results Table (relative retention time)
Sequence number Na + K + Mg 2+ (S)
1 0.564 0.766 1.000
2 0.564 0.766 1.000
3 0.563 0.765 1.000
4 0.564 0.765 1.000
5 0.564 0.766 1.000
6 0.564 0.766 1.000
RSD(％) 0.08 0.06 0.00
Table 9, precision results table of gypsum standard decoction feature pattern (relative peak area)
The results show that the same sample solution is continuously sampled for 6 times by Mg 2+ The peak is a reference peak S, the relative retention time RSD value of each characteristic peak and the S peak is in the range of 0.06% -0.08%, and the relative peak area RSD value is 1.And the range of 27% -1.37% is less than 3.0%, which indicates that the precision of the instrument is good.
(3) Stability investigation
Taking appropriate amount of Gypsum standard decoction (GT 161022), grinding, taking about 0.1g, precisely weighing, preparing sample solution according to sample solution preparation method determined in example 1, and analyzing by sample injection under chromatographic conditions determined in example 1 at 0 hr, 3 hr, 5 hr, 7 hr, 9 hr, and 12 hr, respectively, with Mg 2+ The peak is a reference peak S, the relative retention time and relative peak area of each characteristic peak and S peak are calculated, and RSD values are calculated, and the results are shown in tables 10 and 11.
Table 10, gypsum standard decoction characteristic spectrum stability results table (relative retention time)
Time (h) Na + K + Mg 2+ (S)
0 0.481 0.667 1.000
3 0.481 0.667 1.000
5 0.482 0.667 1.000
7 0.481 0.668 1.000
9 0.481 0.667 1.000
12 0.480 0.667 1.000
RSD(％) 0.08 0.06 0.00
Table 11, characteristic spectrum stability results Table of Gypsum Standard decoction (relative Peak area)
Time (h) Na + K + Mg 2+ (S)
0 0.795 0.427 1.000
3 0.793 0.427 1.000
5 0.802 0.433 1.000
7 0.766 0.395 1.000
9 0.771 0.399 1.000
12 0.792 0.409 1.000
RSD(％) 1.83 3.89 0.00
The results showed that the same testSample solutions were analyzed at 0, 3, 5, 7, 9, 12 hours with Mg 2+ The peak is a reference peak S, the relative retention time RSD value of each characteristic peak and the S peak is in the range of 0.06% -0.08%, the relative peak area RSD value is in the range of 1.83% -3.89%, and the relative retention time RSD value is smaller than 4.0%, which indicates that the relative stability of the sample solution is good in 12 hours.
(4) Repeatability investigation
About 0.1g of the same batch of gypsum standard decoction (GT 161022) is taken, precisely weighed and 6 parts are parallel, 6 parts of sample solution is prepared according to the preparation method of the sample solution determined in the embodiment 1, and the sample solution is analyzed according to the chromatographic condition determined in the embodiment 1 and is prepared by Mg 2+ The peak is a reference peak S, the relative retention time and relative peak area of each characteristic peak and the S peak are calculated, the RSD value is calculated, and the experimental results are shown in tables 12 and 13.
Table 12, characteristic pattern repeatability results table of Gypsum Standard decoction (relative retention time)
TABLE 13, gypsum standard decoction feature pattern repeatability results table (relative peak area)
Sequence number Na + K + Mg 2+ (S)
Repeatability-1 1.128 0.366 1.000
Repeatability-2 1.124 0.364 1.000
Repeatability-3 1.127 0.365 1.000
Repeatability-4 1.127 0.366 1.000
Repeatability-5 1.127 0.364 1.000
Repeatability-6 1.125 0.370 1.000
RSD(％) 0.13 0.58 0.00
The results showed that the same batch of samples was repeatedly assayed 6 times with Mg 2+ The peak is a reference peak S, the relative retention time RSD value of each characteristic peak and the S peak is in the range of 0.02% -0.03%, the relative peak area RSD value is in the range of 0.13% -0.58%, and the relative retention time RSD value is smaller than 3.0%, which shows that the method has good repeatability.
