CN1785306B

CN1785306B - Quality control method of compound Siwei fructus ligustri lucidi preparation

Info

Publication number: CN1785306B
Application number: CN 200510200682
Authority: CN
Inventors: 张芝庭; 郭宗华; 黄�俊; 雷蕾
Original assignee: GUIZHOU MAOJKA GROUP HOLDING CO Ltd
Current assignee: GUIZHOU SHENQI PHARMACEUTICAL CO., LTD.
Priority date: 2005-11-10
Filing date: 2005-11-10
Publication date: 2011-07-27
Anticipated expiration: 2025-11-10
Also published as: CN1785306A

Abstract

A quality control method for the Chinese medicine 'compound Siwei Nuzhenzi' includes such steps as examining its characteristics, checking its capsule, tablet and particle specifications, identifying VB1, VB6 and VE and measuring the contents of VB1, VB6 and VE.

Description

The method of quality control of compound Siwei fructus ligustri lucidi preparation

Technical field: the present invention relates to a kind of method of quality control of compound Siwei fructus ligustri lucidi preparation, belong to technical field of medicaments.

Background technology: in the initial quality standard of compound Siwei fructus ligustri lucidi preparation, because detected parameters is few, quality control standard is low, can't effectively control the quality of medicine, therefore the product quality reliability of producing is not high, quality is unstable, has had a strong impact on the result of treatment of medicine and can't guarantee the safety of medication.

Summary of the invention:

The objective of the invention is to: a kind of method of quality control of compound Siwei fructus ligustri lucidi preparation is provided, and this method of quality control can effectively be controlled the quality of medicine, makes the reliable in quality, stable of medicine, guarantees the result of treatment and the drug safety of medicine.

Compound Siwei fructus ligustri lucidi preparation of the present invention is made all possible compound formulation that comprises capsule, tablet, granule by the medicine of following weight: oryzanol 10～20g, Cobastab ₁5～15g, Cobastab ₆5～15g, vitamin E 5～15g, biotin 1500～2005 μ g, taurine 10～50g, Herba Epimedii extractum 50～100g, Fructus Ligustri Lucidi extractum 80～150g, cultivated land medicinal extract 40～80g, puncturevine caltrop fruit medicinal extract 40～80g.

The present invention is achieved in that method of quality control of the present invention comprises proterties, discriminating, inspection and assay, and wherein proterties comprises: for capsule, the product content thing is that light brown is to brown granular or powder; For tablet, medicine shows light brown to sepia; For granule, product is that light brown is to tan particle; Discriminating comprises Cobastab ₁, Cobastab ₆, vitamin E discriminating; Check: should meet every regulation relevant under capsule, tablet, the granule item; Assay comprises with Cobastab ₁, Cobastab ₆, vitamin E is the content assaying method of reference substance.

Described discriminating is: it is an amount of that (1) gets compound Siwei fructus ligustri lucidi capsule, tablet or granule, removes unlap, porphyrize, add water 5～20ml, warmly make dissolving, filter, filtrate is put in the separating funnel, extract with ethyl acetate 10～30ml jolting, divide and get ethyl acetate liquid, filter, filtrate is put evaporate to dryness in the water-bath, residue adds ethanol 2～10ml makes dissolving, gets ethanol liquid 1～3ml and adds hydrochloric acid 1～3ml, the about 0.1～0.3g of magnesium powder, and solution shows orange or brownish red gradually; (2) get above-mentioned remaining ethanol liquid, be concentrated into about 0.5～2ml, drop on the filter paper, put under the ultraviolet lamp that wavelength is 365nm and inspect, show brown spot; Spray is with the ethanolic solution of 0.5～2% aluminium choride or smokedly then show pale brown color spot point with ammonia, and fluorescence is strengthened; (3) get 1～2 part of compound Siwei fructus ligustri lucidi capsule, tablet or granule, porphyrize adds chloroform 10～30ml, jolting 10 minutes, filter, filtrate is put evaporate to dryness in the water-bath, and residue adds ethanol 2～8ml makes dissolving, 2 of filtrate hydro-oxidation sodium test solutions, promptly show glassy yellow, continuing dropping sodium test solution 0.5～2ml, is to inspect under the ultraviolet lamp of 365nm at wavelength, shows yellow-green fluorescence; (4) get 1～2 part of compound Siwei fructus ligustri lucidi capsule, tablet or granule, porphyrize adds water 10～30ml, stirs and makes dissolving, filters, and gets filtrate 2～8ml, according to Cobastab ₁Differential method (1) item under shows identical reaction from " hydro-oxidation sodium test solution 2.5ml " test; (5) get above-mentioned remaining filtrate 1～3ml, thin up to 5～15ml is according to Cobastab ₆Differential method (1) item is tested from " respectively getting 1ml " in accordance with the law under, shows identical reaction.(6) get 2～6 parts of compound Siwei fructus ligustri lucidi capsule, tablet or granules, porphyrize adds water 10～30ml, stirring makes dissolving, gets filtrate 1～3ml, hydro-oxidation sodium 0.1～0.5g, after making dissolving, the heating evaporate to dryness is after the little live charcoalization, in 500 ℃ blazing 2～4 hours, put coldly, residue adds water 3～8ml, stir evenly, filter, filtrate adds 1 of sodium nitroprusside test solution, i.e. displaing yellow.

