CN102507842A - Testing method for Xueshuan xinmaining tablet - Google Patents
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Abstract
The invention relates to a testing method for a Xueshuan xinmaining tablet, and belongs to the field of traditional Chinese medicine testing. The method comprises an identification method for total saponin in stems and leaves of a ginseng and calculus bovis factitious as well as a content testing method for saponin in the stems and the leaves of a ginseng, normalizes the testing method for the Xueshuan xinmaining tablet, solves the problems of severe spot diffusion and tailing existing in identification of cholic acid in calculus bovis factitious in a raw tablet, and coordinates to identify and determinate the content of total saponin in the stems and the leaves of the ginseng, as a result, the components of a tested adjuvant can be tested more comprehensively, the content and the reliability of the adjuvant can be recognized, and the explanation of material base for drug taking is facilitated.
Description
Technical field
The invention belongs to modern Chinese traditional medicine field, be specifically related to a kind of detection method of XUESHUANXINMAINING sheet.
Background technology
Traditional Chinese medicine ingredients is complicated, and particularly Chinese medicine compound prescription often contains multiclass, multiple composition, has constituted a very complicated system.At present, the new active component Chinese medicine preparation and the research and development of modern Chinese traditional compound medicine are on the increase, and new technology and multiple different extracting mode also are used for the research and the production of medicine.
Along with the fast development of modern analysis measuring technology, equipment and instrument, the assay of Chinese medicine, chromatogram are differentiated the important means that becomes detection.But the assay or the discriminating of generally only setting up a certain index chemical constitution at present, to the simultaneously applied situation of above-mentioned multiple new technology, the assay or the discriminating of only setting up certain single component have bigger one-sidedness to a great extent.
The XUESHUANXINMAINING sheet has qi and activate blood circulation, and the pain-relieving functions of having one's ideas straightened out is used for apoplexy, the obstruction of qi in the chest due to the qi deficiency to blood stasis, and disease opinion is had a dizzy spell, hemiplegia, pained, palpitation uncomfortable in chest; Ishemic stroke convalescence, coronary disease and angina pectoris are seen above-mentioned disease person.
The XUESHUANXINMAINING sheet is a Film coated tablets, removes to show brown behind the film-coating; The little perfume (or spice) of gas, mildly bitter flavor.
Its prescription is: Ligusticum wallichii 500g, red sage root 500g, sophora flower 250g, leech 125g, ilex pubescens 250g, muscone 1.25g, calculus bovis factitius 12.5g, ginsenoside 25g, borneol 2.5g, dried venom of toads 1.25g;
Method for making: above ten flavors, get muscone, borneol, calculus bovis factitius, ginseng stem and leave general saponin, the dried venom of toads and be ground into fine powder; The red sage root, ilex pubescens be with 80% alcohol reflux 2 times, and each 1.5 hours, filter, merge aluminium filtrating, reclaim ethanol, being concentrated into relative density is the thick paste of 1.28～1.30 (60 ℃), the dregs of a decoction are subsequent use.Ligusticum wallichii extracts volatile oil, and it is subsequent use to collect volatile oil, and distillation rear solution device is in addition collected, and the dregs of a decoction are subsequent use.The dregs of a decoction after the leech and the red sage root, the ilex pubescens alcohol extracting merge, and boiling 2 hours filters; Filtrating is placed, and the dregs of a decoction and the above-mentioned Ligusticum wallichii dregs of a decoction merge, boiling 2 hours; Filter, filtrating merges with above-mentioned each decocting liquid, and being concentrated into relative density is the thick paste of 1.28～1.30 (60 ℃); Mix with the red sage root, ilex pubescens alcohol extracting cream,, be ground into fine powder in 60～70 ℃ of dryings; Get sophora flower and add suitable quantity of water, regulate pH value to 8～9, be heated to little boiling, be incubated 30 minutes, filter while hot with saturated calcium hydroxide aqueous solution; The dregs of a decoction as above method extract 2 times again, merging filtrate, and 60～70 ℃ are stirred adding hydrochloric acid, regulate pH value to 4～5, leave standstill 1～2 hour; The suction filtration supernatant, deposition is with an amount of washing 3～4 times, and 60 ℃ of drying under reduced pressure are ground into fine powder, with above-mentioned each fine powder mixing; Granulate, drying adds an amount of talcum powder, dolomol, carboxyrnethyl starch sodium; Spray adds rhizoma chuanxiong volatile oil, and mixing is pressed into 1000, and the bag film-coating promptly gets.Every heavy 0.40g; Oral.One time 2,3 times on the one.