(5) Durability inspection
(1) Investigation of different column temperatures
Comparing the influence of different column temperatures of 28 ℃, 30 ℃ and 32 ℃ on the characteristic spectrum of the standard gypsum decoction.
Taking gypsum standard decoction (GT 161022) about 0.1g, precisely weighing, preparing sample solution according to the sample solution preparation method determined in example 1, analyzing the same with the specification of example 1 except column temperature of 28deg.C, 30deg.C and 32deg.C, and using Mg 2+ The peak is a reference peak S, the relative retention time and relative peak area of each characteristic peak to S peak are calculated, and the RSD value is calculated. The experimental results are shown in Table 14, table 15 and FIG. 17.
Table 14, characteristic spectrum durability results Table of Gypsum Standard decoction (relative retention time)
Column temperature (. Degree. C.) Na + K + Mg 2+ (S)
28 0.483 0.676 1.000
30 0.482 0.668 1.000
32 0.480 0.659 1.000
RSD(％) 0.31 1.33 0.00
Table 15, characteristic spectrum durability results Table of Gypsum Standard decoction (relative Peak area)
Column temperature (. Degree. C.) Na + K + Mg 2+ (S)
28 0.796 0.408 1.000
30 0.776 0.403 1.000
32 0.761 0.405 1.000
RSD(％) 2.26 0.65 0.00
The results show that the relative retention time RSD value of each characteristic peak and the S peak is in the range of 0.31-1.33% and the relative peak area RSD is in the range of 0.65-2.26% at different column temperatures, which shows that the change of the column temperature has less influence on the relative retention time and the relative peak area of each characteristic peak when the column temperature is +/-2 ℃.
(2) Investigation of different flow rates
The different flow rates are compared, and the influence of the different flow rates on the characteristic spectrum of the gypsum standard decoction is respectively 0.6ml/min, 0.8ml/min and 1.0 ml/min.
About 0.1g of gypsum standard decoction (GT 161022) is taken, precisely weighed, and a sample solution is prepared according to the preparation method of the sample solution determined in the embodiment 1, wherein the other chromatographic conditions are the same as those determined in the embodiment 1 except that the flow rates are 0.6ml/min, 0.8ml/min and 1.0ml/min respectively, and the sample solution is analyzed by Mg 2+ The peak is a reference peak S, the relative retention time and relative peak area of each characteristic peak to S peak are calculated, and the RSD value is calculated. The experimental results are shown in Table 16, table 17 and FIG. 18.
Table 16, characteristic spectrum durability results Table of Gypsum Standard decoction (relative retention time)
Flow rate (ml/min) Na + K + Mg 2+ (S)
0.6 0.548 0.749 1.000
0.8 0.481 0.668 1.000
1.0 0.474 0.659 1.000
RSD(％) 8.15 7.17 0.00
Table 17, characteristic spectrum durability results Table of Gypsum Standard decoction (relative Peak area)
Flow rate (ml/min) Na + K + Mg 2+ (S)
0.6 0.746 0.421 1.000
0.8 0.776 0.403 1.000
1.0 0.779 0.429 1.000
RSD(％) 2.37 3.26 0.00
The results show that the relative retention time RSD value of each characteristic peak and the S peak is in the range of 7.16-8.15% and the relative peak area RSD is in the range of 2.37-3.27% at different flow rates, which shows that when the flow rate is +/-0.2 ml/min, the change of the flow rate of the eluent has a larger influence on the relative retention time of each characteristic peak and a smaller influence on the relative peak area, so that the chromatographic peak is recommended to be identified by using a reference substance. The flow rate of the eluent is strictly controlled.
(3) Investigation of different chromatographic columns from the same manufacturer
Comparing the standard decoction Na of different batch chromatographic columns to gypsum of the same manufacturer + 、K + 、Mg 2+ Influence of the characteristic map.
Preparing standard decoction of Gypsum (GT 161022) under the conditions determined in example 1, preparing sample solution by using different chromatographic columns of the same manufacturer, performing sample injection analysis under the same conditions as those determined in example 1, and performing Mg analysis 2+ The peak is a reference peak S, the relative retention time and relative peak area of each characteristic peak to S peak are calculated, and the RSD value is calculated. The experimental results are shown in Table 18, table 19 and FIG. 19.
