CN102068649A - Quality control method for gastric condition-regulating pill as traditional Chinese preparation - Google Patents
Quality control method for gastric condition-regulating pill as traditional Chinese preparation Download PDFInfo
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- CN102068649A CN102068649A CN2009102283606A CN200910228360A CN102068649A CN 102068649 A CN102068649 A CN 102068649A CN 2009102283606 A CN2009102283606 A CN 2009102283606A CN 200910228360 A CN200910228360 A CN 200910228360A CN 102068649 A CN102068649 A CN 102068649A
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Abstract
The invention relates to a quality control method for a gastric condition-regulating pill as traditional Chinese preparation, which is prepared from Chinese herbal medicines as raw materials, belonging to the technical field of Chinese patent medicine; the method comprises the steps that: a microscope is arranged for quality standard to identify whether the formula of the gastric condition-regulating pill contains betel nut, immature bitter orange, magnolia bark and cinnamon; with synephrine, dehydrocostus lactone and liquorice as reference substances, thin-layer chromatography is adopted to identify whether the formula of gastric condition-regulating pill contains the ingredients of synephrine, dehydrocostus lactone and liquorice; and with magnolol and honokiol as reference substances, high efficiency liquid chromatography is adopted to detect the quantitative index and the inspection method of the content of the magnolia bark in the formula of gastric condition-regulating pill, thereby relatively perfecting the quality standard, strictly controlling the quality of medicines, ensuring the improvement of product quality and improving the quality control for medicines owing to the revised quality standard.
Description
Technical field
The invention belongs to the technical field of Chinese patent medicine, relating to the Chinese crude drug is the method for quality control that a kind of Chinese medicine preparation that raw material is made is transferred the stomach pellet.
Background technology
Transferring the stomach pellet is a kind of classical Chinese patent medicine, Ministry of Public Health standard WS
3-B-0395-90 record prescription and quality standard:
Prescription: Radix Aucklandiae 75g Fructus Amomi 75g Radix Glycyrrhizae 75g
Semen Arecae 75g Fructus Aurantii Immaturus (stir-fry) 75g Cortex Magnoliae Officinalis (processed with Rhizoma Zingiberis Recens) 75g
Rhizoma Cyperi (vinegar system) 75g Fructus Amomi Rotundus 75g Oletum Trogopterori (vinegar system) 50g
High langal 25g Flos Caryophylli 25g Cortex Cinnamomi 25g
Method for making: above 12 flavors, be ground into fine powder, sieve, mixing is used water pill, drying, the dried ball of every 500g is used Cinnabar 16g, Resina persicae 3gization water coating, polishing, drying, promptly.
Check: should meet every regulation relevant under the pill item (2 pages of appendix).
More than fill a vacancy
The document of relevant at present accent stomach pellet has a lot, but Shang Weijian and above-mentioned disclosed Ministry of Public Health standard WS
3-B-0395-90 transfers the red prescription of stomach similar or identical, proportioning is identical, the document of the product standard that effect is similar, but also there are shortcomings such as discriminating project, discriminating means, content assaying method imperfection in the standard of prior art, for improving the product quality of Chinese medicine, need improve target level of product quality.
Summary of the invention
The objective of the invention is to overcome the red standard weak point of accent stomach of prior art, a kind of method of quality control of accent stomach pellet that can the qualitative and quantitative analysis ingredient is provided.
In order to reach the technical scheme that purpose of the present invention adopts be: a kind of Chinese medicine preparation is transferred the method for quality control of stomach pellet, and wherein said pharmaceutical formulation is by Chinese crude drug:
Radix Aucklandiae 75g Fructus Amomi 75g Radix Glycyrrhizae 75g
Semen Arecae 75g Fructus Aurantii Immaturus (parched with bran) 75g Cortex Magnoliae Officinalis (processed with Rhizoma Zingiberis Recens) 75g
Rhizoma Cyperi (vinegar system) 75g Fructus Amomi Rotundus 75g Oletum Trogopterori (vinegar system) 50g
Rhizoma Alpiniae Officinarum 25g Flos Caryophylli 25g Cortex Cinnamomi 25g forms,
Method for making; More than 12 the flavor, be ground into fine powder, sieve, mixing, the water pill, polishing, drying promptly, is characterized in that: the step of its method is:
(1) be contrast with Semen Arecae, Fructus Aurantii Immaturus, Cortex Magnoliae Officinalis, cinnamomic microscopic features, microscope differentiates to transfer in the red prescription of stomach whether contain Semen Arecae, Fructus Aurantii Immaturus, Cortex Magnoliae Officinalis, Cortex Cinnamomi; With Neosynephrine, dehydrocostuslactone, Radix Glycyrrhizae is reference substance, and thin layer chromatography differentiates to transfer in the red prescription of stomach whether contain Neosynephrine, dehydrocostuslactone, licorice ingredient;
(2) be reference substance with magnolol, honokiol, high performance liquid chromatography detects the content of transferring Cortex Magnoliae Officinalis in the red prescription of stomach.
