CN113203808A

CN113203808A - Detection method of fire therapy liniment

Info

Publication number: CN113203808A
Application number: CN202110455689.7A
Authority: CN
Inventors: 张志锋; 苏学燕; 阿俄拉子; 沙学忠
Original assignee: Southwest Minzu University
Current assignee: Southwest Minzu University
Priority date: 2021-04-27
Filing date: 2021-04-27
Publication date: 2021-08-03

Abstract

The invention provides a detection method of a fire therapy liniment, and the raw material medicines of the liniment comprise cortex acanthopanacis, myrrh, radix sileris, pawpaw, cortex mori radicis, rhizoma anemones raddeanae, gynura segetum, elderberry, illicium simonsii and borneol; mainly comprises thin layer chromatography and UPLC detection. The method can detect various medicinal materials and main components in the fire therapy liniment, thereby providing a basis for quality standard control of the fire therapy liniment and bringing positive significance to the popularization of the liniment.

Description

Detection method of fire therapy liniment

Technical Field

The invention relates to the field of national medicines and quality detection thereof.

Background

The fire therapy technology is formed by collecting characteristics of the Yi nationality and improving the technology, firstly spraying the medicine of the Yi nationality formula on a treatment part, covering wet gauze, then spraying medicinal liquor on the gauze, igniting the gauze by using a lighter, extinguishing the burning gauze when a patient can not bear high temperature, repeatedly operating for 3-5 times after one minute, and realizing the optimal effect of flushing skin at the treatment part and the effects of activating blood and relieving pain.

The Yi-nationality medical hospital in the Liangshan-Zhou-combined Hospital has a clinical experience prescription for many years, which comprises troops (cortex acanthopanacis), kojiu soil (myrrh), Wabuyou (two hairs), Szechwan-Chinen (ramulus Sambuci Williamsii), radix frontal (illicium verum), Erlesei-ben (radix sileris), Chubu (pawpaw), fructus zanthoxyli (cortex mori radicis), lamogger (gynura segetum) and borneol. The empirical formula is usually soaked to obtain liniment. The liniment has the main functions: promoting blood circulation, removing blood stasis, relieving pain, and treating pain, numbness and swelling of neck, waist, leg and knee joint caused by pathogenic wind-cold-dampness and strain, rheumatism, gout, scapulohumeral periarthritis, fasciitis, and gonarthritis. Spraying the medicinal liquid on affected part, wiping with palm, spraying the medicinal liquid on gauze for wet application of pain part 2-5 times per day, or applying medicinal liquid for Yi medical extraction and suction, and fire therapy. The fire therapy liniment has simple processing technology and convenient use, no toxic or side effect is found, the clinical application in Yi medicine hospitals of Chinese and western medicine combination hospitals in Liangshan is more than 5 years, the number of cases is more than 5000, and the total effective rate reaches 91.02%.

However, the liniment has no related quality research, and has certain restriction on the use and popularization of the product.

Disclosure of Invention

Therefore, the invention aims to provide a detection method capable of effectively controlling a fire therapy liniment.

Specifically, the invention provides a detection method of a fire therapy liniment, and the raw material medicines of the liniment comprise cortex acanthopanacis, myrrh, radix sileris, pawpaw, cortex mori radicis, rhizoma anemones raddeanae, gynura segetum, elderberry, illicium simonsii and borneol; it comprises the UPLC detection content of the cimicidin:

using the cimicidin as a reference substance; the detection wavelength is 325 nm;

a chromatographic column: c18 as stationary phase

Mobile phase: acetonitrile: 0.05 to 0.2 percent of phosphoric acid

Wherein the phosphoric acid concentration may be selected from 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.10%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.20%, and the like.

The raw material medicaments can be prepared from the following components in percentage by weight:

the dimensions of the column were as follows: 2.1X 100mm, 1.8 μm.

Wherein, the preparation of the test solution in the UPLC comprises the following steps: drying liniment, and extracting with methanol.