Cobastab ₁, Cobastab ₆Content assaying method comprise that with octadecylsilane key and silica gel be filling agent, 0.01～0.05mol/L acetonitrile: potassium dihydrogen phosphate is to be the high performance liquid chromatography of moving phase at 1～10: 99～90; The content assaying method of vitamin E comprises with silicone OV-17 being the vapor-phase chromatography of stationary phase.

Specifically, described Cobastab ₁, Cobastab ₆Content assaying method comprise:

(1) chromatographic condition and system suitability test: with octadecylsilane key and silica gel is filling agent; 0.01～0.05mol/L acetonitrile: potassium dihydrogen phosphate=1～10: 99～90 is moving phase; The detection wavelength is 276nm, and number of theoretical plate is pressed Cobastab ₁The peak calculates should be not less than 10～3000.

(2) preparation of reference substance solution: precision takes by weighing Cobastab ₁, Cobastab ₆Reference substance is an amount of, adds water or methyl alcohol and makes every 1ml and contain Cobastab ₁And Cobastab ₆Be respectively the solution of 0.05～0.3mg, promptly.

(3) preparation of need testing solution: it is an amount of that precision takes by weighing compound Siwei fructus ligustri lucidi capsule, tablet or granule, is equivalent to Cobastab ₁5～20mg, Cobastab ₆5～20mg puts in 50～100ml measuring bottle, and it is an amount of to add water, ultrasonicly makes moltenly, puts coldly, adds water to scale, shakes up, and filters with the miillpore filter of 0.45 μ m, promptly.

(4) determination method: accurate respectively reference substance solution and each 5～15 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly.

The assay of vitamin E comprises:

(1) chromatographic condition and system suitability experiment: the silicone with OV-17 is a stationary phase, and coating concentration is 1～5%; Column temperature is 200～300 ℃, and number of theoretical plate calculates by the vitamin E peak should be not less than 300～1000, and the degree of separation of vitamin E peak and internal standard compound mass peak should be greater than more than 1.5.

(2) correction factor is measured: it is an amount of to get n-dotriacontane, add n-hexane dissolution and be diluted to the solution that contains 0.25～1mg among every 1ml, shake up, as inner mark solution, other gets vitamin E reference substance 10～30mg, the accurate title, decide, and puts in the brown tool plug conical flask, the accurate inner mark solution 10ml that adds, close plug, jolting makes dissolving, gets 2 μ l inject gas chromatographs, the calculation correction factor.

(3) determination method: the content of getting compound Siwei fructus ligustri lucidi capsule, tablet or granule, mixing, precision take by weighing in right amount, are equivalent to vitamin e1 0～30mg, put in the brown tool plug conical flask, the accurate inner mark solution 5～20ml that adds, close plug, jolting makes dissolving, get 1～3 μ l inject gas chromatograph, measure, calculate, promptly.

More precisely:

Described discriminating is:

(1) it is an amount of to get compound Siwei fructus ligustri lucidi capsule, tablet or granule, removes unlap, porphyrize, add water 10ml, warmly make dissolving, filter, filtrate is put in the separating funnel, extract with ethyl acetate 15ml jolting, divide and get ethyl acetate liquid, filter, filtrate is put evaporate to dryness in the water-bath, residue adds ethanol 5ml makes dissolving, gets ethanol liquid 2ml and adds hydrochloric acid 2ml, the about 0.2g of magnesium powder, and solution shows orange or brownish red gradually; (2) get above-mentioned remaining ethanol liquid, be concentrated into about 1ml, drop on the filter paper, put under the ultraviolet lamp that wavelength is 365nm and inspect, show brown spot; Spray is with the ethanolic solution of 1% aluminium choride or smokedly then show pale brown color spot point with ammonia, and fluorescence is strengthened; (3) get 1 part of compound Siwei fructus ligustri lucidi capsule, tablet or granule, porphyrize adds chloroform 20ml, jolting 10 minutes, filter, filtrate is put evaporate to dryness in the water-bath, and residue adds ethanol 4ml makes dissolving, 2 of filtrate hydro-oxidation sodium test solutions, promptly show glassy yellow, continuing dropping sodium test solution 1ml, is to inspect under the ultraviolet lamp of 365nm at wavelength, shows yellow-green fluorescence; (4) get 1 part of compound Siwei fructus ligustri lucidi capsule, tablet or granule, porphyrize adds water 20ml, stirs and makes dissolving, filters, and gets filtrate 5ml, according to Cobastab ₁Differential method (1) item under shows identical reaction from " hydro-oxidation sodium test solution 2.5ml " test; (5) get above-mentioned remaining filtrate 2ml, thin up is to 10ml, according to Cobastab ₆Differential method (1) item is tested from " respectively getting 1ml " in accordance with the law under, shows identical reaction.(6) get 4 parts of compound Siwei fructus ligustri lucidi capsule, tablet or granules, porphyrize adds water 20ml, stirring makes dissolving, gets filtrate 2ml, hydro-oxidation sodium 0.3g, after making dissolving, the heating evaporate to dryness is after the little live charcoalization, in 500 ℃ blazing 2 hours, put coldly, residue adds water 5ml, stir evenly, filter, filtrate adds 1 of sodium nitroprusside test solution, i.e. displaing yellow.