Described Chinese medicine preparation is transferred the method for quality control of stomach pellet, it is characterized in that: the described microscope of putting differentiate to be transferred and not to be contained Semen Arecae, Fructus Aurantii Immaturus, Cortex Magnoliae Officinalis, cinnamomic method in the red prescription of stomach and be: get this product, putting microscopically observes: thin ancient piece of jade, round, flat and with a hole in its centre cell contains prism of calcium oxalate around the fibre bundle, forms crystalline cellulose; The endosperm cell debris is colourless, and wall is thicker, and more great similar round stricture of vagina hole is arranged; Prism of calcium oxalate is present in the Bao Bi tissue in flakes; Stone cell divides dendritic, and behind the wall, laminated striation is obvious; Square or the similar round of stone cell class, wall are simultaneously poor;
Described thin layer chromatography is differentiated and transferred the method that whether contains the Neosynephrine composition in the red prescription of stomach to be: get this product 5g, porphyrize adds methanol 15ml, and supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the Neosynephrine reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to thin layer chromatography, draw each 2～5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-glacial acetic acid-water is developing solvent, with n-butyl alcohol: glacial acetic acid: the upper solution of water=4: 1: 5, launch, take out, dry, spray is with 0.5% ethanol solution of ninhydrin, and it is clear to be heated to speckle colour developing; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
Described thin layer chromatography is differentiated and transferred the method that whether contains the dehydrocostuslactone composition in the red prescription of stomach to be: get this product 5g, porphyrize adds chloroform 10ml, and supersound process 30 minutes filters, and filtrate is as need testing solution; Other removes hydrogen constuslactone reference substance, adds chloroform and makes the solution that every 1ml contains 0.5mg, in contrast product solution; According to the thin layer chromatography test, draw each 4～6 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with cyclohexane extraction-acetone, with cyclohexane extraction: acetone=10: 2, launch, take out, dry, spray is with 5% vanillin sulfuric acid solution, and it is clear to be heated to speckle colour developing; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
Described thin layer chromatography is differentiated and is transferred the method that whether contains licorice ingredient in the red prescription of stomach to be: get this product 5g, porphyrize, the 40ml that adds diethyl ether, reflux 20 minutes, filter, discard filtrate, medicinal residues volatilize ether, add methanol 30ml, reflux 1 hour, filter, filtrate evaporate to dryness, residue add water 40ml makes dissolving, with water saturated n-butanol extraction 3 times, each 20ml merges n-butyl alcohol liquid, water 20ml washing, discard water liquid, n-butyl alcohol liquid evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 1g in addition, the 10ml that adds diethyl ether, reflux 20 minutes, filter, discard filtrate, medicinal residues volatilize ether, add methanol 10ml, reflux 1 hour filters, filtrate evaporate to dryness, residue add water 10ml makes dissolving, with water saturated n-butanol extraction 3 times, each 10ml merges n-butyl alcohol liquid, water 10ml washing, discard water liquid, n-butyl alcohol liquid evaporate to dryness shines medical material solution in pairs with legal system.Test according to thin layer chromatography, draw each 2～4 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water is developing solvent, chloroform: methanol: water=40: 10: 1, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to speckle colour developing; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
Described Chinese medicine preparation is transferred the method for quality control of stomach pellet, and it is characterized in that: with magnolol, honokiol is reference substance, and liquid chromatography detects transfers the method for the content of Cortex Magnoliae Officinalis in the red prescription of stomach to be:
(1) chromatographic condition and system suitability test: chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Methanol: water=78: 22 is mobile phase; The detection wavelength is 294nm; Number of theoretical plate calculates by the magnolol peak should be not less than 3800;
(2) preparation of reference substance solution: get the magnolol reference substance, the honokiol reference substance is an amount of, accurately claim surely, add methanol and make the solution that every 1ml contains magnolol 130 μ g, honokiol 45 μ g, promptly;
(3) preparation of need testing solution: it is an amount of to get this product, is ground into fine powder, gets powder 0.5g, accurate claim fixed, the accurate methanol 25ml that adds, claim decide weight, supersound process 30 minutes is put coldly, and weight decided in title again, supplies the weight that subtracts mistake with methanol, shake up, filter, get subsequent filtrate, promptly;
(4) algoscopy: algoscopy is drawn above-mentioned reference substance solution and each 5 μ l of need testing solution respectively, injects chromatograph of liquid, measures, promptly; The every 1g of this product contains Cortex Magnoliae Officinalis with magnolol (C
18H
18O
2) and honokiol (C
18H
18O
2) the total amount meter, must not be less than 1.6mg
Invention beneficial effect: microscope is set in the quality standard differentiates that accent stomach pellet does not contain Semen Arecae, Fructus Aurantii Immaturus, Cortex Magnoliae Officinalis, Cortex Cinnamomi in writing out a prescription; With Neosynephrine, dehydrocostuslactone, Radix Glycyrrhizae is reference substance, and thin layer chromatography differentiates to transfer in the red prescription of stomach whether contain Neosynephrine, dehydrocostuslactone, licorice ingredient;
(2) be reference substance with magnolol, honokiol, high performance liquid chromatography detects the content of transferring Cortex Magnoliae Officinalis in the red prescription of stomach, and the every 1g of this product contains Cortex Magnoliae Officinalis with magnolol (C
18H
18O
2) and honokiol (C
18H
18O
2) the total amount meter, must not be less than the quantitative target and the method for inspection of 1.6mg, make quality standard comparatively perfect, revised quality standard has improved the quality control of medicine.