In the preliminary experiment, various UPLC detection conditions are adopted for detection, but the separation effect of the cimicidin is poor, for example:

a is acetonitrile: 0.1% phosphoric acid:

as a result: is not separated from the rear peak and the front peak (see the attached drawings of the specification)

B. Acetonitrile: 0.1% phosphoric acid

As a result: not separated from the preceding peak

C. Acetonitrile: 0.1% phosphoric acid

As a result: not separated from the following peak

Therefore, the invention finally selects the chromatographic conditions capable of realizing effective separation of the linarin glycoside through a large amount of experimental screening:

acetonitrile: 0.05 to 0.2 percent of phosphoric acid

Wherein, it also includes the thin layer content of elderberry:

(1) collecting liniment, removing solvent, and dissolving the residue with methanol to obtain test solution;

(2) extracting ramulus Sambuci Williamsii reference medicinal material by liniment preparation method, removing solvent from the extractive solution, and extracting with methanol to obtain ramulus Sambuci Williamsii reference medicinal material solution;

(3) the gel was developed on silica gel G thin layer plates with cyclohexane-ethyl acetate-methanol ═ 6:2: 1.

With the method, compound spots can be effectively spread, so that elderberry can be effectively detected in the liniment. In the former period, the inventor also uses other detection methods, such as the standard of traditional Chinese medicinal materials in Yunnan province, cyclohexane-ethyl acetate-methanol is 3: 1: 0.5, however, the sample solution was still at the origin and could not be developed.

Wherein, it also includes the thin layer detection content of the mulberry bark:

(2) extracting cortex Mori with reference to the liniment preparation method, removing solvent from the extractive solution, and extracting with methanol to obtain cortex Mori reference solution;

(3) chloroform-methanol 5:1 was applied to silica gel G thin layer plates.

By using the method, compound spots can be effectively spread, so that the cortex mori radicis can be effectively detected in the liniment. In the former period, the inventor also uses other detection methods, such as Liu Yi culvert, and the like, quality detection and quality standard of the cortex mori radicis medicinal material, Ji head university newspaper (Nature science edition), 2019,40(05):45-52. ", and dichloromethane-ethyl acetate-methanol-glacial acetic acid (1: 6: 2.5: 1), but the test article cannot be effectively developed.

Wherein, it also includes the thin layer detection content of cortex acanthopanacis:

(1) removing solvent from liniment, extracting with water-chloroform solvent system, removing volume of chloroform extractive solution, and dissolving with methanol to obtain test solution;

(2) taking a cortex acanthopanacis reference medicinal material, extracting according to the preparation method of the liniment, and preparing a cortex acanthopanacis reference medicinal material solution according to the content in the step (1);

(3) chloroform-methanol was applied at 21:1 on silica gel G thin layer plates.

Cortex acanthopanacis: pungent and warm in property and bitter in taste. It enters liver and kidney meridians. Has the functions of dispelling wind and eliminating dampness, tonifying liver and kidney, strengthening tendons and bones, inducing diuresis and relieving swelling. Shen nong Ben Cao Jing: "treating hernia and abdominal pain, benefiting qi for , it is impossible for children to treat ulcer and erosion of the yin. "miscellaneous records of famous physicians": the traditional Chinese medicine composition mainly treats impotence of men's yin, dampness under the sac, dribbling urine, pruritus vulvae and pain of waist and back of women, pain and numbness of both feet and wind weakness, tonifying middle-jiao and replenishing vital essence, strengthening muscles and bones, strong will, long-time taking, light weight and aging resistance. "modern pharmacology: cortex Acanthopanacis has antiinflammatory, analgesic, tranquilizing, blood serum antibody concentration increasing, mononuclear macrophage phagocytic function promoting, anti-stress, nucleic acid synthesis promoting, blood sugar lowering, sex hormone-like, anti-tumor, anti-mutagenesis, antiulcer, and anti-rejection effects.