Described Cobastab ₁, Cobastab ₆Content assaying method comprise:

(1) chromatographic condition and system suitability test: with octadecylsilane key and silica gel is filling agent; 0.025mol/L acetonitrile: potassium dihydrogen phosphate=3: 97 is a moving phase; The detection wavelength is 276nm, and number of theoretical plate is pressed Cobastab ₁The peak calculates should be not less than 1500.

(2) preparation of reference substance solution: precision takes by weighing Cobastab ₁, Cobastab ₆Reference substance is an amount of, adds water or methyl alcohol and makes every 1ml and contain Cobastab ₁And Cobastab ₆Be respectively the solution of 0.1mg, promptly.

(3) preparation of need testing solution: it is an amount of that precision takes by weighing compound Siwei fructus ligustri lucidi capsule, tablet or granule, is equivalent to Cobastab ₁10mg, Cobastab ₆10mg puts in the 100ml measuring bottle, and it is an amount of to add water, ultrasonicly makes moltenly, puts coldly, adds water to scale, shakes up, and filters with the miillpore filter of 0.45 μ m, promptly.

(4) determination method: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly.

The assay of vitamin E comprises:

(1) chromatographic condition and system suitability experiment: the silicone with OV-17 is a stationary phase, and coating concentration is 2%; Column temperature is 265 ℃, and number of theoretical plate calculates by the vitamin E peak should be not less than 500, and the degree of separation of vitamin E peak and internal standard compound mass peak should be greater than more than 2.

(2) correction factor is measured: it is an amount of to get n-dotriacontane, add n-hexane dissolution and be diluted to the solution that contains 0.5mg among every 1ml, shake up, as inner mark solution, other gets vitamin E reference substance 20mg, the accurate title, decide, and puts in the brown tool plug conical flask, the accurate inner mark solution 10ml that adds, close plug, jolting makes dissolving, gets 2 μ l inject gas chromatographs, the calculation correction factor.

(3) determination method: the content of getting compound Siwei fructus ligustri lucidi capsule, tablet or granule, mixing, precision take by weighing in right amount, are equivalent to vitamin E 20mg, put in the brown tool plug conical flask, the accurate inner mark solution 10ml that adds, close plug, jolting makes dissolving, get 2 μ l inject gas chromatographs, measure, calculate, promptly.

Generally speaking, the method for quality control of compound Siwei fructus ligustri lucidi preparation of the present invention comprises proterties, discriminating, inspection, assay, wherein:

Described proterties comprises: for capsule, the product content thing is that light brown is to brown granular or powder; For tablet, medicine shows light brown to sepia; For granule, product is that light brown is to tan particle;

Described discriminating comprises: it is an amount of that (1) gets compound Siwei fructus ligustri lucidi capsule, tablet or granule, removes unlap, porphyrize, add water 10ml, warmly make dissolving, filter, filtrate is put in the separating funnel, extract with ethyl acetate 15ml jolting, divide and get ethyl acetate liquid, filter, filtrate is put evaporate to dryness in the water-bath, residue adds ethanol 5ml makes dissolving, gets ethanol liquid 2ml and adds hydrochloric acid 2ml, the about 0.2g of magnesium powder, and solution shows orange or brownish red gradually; (2) get above-mentioned remaining ethanol liquid, be concentrated into about 1ml, drop on the filter paper, put under the ultraviolet lamp that wavelength is 365nm and inspect, show brown spot; Spray is with the ethanolic solution of 1% aluminium choride or smokedly then show pale brown color spot point with ammonia, and fluorescence is strengthened; (3) get 1 part of compound Siwei fructus ligustri lucidi capsule, tablet or granule, porphyrize adds chloroform 20ml, jolting 10 minutes, filter, filtrate is put evaporate to dryness in the water-bath, and residue adds ethanol 4ml makes dissolving, 2 of filtrate hydro-oxidation sodium test solutions, promptly show glassy yellow, continuing dropping sodium test solution 1ml, is to inspect under the ultraviolet lamp of 365nm at wavelength, shows yellow-green fluorescence; (4) get 1 part of compound Siwei fructus ligustri lucidi capsule, tablet or granule, porphyrize adds water 20ml, stirs and makes dissolving, filters, and gets filtrate 5ml, according to Cobastab ₁Differential method (1) item under shows identical reaction from " hydro-oxidation sodium test solution 2.5ml " test; (5) get above-mentioned remaining filtrate 2ml, thin up is to 10ml, according to Cobastab ₆Differential method (1) item is tested from " respectively getting 1ml " in accordance with the law under, shows identical reaction.(6) get 4 parts of compound Siwei fructus ligustri lucidi capsule, tablet or granules, porphyrize adds water 20ml, stirring makes dissolving, gets filtrate 2ml, hydro-oxidation sodium 0.3g, after making dissolving, the heating evaporate to dryness is after the little live charcoalization, in 500 ℃ blazing 2 hours, put coldly, residue adds water 5ml, stir evenly, filter, filtrate adds 1 of sodium nitroprusside test solution, i.e. displaing yellow;