Table 18, characteristic spectrum durability results Table of Gypsum Standard decoction (relative retention time)
Table 19, characteristic spectrum durability results Table of Gypsum Standard decoction (relative Peak area)
In tables 18 and 19 above, the 1 st and 2 nd lines in lot 1 are the detection results corresponding to two parallel samples, and the 1 st and 2 nd lines in lot 2 are the detection results corresponding to two parallel samples.
The results show that different chromatographic columns of the same manufacturer are used for measuring Na + 、K + 、Mg 2+ By Mg 2+ As a control peak, na + 、K + The relative retention time RSD values were 0.17% and 0.20% respectively, and the relative peak area RSD values were 1.87% and 5.82% respectively, indicating that the method has good durability against small variations in different chromatographic columns.
(4) Investigation of different Leaching solution concentrations
Comparing the concentration of the different leachates, the concentration of methanesulfonic acid is (18 mmol/L, pH=1.77) (20 mmol/L, pH=1.66), (22 mmol/L, pH=1.66) to the gypsum standard decoction Na + 、K + 、Mg 2+ Influence of content determination.
Preparing standard decoction of Gypsum (GT 161022), preparing sample solution with reference to the conditions determined in example 1, respectively using different leacheate concentrations, performing sample injection analysis under other chromatographic conditions identical to those determined in example 1, and calculating Na + 、K + 、Mg 2+ The content and RSD values are shown in table 20, table 21 and fig. 20.
Table 20, characteristic spectrum durability results Table of Gypsum Standard decoction (relative retention time)
Table 21, standard decoction of Gypsum characteristic spectrum durability results Table (relative Peak area)
The results show that the concentration of methane sulfonic acid in the leaching solution is +/-2 mmol/L to measure Na + 、K + 、Mg 2+ By Mg 2+ As a control peak, na + 、K + Relative retention time RSD values were 7.52% and 6.57% respectively, and relative peak area RSD values were 0.78% and 4.89% respectively, indicating that the method has good durability against small fluctuations in eluent concentration.
Example 3
Referring to example 1, mirabilite, oyster shell and talc were prepared into standard decoction, respectively, and ion chromatography was performed.
According to FIG. 15 (characteristic pattern of gypsum standard decoction constructed by the present invention), FIG. 21 (characteristic pattern of mirabilite standard decoction), FIG. 22 (characteristic pattern of oyster standard decoction), FIG. 23 (characteristic pattern of talcum standard decoction), the relative areas of the peaks contained in the characteristic patterns were compared, and the results showed that the mirabilite standard decoction, oyster standard decoction, talcum standard decoction were compared with gypsum standard decoction, although Na was detected + 、K + 、Mg 2+ However, the content is obviously different, and can be distinguished from the standard decoction of gypsum.
Table 22, results table of decoction characteristic patterns of different varieties
From the data, na in the standard decoction of mirabilite + The peak area and the relative peak area are obviously higher than those of other standard decoction, and are obviously distinguished from the gypsum standard decoction; oyster standard decoction K + 、Mg 2+ Peak is lower than gypsum standard decoction and K thereof + The peak relative peak area is also lower than the data of the gypsum standard decoction, and can be obviously distinguished from the gypsum standard decoction; the peak areas of the three characteristic peaks in the talcum standard decoction and the relative peak areas are lower than those of the gypsum standard decoction, and can be distinguished from the gypsum standard decoction.
In the invention, the characteristic spectrum of the gypsum standard decoction is constructed based on the ion chromatography, the characteristic spectrum of the gypsum standard decoction comprises 3 characteristic peaks, and peak 1 is Na + Peak, peak 2 is K + Peak, peak 3 is Mg 2+ Peak to peak 3 (Mg 2+ ) The corresponding peak of the reference object is a reference peak S, the relative retention time of the rest characteristic peaks is within + -10% of the corresponding specified value, and the specified values of the characteristic peaks are respectively: peak 1 was 0.48 and peak 2 was 0.66, and each characteristic peak was matched consistent with its control retention time. Also, although the peak-to-peak area varies from batch to batch of gypsum standard decoction, the variation is limited within the gypsum standard decoction variety, and the variation is significant relative to decoction from other mineral drug varieties (e.g., mirabilite, oyster, talc), and the difference in the relative retention areas present in the different varieties is sufficient to distinguish the gypsum standard decoction from the other mineral drug varieties.