Myrrh: pungent, bitter and mild. It enters heart, liver and spleen meridians. Dispel stasis, relieve pain, resolve swelling and promote tissue regeneration. Can be used for treating thoracic obstruction, cardialgia, gastralgia, dysmenorrhea, amenorrhea, puerperal stagnation, abdominal mass, rheumatalgia, traumatic injury, carbuncle, swelling, and pyocutaneous disease. Myrrh comes from "the book of the herbal drawing" (the book of changes of the materia Medica): myrrh, raw Persian, all the countries of the south of the jin Hai and Guangzhou. The roots and the plants of wood are like olive and green and dense leaves, and the old people have the paste liquid dripping underground and congealing into blocks, which are large or small, and like benzoin. The term "marine herbs" is: "follow Xuzhu nan Zhou Ji": shengbosu, pine resin in their place, is like Shenxiang red black. "Kaibao materia Medica" (Kaibao): "Shengbosu, similar to benzoin, has variable block size and is black. The book's herbal drawing is recorded: myrrh, raw Persian. Jin Hai nan all countries and Guangzhou. The roots of wood are like olive and dense leaves, and the old people have the paste and liquid dripping underground and congealing into blocks, large or small, like benzoin, and the collection is not repeated. "compendium" is listed: according to Yi Tong Zhi from Yi Tong Zhi, Mo Yao is as tall as pine, thick as one inch, when digging out the lower part of the tree, it is cut down with an axe, and the resin flows to the bank, the ten heaven. "conform to the imported myrrh as above. Modern pharmacology: the main chemical components of myrrh include monoterpene, sesquiterpene, triterpene, steroid and lignin. Pharmacological research shows that the myrrh has pharmacological activity in many aspects such as defervescence, anti-inflammation, analgesia, neuroprotection and the like, and has no obvious toxic or side effect.

Wind prevention: pungent and sweet with mild temperature. It enters bladder, liver and spleen meridians. Dispel wind and relieve exterior syndrome, subdue dampness and alleviate pain, stop spasm. Can be used for treating common cold, headache, rheumatalgia, rubella, pruritus, and tetanus. It is mainly caused by strong wind, dizziness, headache, aversion to wind, wind pathogen, blindness, wind moving around the body, arthralgia and arthralgia. (Yu Shu (imperial Explanation) with pain), restlessness. Can lighten the body after long-term use. A Chinese name of Cuyun (Miscant from Yu Shu). Shengchuan ze. Wu pu (wu pu) is recorded as follows: divaricate saposhnikovia root, cloudberry, lemongrass, roots, allium tuberosum, hundreds of species, shennong huangdi qi bertong jun leigong magpie, which is sweet and nontoxic, Lishi xian, or Japanese handan shancai, Zhengyue leaf, thin circle, black, yellow and white, May flower yellow, June black, Rohdea chinensis, sun stem, and Lang Han Lian (imperial view). Famous physicians' stand: an anise, a baizhi, a pingfeng, an root, a baichai, a raw flatstem milkvetch, and a handan, an enamel evil, a shancai, a root of May-February, and a fierce dryout. Table "paradigm ran" cloud: fang Feng is good at three auxiliary herbs and Bai is good at. Modern pharmacology: the divaricate saposhnikovia root mainly contains chemical components such as chromone, coumarins, acidic polysaccharide and the like, and can achieve the effects of relieving fever and easing pain by influencing the expression of in-vivo fever inflammatory factors and the release of related pain factors; the antibacterial and anti-inflammatory effects are achieved by inhibiting the generation of related inflammatory factors or inflammatory mediators; the anti-allergic effect is achieved by reducing the expression of related anti-allergic factors; the anti-tumor effect is achieved by inhibiting the growth of related tumor factors; the function of improving the immunity of the organism is achieved by selectively increasing macrophage release related factors.

Pawpaw: acid, warm. It enters liver and spleen meridians. Relaxing tendons and activating collaterals, regulating stomach function and eliminating dampness. Can be used for treating damp arthralgia, spasm, soreness and pain of waist and knee joint, summer dampness vomiting and diarrhea, spasm pain of tendon, loempe, and edema. Modern pharmacology: the chemical components in the pawpaw mainly comprise saccharides, flavonoids, organic acids, terpenoids, volatile oil and biological enzyme substances, and the pawpaw tea has the effects of calming the liver, harmonizing the stomach, relaxing tendons, activating collaterals, protecting the liver, resisting bacteria, diminishing inflammation, dispelling wind-damp, resisting oxidation and the like.