In the side with hot temperature, go into the liver and gall warp, activating QI to alleviate the depression dispels pathogenic wind and remove dampness, the Ligusticum wallichii of promoting blood circulation and stopping pain and bitter tepor are gone into the conscience warp, promoting blood circulation, calming heart and tranquilizing mind be pain, the red sage root that nourishes blood is monarch drug in a prescription altogether.Minister is with ilex pubescens, leech, sophora flower, muscone, and is promoting blood circulation and removing blood stasis, the inducing resuscitation of having one's ideas straightened out, removes obstruction in channels to relieve pain.Assistant is promoting blood circulation and removing blood stasis with borneol, calculus bovis factitius, ginseng stem and leave general saponin three medicines assistant the monarch and his subjects, and the inducing resuscitation of having one's ideas straightened out is removed obstruction in channels to relieve pain, the hard kidney water of the liver that clears away heart-fire, and it is main taking stopgap measures, and takes into account the gesture of this void and Yu Huare.Make with the dried venom of toads, pain relieving is kept fit by the priming sick institute that goes directly, current channels and collaterals.
The discriminating that in the original detection method of XUESHUANXINMAINING sheet only there are pair ginsenoside, borneol in the discriminating and the assay aspect of adjutant; Discriminating to cholic acid in the calculus bovis factitius in the former tablet is to use the extracted by ether need testing solution; Make and contain ether in the development system; Spot spreads, hangover is serious, and identification result has much room for improvement; Also not to the assay of ginsenoside, such survey detecting method has bigger one-sidedness to a great extent, can not comprehensively reflect the content of the important composition of XUESHUANXINMAINING sheet simultaneously.
Summary of the invention
The present invention provides a kind of detection method of XUESHUANXINMAINING sheet, to solve the incomplete problem of detection method that present detection method exists.
The technical scheme that the present invention takes is to comprise following discrimination method and content assaying method:
(1) discrimination method
(1) get 6 of these article, porphyrize adds methyl alcohol 50ml, and reflux 1 hour filters; The filtrating evaporate to dryness, residue adds water 30ml dissolving, with extracted by ether 2 times, each 20ml, merging ether solution; Evaporate to dryness, the WS is subsequent use, and residue adds methyl alcohol 2ml makes dissolving, as need testing solution; Other gets calculus bovis factitius control medicinal material 0.5g, adds methyl alcohol 5ml, and cold soaking spends the night, and filters, and filtrating is as control medicinal material solution; Get the cholic acid reference substance again, add methyl alcohol and process the solution that every 1ml contains 1mg, as reference substance solution; According to the thin-layered chromatography test, draw each 3 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate; With normal hexane-ethyl acetate-acetic acid-methyl alcohol 20: 25: 2: 3 upper solution was a developping agent; Launch, take out, dry; Spray is with 5% phosphomolybdic acid ethanol solution, and it is clear to be heated to the spot colour developing at 105 ℃;
(2) get discriminating (1) the subsequent use WS down, with water-saturated n-butanol jolting extraction 2 times, 20ml at every turn; Merge n-butanol extracting liquid, get n-butanol extracting liquid, with triplication ammonia solution washing 2 times; Normal butyl alcohol liquid evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution; Other gets ginsenoside Re's reference substance, ginsenoside Rg1's reference substance, ginseng stem and leave general saponin reference substance; Add methyl alcohol respectively and process the solution that every 1ml contains ginsenoside Re and ginsenoside Rg1's reference substance 1mg, ginseng stem and leave general saponin reference substance 5mg, as the contrast drug; According to thin-layered chromatography; Each 2 μ l of above-mentioned four kinds of solution are drawn in test, put respectively on same silica gel g thin-layer plate; With methenyl choloride-ethyl acetate-methanol-water 15: 40: 22:: 1,10 ℃ of lower floor's solution with held are developping agent; Launch, take out, dry; Spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the spot colour developing at 105 ℃;