Described inspection: should meet every regulation relevant under capsule, tablet, the granule item;

Described content assaying method comprises:

Cobastab ₁, Cobastab ₆Assay: (1) chromatographic condition and system suitability test: with octadecylsilane key and silica gel is filling agent; 0.025mol/L acetonitrile: potassium dihydrogen phosphate=3: 97 is a moving phase; The detection wavelength is 276nm, and number of theoretical plate is pressed Cobastab ₁The peak calculates should be not less than 1500; (2) preparation of reference substance solution: precision takes by weighing Cobastab ₁, Cobastab ₆Reference substance is an amount of, adds water or methyl alcohol and makes every 1ml and contain Cobastab ₁And Cobastab ₆Be respectively the solution of 0.1mg, promptly; (3) preparation of need testing solution: it is an amount of that precision takes by weighing compound Siwei fructus ligustri lucidi capsule, tablet or granule, is equivalent to Cobastab ₁10mg, Cobastab ₆10mg puts in the 100ml measuring bottle, and it is an amount of to add water, ultrasonicly makes moltenly, puts coldly, adds water to scale, shakes up, and filters with the miillpore filter of 0.45 μ m, promptly; (4) determination method: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly;

The assay of vitamin E comprises: (1) chromatographic condition and system suitability experiment: the silicone with OV-17 is a stationary phase, and coating concentration is 2%; Column temperature is 265 ℃, and number of theoretical plate calculates by the vitamin E peak should be not less than 500, and the degree of separation of vitamin E peak and internal standard compound mass peak should be greater than more than 2; (2) correction factor is measured: it is an amount of to get n-dotriacontane, add n-hexane dissolution and be diluted to the solution that contains 0.5mg among every 1ml, shake up, as inner mark solution, other gets vitamin E reference substance 20mg, the accurate title, decide, and puts in the brown tool plug conical flask, the accurate inner mark solution 10ml that adds, close plug, jolting makes dissolving, gets 2 μ l inject gas chromatographs, the calculation correction factor; (3) determination method: the content of getting compound Siwei fructus ligustri lucidi capsule, tablet or granule, mixing, precision take by weighing in right amount, are equivalent to vitamin E 20mg, put in the brown tool plug conical flask, the accurate inner mark solution 10ml that adds, close plug, jolting makes dissolving, get 2 μ l inject gas chromatographs, measure, calculate, promptly.

For above-mentioned method of quality control is verified, the applicant has carried out a series of test, research, and its result is as follows:

[assay]

(1) Cobastab ₁And Cobastab ₆:

1. instrument and reagent:

Instrument: Tianjin, island LC-10Avp high performance liquid chromatograph; The SPD-10AM diode array detector; The CLASS-VP chromatographic work station; Chromatographic column: Dalian Yi Lite ODS chromatographic column (post E1423925)

Reagent and reagent: Cobastab ₆(Nat'l Pharmaceutical ﹠ Biological Products Control Institute provides assay reference substance, lot number: 0116-9801) to reference substance; Cobastab ₁Reference substance (getting percentage concentration with feed purification is 99.64%); Potassium dihydrogen phosphate (analyzing pure); Acetonitrile (analyzing pure); Hydrochloric acid (analyzing pure); Water is double distilled water.

2. the selection of chromatographic condition: adopt ODS (C ₁₈) 250 * 4.6mm chromatographic column, through experiment, with the 0.025mol/L acetonitrile: potassium dihydrogen phosphate is separating effect the best that 3: 97 (pH=3) is moving phase.The detection wavelength is 276nm, and flow velocity is 0.8ml/min, 30 ℃ of column temperatures, and theoretical cam curve is with Cobastab ₁Note must not be less than 1500.

3. system suitability test: under the standard operation condition, prepare the preparation that all the other each raw materials are made except that vitamin B1 and vitamin B6 in this prescription respectively, detect with diode array detector.Negative control is not seen the absorption peak of vitamin B1, vitamin B6, and the absorption peak of vitamin B1 and vitamin B6 and spectral purity and reference substance are in full accord in the sample.Through repeatedly experiment, theoretical cam curve is with Cobastab ₁Note must not be less than 1500.