White mulberry root-bark: sweet and cold in nature. It enters lung meridian. Purge lung, relieve dyspnea, induce diuresis to alleviate edema. Can be used for treating cough and asthma due to lung heat, edema, fullness, oliguria, and edema of face, eye and skin. Treatise on herb Property: it is indicated for lung qi dyspnea with distention and edema of water and dampness, mainly impairing the function of body fluid, promoting diuresis, relieving edema, consumptive disease, fever, headache and internal deficiency. "compendium of materia Medica": sang Bai Pi is good at inducing diuresis and draining excess syndrome. Therefore, it is advisable for the lung to contain water and qi and the lung to have the rest fire. "modern pharmacology: the product contains various flavonoid derivatives, such as sanguirin, sanguichromene, sanguirin, etc.; umbelliferone, scopoletin, and antihypertensive component similar to acetylcholine; in addition, mulberry bark furan A is extracted. The product has effects in relieving cough, promoting urination, and increasing urine output and sodium, potassium and chloride output; the decoction and the extracts of ethanol, ether and methanol thereof have the function of reducing blood pressure to different degrees; has tranquilizing, anticonvulsive, analgesic, and cooling effects on nervous system; has exciting effect on intestine and uterus. The decoction has effect in inhibiting Staphylococcus aureus, Bacillus typhi and Bacillus dysenteriae. The product has inhibitory effect on cervical cancer JTC28 and lung cancer cells, and recent research also shows that the product also has anti-HIV effect.

The two-head hair contains a plurality of chemical components, and compounds such as monoterpene, iridoid, sesquiterpene, triterpene, steroid, alkaloid, flavone, amide, phenylpropanoid and the like are separated from the root stem or the overground part of the two-head hair at present. The extractive or the chemical components contained in the extractive have the functions of promoting cell differentiation, resisting inflammation, resisting bacteria and oxidation, preventing and treating cholelithiasis, and the like, and has obvious curative effect when being clinically used for treating hepatitis, lithiasis and the like. The chloroform extract of the double-head hair has anti-inflammatory effect, which may be the reason for the double-head hair used for treating rheumatic bone pain, swelling, furuncle and various infectious diseases, so the chloroform extract may be the effective part of the anti-inflammatory and antipyretic effects of the double-head hair. The two hairs have the obvious effect of reducing serum transaminase and have the protective effect on liver injury caused by carbon tetrachloride, so the toilet flower tablet (containing ursolic acid 75%) used by the liberation military 45 hospital and other units has the obvious curative effect of treating acute viral hepatitis and high cure rate. The two-head hair has bacteriostasis to various pathogenic bacteria, especially to staphylococcus aureus, and has obvious curative effect on upper respiratory tract, urinary system, gastrointestinal tract and other infectious diseases clinically. The antibacterial active ingredient is alkaloid component

The pseudo-ginseng has various chemical components, and the research parts are mainly concentrated on stems and leaves. The separated and identified chemical components comprise flavonoids, phenolic acids, alkaloids, terpenes, phenylpropanoids, fatty acids, steroids and glycosides thereof. The gynura segetum has the effect of resisting gout, and a mouse hyperuricemia experiment, a mouse writhing experiment and a mouse ear swelling experiment show that the gynura segetum can obviously reduce the mouse serum uric acid level, the writhing frequency caused by glacial acetic acid stimulation and the mouse ear swelling degree caused by dimethylbenzene, and the gynura segetum alcohol extract has the obvious effect of resisting gout

Ramulus Sambuci Williamsii contains ursolic acid, oleanolic acid, gallic acid, rutin, quercetin, protocatechuic acid, and morroniside. Is prepared from Chuan-xiong rhizome, 3-carboxychuanxiong rhizome, 1,2,3, 4-tetrahydro-1-methyl-beta-carboline-3-carboxylic acid, salidroside, 4-hydroxybenzyl-beta-D-glucoside, 4-hydroxy-1- (2-hydroxyethyl) -phenyl-3-O-beta-D-glucopyranoside, coniferin, vanillic acid-4-O-beta-D-glucopyranoside, isorhamnetin-3-O-beta-D-rutinoside, etc. The elderberry has obvious effect on repairing bone density, is beneficial to bone growth and bone content increase of thighbone and shinbone, has obvious inhibition effect on permeability increase of capillary vessels in abdominal cavity of the elderberry, and prompts that stem branches of the elderberry have obvious analgesic and anti-inflammatory effects. The research result shows that the lariciresinol separated from the methanol extract of the elderberry stem and branch shows the antifungal activity by interfering the cell membrane of fungi, and can be used as a novel antifungal agent for treating fungal infection diseases of human beings.

