CN105287732A - Pharmaceutical formulation for treating gastric ulcer and preparation method of pharmaceutical formulation - Google Patents
Pharmaceutical formulation for treating gastric ulcer and preparation method of pharmaceutical formulation Download PDFInfo
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- CN105287732A CN105287732A CN201510834617.8A CN201510834617A CN105287732A CN 105287732 A CN105287732 A CN 105287732A CN 201510834617 A CN201510834617 A CN 201510834617A CN 105287732 A CN105287732 A CN 105287732A
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- radix tinosporae
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- palmatine hydrochloride
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Abstract
The invention discloses a pharmaceutical formulation for treating gastric ulcer and a preparation method of the pharmaceutical formulation. The pharmaceutical formulation comprises main materials of 4 parts of radix tinosporae and 1 part of Chinese olive, and comprises agents of ethanol, methanol, a standard substance of palmatine hydrochloride, and a standard substance of gallic acid. The preparation method comprises: extraction of active ingredients of radix tinosporae and Chinese olive, measurement of the content of the active ingredients of radix tinosporae and Chinese olive, preparation of a solution of a reference substance, exploration of a linear relation, and preparation of a test solution. In comparison with a medicine for treating related diseases in the prior art, the pharmaceutical formulation is relatively pure, is accurate in curative effect and short in course of treatment, and has no toxic or side effect. In comparison with the prior art, the preparation method takes 70% of ethanol as an extraction solvent, has no toxic effect on the body, is low in cost, and is suitable for industrial mass production. The main anti-ulcer component of palmatine hydrochloride in radix tinosporae is subjected to optimization of an extraction process and quantitative analysis through high performance liquid chromatography. The pharmaceutical formulation and the preparation method provide a theoretical reference for promoting development and utilization of radix tinosporae, and have promotion and application value.
Description
Technical field
The present invention relates to a kind of plant component extraction technique, particularly relate to a kind of pharmaceutical formulation for the treatment of gastric ulcer and preparation method thereof.
Background technology
Radix Tinosporae, Chinese medicine name.The dried root of menispermaceous plants limacia sagittata or Radix Tinosporae.Autumn, season in winter two excavate, and removing fibrous root, cleans, dry.Nature and flavor bitter cold, has heat-clearing and toxic substances removing, sore-throat relieving, effect of pain relieving.For laryngopharynx swelling and pain, carbuncle furunculosis, has loose bowels, dysentery, epigastric pain, and Radix Tinosporae has now been widely used in anti-inflammation widely, viral infection resisting, paroxysmal pain, and enhancing human body immunity ability.The extracting method of Radix Tinosporae effective ingredient has water extraction, steam distillation and organic solvent extractionprocess etc.There are research reflux extraction, ultrasonic extraction, Soakage extraction method three kinds of methods in early stage to extract the effective ingredient of Radix Tinosporae, but for the extraction process of Radix Tinosporae, at present, has no relevant report, therefore, there is room for improvement.
Summary of the invention
Object of the present invention is just to provide a kind of pharmaceutical formulation for the treatment of gastric ulcer and preparation method thereof to solve the problem.
The present invention is achieved through the following technical solutions above-mentioned purpose:
A kind of pharmaceutical formulation for the treatment of gastric ulcer of the present invention, comprise main material: Radix Tinosporae 4 parts, Fructus Canarii 1 point, reagent: ethanol, methanol, palmatine hydrochloride standard substance, gallic acid standard substance, further, it is for subsequent use that described Radix Tinosporae, Fructus Canarii cross No. 4 sieves after all adopting and drying dozen powder, described ethanol is analytical pure, and described methanol is chromatographically pure.