Description of drawings
Fig. 1 Semen Arecae microscopical identification figure.
Fig. 2 Fructus Aurantii Immaturus microscopical identification figure.
Fig. 3 Cortex Magnoliae Officinalis microscopical identification figure.
Fig. 4 Cortex Cinnamomi microscopical identification figure.
Fig. 5, Fig. 6 are the thin-layer chromatograms of transferring Fructus Aurantii Immaturus in the stomach pellet.
Fig. 7 is a thin-layer chromatogram of transferring the Radix Aucklandiae in the stomach pellet.
Fig. 8 is a thin-layer chromatogram of transferring Radix Glycyrrhizae in the stomach pellet.
Fig. 9 is honokiol and magnolol reference substance curve chart.
Figure 10 transfers the red curve chart of stomach.
Figure 11 is the negative sample curve chart.
Figure 12 is honokiol and magnolol reference substance curve chart (response rate).
Figure 13 transfers the red sample mark-on curve chart (response rate) of stomach.
Figure 14 magnolol linear graph.
Figure 15 honokiol linear graph.
Description of reference numerals:
Fig. 5: [left side → right side: Neosynephrine (0727-200105), Fructus Aurantii Immaturus medical material, accent stomach pellet (060301) test sample, feminine gender]
Fig. 6: [left side → right side: Neosynephrine (0727-200105), accent stomach pellet (060301), accent stomach pellet (060302), accent stomach pellet (060303), feminine gender]
Fig. 7: { left side → right side: reference substance [dehydrocostuslactone (111525-200404)], accent stomach pellet (060301), accent stomach pellet (060302), accent stomach pellet (060303), feminine gender }
Fig. 8: [left side → right side: Radix Glycyrrhizae control medicinal material (120904-200511), accent stomach pellet (060301), accent stomach pellet (0603002), accent stomach pellet (060303), feminine gender]
The specific embodiment
Below in conjunction with embodiment, the present invention is further described, the red quality standard of accent stomach after embodiment 1 improves: prescription:
Radix Aucklandiae 75g Fructus Amomi 75g Radix Glycyrrhizae 75g
Semen Arecae 75g Fructus Aurantii Immaturus (parched with bran) 75g Cortex Magnoliae Officinalis (processed with Rhizoma Zingiberis Recens) 75g
Rhizoma Cyperi (vinegar system) 75g Fructus Amomi Rotundus 75g Oletum Trogopterori (vinegar system) 50g
Rhizoma Alpiniae Officinarum 25g Flos Caryophylli 25g Cortex Cinnamomi 25g
Method for making: above 12 flavors, be ground into fine powder, sieve, mixing, the water pill, polishing, drying, promptly.
Character: this product be yellowish-brown to the sepia watered pill, gas fragrance, acrid in the mouth hardship.
Differentiate: (1) gets this product, puts microscopically and observes: thin ancient piece of jade, round, flat and with a hole in its centre cell contains prism of calcium oxalate around the fibre bundle, forms crystalline cellulose.The endosperm cell debris is colourless, and wall is thicker, and more great similar round pit is arranged.Prism of calcium oxalate is present in the Bao Bi tissue in flakes.Stone cell divides dendritic, and behind the wall, laminated striation is obvious.Square or the similar round of stone cell class, wall are simultaneously poor.