The wild anise contains de-4 '-O-methylylangambin, fargesin, 3' -O-demethyllepiperonesoxim, lariciresinioldimethyl ether, dehydrodiconiferol, myricetin-3-O-alpha-L-rhamnoside, kaempferol, quercetin, kaempferin 3-O-alpha-L-rhamnoside, quercetin-3-O-alpha-L-rhamnopyranosyl (1 → 6) -beta-D-glucopyranoside, etc. the modern research shows that the wild anise contains the compound of de-4 '-O-methylylangamin, 3' -O-demethyl ionol, laricinoldimethyl ether, and the compound of dehydrodicentriperonol, myricetin-3-O-alpha-L-rhamnoside. Is widely applied to traumatic hemorrhage, abdominal distention and emesis, traumatic injury, rheumatoid arthritis and other diseases, and has high medical value. The aniseed medicinal plant has strong bacteriostatic activity and has bacteriostatic effects on various strains: typhoid bacillus, diphtheria bacillus, dysentery bacillus, aspergillus flavus, penicillium citrinum, aspergillus niger, bacillus subtilis, escherichia coli and staphylococcus aureus. In addition, the illicium genus medicinal plant has anti-inflammatory, analgesic and antioxidant activities.

The method can detect various medicinal materials and main components in the fire therapy liniment, thereby providing a basis for quality standard control of the liniment and bringing positive significance to popularization of the liniment.

Drawings

FIG. 1 UPLC chromatogram of control

FIG. 2 UPLC chromatogram of test sample

FIG. 3 negative control UPLC chromatogram

FIG. 4 is a thin layer chromatogram of Acanthopanax senticosus, in which 1: sample 2: acanthopanax contrast medicinal material 3: isofraxidin control 4: negative control

FIG. 5 Saposhnikovia divaricata thin layer chromatogram, in which 1-3: sample 4: radix sileris reference medicinal material 5: cimicidin control 6: 5-0-methylvisammioside control 7: negative control

FIG. 6 is a thin layer chromatogram of cortex Mori, wherein 1-3: sample 4: cortex Mori control 5 negative control

FIG. 7 Sambucus nigra thin layer chromatogram, wherein 1-3: sample 4: elderberry control medicinal material 5 negative control

FIG. 8 UPLC chromatogram of mobile phase Condition A

Detailed Description

The raw materials used in the examples were as follows:

cortex Acanthopanacis Radicis is the dried root bark of Acanthopanax gracilistylus (Dunn) S.Y.Hu of Araliaceae. The quality standard of the raw materials (medicinal materials) for research and medication is checked to be in accordance with relevant regulations under 67 pages of Chinese pharmacopoeia (2020 edition I).

Myrrha, this product is dried resin of Comiphora myrrha Engl. or Comiphora molmol Engl. of Burseraceae. It should comply with the relevant regulations under 199 pages of the Chinese pharmacopoeia (2020 edition I). The quality standard of the raw materials (medicinal materials) for research and medication is met through inspection.

The rhizoma anemones Raddeanae is dry whole plant of Lagerstroemia indica Incarvillea arguta (Royle) Royle. It should meet the relevant regulations under 55 pages of the standards for Chinese medicinal materials in Yunnan province (2005 edition, fourth volume, Yi nationality medicine II). The quality standard of the raw materials (medicinal materials) for research and medication is met through inspection.

Sambucus williamsii Hance is dried stem and leaf of Sambucus williamsii Hance of Caprifoliaceae. It should meet the relevant regulations under page 91 of the standards for Chinese medicinal materials in Yunnan province (2005 edition of the second book Yi nationality medicine). The quality standard of the raw materials (medicinal materials) for research and medication is met through inspection.

Wild anise, which is the dried root bark and stem bark of the Magnoliaceae family, Illicium angustifolium lancelebratum A.C. Smith. All the year round, plucked and dried. It should meet the relevant regulations under page 13 of Zhejiang province Chinese medicinal material Standard (2017 edition-first book). The quality standard of the raw materials (medicinal materials) for research and medication is met through inspection.