A kind of preparation method for the treatment of the Chinese medicine formula of gastric ulcer of the present invention, comprises the following steps:
(1) extraction of Radix Tinosporae effective ingredient: 70% alcoholic solution of 10 times of volumes is added in Radix Tinosporae medicinal powder, reflux, extract, twice, each 0.5h, namely Radix Tinosporae extracting solution is obtained after filtration, vacuum drying oven is put into by after extracting solution rotary evaporation to 30mL, 105 DEG C to be dried to after constant weight to obtain Radix Tinosporae dry extract, for subsequent use;
(2) mensuration of Radix Tinosporae active constituent content: chromatographic condition: DiamonsilTMC18 chromatographic column, is specially 4.6mm × 250mm, 5 μm; Mobile phase: methanol and water ratio are 1:1, wherein containing tetramethylethylenediamine 3.5mLL
-1, adjust pH to 7.5 with phosphoric acid; Determined wavelength: 345nm; Flow velocity: 1.0mLmin
-1; Column temperature: 25 DEG C; Sensitivity: 0.1AUFS; Theoretical cam curve calculates by palmatine hydrochloride and is not less than 1500,
(3) preparation of reference substance solution: get palmatine hydrochloride standard substance, accurately weighed, dissolve with mobile phase and dilute, making the solution of hydrochloric palmatine 60 μ g in every 1mL, shake up;
(4) linear relationship is investigated: precision takes palmatine hydrochloride standard substance 3mg, be placed in 50mL measuring bottle, be diluted to scale with dissolve with methanol, shake up, precision measures in solution 1mL to 10mL measuring bottle, add methanol dilution respectively to scale, dilute 5 concentration successively, measure peak area by above-mentioned chromatographic condition, take peak area as vertical coordinate, content is abscissa, drawing standard curve, and regression equation is Y=372510X+99044.Result shows, palmatine hydrochloride is at 1.875 ~ 60mgL
-1in scope, r=0.99996, concentration and peak area are good linear relation;
(5) preparation of need testing solution: get dry extract 0.1g, adds mobile phase appropriate, with mobile phase standardize solution in 5mL volumetric flask, shakes up, as need testing solution;
(6) reference substance solution in the accurate aspiration step of difference (3) and each 5uL of need testing solution in step (5), injection liquid chromatography, measure by above-mentioned chromatographic condition, record chromatogram is to 2 times of main peak retention time, with external standard method, calculate the content of palmatine hydrochloride in need testing solution;
(7) employing step (1) carries out same method processing to the method for step (6) to Fructus Canarii, difference is, step (1) is replaced with add 8 times of volumes 70% alcoholic solution in Fructus Canarii medicinal powder, reflux, extract, three times, each 1.5h, namely obtains Fructus Canarii extracting solution after filtration, puts into vacuum drying oven by after extracting solution rotary evaporation to 30mL, 105 DEG C to be dried to after constant weight to obtain Fructus Canarii dry extract, for subsequent use; Reference substance palmatine hydrochloride standard substance in step (4) are replaced by gallic acid standard substance, and Radix Tinosporae dry extract and Fructus Canarii dry extract can obtain finished product according to the ratio mixing of crude drug content 4:1.
Beneficial effect of the present invention is:
The present invention is a kind of pharmaceutical formulation for the treatment of gastric ulcer and preparation method thereof, and compared with the medicine of existing treatment relevant disease, present component is relatively simple, determined curative effect, and short treating period, has no side effect; Compared with prior art, the present invention for Extraction solvent, to body nonhazardous effect, and with low cost again, is applicable to the production of industrial mass with 70% ethanol to preparation method.Adopting high performance liquid chromatography antiulcer composition palmatine hydrochloride main in Radix Tinosporae to be carried out to optimization and the quantitative analysis of extraction process, for promoting that the exploitation of Radix Tinosporae provide theoretical reference, there is the value applied.
Accompanying drawing explanation
Fig. 1 is palmatine hydrochloride canonical plotting of the present invention;
Fig. 2 is reference substance of the present invention and sample contrast color spectrogram.
Detailed description of the invention
Below in conjunction with accompanying drawing, the invention will be further described:
As depicted in figs. 1 and 2: a kind of pharmaceutical formulation for the treatment of gastric ulcer of the present invention, comprise main material: Radix Tinosporae 4 parts, Fructus Canarii 1 point, reagent: ethanol, methanol, palmatine hydrochloride standard substance, gallic acid standard substance, further, it is for subsequent use that described Radix Tinosporae, Fructus Canarii cross No. 4 sieves after all adopting and drying dozen powder, described ethanol is analytical pure, and described methanol is chromatographically pure.
