CN103512998A - Chinese ephedra medicinal material, and content determination method of three alkaloids in preparation thereof - Google Patents
Chinese ephedra medicinal material, and content determination method of three alkaloids in preparation thereof Download PDFInfo
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Abstract
The invention relates to a Chinese ephedra medicinal material, and a content determination method of three alkaloids in a preparation thereof. The method has the characteristics that: a common reversed-phase column and a non-buffer salt which is a mobile phase of acetonitrile-methanol-0.1% phosphoric acid with a volume ratio of 1-1.5:4-4.2:94.5-95.0 are adopted; at 207+/-2nm, contents of ephedrine hydrochloride, pseudoephedrine hydrochloride, and ephedrine methyl hydrochloride in the Chinese ephedra medicinal material and the preparation thereof are simultaneously determined. A sample is only processed through water-containing methanol ultrasonic extraction and alumina impurity removing, such that a near-colorless transparent solution is obtained. The method is simple, fast, reproducible, and accurate. According to the quantitative chromatogram of the three alkaloids, a baseline is stable, peak separation is good, and the peak appearing is completed within 20min. Compared with a traditional method with a buffer-salt solution, a specific chromatographic column, and unconventional organic phase ratio and flow rate, with the method provided by the invention, instrument and chromatographic column service lives are prolonged, cost is reduced, and detection conditions are communized. The method is suitable for popularization and application of basic units.
Description
Technical field
The invention belongs to traditional Chinese medicine quality control field, relate to the quality determining method of a kind of Herba Ephedrae and preparation thereof, particularly, relate to three kinds of alkaloidal content assaying methods in a kind of Herba Ephedrae and preparation thereof.
Background technology
Chinese ephedra is the dry herbaceous stem stem of Ephedraceae plant plait Chinese ephedra Ephedra sinica Stapf, epheday intermedia Ephedra intermedia Schrenk et C.A.Mey. or ephedra equisetina Ephedra equisetina Bge..Be the conventional Chinese medicine in a kind of cough-relieving compound preparation, tool Pulvis Diaphoreticus is cold, freeing lung and relieving asthma, and the effect of inducing diuresis for removing edema.For anemofrigid cold, uncomfortable in chest breathing with cough, geomantic omen edema.Honey Chinese ephedra is moistened the lung and relieve the cough.Preparation means Chinese ephedra and almond containing cough stopping sheet and golden Chinese ephedra and almond containing cough stopping sheet, is all the compound preparation of cough-relieving.The prescription of Chinese ephedra and almond containing cough stopping sheet, method for making and function cure mainly as follows:
Prescription: Chinese ephedra 600g semen armeniacae amarae 400g gypsum 1200g honey-fried licorice root 300g
Method for making: above four tastes, get gypsum 60g, be ground into fine powder; Residue gypsum and Chinese ephedra, honey-fried licorice root are ground into meal, and boiling secondary each 2 hours, filters merging filtrate; After hardship is deoiled containing benevolence, boiling 2 hours, filters, filtrate and above-mentioned filtrate merge, and being evaporated to relative density is the clear cream of 1.10～1.15 (70 ℃), is evaporated to dry cream, pulverize, appropriate with gypsum fine powder and starch, mix, granulation, dry, add dolomol appropriate, be pressed into 1000, sugar coating or film-coating, obtain.
Function cures mainly: antibechic, eliminate the phlegm, and relieving asthma.For acute and chronic bronchitis and pant etc.
Prescription, method for making and the function of gold Chinese ephedra and almond containing cough stopping sheet cure mainly as follows:
Prescription: golden yellow a kind of reed mentioned in ancient books 150-190 part honey honey-fried FOLIUM ERIABOTRYAE 280-350 part honey is processed Chinese ephedra 150-200 part
Semen armeniacae amarae 150-200 part root of purple-flowered peucedanum 250-320 part root bark of white mulberry 250-320 part
Melon 280-310 part balloonflower root 160-200 part honeysuckle 250-300 part of withering
Caulis bambusae in taenian 250-320 part
Method for making: above ten tastes, Pi is 10 times of amount 60% alcohol extract secondaries for leaf (honey is processed), Chinese ephedra (honey is processed), semen armeniacae amarae, the root of purple-flowered peucedanum, and each 2 hours, merging extract, filtered, and filtrate recycling ethanol is to without alcohol taste, standby; The root bark of white mulberry, caulis bambusae in taenian, Snakegourd Fruit, balloonflower root, honeysuckle, add 10 times of water gagings and decoct to boiling, and adds the root of large-flowered skullcap, decoct secondary, each 2 hours, merge extract, filter, filtrate is condensed into the clear cream of relative density 1.05-1.10 (60 ℃ of heat are surveyed), adds 95%7 alcohol, limit edged stirs, and to determining alcohol 70%, refrigeration is placed 24 hours, filter, filtrate recycling ethanol, to without alcohol taste, merges with alcohol extract, be concentrated into the clear cream of relative density 1.10 one 1.15 (60 ℃ of heat are surveyed), spraying is dry.Get spray powder and appropriate amount of starch, mix, particle processed, is pressed into 1000, and film coating, obtains.
Function cures mainly: clearing lung and eliminating phlegm, and relieving cough and asthma.The acute tracheobronchitis and the AECB that cause for phlegm heat, disease is seen cough, expectoration, asthma, phlegm is yellow or sticky in vain, and body heat is uncomfortable in chest, dry, pharyngalgia, urine be yellow, just do etc., red tongue with yellowish and greasy fur, slippery and rapid pulse.