(2) content assaying method:
Ginsenoside:
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; With acetonitrile-water is moving phase at 20: 80; The detection wavelength is 203nm, and number of theoretical plate calculates by the ginsenoside Re peak should be not less than 6000;
The preparation of reference substance solution: get ginsenoside Rg1's reference substance and ginsenoside Re's reference substance is an amount of, accurately claim surely, add methyl alcohol and process the mixed solution that every 1ml contains ginsenoside Rg1 0.15mg, ginsenoside Re 0.50mg, promptly get;
The preparation of need testing solution: get 20 of these article, the accurate title, decide, and porphyrize is got 0.4～0.6g, and accurate title is fixed, puts in the tool plug conical flask, adds methenyl choloride 50ml, and ultrasonic Extraction 30 minutes filters, and discards filtrating; The dregs of a decoction volatilize solvent together with filter paper, and the accurate methyl alcohol 50ml that adds claims decide weight, and refluxing extraction 1 hour is put coldly, and weight decided in title again; Supply the weight that subtracts mistake with methyl alcohol, shake up, filter, get subsequent filtrate 25ml, evaporate to dryness; Residue adds water-saturated n-butanol solution 20ml dissolving, with the washing of 50ml ammonia solution once, discards the ammonia solution layer, and normal butyl alcohol liquid evaporate to dryness, residue add methyl alcohol makes dissolving in right amount; Be transferred in the 10ml measuring bottle, add methyl alcohol to scale and make dissolving, shake up, filter, promptly get.
Advantage of the present invention is: standard the detection method of XUESHUANXINMAINING sheet; Solve the cholic acid discriminating exists in the calculus bovis factitius in the former tablet spot diffusion, the serious problem of hangover; And cooperation is to the discriminating and the assay of ginseng stem and leave general saponin; Can more comprehensively detect the composition of adjutant, understand its content and stability, also help to explain the medical substance basis of medicine.
Embodiment
The XUESHUANXINMAINING sheet is provided by prescription and explained hereafter in the background technology by Jilin Huakang Pharmaceutical Co., Ltd.
Embodiment 1
Comprise following discrimination method and content assaying method:
(1) discrimination method
(1) get 6 of these article, porphyrize adds methyl alcohol 50ml, and reflux 1 hour filters; The filtrating evaporate to dryness, residue adds water 30ml dissolving, with extracted by ether 2 times, each 20ml, merging ether solution; Evaporate to dryness, the WS is subsequent use, and residue adds methyl alcohol 2ml makes dissolving, as need testing solution; Other gets calculus bovis factitius control medicinal material 0.5g, adds methyl alcohol 5ml, and cold soaking spends the night, and filters, and filtrating is as control medicinal material solution; Get the cholic acid reference substance again, add methyl alcohol and process the solution that every 1ml contains 1mg, as reference substance solution; According to the thin-layered chromatography test, draw each 3 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate; With normal hexane-ethyl acetate-acetic acid-methyl alcohol 20: 25: 2: 3 upper solution was a developping agent; Launch, take out, dry; Spray is with 5% phosphomolybdic acid ethanol solution, and it is clear to be heated to the spot colour developing at 105 ℃;
(2) get discriminating (1) the subsequent use WS down, with water-saturated n-butanol jolting extraction 2 times, 20ml at every turn; Merge n-butanol extracting liquid, get n-butanol extracting liquid, with triplication ammonia solution washing 2 times; Normal butyl alcohol liquid evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution; Other gets ginsenoside Re's reference substance, ginsenoside Rg1's reference substance, ginseng stem and leave general saponin reference substance; Add methyl alcohol respectively and process the solution that every 1ml contains ginsenoside Re and ginsenoside Rg1's reference substance 1mg, ginseng stem and leave general saponin reference substance 5mg, as the contrast drug; According to thin-layered chromatography; Each 2 μ l of above-mentioned four kinds of solution are drawn in test, put respectively on same silica gel g thin-layer plate; With methenyl choloride-ethyl acetate-methanol-water 15: 40: 22:: 1,10 ℃ of lower floor's solution with held are developping agent; Launch, take out, dry; Spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the spot colour developing at 105 ℃;