4. the investigation of linear relationship: (1) Cobastab ₁: precision takes by weighing Cobastab ₁Reference substance 0.0502g in the 50ml volumetric flask, water or methanol constant volume.Precision is measured 10ml in the 100ml volumetric flask, the water constant volume.Obtain the reference substance solution that concentration is 0.1004mg/ml.Draw 4,8,12,16,20 μ l sample introductions respectively, the record chromatogram.To sample size (μ g) regression Calculation, the straight-line equation that obtains, regression equation are C=9.123 * 10 with peak area (mAU.s) ^-7A-0.01456, correlation coefficient r=0.99959.Illustrate that sample size is good in 0.40 μ g～2.0 μ g scope internal linear relation.The results are shown in Table 1.

Table 1 Cobastab ₁Typical curve

Volume (μ l)	4 8 12 16 20
		Area (mAU.s) sample size (μ g)	446023 890751 1356977 1797034 21921200.4016 0.8032 1.2048 1.6064 2.008

Record the average peak area value by the one point method calculation correction factor with the precision test, getting the one point method equation is C=8.57253 * 10 ^-7A.Peak area measured value 446023 difference substitution regression equation C=9.123 * 10 with linearity range low concentration point ^-7A-0.01456 and former point equation C=8.57253 * 10 ^-7A calculates, and gets 0.3923 and 0.3824 μ g respectively, and its mean relative deviation is 1.3%, illustrates that the intercept of regression equation is less, can ignore, and an available point calibration method calculates content.

(2) Cobastab ₆: precision takes by weighing Cobastab ₆Reference substance 0.0501g in the 50ml volumetric flask, water or methanol constant volume.Precision is measured in 10ml to the 100ml volumetric flask, and the water constant volume obtains the reference substance solution that concentration is 0.1002mg/ml.Draw 4,8,12,16,20 μ l sample introductions respectively, the record chromatogram.To sample size (μ g) regression Calculation, obtain straight-line equation with peak area (mAU.s), regression equation is C=5.89324 * 10 ^-7A-0.03438, correlation coefficient r=0.9996.Illustrate that sample size is good in 0.40 μ g～2.0 μ g scope internal linear relation.The results are shown in Table 2.

Table 2 Cobastab ₆Typical curve

Volume (μ l)	4 8 12 16 20
		Area (mAU.s) sample size (μ g)	711271 1428382 2126499 2808132 34189090.4008 0.8016 1.2024 1.6032 2.004

Record the average peak area value by the one point method calculation correction factor with the precision test, getting the one point method equation is C=5.49001 * 10 ^-7A.Peak area measured value 711271 difference substitution regression equation C=5.89324 * 10 with linearity range low concentration point ^-7A-0.03438 and former point equation C=5.49001 * 10 ^-7A calculates, and gets 0.3848 and 0.3905 μ g respectively, and its mean relative deviation is 0.73%, illustrates that the intercept of regression equation is less, can ignore, and an available point calibration method calculates content.

5. precision test: get Cobastab ₁, vitamin B6 mixes reference substance solution 10 μ l, each sample introduction 5 times, and the record chromatogram, relative standard deviation (RSD) is respectively 0.35%, 0.47%, can find out that precision is good, the results are shown in Table 3, table 4.

Table 3 Cobastab ₁The precision test

Test number (TN)	1	2	3	4	5	Mean value	RSD(％)
								Peak area (A)	1174495	1173873	1164517	1170017	1173011	1171182.6	0.35

Table 4 Cobastab ₆The precision test

Test number (TN)	1	2	3	4	5	Mean value	RSD(％)
								Peak area (A)	1838483	1824407	1814253	1823904	1824616	1825132.6	0.47

6. replica test: (1) Cobastab ₁Repeatability: get 5 parts in same lot number sample, (be equivalent to Cobastab ₁10mg), handle in accordance with the law, sample introduction, the record chromatogram calculates Cobastab ₁Mean value is 29.06mg/g, and RSD% is 1.23%, shows that repeatability is good.The results are shown in Table 5.

Table 5 Cobastab ₁Replica test

Test number (TN)	1	2	3	4	5	Mean value	RSD％
								Content (mg/g)	29.59	28.62	29.12	28.87	29.09	29.06	1.23

(2) Cobastab ₆Repeatability: get 5 parts in same lot number sample, (be equivalent to Cobastab ₆10mg), handle in accordance with the law, sample introduction, the record chromatogram calculates Cobastab ₆Mean value is 29.20mg/g, and RSD% is 2.27%, shows that repeatability is good.The results are shown in Table 6.