Bibliographical information, the main effective constituent of Chinese ephedra is alkaloids, substantially has six kinds of alkaloids, i.e. ephedrine, d-pseudo-ephedrine, demethyl ephedrine, demethyl pseudoephedrine, methylephedrine, methyl pseudoephedrine.What wherein content was higher is ephedrine, d-pseudo-ephedrine and methylephedrine, and its sequence is ephedrine > pseudoephedrine > methylephedrine, three's sum accounts for the more than 80% of six kinds of ephedrine total amounts.Remaining methyl pseudoephedrine, demethyl ephedrine and demethyl Determination of Pseudoephedrine are very low, generally in compound extraction preparation, dosage is humble, so in the statutory standards of compound preparation, be all to control content limit with the content summation of ephedrine and pseudoephedrine at present.
In compound preparation, except ephedrine and pseudoephedrine, the methylephedrine that also has of content can be detected, and pharmacodynamic study shows: ephedrine has nervous centralis and sympathetic activation effect, its sweating is relievingd asthma, cholagogic, the effects such as blood pressure, vasoconstriction that rise are all more intense; Pseudoephedrine has very strong anti-inflammatory and diuresis, and the snuffles aspect curative effect that swelling causes to schneiderian membrane is certain; Methylephedrine is suitable with ephedrine to the dilating effect of tracheal smooth muscle and antitussive effect, and has good anti-allergy action.So with the total amount of three kinds of Herba Ephedrae alkaloids, control the content of medicinal material and its compound preparation, can react more all sidedly the quality of medicinal material and preparation.
By consulting literatures is learnt, Chinese Pharmacopoeia Herba Ephedrae recording of version in 2010 and infantile lung clearing phlegm transforming oral liquid, little Green Dragon particle, the chill ball etc. of only coughing, it is all the content summation of controlling ephedrine and pseudoephedrine, the chromatographic column of Herba Ephedrae assay must be equipped with specially, and its filling agent is that the polarity ether that the utmost point is of little use connects phenyl bonded silica gel.And the mobile phase of these medicinal materials and preparation is (3) lower phosphoric acid buffer salt of not advising use of an appendix VI D of Chinese Pharmacopoeia version in 2010 entirely, and organic phase ratio is many lower than 5%, not only give serviceable life of instrument and chromatographic column and brought harm, also, to after measuring, the flushing of chromatographic column has increased operation steps.
Although also there is the report of measuring five kinds of alkaloids (ephedrine, d-pseudo-ephedrine, methylephedrine, demethyl ephedrine, demethyl pseudoephedrine) in Herba Ephedrae, four kinds of alkaloids (ephedrine, d-pseudo-ephedrine, methylephedrine, demethyl ephedrine) and three kinds of alkaloids (ephedrine, d-pseudo-ephedrine, methylephedrine) simultaneously, but its five kinds alkaloidal mensuration need special phenyl post, column length is 250mm, mobile phase is phosphoric acid buffer salt system, the ratio of organic phase and buffer salt water system is 4: 96, and flow velocity is 0.6ml/min; Though four kinds of alkaloidal mensuration are common Waters-ODS post, its column length is 250mm, and mobile phase is phosphoric acid buffer salt system, and the ratio of organic phase and buffer salt water system is 3: 97.25, and flow velocity is 0.8ml/min; And three kinds of alkaloidal Phenomenex synersi posts that are determined as, its length is 250mm, and mobile phase is phosphoric acid buffer salt system, and the ratio of organic phase and buffer salt water system is 2: 98, and flow velocity is 1ml/min.In view of this, in the time of multiple ephedrines composition, measure, the setting of its various parameters is all outside pharmacopeia appendix VI D high performance liquid chromatography regulation parameter, all needs brokenly conventional independent setting.
In above-mentioned background situation, quality for easy, quick, science, standard, many indexs control medicinal material and its preparation, having invented and having adopted common chromatographic column, non-buffer salt is mobile phase, in pharmacopeia appendix VI D high performance liquid chromatography defined parameter area, measure three kinds of alkaloidal content in Herba Ephedrae and preparation thereof simultaneously.
Summary of the invention
First with common reverse-phase chromatographic column, non-buffer salt mobile phase, its volume ratio is 1～1.5: 4～4.2: 94.5～95.0 acetonitrile-methyl alcohol-0.1% phosphoric acid, at 207 ± 2nm place, measured the content of ephedrine hydrochloride, pseudoephedrine hydrochloride and mephedrine in Herba Ephedrae and preparation thereof simultaneously.According to three kinds of alkaloidal quantitative chromatograms, crest is separated good, and baseline was steady, at 20 minutes Nei Chu peaks complete (Fig. 1,2,3,4,5).