The above examples merely represent a few embodiments of the present invention, which facilitate a specific and detailed understanding of the technical solutions of the present invention, but are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. It should be understood that, based on the technical solutions provided by the present invention, those skilled in the art obtain technical solutions through logical analysis, reasoning or limited experiments, all of which are within the scope of protection of the appended claims. The scope of the patent is therefore intended to be covered by the appended claims, and the description and drawings may be interpreted as illustrative of the contents of the claims.

Claims (6)

1. The construction method of the characteristic spectrum of the gypsum standard decoction is characterized by comprising the following steps:
providing a reference solution, preparing a sample solution, detecting the reference solution by adopting an ion chromatography, detecting the sample solution by adopting an ion chromatography, introducing a chromatogram obtained by detecting the reference solution and a chromatogram obtained by detecting the sample solution into a traditional Chinese medicine chromatographic fingerprint similarity evaluation system, and making a characteristic map of a gypsum standard decoction;
The reference object solution is a solution obtained by dissolving a reference object in a solvent, wherein the reference object comprises sodium ions, potassium ions and magnesium ions;
the preparation of the test solution comprises the following steps: extracting gypsum standard decoction by using an extraction solvent, and collecting an extracting solution to obtain a sample solution, wherein the extraction solvent is a methane sulfonic acid solution; the mass percentage of methane sulfonic acid in the extraction solvent is 0.54%; the dosage of the extraction solvent is 20ml corresponding to each 0.1g gypsum standard decoction;
the conditions detected include:
the guard column is IonPac CG12A, the inner diameter of the guard column is 4mm, the column length is 50mm, and the particle size of the filler is 8 mu m;
the analytical column is IonPac CS12A, the inner diameter of the analytical column is 4mm, the column length is 250mm, and the particle size of the filler is 8 mu m;
the suppressor is CSRS 4mm;
the temperature of the detection tank is 35 ℃;
the suppression current was 47mA;
the detector sensitivity was 5. Mu.S/cm;
the sample injection amount is 5 μl;
the detection method comprises the steps that the eluent adopted by detection is a methane sulfonic acid solution, and the concentration of methane sulfonic acid in the eluent is 20mmol/L;
the flow rate of the eluent adopted in the detection is 0.8mL/min;
the column temperature adopted for detection is 30 ℃;
the characteristic spectrum of the gypsum standard decoction comprises 3 characteristic peaks, wherein peak 1 is Na + Peak, peak 2 is K + Peak, peak 3 is Mg 2+ Peak in Mg 2+ The corresponding peak 3 of the reference object is a reference peak S, the relative retention time of each other characteristic peak is within + -10% of the corresponding specified value, and the specified values of the characteristic peaks are respectively: peak 1 was 0.48 and peak 2 was 0.66.
2. The method for constructing a characteristic spectrum of gypsum standard decoction according to claim 1, wherein the solvent is water.
3. The method for constructing a characteristic spectrum of a gypsum standard decoction according to claim 1, wherein the concentration of sodium ions is 0.8-1.2 μg/mL, the concentration of magnesium ions is 0.8-1.2 μg/mL, and the concentration of potassium ions is 0.8-1.2 μg/mL.
4. The method for constructing a characteristic spectrum of a gypsum standard decoction according to claim 1, wherein the gypsum standard decoction is extracted by vibration.
5. The method for constructing a characteristic spectrum of gypsum standard decoction according to claim 1, wherein the extraction solution is collected by filtration.
6. Use of the method for constructing a characteristic map of a gypsum standard decoction according to any one of claims 1 to 5 for detecting a mineral drug decoction, wherein the mineral drug decoction is a mirabilite standard decoction, an oyster standard decoction or a talc standard decoction.
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