(2) get this product 5g, porphyrize adds methanol 15ml, and supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Other gets the Neosynephrine reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution.According to thin layer chromatography (" first appendix VI of Chinese pharmacopoeia version in 2005 B) test, draw each 2～5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, upper solution with n-butyl alcohol-glacial acetic acid-water=4: 1: 5 is developing solvent, launch, take out, dry, spray is with 0.5% ethanol solution of ninhydrin, and it is clear to be heated to the speckle colour developing.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
(3) get this product 5g, porphyrize adds chloroform 10ml, and supersound process 30 minutes filters, and filtrate is as need testing solution.Other removes hydrogen constuslactone reference substance, adds chloroform and makes the solution that every 1ml contains 0.5mg, in contrast product solution.According to thin layer chromatography (" an appendix VI of Chinese pharmacopoeia version in 2005 B) test, draw each 4～6 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with cyclohexane extraction-acetone=10: 2 was developing solvent, launch, take out, dry, spray is with 5% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
(4) get this product 5g, porphyrize, the 40ml that adds diethyl ether, reflux 20 minutes, filter, discard filtrate, medicinal residues volatilize ether, add methanol 30ml, reflux 1 hour filters the filtrate evaporate to dryness, residue adds water 40ml makes dissolving, with water saturated n-butanol extraction 3 times, and each 20ml, merge n-butyl alcohol liquid, water 20ml washing discards water liquid, n-butyl alcohol liquid evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution.Extracting liquorice control medicinal material 1g in addition, the 10ml that adds diethyl ether, reflux 20 minutes, filter, discard filtrate, medicinal residues volatilize ether, add methanol 10ml, reflux 1 hour filters, filtrate evaporate to dryness, residue add water 10ml makes dissolving, with water saturated n-butanol extraction 3 times, each 10ml merges n-butyl alcohol liquid, water 10ml washing, discard water liquid, n-butyl alcohol liquid evaporate to dryness shines medical material solution in pairs with legal system.According to thin layer chromatography (" an appendix VI of Chinese pharmacopoeia version in 2005 B) test, draw each 2～4 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water=40: 10: 1 was developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
Check: should meet every regulation relevant under the pill item (" an appendix I of Chinese pharmacopoeia version in 2005 A).
Assay: the photograph high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2005 D) measure.
Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Methanol-water=78: 22 is a mobile phase; The detection wavelength is 294nm.Number of theoretical plate calculates by the magnolol peak should be not less than 3800.
The magnolol reference substance is got in the preparation of reference substance solution, the honokiol reference substance is an amount of, and accurate the title decides, and adds methanol and makes the solution that every 1ml contains magnolol 130 μ g, honokiol 45 μ g, promptly.
It is an amount of that this product is got in the preparation of need testing solution, is ground into fine powder, gets powder 0.5g, and accurate the title decides, the accurate methanol 25ml that adds, claim decide weight, supersound process 30 minutes is put coldly, and weight decided in title again, supplies the weight that subtracts mistake with methanol, shake up, filter, get subsequent filtrate, promptly.
Algoscopy is drawn above-mentioned reference substance solution and each 5 μ l of need testing solution respectively, injects chromatograph of liquid, measures, promptly.The every 1g of this product contains Cortex Magnoliae Officinalis with magnolol (C
18H
18O
2) and honokiol (C
18H
18O
2) the total amount meter, must not be less than 1.6mg.
Function with cure mainly: the alleviating distention in middle-JIAO that is good for the stomach, soothing liver-QI is pleasant.Be used for the gastric acid cold syndrome of the stomach, the chest distension, it is noisy to fall to satisfy, appetite pain.
Usage and consumption; Oral.One time 20 ball, 2 times on the one.Attention: be not taken by pregnant women.Specification: per 10 balls weigh 1.45 grams.Storage: sealing.
Differentiate:
One, instrument and reagent
High performance liquid chromatograph: Tianjin, island LC-2010A
HT
Ultrasound Instrument: KUDOS SK8200LH (Shanghai High Kudos Science Instrument Co., Ltd.);
Electronic balance: Sartorius CP225D/CP224S (Beijing Sai Duolisi instrument system company limited);
Silica gel g thin-layer plate, silica gel H lamellae, high-efficient silica gel G lamellae (subsidiary factory of Haiyang Chemical Plant, Qingdao);
Reference substance: dehydrocostuslactone (lot number: 1525-200102)
Neosynephrine (lot number: 110727-200105)
Above reference substance is all available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute;
Agents useful for same is an analytical pure, and water is double distilled deionized water;
Transfer stomach pellet (lot number: 060301,060302,060303)
Provide by Lerentang Pharmaceutical Factory, Zhongxin Pharmaceutical Group Co., Ltd., Tianj.
Two, differentiate
2.1 transfer the red microscopical identification of stomach
Get this product, put microscopically and observe: thin ancient piece of jade, round, flat and with a hole in its centre cell contains prism of calcium oxalate around the fibre bundle, forms crystalline cellulose (Fructus Aurantii Immaturus).The endosperm cell debris is colourless, and wall is thicker, and more great similar round pit (Semen Arecae) is arranged.Prism of calcium oxalate is present in the Bao Bi tissue in flakes.Stone cell divides dendritic, and behind the wall, laminated striation is (Cortex Magnoliae Officinalis) obviously.Square or the similar round of stone cell class, wall simultaneously poor (Cortex Cinnamomi), (the results are shown in Figure 1, Fig. 2, Fig. 3, Fig. 4)
2.2 transfer the qualitative identification of Fructus Aurantii Immaturus in the stomach pellet
The preparation of medical material solution: get Fructus Aurantii Immaturus medicinal powder 0.5g, add methanol 10mL, supersound process 20 minutes filters, and filtrate evaporate to dryness, residue add methanol 0.5mL, make dissolving, as the medical material need testing solution.