By methodological study, the sample size of ephedrine hydrochloride is at 0.1029-0.686 μ g, is good linear relationship (seeing Fig. 6) with peak area, and regression equation is: Y=2303175.3X-1526, r=0.9999; Pseudoephedrine hydrochloride sample size is at 0.02511-0.1674 μ g, is good linear relationship (seeing Fig. 7) with peak area, and regression equation is Y=2307131X-4611, r=0.9996; Mephedrine sample size is at 0.02223～0.1482 μ g, is good linear relationship (seeing Fig. 8) with peak area, and regression equation is: Y=2054332.3X-1016, r=0.9999.Adopt application of sample recovery experiment, result shows: the average recovery rate of ephedrine hydrochloride is 100.56% (n=6), and RSD is 0.20% (in Table 1); The average recovery rate of pseudoephedrine hydrochloride is 97.84% (n=6), and RSD is 0.94% (in Table 2).The average recovery rate of mephedrine is 98.45% (n=6), and RSD is 1.05% (in Table 3).Precision (in Table 4), stability (in Table 5), repeatability (in Table 6), specificity (seeing Fig. 3,4,5) etc., empirical tests all meets methodology requirement.Be applicable to three kinds of alkaloidal quantitative measurements in Herba Ephedrae and preparation thereof.
The technical solution adopted for the present invention to solve the technical problems is:
(1) chromatographic condition and system suitability be take octadecylsilane chemically bonded silica as filling agent; Take volume ratio as 1～1.5: 4～4.2: 94.5～95.0 acetonitrile-methyl alcohol-0.1% phosphoric acid is mobile phase; Column temperature: 40 ℃; Detection wavelength is 207nm; By ephedrine hydrochloride peak, calculate and should be not less than 3000;
(2) it is appropriate that ephedrine hydrochloride, pseudoephedrine hydrochloride and mephedrine are got in the preparation of reference substance solution, accurately weighed, add volume ratio and be hydrochloric acid-70% methanol solution of 1: 200, make the solution of hydrochloric ephedrine 30～15 μ g of every 1ml, pseudoephedrine hydrochloride 10～5 μ g, mephedrine 6～3 μ g, in contrast product solution;
(3) Herba Ephedrae fine powder 0.2～0.3g is got in the preparation of need testing solution, accurately weighed, or Chinese ephedra preparation 0.5～0.8g, accurately weighed, put respectively in tool plug conical flask, precision adds 70% methyl alcohol 25ml, close plug, weighed weight, with power 250W, the ultrasonic processing of frequency 33kHz 10～30 minutes, let cool, weighed weight again, with 70% methyl alcohol, supply the weight of less loss, shake up, , filter, precision measures subsequent filtrate 2ml, put in the chromatographic column of the internal diameter 1cm that 100-120 order neutral alumina 1～2g is housed, with 70% methanol-eluted fractions in the measuring bottle of 10ml to about 9ml, add 1 phosphoric acid, with 70% methyl alcohol, be diluted to scale, shake up, with the miillpore filter of 0.45 μ m, filter, get subsequent filtrate, as need testing solution,
(4) accurate above-mentioned reference substance solution and each 10 μ l of need testing solution, injection liquid chromatography, the three kinds of alkaloidal content in working sample drawn of determination method difference.
For the preferred determination techniques scheme of Herba Ephedrae, be:
(1) chromatographic condition and system suitability be take octadecylsilane chemically bonded silica as filling agent; Acetonitrile-methyl alcohol-0.1% phosphoric acid that the volume ratio of take is 1.5: 4: 94.5 is mobile phase; Column temperature: 40 ℃; Detection wavelength is 207nm; By ephedrine hydrochloride peak, calculate and should be not less than 3000;
(2) it is appropriate that ephedrine hydrochloride, pseudoephedrine hydrochloride and mephedrine are got respectively in the preparation of reference substance solution, accurately weighed, add volume ratio and be hydrochloric acid-70% methanol solution of 1: 200, the solution of making the hydrochloric ephedrine 20 μ g of every 1ml, pseudoephedrine hydrochloride 10 μ g, mephedrine 3 μ g, obtains;
(3) Herba Ephedrae fine powder 0.2g is got in the preparation of need testing solution, accurately weighed, put in tool plug conical flask, precision adds 70% methyl alcohol 25ml, close plug, weighed weight, with power 250W, the ultrasonic processing of frequency 33kHz 30 minutes, let cool, weighed weight again, with 70% methyl alcohol, supply the weight of less loss, shake up, filter paper filters, precision measures subsequent filtrate 2ml, put in the chromatographic column of the internal diameter 1cm that 100-120 order neutral alumina 1.5g is housed, with 70% methanol-eluted fractions in the measuring bottle of 10ml to about 9ml, add 1 phosphoric acid, with 70% methyl alcohol, be diluted to scale, shake up, with 0.45 μ m miillpore filter, filter, get subsequent filtrate, as need testing solution,
(4) accurate above-mentioned reference substance solution and each 10 μ l of need testing solution of drawing of determination method difference, injection liquid chromatography, measures three kinds of alkaloidal content in Herba Ephedrae.