Radix Saposhnikoviae, which is dried root of SaposhnikobaPiperivacatala (Turcz.) Schischk. It should comply with the relevant regulations under 159 pages of the "Chinese pharmacopoeia" (2020 edition I). The quality standard of the raw materials (medicinal materials) for research and medication is met through inspection.

Cortex Mori, which is the dried root bark of Morus alba L. The quality standard of the raw materials (medicinal materials) for research and medication is determined to be in accordance with the relevant regulations under the Chinese pharmacopoeia (2020 edition, part I319).

Pawpaw, a dry nearly mature fruit of chaenomeles speciosa (sweet) Nakai of Rosaceae, should meet the requirements of the Chinese pharmacopoeia (2020 edition one) at page 63. The quality standard of the raw materials (medicinal materials) for research and medication is met through inspection.

Gynura procumbens (lour.) Merr, dried root tuber of Gynura japonica (Thunb) Juel of Compositae. It should meet the relevant regulations under 87 pages of the standards for Chinese medicinal materials in Yunnan province (2005 edition of the second book Yi nationality medicine). The quality standard of the raw materials (medicinal materials) for research and medication is met through inspection.

Borneol, RNEOLUM SYNTHETIC

CioHisO 154.2

It should comply with the relevant regulations under 199 pages of the Chinese pharmacopoeia (2020 edition I). The quality standard of the raw materials (medicinal materials) for research and medication is met through inspection.

Example 1

Cutting other medicinal materials except the borneol into pieces for later use; adding Borneolum Syntheticum, heating and refluxing with 60% ethanol for 5 hr, filtering to remove the extractive solution, shaking, and standing in the shade to obtain liniment.

Example 2

Cutting other medicinal materials except the borneol into pieces for later use; adding Borneolum Syntheticum, extracting with 56% (v/v) Chinese liquor for 2 hr under ultrasonic assistance, extracting for 3 times, filtering to remove the extractive solution, shaking, and standing in the shade to obtain liniment.

EXAMPLE 3 Ramarioside content determination

1. Chromatographic conditions and System suitability test

Chromatographic Column ACQUITY UPLCHSS C18 chromatographic Column (2.1X 100mm Column, 1.8 μm); mobile phase acetonitrile-0.1% phosphoric acid solution (V/V); the detection wavelength is 325 nm; the flow rate is 0.2 mL/min; the column temperature is 30 ℃; the sample injection amount is 2 muL. Under the chromatographic condition, the number of theoretical plates is not less than 2000 in terms of chlorogenic acid peak.

Mobile phase conditions:

the chromatogram is shown in FIGS. 1-3.

2. Preparation of test solution

Collecting liniment, precisely weighing, removing solvent, drying, adding methanol, extracting under supercritical reflux for 30min, and filtering to obtain extractive solution as test solution.

3. Preparation of control solutions

7.02mg of the linarin control was accurately weighed and dissolved in methanol to prepare a 0.702mg/mL standard solution as a control solution.

4 linear relationship investigation

Precisely sucking 1mL of reference stock solution (mass concentration of 0.702mg/mL) respectively, placing in a 5mL volumetric flask, adding methanol to a constant volume, shaking up, sequentially taking 1mL of constant volume step by step, wherein the mass concentrations are 0.7020mg/mL, 0.1404mg/mL, 0.0280mg/mL, 0.0056mg/mL and 0.0011mg/mL respectively, filtering through a microporous membrane, carrying out sample injection determination according to a proposed chromatographic condition, and recording peak area. Drawing a standard curve by taking the l-ephedrine glycoside sample injection concentration (C) as an abscissa and taking a chromatographic peak area value (Y) as an ordinate to obtain a regression equation Y which is 12,376,023x +24,720R²0.9999(n 5). The result shows that the linear relation between the area of the peak and the area of the linarin glycoside is good in the range of 0.0011mg/ml to 0.7020 mg/ml.

5 precision test

Precisely sucking a proper amount of 2.6.3 reference substance solution, continuously and repeatedly injecting sample for 6 times according to the drawn chromatographic conditions, measuring by the method, and recording the peak area. As a result, the RSD of the area of the region of the L-ephedrine glycoside peak was 0.19% (n-6), indicating good precision of the apparatus.