Table 6 Cobastab ₆Replica test

Test number (TN)	1	2	3	4	5	Mean value	RSD％
								Content (mg/g)	29.63	29.97	28.23	29.00	29.16	29.20	2.27

7. recovery test:

(1) Cobastab ₁Get the sample with the same lot number of replica test, accurately claim surely, (getting concentration is the 2.011mg/ml Cobastab to add a certain amount of reference substance respectively ₁Reference substance solution 2.5ml, 4ml, 5ml), handle in accordance with the law, sample introduction, the record chromatogram is calculated and is recorded the recovery.Average recovery rate is 99.34%, and RSD is 0.54%, illustrates to reclaim well, the results are shown in Table 7.

Table 7 Cobastab ₁The application of sample recovery test

(2) Cobastab ₆Get the sample of testing same lot number with reappearance, the accurate title, decide, and adds a certain amount of reference substance (getting concentration is 2.153mg/ml vitamin B6 reference substance solution 2.5ml, 4ml, 5ml) respectively, handle in accordance with the law, and sample introduction, the record chromatogram calculates the recovery.Average recovery rate is 100.88, and RSD% is 0.60%, illustrates to reclaim well, the results are shown in Table 8.

Table 8 Cobastab ₆The application of sample recovery test

8. sample size is measured computing method: this determination method employing external standard method (" two appendix VI of Chinese pharmacopoeia version in 2005 D) calculate content, its formula is as follows:

(1) content (Mr)=Ax * Mr/Ar, when reference substance solution was represented with mg/ml, Mr represented that promptly reference substance is the concentration unit of mg/ml, the concentration that records sample simultaneously also is mg/ml.Draft every and contain Cobastab ₁, Cobastab ₆With the mg unit representation, i.e. mg/ grain, the substitution following formula calculates the content (mg/ grain) that is sample:

(2) content (mg/ grain)=Mx * P/ (Wx/V)

Wx is the weighing (mg) of sample in the formula, and V is average particle heavy (mg) for quantity of solvent (ml) P that extracts sample.Merge (1), (2) formula, obtain the cubage formula:

Content (mg/ grain)=Ax * Mr * V * P/ (Ar * Wx)

Table 9 Cobastab ₁, Cobastab ₂10 batch sample assays

Mean value according to 10 batch sample assay results: Cobastab ₁Be the 9.94mg/ grain, be 99.4% of labelled amount, Cobastab ₆Be the 9.82mg/ grain, be 98.2% of labelled amount.Each time result is all in the limit of labelled amount 90.0%～110.0%.

(2) vitamin E:

1. instrument and reagent:

Instrument: day island proper Tianjin GC-8A gas chromatograph.

Reagent and reagent: the vitamin E reference substance (Nat'l Pharmaceutical ﹠ Biological Products Control Institute provides the assay reference substance, lot number: 10062-0007), and n-dotriacontane (analyzing pure); Normal hexane (analyzing pure).

2. the selection of chromatographic condition: with silicone (OV-17) is stationary phase, and coating concentration is 2%, and column temperature 265, detecting device are flame ionization ditector, 290 ℃ of temperature; Theoretical cam curve must not remember with the vitamin E peak and is less than 500.

3. the investigation of linear relationship: precision takes by weighing n-dotriacontane 50.50mg in the 100ml volumetric flask, adds normal hexane and makes dissolving and be diluted to scale, shakes up, as inner mark solution.Hexane solution (6.114mg/ml) 1.0,2.0,3.0,4.0,5.0ml that precision is measured the vitamin E reference substance split in the volumetric flask, add inner mark solution 10ml, shake up, and draw 2 μ l inject gas chromatographs respectively, the record chromatogram.Vertical with sample size (μ g) be coordinate, and reference substance and interior mark peak area integration be than being horizontal ordinate, the drawing standard curve, and to obtain straight-line equation be C=5.351A+0.02901, correlation coefficient r=0.99930.Illustrate that sample size is good in 6.114 μ g～30.570 μ g scope internal linear.

Table 10 vitamin E typical curve

Sample size (μ g)	6.114 12.228 18.342 24.456 30.570
		Mark peak area in the reference substance peak area	125845 238378 382145 460703 644704110324 104905 113096 98533 114230
Peak area ratio	1.141 2.272 3.379 4.676 5.644

The linear equation that fitted to initial point is C=5.358A.Peak area ratio 1.141 difference substitution regression equation C=5.351A+0.02901 and former point equation C=5.358A that linearity range low concentration point is recorded calculate, get 6.1345 and 6.1135 μ g respectively, its mean relative deviation is 0.17%, the intercept that regression equation is described is less, can ignore, an available point calibration method calculates content.

4. precision test: get vitamin E reference substance solution 2 μ l, repeat sample introduction 5 times, the record chromatogram.Calculating RSD% with peak area ratio is 0.88%, can find out that precision is good.

The test of table 11 vitamin E precision

5. replica test: get 5 parts in same lot number sample, (being equivalent to vitamin E 20mg), and press content assaying method and handle, sample introduction, the record chromatogram, calculating vitamin E mean value is 32.15mg/g, RSD% is 2.12%, shows that repeatability is good.Table 12 as a result.