The preparation of need testing solution: get different lot numbers and transfer the red powder of stomach (crossing 80 mesh sieves) 5g, add methanol 15mL, supersound process 20 minutes filters, as need testing solution.
The preparation of reference substance solution: get the Neosynephrine reference substance, add methanol and make the solution that every 1mL contains 0.5mg, in contrast product solution.
The preparation of negative solution: except that Fructus Aurantii Immaturus,,, get 5g,, be prepared as negative solution according to the operation down of need testing solution preparation method item according to the operation of test sample preparation process by all medicines of prescription preparation.
Differential method: according to thin layer chromatography (first appendix VIB of Chinese Pharmacopoeia version in 2005) test, draw each 2～5 μ L of above-mentioned solution, put respectively on same silica gel H lamellae, upper solution with n-butyl alcohol-glacial acetic acid-water=4: 1: 5 is developing solvent, launch, take out, dry, spray is with 0.5% ethanol solution of ninhydrin, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.Then there is not this speckle in the negative control chromatograph.Illustrate that this method is exclusive, reliable.(the results are shown in Figure 5, Fig. 6)
2.3 transfer the qualitative identification of the Radix Aucklandiae in the stomach pellet: the preparation of medical material solution: get Radix Aucklandiae medicinal powder 0.5g, add chloroform 10mL, supersound process 30 minutes filters, and filtrate is as the medical material need testing solution.
The preparation of need testing solution: red each 5g of accent stomach that gets different lot numbers pulverizes, and adds chloroform 10mL, and supersound process 30 minutes filters, and filtrate is as need testing solution.
The preparation of reference substance solution: it is an amount of to remove hydrogen constuslactone reference substance, adds chloroform and makes the solution that every 1mL contains 0.5mg, in contrast product solution.
The preparation of negative solution: except that the Radix Aucklandiae,,, get 5g,, be prepared as negative solution according to the operation down of need testing solution preparation method item according to the operation of test sample preparation process by all medicines of prescription preparation.
Differential method: the photograph thin layer chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 4～6 μ L of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with cyclohexane-acetone=10: 2 was developing solvent, launch, take out, dry, spray is with 5% vanillin sulfuric acid solution, and it is clear to be heated to the speckle colour developing.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.Then there is not this speckle in the negative control chromatograph.Illustrate that this method is exclusive, reliable.(the results are shown in Figure 7)
2.4 transfer the qualitative identification of Radix Glycyrrhizae in the stomach pellet
The preparation of need testing solution: red each 5g of accent stomach that gets different lot numbers pulverizes, the 40mL that adds diethyl ether, and reflux 20 minutes filters, filtrate discards, and medicinal residues volatilize ether, adds methanol 30mL, reflux 1 hour filters the filtrate evaporate to dryness, residue adds water 40mL makes dissolving, with water saturated n-butanol extraction 3 times, and each 20mL, merge n-butyl alcohol liquid, water 20mL washing discards water liquid, n-butyl alcohol liquid evaporate to dryness, residue add methanol 2mL makes dissolving, as need testing solution.
The preparation of control medicinal material solution: extracting liquorice control medicinal material 1g, according to the operation down of need testing solution preparation method item, make control medicinal material solution.
The preparation of negative solution: except that Radix Glycyrrhizae,,, get 5g,, make negative solution according to the operation down of need testing solution preparation method item according to the operation of test sample preparation process by all medicines of prescription preparation.
Differential method: the photograph thin layer chromatography (" appendix VIB of Chinese pharmacopoeia version in 2005) test, draw each 2～4 μ L of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water=40: 10: 1 was developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.Then there is not this speckle in the negative control chromatograph.(the results are shown in Figure 8) assay:
One, instrument and reagent:
1. instrument condition
Instrument: high performance liquid chromatograph: Tianjin, island LC-2010AHT detector: UV/VIS; Detect wavelength: 294nm.
2. chromatographic condition:
Chromatographic column:
C18 5um 4.6 * 150mm Column; Mobile phase: methanol-water (78: 22); Flow velocity: 0.8mL/min; Column temperature: 25 ℃; Sample size: 5 μ L
3. reagent and reagent
Reference substance: magnolol (lot number: 110729-200309), honokiol (lot number: 110730-200206) provide by the biological calibrating institute that makes of Chinese medicine;
Reagent: methanol (chromatographically pure)
Test sample: stomach pellet (lot number: 060301,060302,060303) provide by Lerentang Traditional Chinese Medicine Factory, Tianjin is provided.
Two, the investigation of experimental technique
1. the investigation of extraction conditions
1.1 the selection of extraction solvent: this experiment is to 30%, 50%, 70%, 100% methanol;
1.2 the selection of extracting method: hot reflux twice 45 minutes/time, dipping 24 hours, ultrasonic.