For the preferred determination techniques scheme of Chinese ephedra and almond containing cough stopping sheet, be:
(1) chromatographic condition and system suitability be take octadecylsilane chemically bonded silica as filling agent; Acetonitrile-methyl alcohol-0.1% phosphoric acid that the volume ratio of take is 1: 4.2: 94.8 is mobile phase; Column temperature: 40 ℃; Detection wavelength is 207nm; By ephedrine hydrochloride peak, calculate and should be not less than 3000;
(2) it is appropriate that ephedrine hydrochloride, pseudoephedrine hydrochloride and mephedrine are got respectively in the preparation of reference substance solution, accurately weighed, add volume ratio and be hydrochloric acid-70% methanol solution of 1: 200, the solution of making the hydrochloric ephedrine 30 μ g of every 1ml, pseudoephedrine hydrochloride 10 μ g, mephedrine 5 μ g, obtains;
(3) 20 of Chinese ephedra and almond containing cough stopping sheets are got in the preparation of need testing solution, remove dressing, accurately weighed, porphyrize, cross 50 mesh sieves, get 0.8g, accurately weighed, put in tool plug conical flask, precision adds 70% methyl alcohol 25ml, close plug, weighed weight, with power 250W, the ultrasonic processing of frequency 33kHz 10 minutes, let cool, weighed weight again, with methyl alcohol, supply the weight of less loss, shake up, filter, precision measures subsequent filtrate 2ml, put in the chromatographic column of the internal diameter 1cm that 100-120 order neutral alumina 1.5g is housed, with 70% methanol-eluted fractions in the measuring bottle of 10ml to about 9ml, add 1 phosphoric acid, with 70% methyl alcohol, be diluted to scale, shake up, with the miillpore filter of 0.45 μ m, filter, get subsequent filtrate, as need testing solution,
(4) accurate above-mentioned reference substance solution and each 10 μ l of need testing solution of drawing of determination method difference, injection liquid chromatography, measures three kinds of alkaloidal content in Chinese ephedra and almond containing cough stopping sheet.
For gold fiber crops, containing the preferred determination techniques scheme of cough stopping tablet, be:
(1) chromatographic condition and system suitability be take octadecylsilane chemically bonded silica as filling agent; Acetonitrile-methyl alcohol-0.1% phosphoric acid that the volume ratio of take is 1: 4: 95 is mobile phase; Column temperature: 40 ℃; Detection wavelength is 207nm; By ephedrine hydrochloride peak, calculate and should be not less than 3000;
(2) it is appropriate that ephedrine hydrochloride, pseudoephedrine hydrochloride and mephedrine are got respectively in the preparation of reference substance solution, accurately weighed, adding volume ratio is ］: 200 hydrochloric acid-70% methanol solution, the solution of making the hydrochloric ephedrine 15 μ g of every 1ml, pseudoephedrine hydrochloride 5 μ g, mephedrine 3 μ g, obtains;
(3) 20 of golden Chinese ephedra and almond containing cough stopping sheets are got in the preparation of need testing solution, remove dressing, accurately weighed, porphyrize, cross 50 mesh sieves, get 0.8g, accurately weighed, put in tool plug conical flask, precision adds 70% methyl alcohol 25ml, close plug, weighed weight, with power 250W, the ultrasonic processing of frequency 33kHz 10 minutes, let cool, weighed weight again, with methyl alcohol, supply the weight of less loss, shake up, filter, precision measures subsequent filtrate 2ml, put in the chromatographic column of the internal diameter 1cm that 100-120 order neutral alumina 1.8g is housed, with 70% methanol-eluted fractions in the measuring bottle of 10ml to about 9ml, add 1 phosphoric acid, with 70% methyl alcohol, be diluted to scale, shake up, with the miillpore filter of 0.45 μ m, filter, get subsequent filtrate, as need testing solution,
(4) accurate above-mentioned reference substance solution and each 10 μ l of need testing solution of drawing of determination method difference, injection liquid chromatography, measures three kinds of alkaloidal content in golden Chinese ephedra and almond containing cough stopping sheet.
Principle of the present invention is as follows:
Utilize ephedrine hydrochloride, pseudoephedrine hydrochloride and mephedrine, at 205 ± 2nm, all present maximum absorption band (seeing Fig. 9,10,11), take 207nm as detecting wavelength, these three kinds of alkaloidal sample sizes within the specific limits, present good linear relationship with its peak area respectively, and for quantitative measurement.
Innovative point of the present invention and beneficial effect are as follows:
(1) first with common reverse-phase chromatographic column, non-buffer salt mobile phase, be that volume ratio is 1～1.5: 4～4.2: 94.5～95.0 acetonitrile-methyl alcohol-0.1% phosphoric acid, at 207 ± 2nm place, measured the content of ephedrine hydrochloride, pseudoephedrine hydrochloride and mephedrine in Herba Ephedrae and preparation thereof simultaneously.Measurement result prompting, these three kinds of alkaloidss, in Herba Ephedrae and preparation, content belongs to higher, and the mensuration with evaluation quality is worth.
(2) By consulting literatures is learnt, current Herba Ephedrae has measures three kinds of above alkaloidal content reports, in the preparation forming at compound, due to increasing of flavour of a drug, the interference of composition, is all to measure ephedrine at present, or the report of ephedrine and pseudoephedrine total amount, its mobile phase is also is mostly buffering salts, the optics of perishable instrument and damage chromatographic column.
(3) non-buffer salt mobile phase of the present invention, has effectively protected instrument performance, has extended the chromatographic column life-span, after having simplified and detecting and to finish, and the cleaning procedure of chromatographic column.
(4) common reverse-phase chromatographic column of the present invention is compared with the specific chromatographic column of report, first the length range of chromatographic column expands, the long chromatographic column of 150～250nm can be applied, and crest separation all meets the requirements, no longer be subject to the restriction of chromatographic column brand, make testing conditions generalization, cost, is beneficial to the universal of grass-roots unit and applies.