Table 12 replica test

Test number (TN)	1	2	3	4	5	Mean value	RSD％
								Content (mg/g)	32.43	31.82	33.21	31.79	31.50	32.15	2.12

6. recovery test: get the sample of testing same lot number with reappearance, the accurate title, decide, add a certain amount of reference substance (getting concentration is 6.114mg/ml vitamin E reference substance solution 1.0ml, 1.5ml, 2.0ml) respectively, pressing content assaying method handles, sample introduction, the record chromatogram is calculated and is recorded the recovery.Average recovery rate is 99.03%, and RSD is 2.64%, illustrates to reclaim well, the results are shown in Table 13.

Table 13 vitamin E application of sample recovery test

7. sample size is measured computing method: this determination method employing external standard method (" two appendix VI of Chinese pharmacopoeia version in 2005 D) calculate content, its formula is as follows:

Content (Mr)=Ax * Mr/Ar, when reference substance solution was represented with mg/ml, Mr represented that promptly reference substance is the concentration unit of mg/ml, the concentration that records sample simultaneously also is mg/ml.Draft every and contain vitamin E with the mg unit representation, i.e. mg/ grain, the substitution following formula calculates the content (mg/ grain) that is sample:

Content (mg/ grain)=Mx * P/ (Wx/V),

Wx is the weighing (mg) of sample in the formula, and V is average particle heavy (mg) for quantity of solvent (ml) P that extracts sample.Merge (1), (2) formula, obtain the cubage formula: content (mg/ grain)=Ax * Mr * V * P/ (Ar * Wx)

Table 14 vitamin E 10 batch sample assays

Mean value according to 10 batch sample assay results: vitamin E is the 10.03mg/ grain, is 100.3% of labelled amount.Each time result is all in the limit of labelled amount 90.0%～110.0%.

The present invention utilizes Cobastab ₁, B ₆The characteristic absorption that has adopts high performance liquid chromatography control Cobastab ₁, B ₆Content, content with vapor-phase chromatography control vitamin E, compare with the ultraviolet spectrophotometry that prior art adopts, more effective to Determination on content, the accuracy and the advance of quality inspection standard are higher, can effectively control the quality of medicine, make the reliable in quality, stable of the medicine of producing, guarantee the result of treatment and the drug safety of medicine.

Embodiment:

Embodiments of the invention 1: the method for quality control of compound Siwei fructus ligustri lucidi capsule

[proterties] content is a chocolate brown powder.

2 of compound Siwei fructus ligustri lucidi capsules are got in [discriminating] (1), remove capsule, porphyrize, add water 10ml, warmly make dissolving, filter, filtrate is put in the separating funnel, extract with ethyl acetate 15ml jolting, divide and get ethyl acetate liquid, filter, filtrate is put evaporate to dryness in the water-bath, residue adds ethanol 5ml makes dissolving, gets ethanol liquid 2ml and adds hydrochloric acid 2ml, the about 0.2g of magnesium powder, and solution shows orange or brownish red gradually.

(2) get discriminating (1) item remaining ethanol liquid down, be concentrated into about 1ml, drop on the filter paper, put and inspect under the ultraviolet lamp (365nm), show brown spot; Spray is with the ethanolic solution of 1% aluminium choride or smokedly then show pale brown color spot point with ammonia, and fluorescence is strengthened.

(3) get 1 of compound Siwei fructus ligustri lucidi capsule, remove capsule, porphyrize, add chloroform 20ml, jolting 10 minutes filters, and filtrate is put evaporate to dryness in the water-bath, residue adds ethanol 4ml makes dissolving, 2 of filtrate hydro-oxidation sodium test solutions promptly show glassy yellow, continue the about 1ml of dropping sodium test solution, under ultraviolet lamp (365nm), inspect, show yellow-green fluorescence.

(4) get 1 of compound Siwei fructus ligustri lucidi capsule, remove capsule, porphyrize adds water 20ml, stirs to make dissolving, filters, and gets filtrate 5ml, according to Cobastab ₁Differential method (1) down (" Chinese pharmacopoeia 2005 version two ones 784 pages) is tested apparent identical reaction from " hydro-oxidation sodium test solution 2.5ml ".

(5) get discriminating (a 4) remaining filtrate 2ml, thin up is to 10ml, according to Cobastab ₆Down differential method (1) (" Chinese pharmacopoeia 2005 version two ones 788 pages) is tested apparent identical reaction in accordance with the law from " respectively getting 1ml ".

(6) get 4 of compound Siwei fructus ligustri lucidi capsules, remove capsule, porphyrize, add water 20ml, stir and make dissolving, get filtrate 2ml, hydro-oxidation sodium 0.3g, make dissolving after, the heating evaporate to dryness, after the little live charcoalization, in 500 ℃ blazing 2 hours, put cold, residue adds water 5ml, stirs evenly, and filters, filtrate adds 1 of sodium nitroprusside test solution, i.e. displaing yellow.

Check: should meet every regulation relevant under the capsule item (" two appendix IE of Chinese pharmacopoeia version in 2005).