(2) content assaying method:
Ginsenoside:
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; With acetonitrile-water is moving phase at 20: 80; The detection wavelength is 203nm, and number of theoretical plate calculates by the ginsenoside Re peak should be not less than 6000;
The preparation of reference substance solution: get ginsenoside Rg1's reference substance and ginsenoside Re's reference substance is an amount of, accurately claim surely, add methyl alcohol and process the mixed solution that every 1ml contains ginsenoside Rg1 0.15mg, ginsenoside Re 0.50mg, promptly get;
The preparation of need testing solution: get 20 of these article, the accurate title, decide, and porphyrize is got 0.4g, and accurate title is fixed, puts in the tool plug conical flask, adds methenyl choloride 50ml, and ultrasonic Extraction 30 minutes filters, and discards filtrating; The dregs of a decoction volatilize solvent together with filter paper, and the accurate methyl alcohol 50ml that adds claims decide weight, and refluxing extraction 1 hour is put coldly, and weight decided in title again; Supply the weight that subtracts mistake with methyl alcohol, shake up, filter, get subsequent filtrate 25ml, evaporate to dryness; Residue adds water-saturated n-butanol solution 20ml dissolving, with the washing of 50ml ammonia solution once, discards the ammonia solution layer, and normal butyl alcohol liquid evaporate to dryness, residue add methyl alcohol makes dissolving in right amount; Be transferred in the 10ml measuring bottle, add methyl alcohol to scale and make dissolving, shake up, filter, promptly get.
Embodiment 2
In content assaying method, the preparation of need testing solution: get 20 of these article, the accurate title, decide, and porphyrize is got 0.5g, and accurate title is fixed, and all the other are with embodiment 1.
Embodiment 3
In content assaying method, the preparation of need testing solution: get 20 of these article, the accurate title, decide, and porphyrize is got 0.6g, and accurate title is fixed, and all the other are with embodiment 1.
Every of these article contain ginsenoside with ginsenoside Rg1 (C
42H
72O
14) and ginsenoside Re (C
48H
82O
18) the total amount meter, be no less than 4.0mg.
Test Example 1:
Embodiment 2 is seen in the preparation of test sample, control medicinal material and reference substance, and other gets the negative control that does not contain calculus bovis factitius, processes negative control solution with method.According to thin-layered chromatography (an appendix VI of Chinese Pharmacopoeia version in 2010 B) test, draw each 3 μ l of above-mentioned four kinds of solution, put respectively in same with silica gel g thin-layer plate on; With normal hexane-ethyl acetate-acetic acid-methyl alcohol (20: 25: 2: 3) be developping agent; Launch, take out, dry; Spray is with 5% phosphomolybdic acid ethanol solution, and it is clear to be heated to the spot colour developing at 105 ℃.In the test sample chromatogram, with reference substance, the corresponding position of control medicinal material chromatogram on, show the spot of same color.Negative control is noiseless.Tested 5 lot sample article, all can reappear.
Test Example 2: face further specifies inventor's ginseng stem leaf saponin content assaying method through Test Example.
1, stability test
To with a need testing solution (Jilin Huakang Pharmaceutical Co., Ltd's lot number: 110201); By chromatographic condition of the present invention, measure once at regular intervals, investigate 25 hours altogether; Measured 6 times; Measure ginsenoside Rg1 and ginsenoside Re's content and average out to 4.3mg/ sheet 6 times as a result, RSD=0.63% (n=6) sees table 1 for details.
Table 1 ginsenoside stability test result
The result shows that need testing solution is stable in 25 hours.
2, precision test
Instrument precision test is to a need testing solution (Jilin Huakang Pharmaceutical Co., Ltd's lot number: 110201); By chromatographic condition of the present invention; Continuous sample introduction 6 times; Ginsenoside Rg1 and ginsenoside Re's peak area in the working sample, the RSD value is respectively 1.00% and 0.47%, and the result sees table 2.
Table 2 instrument precision test findings
The result shows that the precision of instrument is good.