(5) in the Herba Ephedrae of having reported, five kinds of alkaloidal mensuration, the ratio of organic phase is 4, flow velocity is 0.6ml/min; Four kinds of alkaloidal mensuration, the ratio 3 of organic phase, flow velocity is 0.8ml/min; Three kinds of alkaloidal mensuration, the ratio 2 of organic phase, flow velocity is 1ml/min.Not only its organic phase ratio is less than 5, and flow velocity is less than 1ml/min more, like this, will extend the working time of mensuration, the column length time is in a high proportion of buffer salt water system that contains, not only there will be the random curling component retention causing of C18 chain to change, the chromatographic system causing is unstable, also there will be the theoretical cam curve straight line of chromatographic column to decline, crest alteration of form, do not meet mensuration requirement, chromatographic column is scrapped.Therefore, mobile phase of the present invention, has fallen into by the ratio of organic phase the organic phase ratio of stipulating under appendix VI D item of Chinese Pharmacopoeia version in 2010 and conventionally should be not less than 5% regulation, has avoided C
18chain random curling and cause adverse consequences, has kept the normal service life of chromatographic column.The six kind alkaloids (see Fig. 2) of mobile phase of the present invention in can also effective separated Herba Ephedrae, but be limited to the consistance with preparation, its evaluation index just has quality control meaning, three kinds of alkaloids that Selective determination content is higher.
(6) appearance of the inventive method, solved in Herba Ephedrae and preparation Chinese ephedra and almond containing cough stopping sheet thereof and golden Chinese ephedra and almond containing cough stopping sheet multiple alkaloid easy, quick, a low-cost difficult problem of measuring simultaneously, realized multicomponent, many indexs, comprehensively judged the object of drug quality.Be also other preparations containing Herba Ephedrae, provide many index quantifications to measure new way, tool example and exemplary role.
Accompanying drawing explanation
Tri-kinds of alkaloid reference substance HPLC chromatograms of Fig. 1
Fig. 2 Herba Ephedrae HPLC chromatogram
Fig. 3 gold Chinese ephedra and almond containing cough stopping sheet HPLC chromatogram
Fig. 4 Chinese ephedra and almond containing cough stopping sheet HPLC chromatogram
The blank HPLC chromatogram of Fig. 5 Chinese ephedra and almond containing cough stopping sheet
The linear relationship chart of Fig. 6 ephedrine hydrochloride
The linear relationship chart of Fig. 7 pseudoephedrine hydrochloride
The linear relationship chart of Fig. 8 mephedrine
Fig. 9 ephedrine hydrochloride spectral scan figure
Figure 10 pseudoephedrine hydrochloride spectral scan figure
Figure 11 mephedrine spectral scan figure
In Fig. 6, Fig. 7, Fig. 8, ordinate is peak area; Horizontal ordinate is sample size (μ g)
In Fig. 9, Figure 10, Figure 11, ordinate is absorbance unit (mAU); Horizontal ordinate is wavelength (nm)
In Fig. 1, Fig. 2, Fig. 3, Fig. 4, being 1. ephedrine hydrochloride peak, is 2. pseudoephedrine hydrochloride peak, is 3. mephedrine peak, 4. hydrochloride methyl pseudoephedrine 5. hydrochloric acid demethyl ephedrine 6. hydrochloric acid demethyl pseudoephedrines
The specific embodiment of the invention
Embodiment 1:
(1) chromatographic condition and system suitability be take octadecylsilane chemically bonded silica as filling agent; Take volume ratio as 1～1.5: 4～4.2: 94.5～95.0 acetonitrile-methyl alcohol-0.1% phosphoric acid is mobile phase; Column temperature: 40 ℃; Detection wavelength is 207nm; By ephedrine hydrochloride peak, calculate and should be not less than 3000;
(2) it is appropriate that ephedrine hydrochloride, pseudoephedrine hydrochloride and mephedrine are got in the preparation of reference substance solution, accurately weighed, add volume ratio and be hydrochloric acid-70% methanol solution of 1: 200, make the solution of hydrochloric ephedrine 30～15 μ g of every 1ml, pseudoephedrine hydrochloride 10～5 μ g, mephedrine 6～3 μ g, in contrast product solution;
(3) Herba Ephedrae fine powder 0.2～0.3g is got in the preparation of need testing solution, accurately weighed, or Chinese ephedra preparation 0.5～0.8g, accurately weighed, put respectively in tool plug conical flask, precision adds 70% methyl alcohol 25ml, close plug, weighed weight, with power 250W, the ultrasonic processing of frequency 33kHz 10～30 minutes, let cool, weighed weight again, with 70% methyl alcohol, supply the weight of less loss, shake up, , filter, precision measures subsequent filtrate 2ml, put in the chromatographic column of the internal diameter 1cm that 100-120 order neutral alumina 1～2g is housed, with 70% methanol-eluted fractions in the measuring bottle of 10ml to about 9ml, add 1 phosphoric acid, with 70% methyl alcohol, be diluted to scale, shake up, with the miillpore filter of 0.45 μ m, filter, get subsequent filtrate, as need testing solution,
(4) accurate above-mentioned reference substance solution and each 5～10 μ l of need testing solution, injection liquid chromatography, the content of three kinds of ephedrines in working sample drawn of determination method difference.Measure commercially available 3 batches of Chinese ephedra content, the results are shown in Table 7.