1.3 the preparation of reference substance solution: precision takes by weighing the magnolol reference substance, the honokiol reference substance is an amount of, adds methanol and makes the solution that every 1mL contains magnolol 135.68 μ g, honokiol 44.896 μ g respectively, promptly.
1.4 the preparation of need testing solution: get and transfer stomach pellet (lot number: 060301) powder (crossing 80 mesh sieves), about 1.0 grams, the accurate title, decide, put in the tool plug conical flask, the accurate methanol 25mL that adds above-mentioned four kinds of concentration shakes up respectively, close plug, weigh respectively and to handle, mend heavy back and filter (hot reflux handle after, the filtration standardize solution is 50mL) as need testing solution with above-mentioned three kinds of methods.The results are shown in Table 1,2,3:
Table 1 hot reflux method is investigated the result
Table 2 dipping method is investigated the result
2. the selection of condition determination
2.1 the selection of wavelength: select 294 down for detecting wavelength according to first the 176th page of Cortex Magnoliae Officinalis assay item of Chinese Pharmacopoeia version in 2005.
The investigation result of table 3 ultrasonic method
Experimental result shows the ultrasonic optimum extracting method that is, 100% methanol is the optimum extraction solvent.
2.2 the selection of mobile phase
Selecting mobile phase down according to first the 176th page of Cortex Magnoliae Officinalis assay item of Chinese Pharmacopoeia version in 2005 is methanol-water (78: 22)
3. blank experiment
An amount of by the flavour of a drug that sample prescription takes by weighing except that Cortex Magnoliae Officinalis, make blank by the test sample preparation method, use the chromatographic condition identical to measure with sample, the while working sample, the result shows that negative sample goes out the place, peak at magnolol and honokiol and do not have the peak.See accompanying drawing
4. linear relationship is investigated
The preparation of magnolol and honokiol reference substance solution: precision takes by weighing magnolol and the honokiol reference substance is an amount of, makes the reference substance solution that every 1mL contains magnolol 135.68 μ g, honokiol 44.896 μ g with methanol.Draw above-mentioned solution 1.0,2.0,4.0,5.0,6.0,8.0,10ul, peak area is measured in sample introduction analysis successively, the results are shown in Table 4-5:
Table 4 magnolol linear relationship is investigated the result
With the sample size is abscissa, and peak area is a vertical coordinate, drawing standard curve, Figure 14 magnolol linearity as follows.Learn by statistics and calculate regression equation: Y=1.396 * 10
7X-3.293 * 10
4R=0.9998
Table 5 honokiol linear relationship is investigated the result
With the sample size is abscissa, and peak area is a vertical coordinate, drawing standard curve, Figure 15 honokiol linearity as follows.Learn by statistics and calculate regression equation: Y=2.191 * 10
6X-8.636 * 10
3R=0.9998
5. precision test
The sample of the same lot number of accurate absorption (lot number: 060301) 5 μ L, repeat sample introduction 6 times, measure the peak area integrated value by above-mentioned chromatographic condition, calculating relative standard deviation is 0.95%, 0.92%.The results are shown in Table 6:
Table 6 precision experimental result
6. repeatability test
Get same lot number sample (lot number: 060301) powder (cross 80 mesh sieves), take by weighing 6 parts, every part of about 0.5g, the content assaying method by drafting extracts, and analyzes, and records the peak area integrated value, the calculating relative standard deviation is 0.71%, 1.13%.The results are shown in Table 7:
Table 7 repeatability experimental result
7. recovery test
Take by weighing sample (transferring stomach pellet 060301) the about 0.25g of powder, 6 parts, the accurate title, decide, put in the 50mL tool plug triangular flask, (2mL is 0.848mg/mL) with honokiol (1mL to add the magnolol reference substance respectively, 0.5612mg/mL), press the test sample preparation and measure an operation down, calculate recovery rate the results are shown in Table 8-9:
8. stability experiment
With the need testing solution sample introduction analysis for preparing, every 1 hour sample introduction, measure the peak area integrated value by above-mentioned chromatographic condition, calculating relative standard deviation is 1.43%, 1.40%.Show that sample is basicly stable in 8h.The results are shown in Table 10:
9. different lot numbers are transferred magnolol and honokiol assay in the stomach pellet:
9.1 the preparation precision of reference substance solution takes by weighing the magnolol reference substance, the honokiol reference substance is an amount of, adds methanol and makes the solution that every 1mL contains magnolol 135.68ug, honokiol 44.896ug respectively, promptly.
Table 8 honokiol response rate experimental result
Table 9 magnolol response rate experimental result
Table 10 stability experiment result
9.2 the about 0.5g of different lot number this product powder is got in the preparation of need testing solution, the accurate title, decide, and to 50mL tool plug triangular flask, adds methanol (100%) 25mL, weigh, supersound process 30 minutes is taken out, and is placed to room temperature, weigh again, mend heavy after microporous filter membrane filters, promptly.