6 stability test

Precisely sucking the same sample solution, respectively carrying out sample injection measurement at 0, 1,2, 4, 8, 12 and 24h, and recording peak area. As a result, the RSD of the area of the l-ephedrine glycoside peak was 0.27% (n-6), indicating that the test solution was stable within 24 hours.

7 repeatability test

Taking a proper amount of samples of the same batch, precisely weighing, preparing 6 parts of test sample solutions in parallel according to a method, and injecting samples according to drawn-up chromatographic conditions for determination. RSD was 1.49% (n ═ 6), indicating good reproducibility of the method.

8 sample recovery test

Taking a proper amount of 6 samples with known content, respectively and precisely adding reference substances, preparing a sample solution according to a method, carrying out sample injection measurement according to drawn-up chromatographic conditions, recording peak areas, and calculating the sample injection recovery rate. The results are shown in Table 1.

TABLE 1 recovery test

9 sample content determination

Taking 6 batches of samples, preparing a test solution according to a method, carrying out sample injection measurement according to drawn-up chromatographic conditions, recording peak areas, and calculating the content of the glucoma-roside. As a result, the content of the linarin in 6 samples was 0.0576mg/g, 0.0526mg/g, 0.0603mg/g, 0.0589mg/g, 0.0580mg/g, 0.0602mg/g, and RSD was 4.88%, respectively.

Example 4 thin layer identification of Acanthopanax gracilistylus

Preparing a test solution by taking 100ml of fire therapy Yi formulation liniment, evaporating to dryness, adding 10ml of water into residue for dissolving, shaking and extracting with chloroform for 2 times (5 ml each time), combining chloroform solutions, evaporating to dryness, adding 1ml of methanol into residue for dissolving, taking 2.1g of acanthopanax root as a reference medicinal material for preparing a reference medicinal material solution, preparing a preparation according to a fire therapy Yi formulation liniment preparation process, and preparing the reference medicinal material solution according to a test solution preparation method.

Control solution preparation method comprises collecting control of Isoxazine sneeze, adding methanol to obtain solution containing lmg per 1ml

The negative control solution is prepared by taking the rest medicinal materials except for Acanthopanax gracilistylus according to the formula components, preparing YIFANG liniment without Acanthopanax gracilistylus for fire therapy according to the process requirements, and preparing the negative control solution according to the preparation method of test solution

Silica gel G thin layer plate under thin layer condition

Dispensing 10ul of each of the negative control solutions of the reference medicinal material solution and the reference solution of the sample solution, developing with chloroform-methanol (21:1), and inspecting under an ultraviolet lamp (365nm)

As a result, in the chromatogram of the test sample 4, spots of the same color appear in the chromatogram of the test sample at the positions corresponding to the chromatograms of the reference drug and the reference substance, and the negative reference chromatogram of acanthopanax has no interference at the corresponding positions, so the income standard is met.

Example 5 Ledebouriella seseloides layer authentication

Preparing test solution by taking 90ml of Yi liniment for fire therapy, evaporating to dryness, and dissolving residue with 10ml of ethanol; the reference medicinal solution is prepared by taking 1.5g of radix Saposhnikoviae as reference medicinal material, making into preparation according to the process for preparing liniment for fire therapy, and making into reference medicinal solution according to the preparation method of test solution.

Preparing negative control solution by taking the rest medicinal materials except radix Saposhnikoviae according to prescription composition, making into YIFANG liniment without Acanthopanax fire therapy according to process requirement, and making into negative control solution according to test solution preparation method

Thin layer condition silica gel G thin layer plate, sample amount sample solution 10ul each of negative control solution of control medicinal material solution

Developing solvent trichloromethane-methanol (6:1)

The inspection device is arranged under an ultraviolet lamp (365nm) (sprayed with 10 percent ethanol sulfate solution)

In the result chromatogram of the test sample 5, spots with the same color appear at the corresponding positions of the chromatogram of the control drug, and the windproof negative control chromatogram has no interference at the corresponding positions, so the income standard is met.