Embodiment 2:
(1) chromatographic condition and system suitability be take octadecylsilane chemically bonded silica as filling agent; Acetonitrile-methyl alcohol-0.1% phosphoric acid that the volume ratio of take is 1: 4.2: 94.8 is mobile phase; Column temperature: 40 ℃; Detection wavelength is 207nm; By ephedrine hydrochloride peak, calculate and should be not less than 3000;
(2) it is appropriate that ephedrine hydrochloride, pseudoephedrine hydrochloride and mephedrine are got respectively in the preparation of reference substance solution, accurately weighed, add volume ratio and be hydrochloric acid-70% methanol solution of 1: 200, the solution of making the hydrochloric ephedrine 30 μ g of every 1ml, pseudoephedrine hydrochloride 10 μ g, mephedrine 5 μ g, obtains;
(3) 20 of Chinese ephedra and almond containing cough stopping sheets are got in the preparation of need testing solution, remove dressing, accurately weighed, porphyrize, cross 50 mesh sieves, get 0.8g, accurately weighed, put in tool plug conical flask, precision adds 70% methyl alcohol 25ml, close plug, weighed weight, with power 250W, the ultrasonic processing of frequency 33kHz 10 minutes, let cool, weighed weight again, with methyl alcohol, supply the weight of less loss, shake up, filter, precision measures subsequent filtrate 2ml, put in the chromatographic column of the internal diameter 1cm that 100-120 order neutral alumina 1.5g is housed, with 70% methanol-eluted fractions in the measuring bottle of 10ml to about 9ml, add 1 phosphoric acid, with 70% methyl alcohol, be diluted to scale, shake up, with the miillpore filter of 0.45 μ m, filter, get subsequent filtrate, as need testing solution,
(4) accurate above-mentioned reference substance solution and each 10 μ l of need testing solution of drawing of determination method difference, injection liquid chromatography, the content of three kinds of ephedrines in mensuration test sample.Measure commercially available 3 batches of fiber crops containing cough stopping tablet content, the results are shown in Table 7.
Embodiment 3:
(1) chromatographic condition and system suitability be take octadecylsilane chemically bonded silica as filling agent; Acetonitrile-methyl alcohol-0.1% phosphoric acid that the volume ratio of take is 1: 4: 95 is mobile phase; Column temperature: 40 ℃; Detection wavelength is 207nm; By ephedrine hydrochloride peak, calculate and should be not less than 3000;
(2) it is appropriate that ephedrine hydrochloride, pseudoephedrine hydrochloride and mephedrine are got respectively in the preparation of reference substance solution, accurately weighed, add volume ratio and be hydrochloric acid-70% methanol solution of 1: 200, the solution of making the hydrochloric ephedrine 15 μ g of every 1ml, pseudoephedrine hydrochloride 5 μ g, mephedrine 3 μ g, obtains;
(3) 20 of golden Chinese ephedra and almond containing cough stopping sheets are got in the preparation of need testing solution, remove dressing, accurately weighed, porphyrize, cross 50 mesh sieves, get 0.8g, accurately weighed, put in tool plug conical flask, precision adds 70% methyl alcohol 25ml, close plug, weighed weight, with power 250W, the ultrasonic processing of frequency 33kHz 10 minutes, let cool, weighed weight again, with methyl alcohol, supply the weight of less loss, shake up, filter, precision measures subsequent filtrate 2ml, put in the chromatographic column of the internal diameter 1cm that 100-120 order neutral alumina 1.8g is housed, with 70% methanol-eluted fractions in the measuring bottle of 10ml to about 9ml, add 1 phosphoric acid, with 70% methyl alcohol, be diluted to scale, shake up, with the miillpore filter of 0.45 μ m, filter, get subsequent filtrate, as need testing solution,
(4) accurate above-mentioned reference substance solution and each 10 μ l of need testing solution of drawing of determination method difference, injection liquid chromatography, the content of three kinds of ephedrines in mensuration test sample.Measure 3 batches of golden Chinese ephedra and almond containing cough stopping sheet content, the results are shown in Table 7.
The recovery test result of ephedrine hydrochloride in table 1 sample
The recovery test result of pseudoephedrine hydrochloride in table 2 sample
The recovery test result of mephedrine in table 3 sample
Note: for saving reference substance, the cumulative volume of sample is not 25ml, is 15ml, in the consistent situation of concentration, sampling amount reduces.
Table 4 reference substance Precision Experiment result (peak area)
Table 5 reference substance stability experiment result (peak area)
Table 6 replica test result (mg/g)
The assay result (mg/g) of table 7 Herba Ephedrae and preparation
Claims (5)
1. three kinds of alkaloidal content assaying methods in Herba Ephedrae and preparation thereof, is characterized in that:
(1) chromatographic condition and system suitability be take octadecylsilane chemically bonded silica as filling agent; Take volume ratio as 1～1.5: 4～4.2: 94.5～95.0 acetonitrile-methyl alcohol-0.1% phosphoric acid is mobile phase; Column temperature: 40 ℃; Detection wavelength is 207nm; By ephedrine hydrochloride peak, calculate and should be not less than 3000;
(2) it is appropriate that ephedrine hydrochloride, pseudoephedrine hydrochloride and mephedrine are got in the preparation of reference substance solution, accurately weighed, add volume ratio and be hydrochloric acid-70% methanol solution of 1: 200, make the solution of hydrochloric ephedrine 30～15 μ g of every 1ml, pseudoephedrine hydrochloride 10～5 μ g, mephedrine 6～3 μ g, in contrast product solution;
(3) Herba Ephedrae fine powder 0.2～0.3g is got in the preparation of need testing solution, accurately weighed, or Chinese ephedra preparation 0.5～0.8g, accurately weighed, put respectively in tool plug conical flask, precision adds 70% methyl alcohol 25m], close plug, weighed weight, with power 250W, the ultrasonic processing of frequency 33kHz 10～30 minutes, let cool, weighed weight again, with 70% methyl alcohol, supply the weight of less loss, shake up, filter, precision measures subsequent filtrate 2ml, put in the chromatographic column of the internal diameter 1cm that 100-120 order neutral alumina 1～2g is housed, with 70% methanol-eluted fractions in the measuring bottle of 10ml to about 9ml, add 1 phosphoric acid, with 70% methyl alcohol, be diluted to scale, shake up, with the miillpore filter of 0.45 μ m, filter, get subsequent filtrate, as need testing solution,
(4) accurate above-mentioned reference substance solution and each 5～10 μ l of need testing solution, injection liquid chromatography, the three kinds of alkaloidal content in working sample drawn of determination method difference.