A kind of preparation method for the treatment of the Chinese medicine formula of gastric ulcer of the present invention, comprises the following steps:
(1) extraction of Radix Tinosporae effective ingredient: 70% alcoholic solution of 10 times of volumes is added in Radix Tinosporae medicinal powder, reflux, extract, twice, each 0.5h, namely Radix Tinosporae extracting solution is obtained after filtration, vacuum drying oven is put into by after extracting solution rotary evaporation to 30mL, 105 DEG C to be dried to after constant weight to obtain Radix Tinosporae dry extract, for subsequent use;
(2) mensuration of Radix Tinosporae active constituent content: chromatographic condition: DiamonsilTMC18 chromatographic column, is specially 4.6mm × 250mm, 5 μm; Mobile phase: methanol and water ratio are 1:1, wherein containing tetramethylethylenediamine 3.5mLL
-1, adjust pH to 7.5 with phosphoric acid; Determined wavelength: 345nm; Flow velocity: 1.0mLmin
-1; Column temperature: 25 DEG C; Sensitivity: 0.1AUFS; Theoretical cam curve calculates by palmatine hydrochloride and is not less than 1500,
(3) preparation of reference substance solution: get palmatine hydrochloride standard substance, accurately weighed, dissolve with mobile phase and dilute, making the solution of hydrochloric palmatine 60 μ g in every 1mL, shake up;
(4) linear relationship is investigated: precision takes palmatine hydrochloride standard substance 3mg, be placed in 50mL measuring bottle, be diluted to scale with dissolve with methanol, shake up, precision measures in solution 1mL to 10mL measuring bottle, add methanol dilution respectively to scale, dilute 5 concentration successively, measure peak area by above-mentioned chromatographic condition, take peak area as vertical coordinate, content is abscissa, drawing standard curve, and regression equation is Y=372510X+99044.Result shows, palmatine hydrochloride is at 1.875 ~ 60mgL
-1in scope, r=0.99996, concentration and peak area are good linear relation;
(5) preparation of need testing solution: get dry extract 0.1g, adds mobile phase appropriate, with mobile phase standardize solution in 5mL volumetric flask, shakes up, as need testing solution;
(6) reference substance solution in the accurate aspiration step of difference (3) and each 5uL of need testing solution in step (5), injection liquid chromatography, measure by above-mentioned chromatographic condition, record chromatogram is to 2 times of main peak retention time, with external standard method, calculate the content of palmatine hydrochloride in need testing solution;
(7) employing step (1) carries out same method processing to the method for step (6) to Fructus Canarii, difference is, step (1) is replaced with add 8 times of volumes 70% alcoholic solution in Fructus Canarii medicinal powder, reflux, extract, three times, each 1.5h, namely obtains Fructus Canarii extracting solution after filtration, puts into vacuum drying oven by after extracting solution rotary evaporation to 30mL, 105 DEG C to be dried to after constant weight to obtain Fructus Canarii dry extract, for subsequent use; Reference substance palmatine hydrochloride standard substance in step (4) are replaced by gallic acid standard substance, and Radix Tinosporae dry extract and Fructus Canarii dry extract can obtain finished product according to the ratio mixing of crude drug content 4:1.
Interpretation:
Extraction time is on the impact of extraction ratio
Choose 3 temperature values to test, the results are shown in Table 1.Experimental result shows, along with the increase of time, extraction ratio improves gradually; Time reaches 1.5 hours, and extraction ratio is maximum; After this along with the increase extraction ratio of time slowly declines.Its reason may be owing to causing extraction ratio to decline along with the rapid stripping of impurity in the increase medical material of time.Therefore 1.5 hours is this experiment the suitableeest extraction time.
The relation (n=3) of table 1 extraction ratio and extraction time
Table1Therelationshipbetweenextractionrateandtime(n＝3)
Ethanol mass fraction is on the impact of extraction ratio
Choose 3 concentration of alcohol values to test, the results are shown in Table 2.Experimental result shows, along with the rising of concentration of alcohol, extraction ratio raises gradually; When concentration of alcohol reaches 70%, extraction ratio is the highest; After this along with the rising extraction ratio of methanol concentration declines.Because palmatine hydrochloride is water miscible alkaloid, therefore along with the rising of methanol concentration, extraction ratio presents the trend reduced gradually.Thus, the suitableeest ethanol mass fraction of this experiment is 70%.
The relation (n=3) of table 2 extraction ratio and ethanol mass fraction
Table2Therelationshipbetweenextractionrateandmassfractionofmethanol(n＝3)
Solid-to-liquid ratio is on the impact of extraction ratio
Choose 3 solid-to-liquid ratios to test, the results are shown in Table 3.Experimental result shows, along with increasing of solid-to-liquid ratio, extraction ratio raises gradually; When solid-to-liquid ratio reaches 1:8, extraction ratio is the highest; Solid-to-liquid ratio increases extraction ratio more afterwards then a small amount of minimizing.The raising of large solid-to-liquid ratio to extraction ratio is not obvious.