According to drafting method, draw each 5 μ L sample introduction of reference substance solution and need testing solution, measure.Measure 3 batches altogether and transferred magnolol and honokiol content in the stomach pellet, the results are shown in Table 11:
Magnolol and honokiol assay result in table 11 sample
Magnolol and honokiol assay result in table 11 sample
Limit: the every gram of this product contains Cortex Magnoliae Officinalis with magnolol (C
18H
18O
2) and honokiol (C
18H
18O
2) the total amount meter, must not be less than 1.6mg.
Claims (3)
1. a Chinese medicine preparation is transferred the method for quality control of stomach pellet, and wherein said pharmaceutical formulation is by Chinese crude drug:
Radix Aucklandiae 75g Fructus Amomi 75g Radix Glycyrrhizae 75g
Semen Arecae 75g Fructus Aurantii Immaturus (parched with bran) 75g Cortex Magnoliae Officinalis (processed with Rhizoma Zingiberis Recens) 75g
Rhizoma Cyperi (vinegar system) 75g Fructus Amomi Rotundus 75g Oletum Trogopterori (vinegar system) 50g
Rhizoma Alpiniae Officinarum 25g Flos Caryophylli 25g Cortex Cinnamomi 25g forms, method for making; More than 12 the flavor, be ground into fine powder, sieve, mixing, the water pill, polishing, drying promptly, is characterized in that: the step of its method is:
(1) be contrast with Semen Arecae, Fructus Aurantii Immaturus, Cortex Magnoliae Officinalis, cinnamomic microscopic features, microscope differentiates to transfer in the red prescription of stomach whether contain Semen Arecae, Fructus Aurantii Immaturus, Cortex Magnoliae Officinalis, Cortex Cinnamomi; With Neosynephrine, dehydrocostuslactone, Radix Glycyrrhizae is reference substance, and thin layer chromatography differentiates to transfer in the red prescription of stomach whether contain Neosynephrine, dehydrocostuslactone, licorice ingredient;
(2) be reference substance with magnolol, honokiol, high performance liquid chromatography detects the content of transferring Cortex Magnoliae Officinalis in the red prescription of stomach.
2. Chinese medicine preparation according to claim 1 is transferred the method for quality control of stomach pellet, it is characterized in that: the described microscope of putting differentiate to be transferred and not to be contained Semen Arecae, Fructus Aurantii Immaturus, Cortex Magnoliae Officinalis, cinnamomic method in the red prescription of stomach and be: get this product, putting microscopically observes: thin ancient piece of jade, round, flat and with a hole in its centre cell contains prism of calcium oxalate around the fibre bundle, forms crystalline cellulose; The endosperm cell debris is colourless, and wall is thicker, and more great similar round stricture of vagina hole is arranged; Prism of calcium oxalate is present in the Bao Bi tissue in flakes; Stone cell divides dendritic, and behind the wall, laminated striation is obvious; Square or the similar round of stone cell class, wall are simultaneously poor;
Described thin layer chromatography is differentiated and transferred the method that whether contains the Neosynephrine composition in the red prescription of stomach to be: get this product 5g, porphyrize adds methanol 15ml, and ultrasonic place 20 minutes filters, and filtrate evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Other gets the Neosynephrine reference substance, adds methanol and makes the solution that every 1ml contains 0.5mg, in contrast product solution; Test according to thin layer chromatography, draw each 2～5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-glacial acetic acid-water is developing solvent, with n-butyl alcohol: glacial acetic acid: the upper solution of water=4: 1: 5, launch, take out, dry, spray is with 0.5% ethanol solution of ninhydrin, and it is clear to be heated to speckle colour developing; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
Described thin layer chromatography is differentiated and transferred the method that whether contains the dehydrocostuslactone composition in the red prescription of stomach to be: get this product 5g, porphyrize adds chloroform 10ml, and ultrasonic place 30 minutes filters, and filtrate is as need testing solution; Other removes hydrogen constuslactone reference substance, adds chloroform and makes the solution that every 1ml contains 0.5mg, in contrast product solution; According to the thin layer chromatography test, draw each 4～6 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with cyclohexane extraction-acetone, with cyclohexane extraction: acetone=10: 2, launch, take out, dry, spray is with 5% vanillin sulfuric acid solution, and it is clear to be heated to speckle colour developing; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