2. three kinds of alkaloidal content assaying methods in a kind of Herba Ephedrae according to claim 1 and preparation thereof, is characterized in that described preparation refers to Chinese ephedra and almond containing cough stopping sheet and golden Chinese ephedra and almond containing cough stopping sheet.
3. three kinds of alkaloidal content assaying methods in a kind of Herba Ephedrae according to claim 1 and 2 and preparation thereof, are further characterized in that:
(1) chromatographic condition and system suitability be take octadecylsilane chemically bonded silica as filling agent; Acetonitrile-methyl alcohol-0.1% phosphoric acid that the volume ratio of take is 1.5: 4: 94.5 is mobile phase; Column temperature: 40 ℃; Detection wavelength is 207nm; By ephedrine hydrochloride peak, calculate and should be not less than 3000;
(2) it is appropriate that ephedrine hydrochloride, pseudoephedrine hydrochloride and mephedrine are got respectively in the preparation of reference substance solution, accurately weighed, add volume ratio and be hydrochloric acid-70% methanol solution of 1: 200, the solution of making the hydrochloric ephedrine 20 μ g of every 1ml, pseudoephedrine hydrochloride 10 μ g, mephedrine 3 μ g, obtains;
(3) Herba Ephedrae fine powder 0.2g is got in the preparation of need testing solution, accurately weighed, put in tool plug conical flask, precision adds 70% methyl alcohol 25ml, close plug, weighed weight, with power 250W, the ultrasonic processing of frequency 33kHz 30 minutes, let cool, weighed weight again, with 70% methyl alcohol, supply the weight of less loss, shake up, filter paper filters, precision measures subsequent filtrate 2ml, put in the chromatographic column of the internal diameter 1cm that 100-120 order neutral alumina 1.5g is housed, with 70% methanol-eluted fractions in the measuring bottle of 10m1 to about 9m1, add 1 phosphoric acid, with 70% methyl alcohol, be diluted to scale, shake up, with 0.45 μ m miillpore filter, filter, get subsequent filtrate, as need testing solution,
(4) accurate above-mentioned reference substance solution and each 10 μ l of need testing solution of drawing of determination method difference, injection liquid chromatography, measures three kinds of alkaloidal content in Herba Ephedrae.
4. three kinds of alkaloidal content assaying methods in a kind of Herba Ephedrae according to claim 1 and 2 and preparation thereof, are further characterized in that:.
(1) chromatographic condition and system suitability be take octadecylsilane chemically bonded silica as filling agent; Acetonitrile-methyl alcohol-0.1% phosphoric acid that the volume ratio of take is 1: 4.2: 94.8 is mobile phase; Column temperature: 40 ℃; Detection wavelength is 207nm; By ephedrine hydrochloride peak, calculate and should be not less than 3000;
(2) it is appropriate that ephedrine hydrochloride, pseudoephedrine hydrochloride and mephedrine are got respectively in the preparation of reference substance solution, accurately weighed, add volume ratio and be hydrochloric acid-70% methanol solution of 1: 200, the solution of making the hydrochloric ephedrine 30 μ g of every 1ml, pseudoephedrine hydrochloride 10 μ g, mephedrine 5 μ g, obtains;
(3) 20 of Chinese ephedra and almond containing cough stopping sheets are got in the preparation of need testing solution, remove dressing, accurately weighed, porphyrize, cross 50 mesh sieves, get 0.8g, accurately weighed, put in tool plug conical flask, precision adds 70% methyl alcohol 25ml, close plug, weighed weight, with power 250W, the ultrasonic processing of frequency 33kHz 10 minutes, let cool, weighed weight again, with methyl alcohol, supply the weight of less loss, shake up, filter, precision measures subsequent filtrate 2ml, put in the chromatographic column of the internal diameter 1cm that 100-120 order neutral alumina 1.5g is housed, with 70% methanol-eluted fractions in the measuring bottle of 10ml to about 9ml, add 1 phosphoric acid, with 70% methyl alcohol, be diluted to scale, shake up, with the miillpore filter of 0.45 μ m, filter, get subsequent filtrate, as need testing solution,
(4) accurate above-mentioned reference substance solution and each 10 μ l of need testing solution of drawing of determination method difference, injection liquid chromatography, measures three kinds of alkaloidal content in Chinese ephedra and almond containing cough stopping sheet.