3, replica test is to same sample (Jilin Huakang Pharmaceutical Co., Ltd's lot number: 110201); By 6 parts of method replicate determinations of the present invention, calculate these article contain ginsenoside Rg1 and ginsenoside Re and, mean value is the 4.3mg/ sheet; The RSD value is 1.39%, and the result sees table 3.
Table 3 ginsenoside reappearance experimental result
The result shows that the repeatability of this law is good.
4, specificity test
Be further to investigate the specificity of test, do not contain the negative control solution of ginsenoside according to the recipe quantity preparation, measure in accordance with the law, negative control solution is not having chromatographic peak with the corresponding retention time of reference substance chromatographic peak place as a result, shows noiselessly, and specificity is good.
5, linear relationship and scope
Accurate respectively ginsenoside Rg1's reference substance solution (lot number: 110703-200425 of drawing; Concentration 0.159mg/ml) and ginsenoside Re's reference substance solution (lot number: 110754-200421; Concentration 0.2406mg/ml) inject liquid chromatograph, measure by chromatographic condition of the present invention, the result sees the following form.Sample size (μ g) with reference substance is a horizontal ordinate, and chromatographic peak area is an ordinate drawing standard curve.The result sees table 4, table 5.
Table 4 ginsenoside Rg1 linear relationship is investigated the result
| Sample size (μ g) | 0.159 | 0.477 | 0.795 | 1.113 | 1.431 |
| The Rg1 peak area | 66605 | 213226 | 349093 | 496380 | 633770 |
Regression equation is: Y=-2556.2+445749.7X, and R=0.9999, the result shows that the ginsenoside Rg1 is good log-linear relation (ginsenoside Rg1's linear relationship is seen assay Fig. 4 .12) in 0.159 μ g～1.431 μ g scopes
Table 5 ginsenoside Re linear relationship is investigated the result
| Sample size (μ g) | 0.7218 | 1.203 | 2.406 | 3.609 | 4.812 |
| The Re peak area | 250029 | 421690 | 816128 | 1220516 | 1638418 |
Regression equation is: Y=8238.5+337645.5X, and R=0.9999, the result shows that the ginsenoside Re is good linear relation (ginsenoside Re's linear relationship is seen assay Fig. 4 .13) in 0.7218 μ g～4.812 μ g scopes.
6, accuracy test
6.1 ginsenoside Rg1's accuracy test
Precision takes by weighing test sample (Jilin Huakang Pharmaceutical Co., Ltd's lot number: 110201) 6 parts of known content; Every part of 0.25g, accurate ginsenoside Rg1's reference substance solution (0.0324mg/ml, purity 96.3%) 20ml that adds; Extract, measure by method of the present invention; Calculate recovery rate, average recovery rate=96.5%, RSD=2.32% (n=6).Meet the requirement of quantitative test.The result sees table 6.
Table 6 ginsenoside Rg1 accuracy test result
6.2 ginsenoside Re's accuracy test
Precision takes by weighing test sample (Jilin Huakang Pharmaceutical Co., Ltd's lot number: 110201) 6 parts of measuring known content with method; Every part of 0.25g, accurate ginsenoside Re's reference substance solution (0.2256mg/ml, purity 88.8%) 20ml that adds; Extract, measure by method of the present invention; Calculate recovery rate, average recovery rate=96.4%, RSD=1.34% (n=6).Meet the requirement of quantitative test.The result sees table 7.