Described thin layer chromatography is differentiated and is transferred the method that whether contains licorice ingredient in the red prescription of stomach to be: get this product 5g, porphyrize, the 40ml that adds diethyl ether, reflux 20 minutes, filter, discard filtrate, medicinal residues volatilize ether, add methanol 30ml, reflux 1 hour, filter, filtrate evaporate to dryness, residue add water 40ml makes dissolving, with water saturated n-butanol extraction 3 times, each 20ml merges n-butyl alcohol liquid, water 20ml washing, discard water liquid, n-butyl alcohol liquid evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 1g in addition, the 10ml that adds diethyl ether, reflux 20 minutes, filter, discard filtrate, medicinal residues volatilize ether, add methanol 10ml, reflux 1 hour filters, filtrate evaporate to dryness, residue add water 10ml makes dissolving, with water saturated n-butanol extraction 3 times, each 10ml merges n-butyl alcohol liquid, water 10ml washing, discard water liquid, n-butyl alcohol liquid evaporate to dryness shines medical material solution in pairs with legal system.Test according to thin layer chromatography, draw each 2～4 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with chloroform-methanol-water is developing solvent, chloroform: methanol: water=40: 10: 1, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to speckle colour developing; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
3. Chinese medicine preparation according to claim 1 is transferred the method for quality control of stomach pellet, and it is characterized in that: with magnolol, honokiol is reference substance, and liquid chromatography detects transfers the method for the content of Cortex Magnoliae Officinalis in the red prescription of stomach to be:
(1) chromatographic condition and system suitability test: chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Methanol: water=78: 22 is mobile phase; The detection wavelength is 294nm; Number of theoretical plate calculates by the magnolol peak should be not less than 3800;
(2) preparation of reference substance solution: get the magnolol reference substance, the honokiol reference substance is an amount of, accurately claim surely, add methanol and make the solution that every 1ml contains magnolol 130 μ g, honokiol 45 μ g, promptly;
(3) preparation of need testing solution: it is an amount of to get this product, is ground into fine powder, gets powder 0.5g, accurate claim fixed, the accurate methanol 25ml that adds, claim decide weight, supersound process 30 minutes is put coldly, and weight decided in title again, supplies the weight that subtracts mistake with methanol, shake up, filter, get subsequent filtrate, promptly;
(4) algoscopy: algoscopy is drawn above-mentioned reference substance solution and each 5 μ l of need testing solution respectively, injects chromatograph of liquid, measures, promptly; The every 1g of this product contains Cortex Magnoliae Officinalis with magnolol (C
18H
18O
2) and honokiol (C
18H
18O
2) the total amount meter, must not be less than 1.6mg.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102520112A (en) * | 2011-12-16 | 2012-06-27 | 重庆天生药业有限公司 | Quality control method of mangnolia officinalis refined medicinal slice |
| CN106404939A (en) * | 2016-08-30 | 2017-02-15 | 天津中新药业集团股份有限公司乐仁堂制药厂 | Content determination method of stomach conditioning pill |
| CN107202856A (en) * | 2017-05-23 | 2017-09-26 | 四川逢春制药有限公司 | A kind of detection method for the Chinese medicine preparation for treating flu |
| CN114487251A (en) * | 2021-12-02 | 2022-05-13 | 天津中新药业集团股份有限公司隆顺榕制药厂 | Thin-layer chromatography detection method of agilawood depression relieving tablets |
| CN115144251A (en) * | 2021-03-30 | 2022-10-04 | 山东新时代药业有限公司 | Method for processing biological sample |
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2009
- 2009-11-19 CN CN200910228360A patent/CN102068649B/en active Active
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102520112A (en) * | 2011-12-16 | 2012-06-27 | 重庆天生药业有限公司 | Quality control method of mangnolia officinalis refined medicinal slice |
| CN102520112B (en) * | 2011-12-16 | 2016-01-20 | 重庆天生药业有限公司 | A kind of method of quality control of mangnolia officinalis refined medicinal slice |
| CN106404939A (en) * | 2016-08-30 | 2017-02-15 | 天津中新药业集团股份有限公司乐仁堂制药厂 | Content determination method of stomach conditioning pill |
| CN107202856A (en) * | 2017-05-23 | 2017-09-26 | 四川逢春制药有限公司 | A kind of detection method for the Chinese medicine preparation for treating flu |
| CN115144251A (en) * | 2021-03-30 | 2022-10-04 | 山东新时代药业有限公司 | Method for processing biological sample |
| CN114487251A (en) * | 2021-12-02 | 2022-05-13 | 天津中新药业集团股份有限公司隆顺榕制药厂 | Thin-layer chromatography detection method of agilawood depression relieving tablets |
| CN114487251B (en) * | 2021-12-02 | 2023-11-03 | 津药达仁堂集团股份有限公司隆顺榕制药厂 | Thin-layer chromatography detection method for agilawood Shu Yupian |
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|---|---|
| CN102068649B (en) | 2012-10-24 |
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Address after: 300380 Yumen Road, Xiqing District, Tianjin Patentee after: Lerentang Pharmaceutical Factory of Jinyao Darentang Group Co.,Ltd. Address before: 300380 Yumen Road, Xiqing District, Tianjin Patentee before: Letrentang pharmaceutical factory of Tianjin Zhongxin Pharmaceutical Group Co.,Ltd. |