5. three kinds of alkaloidal content assaying methods in a kind of Herba Ephedrae according to claim 1 and 2 and preparation thereof, are further characterized in that:
(1) chromatographic condition and system suitability be take octadecylsilane chemically bonded silica as filling agent; Acetonitrile-methyl alcohol-0.1% phosphoric acid that the volume ratio of take is 1: 4: 95 is mobile phase; Column temperature: 40 ℃; Detection wavelength is 207nm; By ephedrine hydrochloride peak, calculate and should be not less than 3000;
(2) it is appropriate that ephedrine hydrochloride, pseudoephedrine hydrochloride and mephedrine are got respectively in the preparation of reference substance solution, accurately weighed, add volume ratio and be hydrochloric acid-70% methanol solution of 1: 200, the solution of making the hydrochloric ephedrine 15 μ g of every 1ml, pseudoephedrine hydrochloride 5 μ g, mephedrine 3 μ g, obtains;
(3) 20 of golden Chinese ephedra and almond containing cough stopping sheets are got in the preparation of need testing solution, remove dressing, accurately weighed, porphyrize, cross 50 mesh sieves, get 0.8g, accurately weighed, put in tool plug conical flask, precision adds 70% methyl alcohol 25ml, close plug, weighed weight, with power 250W, the ultrasonic processing of frequency 33kHz 10 minutes, let cool, weighed weight again, with methyl alcohol, supply the weight of less loss, shake up, filter, precision measures subsequent filtrate 2ml, put in the chromatographic column of the internal diameter 1cm that 100-120 order neutral alumina 1.8g is housed, with 70% methanol-eluted fractions in the measuring bottle of 10ml to about 9ml, add 1 phosphoric acid, with 70% methyl alcohol, be diluted to scale, shake up, with the miillpore filter of 0.45 μ m, filter, get subsequent filtrate, as need testing solution,
(4) accurate above-mentioned reference substance solution and each 10 μ l of need testing solution of drawing of determination method difference, injection liquid chromatography, measures three kinds of alkaloidal content in golden Chinese ephedra and almond containing cough stopping sheet.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105954379A (en) * | 2016-04-22 | 2016-09-21 | 广西壮族自治区梧州食品药品检验所 | Method for extracting ephedrine hydrochloride in lung ventilating and regulating pill |
| CN106841405A (en) * | 2015-12-03 | 2017-06-13 | 康普药业股份有限公司 | A kind of relevant substance detecting method of ephedrine hydrochloride piece |
| CN106950292A (en) * | 2017-01-17 | 2017-07-14 | 天津中新药业集团股份有限公司达仁堂制药厂 | A kind of lung-clearing anti-inflammatory ball quality standard detecting method |
| CN110297043A (en) * | 2018-03-21 | 2019-10-01 | 上海医药工业研究院 | A kind of analysis method of 2- methylamino -1- phenyl-1-propanol |
| CN113288922A (en) * | 2021-05-13 | 2021-08-24 | 海南葫芦娃药业集团股份有限公司 | Ephedra extract and preparation method thereof |
| CN114544786A (en) * | 2020-11-25 | 2022-05-27 | 中国科学院大连化学物理研究所 | Chromatographic separation method of organic amine alkaloid in relative ephedra by utilizing fluorine-containing fixation |
| CN114949053A (en) * | 2022-07-08 | 2022-08-30 | 太极集团四川绵阳制药有限公司 | Preparation method of ephedra and apricot cough relieving tablet |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5857383A (en) * | 1981-10-01 | 1983-04-05 | Suntory Ltd | Novel flavonoid |
| CN102662024A (en) * | 2012-04-23 | 2012-09-12 | 韩桂茹 | Method for simultaneously determining three alkaloids in granules for eliminating phlegm and stopping cough for children |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SU830238A1 (en) * | 1979-07-17 | 1981-05-15 | Иркутский Государственный Медицинскийинститут | Method of determining papaverine hydrochloride of medicinal mixtures |
-
2013
- 2013-09-10 CN CN201310406568.9A patent/CN103512998B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5857383A (en) * | 1981-10-01 | 1983-04-05 | Suntory Ltd | Novel flavonoid |
| CN102662024A (en) * | 2012-04-23 | 2012-09-12 | 韩桂茹 | Method for simultaneously determining three alkaloids in granules for eliminating phlegm and stopping cough for children |
Non-Patent Citations (4)
| Title |
|---|
| 赵志军等: "麻杏止咳胶囊质量标准的研究", 《中国药品标准》, vol. 7, no. 6, 31 December 2006 (2006-12-31) * |
| 陈莹等: "HPLC法同时测定麻黄中盐酸麻黄碱、盐酸伪麻黄碱及盐酸甲基麻黄碱的含量", 《食品工业科技》, vol. 29, no. 12, 25 December 2008 (2008-12-25) * |
| 韩归茹等: "药材、制剂、半成品中麻黄碱、伪麻黄碱和苦杏仁干的同时定量研究", 《中成药》, vol. 27, no. 4, 30 April 2005 (2005-04-30) * |
| 韩桂茹等: "金麻杏止咳片快速薄层色谱鉴别与3成分同时定量测定研究", 《中国药业》, vol. 21, no. 14, 20 July 2012 (2012-07-20) * |
Cited By (9)
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|---|---|---|---|---|
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| CN110297043A (en) * | 2018-03-21 | 2019-10-01 | 上海医药工业研究院 | A kind of analysis method of 2- methylamino -1- phenyl-1-propanol |
| CN114544786A (en) * | 2020-11-25 | 2022-05-27 | 中国科学院大连化学物理研究所 | Chromatographic separation method of organic amine alkaloid in relative ephedra by utilizing fluorine-containing fixation |
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