Table 7 ginsenoside Re accuracy test result
Claims (1)
1. the detection method of an XUESHUANXINMAINING sheet comprises the ginsenoside discrimination method:
6 of these article of getting, porphyrize adds methyl alcohol 50ml, and reflux 1 hour filters; The filtrating evaporate to dryness, residue adds water 30ml dissolving, with extracted by ether 2 times, each 20ml, merging ether solution; The WS is subsequent use, gets this subsequent use WS, extracts 2 times with the water-saturated n-butanol jolting, and each 20ml merges n-butanol extracting liquid; Get n-butanol extracting liquid, with triplication ammonia solution washing 2 times, normal butyl alcohol liquid evaporate to dryness, residue add methyl alcohol 1ml makes dissolving, as need testing solution; Other gets ginsenoside Re's reference substance, ginsenoside Rg1's reference substance, ginseng stem and leave general saponin reference substance; Add methyl alcohol respectively and process the solution that every 1ml contains ginsenoside Re and ginsenoside Rg1's reference substance 1mg, ginseng stem and leave general saponin reference substance 5mg, as the contrast drug; According to thin-layered chromatography; Each 2 μ l of above-mentioned four kinds of solution are drawn in test, put respectively on same silica gel g thin-layer plate; With methenyl choloride-ethyl acetate-methanol-water 15:40:22::1,10 ℃ of lower floor's solution with held is developping agent; Launch, take out, dry; Spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the spot colour developing at 105 ℃;
It is characterized in that, also comprise the discrimination method of calculus bovis factitius and the content assaying method of ginsenoside:
The discrimination method of calculus bovis factitius: get 6 of these article, porphyrize adds methyl alcohol 50ml, and reflux 1 hour filters; The filtrating evaporate to dryness, residue adds water 30ml dissolving, with extracted by ether 2 times, each 20ml, merging ether solution; The WS is subsequent use, and evaporate to dryness, residue add methyl alcohol 2ml makes dissolving, as need testing solution; Other gets calculus bovis factitius control medicinal material 0.5g, adds methyl alcohol 5ml, and cold soaking spends the night, and filters, and filtrating is as control medicinal material solution; Get the cholic acid reference substance again, add methyl alcohol and process the solution that every 1ml contains 1mg, as reference substance solution; According to the thin-layered chromatography test, draw each 3 μ l of above-mentioned three kinds of solution, put respectively on same silica gel g thin-layer plate; Upper solution with normal hexane-ethyl acetate-acetic acid-methyl alcohol 20:25:2:3 is a developping agent; Launch, take out, dry; Spray is with 5% phosphomolybdic acid ethanol solution, and it is clear to be heated to the spot colour developing at 105 ℃;
The ginsenoside content assaying method:
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filling agent; With acetonitrile-water 20:80 is moving phase; The detection wavelength is 203nm, and number of theoretical plate calculates by the ginsenoside Re peak should be not less than 6000;
The preparation of reference substance solution: get ginsenoside Rg1's reference substance and ginsenoside Re's reference substance is an amount of, accurately claim surely, add methyl alcohol and process the mixed solution that every 1ml contains ginsenoside Rg1 0.15mg, ginsenoside Re 0.50mg, promptly get;
The preparation of need testing solution: get 20 of these article, the accurate title, decide, and porphyrize is got 0.4 ~ 0.6g, and accurate title is fixed, puts in the tool plug conical flask, adds methenyl choloride 50ml, and ultrasonic Extraction 30 minutes filters, and discards filtrating; The dregs of a decoction volatilize solvent together with filter paper, and the accurate methyl alcohol 50ml that adds claims decide weight, and refluxing extraction 1 hour is put coldly, and weight decided in title again; Supply the weight that subtracts mistake with methyl alcohol, shake up, filter, get subsequent filtrate 25ml, evaporate to dryness; Residue adds water-saturated n-butanol solution 20ml dissolving, with the washing of 50ml ammonia solution once, discards the ammonia solution layer, and normal butyl alcohol liquid evaporate to dryness, residue add methyl alcohol makes dissolving in right amount; Be transferred in the 10ml measuring bottle, add methyl alcohol to scale and make dissolving, shake up, filter, promptly get.
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| CN116570645A (en) * | 2023-07-11 | 2023-08-11 | 吉林华康药业股份有限公司 | Pharmaceutical composition for preventing and treating vascular dementia |
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| CN110542734A (en) * | 2019-09-11 | 2019-12-06 | 内蒙古康恩贝药业有限公司 | Method for detecting total saponin content of ginseng stem leaves of compound traditional Chinese medicine |
| CN110542734B (en) * | 2019-09-11 | 2022-03-08 | 内蒙古康恩贝药业有限公司 | Method for detecting total saponin content of ginseng stem leaves of compound traditional Chinese medicine |
| CN116570645A (en) * | 2023-07-11 | 2023-08-11 | 吉林华康药业股份有限公司 | Pharmaceutical composition for preventing and treating vascular dementia |
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