Example 6 thin layer identification of cortex Mori

The test solution is prepared by collecting 100ml of Yi formulation liniment for fire therapy, and steaming to 2 ml.

The reference medicinal solution is prepared by preparing 1g of cortex Mori reference medicinal material, making into preparation according to YI prescription liniment preparation process for fire therapy, and making into reference medicinal solution according to test solution preparation method.

The cortex mori negative control solution is prepared according to the formula composition, the rest medicines except the cortex mori are taken to prepare the Yi prescription liniment without the cortex mori for fire therapy according to the process requirements, and the negative control solution is prepared according to the preparation method of the test solution.

Silica gel G thin layer plate under thin layer condition

The sample amount of the test solution is 10ul each of the negative control solutions of the control medicinal material solution, and the developing agent chloroform-methanol (5:1)

Ultraviolet lamp of inspection device (365nm)

In the result chromatogram of the test sample 6, spots with the same color appear at the corresponding positions of the chromatogram of the reference drug, and the cortex mori negative reference chromatogram has no interference at the corresponding positions, so the income standard is met.

Example 7 elderberry thin layer identification

Preparing test solution by taking 100ml of Yi prescription liniment for fire therapy, evaporating to dryness, dissolving the residue with 1ml of methanol

The reference medicinal solution is prepared by preparing 1g of pawpaw reference medicinal material according to the Yi prescription liniment preparation process for fire therapy, and preparing the reference medicinal solution according to the test solution preparation method.

Preparing fructus Chaenomelis negative control solution according to formula composition, taking the rest medicinal materials except fructus Chaenomelis, making into YIFANG liniment without fructus Chaenomelis for fire therapy according to process requirements, and making into negative control solution according to test solution preparation method

Silica gel G thin layer plate under thin layer condition

The sample amount of the test solution is 10ul each of the negative control solutions of the control medicinal material solution, and the developing agent cyclohexane-ethyl acetate-methanol (6:2:1)

Ultraviolet lamp of inspection device (365nm)

In the result chromatogram 7 of the test sample, spots with the same color appear at the corresponding positions of the chromatogram of the reference drug, and the negative reference chromatogram of elderberry has no interference at the corresponding positions, so the income standard is met.

Claims

1. A detection method of a fire therapy liniment is characterized by comprising the following steps: the raw material medicines of the liniment comprise cortex acanthopanacis, myrrh, radix sileris, pawpaw, cortex mori radicis, rhizoma anemones Raddeanae, gynura segetum, elderberry, illicium simonsii and borneol, and the preparation method of the fire therapy liniment comprises the following steps: pulverizing the raw materials except the borneol to obtain medicinal powder, medicinal powder and borneol, and extracting with 50-70% ethanol to obtain a liniment; it comprises the UPLC detection content of the cimicidin:

a chromatographic column: c18 as stationary phase

Mobile phase: acetonitrile: 0.05 to 0.2 percent of phosphoric acid

2. The detection method according to claim 1, characterized in that: the phosphoric acid concentration is selected from 0.1%.

3. The detection method according to claim 1, characterized in that: the dimensions of the column were as follows: 2.1X 100mm, 1.8 μm.

4. The detection method according to claim 1, characterized in that: preparation of test solution in UPLC: drying liniment, and extracting with methanol.

5. The detection method according to claim 1, characterized in that: the method comprises the following thin-layer detection contents of elderberry:

6. The detection method according to claim 1, characterized in that: the method comprises the following thin-layer detection contents of the white mulberry root-bark:

(3) chloroform-methanol 5:1 was applied to silica gel G thin layer plates.

7. The detection method according to claim 1, characterized in that: the method comprises the following thin-layer detection contents of cortex acanthopanacis:

(3) chloroform-methanol was applied at 21:1 on silica gel G thin layer plates.

8. The detection method according to claim 1, characterized in that: the method comprises the following detection contents of a windproof thin layer:

(1) taking liniment, removing solvent, dissolving with ethanol, and making into test solution;

(2) taking a radix sileris reference medicinal material, extracting according to the preparation method of the liniment, and preparing a radix sileris reference medicinal material solution according to the content in the step (1);

(3) chloroform-methanol 6:1 was developed on silica gel G thin layer plates.