The relation (n=3) of table 3 extraction ratio and solid-to-liquid ratio
Table3Therelationshipbetweenextractionrateandsolid-liquidratio(n＝3)
The selection of each factor level
The mass fraction of extraction time, solvent, solid-to-liquid ratio and extraction time are the key factors determining extraction ratio.On the basis of single factor test screening, intend by 4 factor 3 level design experiment (table 4).
Table 4 orthogonal test factor level table
Table4thedesignofL
9(3
4)orthogonaltest
Orthogonal
According to L
9(3
4) orthogonal table experiment arrangement.Sample thief powder 30g, puts in round-bottomed flask, adds the ethanol of respective quality mark and solid-to-liquid ratio respectively, fully leaves standstill 1 hour, reflux, extract, after mixing, filters, obtain filtrate.Put into vacuum drying oven by after filtrate rotary evaporation to 30mL, 105 DEG C are dried to constant weight and obtain sample.With the palmatine hydrochloride content of high effective liquid chromatography for measuring orthogonal experiment gained, and pass through the yield of extract (table 5) of orthogonal test gained.
Comprehensive grading=(palmatine hydrochloride extraction ratio/maximum palmatine hydrochloride extraction ratio × 0.5+ yield of extract/maximum yield of extract × 0.5) × 100
The intuitive analysis result of comprehensive grading is D>B>A>C, and intuitive analysis optimised process is D
2b
3a
2c
1, be namely 1:10 by solid-to-liquid ratio, with 70% ethanol for Extraction solvent, extraction time is 0.5 hour, extracts 2 times.
Table 5 orthogonal and result
Table5Resultsoforthogonaltest
A: the mass fraction of solvent; B: solid-to-liquid ratio; C: time; D: extraction time
A:Massfractionofethanol；B:Solid-liquidratio；C:Time；D:Extractiontime
Confirmatory experiment
Take 3 parts of each 30g of sample powder respectively, add 10 times amount crude drug powders, 70% ethanol of respective volume, extract 0.5 hour, extract 2 times.Reflux, extract, filters, obtains filtrate.Put into vacuum drying oven by after filtrate rotary evaporation to 30mL, 105 DEG C are dried to constant weight, obtain sample.Measure under the same conditions.The results are shown in Table 6.
Table 6 demonstration test result
Table6Resultsofvalidation
More than show and describe ultimate principle of the present invention and principal character and advantage of the present invention.The technical staff of the industry should understand; the present invention is not restricted to the described embodiments; what describe in above-described embodiment and description just illustrates principle of the present invention; without departing from the spirit and scope of the present invention; the present invention also has various changes and modifications, and these changes and improvements all fall in the claimed scope of the invention.Application claims protection domain is defined by appending claims and equivalent thereof.
Claims (3)
1. treat a pharmaceutical formulation for gastric ulcer, it is characterized in that: comprise main material: Radix Tinosporae 4 parts, Fructus Canarii 1 point, reagent: ethanol, methanol, palmatine hydrochloride standard substance, gallic acid standard substance.
2. the pharmaceutical formulation for the treatment of gastric ulcer according to claim 1, is characterized in that: it is for subsequent use that described Radix Tinosporae, Fructus Canarii cross No. 4 sieves after all adopting and drying dozen powder, and described ethanol is analytical pure, and described methanol is chromatographically pure.
3. treat a preparation method for the Chinese medicine formula of gastric ulcer, it is characterized in that, comprise the following steps:
(1) extraction of Radix Tinosporae effective ingredient: 70% alcoholic solution of 10 times of volumes is added in Radix Tinosporae medicinal powder, reflux, extract, twice, each 0.5h, namely Radix Tinosporae extracting solution is obtained after filtration, vacuum drying oven is put into by after extracting solution rotary evaporation to 30mL, 105 DEG C to be dried to after constant weight to obtain Radix Tinosporae dry extract, for subsequent use;
(2) mensuration of Radix Tinosporae active constituent content: chromatographic condition: DiamonsilTMC18 chromatographic column, is specially 4.6mm × 250mm, 5 μm; Mobile phase: methanol and water ratio are 1:1, wherein containing tetramethylethylenediamine 3.5mLL
-1, adjust pH to 7.5 with phosphoric acid; Determined wavelength: 345nm; Flow velocity: 1.0mLmin
-1; Column temperature: 25 DEG C; Sensitivity: 0.1AUFS; Theoretical cam curve calculates by palmatine hydrochloride and is not less than 1500,
(3) preparation of reference substance solution: get palmatine hydrochloride standard substance, accurately weighed, dissolve with mobile phase and dilute, making the solution of hydrochloric palmatine 60 μ g in every 1mL, shake up;
(4) linear relationship is investigated: precision takes palmatine hydrochloride standard substance 3mg, be placed in 50mL measuring bottle, be diluted to scale with dissolve with methanol, shake up, precision measures in solution 1mL to 10mL measuring bottle, add methanol dilution respectively to scale, dilute 5 concentration successively, measure peak area by above-mentioned chromatographic condition, take peak area as vertical coordinate, content is abscissa, drawing standard curve, and regression equation is Y=372510X+99044.Result shows, palmatine hydrochloride is at 1.875 ~ 60mgL
-1in scope, r=0.99996, concentration and peak area are good linear relation;
(5) preparation of need testing solution: get dry extract 0.1g, adds mobile phase appropriate, with mobile phase standardize solution in 5mL volumetric flask, shakes up, as need testing solution;
(6) reference substance solution in the accurate aspiration step of difference (3) and each 5uL of need testing solution in step (5), injection liquid chromatography, measure by above-mentioned chromatographic condition, record chromatogram is to 2 times of main peak retention time, with external standard method, calculate the content of palmatine hydrochloride in need testing solution;
(7) employing step (1) carries out same method processing to the method for step (6) to Fructus Canarii, difference is, step (1) is replaced with add 8 times of volumes 70% alcoholic solution in Fructus Canarii medicinal powder, reflux, extract, three times, each 1.5h, namely obtains Fructus Canarii extracting solution after filtration, puts into vacuum drying oven by after extracting solution rotary evaporation to 30mL, 105 DEG C to be dried to after constant weight to obtain Fructus Canarii dry extract, for subsequent use; Reference substance palmatine hydrochloride standard substance in step (4) are replaced by gallic acid standard substance, and Radix Tinosporae dry extract and Fructus Canarii dry extract can obtain finished product according to the ratio mixing of crude drug content 4:1.
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| CN1857659A (en) * | 2006-04-03 | 2006-11-08 | 夏书桂 | Gastropathy treating aromatic Chinese medicine |
| CN1939414A (en) * | 2005-09-26 | 2007-04-04 | 蔡军 | Medicinal composition with antibacterial and anti-inflammation functions |
| CN101780161A (en) * | 2009-09-02 | 2010-07-21 | 贵州百灵企业集团制药股份有限公司 | Capsule quality detection method |
| CN102379949A (en) * | 2011-11-17 | 2012-03-21 | 福州海王金象中药制药有限公司 | Preparation process of sini powder |
| CN103768564A (en) * | 2014-01-14 | 2014-05-07 | 唐启裕 | Traditional Chinese medicine for treating acute and chronic gastritis |
-
2015
- 2015-11-25 CN CN201510834617.8A patent/CN105287732A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1939414A (en) * | 2005-09-26 | 2007-04-04 | 蔡军 | Medicinal composition with antibacterial and anti-inflammation functions |
| CN1857659A (en) * | 2006-04-03 | 2006-11-08 | 夏书桂 | Gastropathy treating aromatic Chinese medicine |
| CN101780161A (en) * | 2009-09-02 | 2010-07-21 | 贵州百灵企业集团制药股份有限公司 | Capsule quality detection method |
| CN102379949A (en) * | 2011-11-17 | 2012-03-21 | 福州海王金象中药制药有限公司 | Preparation process of sini powder |
| CN103768564A (en) * | 2014-01-14 | 2014-05-07 | 唐启裕 | Traditional Chinese medicine for treating acute and chronic gastritis |
Non-Patent Citations (3)
| Title |
|---|
| 南京中医药大学: "《中药大辞典》", 31 March 2006 * |
| 许尤琪等: "《防癌抗癌宜吃的食物》", 30 September 2015 * |
| 陈黎: "湖北金果榄物质基础研究", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
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Application publication date: 20160203 |