Assay: Cobastab ₁, Cobastab ₆: the photograph high performance liquid chromatography (" two appendix V of Chinese pharmacopoeia version in 2005 D) measure.

Chromatographic condition and system suitability test: with octadecylsilane key and silica gel is filling agent; 0.025mol/L acetonitrile: potassium dihydrogen phosphate=3: 97 is moving phase (pH=3); The detection wavelength is 276nm.Number of theoretical plate is pressed Cobastab ₁The peak calculates should be not less than 1500.

The preparation of reference substance solution: precision takes by weighing Cobastab ₁, Cobastab ₆Reference substance is an amount of, adds water or methyl alcohol and makes every 1ml and contain Cobastab ₁And Cobastab ₆Be respectively the solution of 0.1mg, promptly.

The preparation of need testing solution: precision takes by weighing this product and (is equivalent to Cobastab approximately in right amount ₁10mg, Cobastab ₆10mg) put in the 100ml measuring bottle, it is an amount of to add water, ultrasonicly makes moltenly, puts coldly, adds water to scale, shakes up, and filters with the miillpore filter of 0.45 μ m, promptly.

Determination method: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly.

Vitamin E: the photograph vapor-phase chromatography (" two appendix V of Chinese pharmacopoeia version in 2005 E) measure.

Chromatographic condition and system suitability experiment: with silicone (OV-17) is stationary phase, and coating concentration is 2%; Column temperature is 265 ℃.Number of theoretical plate calculates by the vitamin E peak should be not less than 500, and the degree of separation of vitamin E peak and internal standard compound mass peak should be greater than 2.

Correction factor is measured: it is an amount of to get n-dotriacontane, adds n-hexane dissolution and is diluted to the solution that contains 0.5mg among every 1ml, shakes up, as inner mark solution.Other gets the about 20mg of vitamin E reference substance, and accurate the title decides, and puts in the brown tool plug conical flask, the accurate inner mark solution 10ml that adds, and close plug, jolting makes dissolving, gets 2 μ l inject gas chromatographs, the calculation correction factor.

Determination method: get the content under the content uniformity item, mixing, precision takes by weighing in right amount (being equivalent to vitamin E2 0mg approximately), puts in the brown tool plug conical flask, the accurate inner mark solution 10ml that adds, close plug, jolting makes dissolving, gets 2 μ l inject gas chromatographs, measure, calculate, promptly.

Embodiments of the invention 2: the method for quality control of compound Siwei fructus ligustri lucidi capsule

Proterties: content is a chocolate brown powder.

Differentiate: (1) gets 1 of compound Siwei fructus ligustri lucidi capsule, removes capsule, porphyrize, add water 5ml, warmly make dissolving, filter, filtrate is put in the separating funnel, extract with ethyl acetate 10ml jolting, divide and get ethyl acetate liquid, filter, filtrate is put evaporate to dryness in the water-bath, residue adds ethanol 2ml makes dissolving, gets ethanol liquid 1ml and adds hydrochloric acid 1ml, the about 0.1g of magnesium powder, and solution shows orange or brownish red gradually.

(2) get discriminating (1) item remaining ethanol liquid down, be concentrated into about 0.5ml, drop on the filter paper, put and inspect under the ultraviolet lamp (365nm), show brown spot; Spray is with the ethanolic solution of 0.5% aluminium choride or smokedly then show pale brown color spot point with ammonia, and fluorescence is strengthened.

(3) get 1 of compound Siwei fructus ligustri lucidi capsule, remove capsule, porphyrize, add chloroform 10ml, jolting 10 minutes filters, and filtrate is put evaporate to dryness in the water-bath, residue adds ethanol 2ml makes dissolving, 2 of filtrate hydro-oxidation sodium test solutions promptly show glassy yellow, continue the about 0.5ml of dropping sodium test solution, under ultraviolet lamp (365nm), inspect, show yellow-green fluorescence.

(4) get 1 of compound Siwei fructus ligustri lucidi capsule, remove capsule, porphyrize adds water 10ml, stirs to make dissolving, filters, and gets filtrate 2ml, according to Cobastab ₁Differential method (1) down (" Chinese pharmacopoeia 2005 version two ones 784 pages) is tested apparent identical reaction from " hydro-oxidation sodium test solution 2.5ml ".

(5) get discriminating (a 4) remaining filtrate 1ml, thin up is to 5ml, according to Cobastab ₆Down differential method (1) (" Chinese pharmacopoeia 2005 version two ones 788 pages) is tested apparent identical reaction in accordance with the law from " respectively getting 1ml ".

(6) get 2 of compound Siwei fructus ligustri lucidi capsules, remove capsule, porphyrize, add water 10ml, stir and make dissolving, get filtrate 1ml, hydro-oxidation sodium 0.1g, make dissolving after, the heating evaporate to dryness, after the little live charcoalization, in 500 ℃ blazing 2 hours, put cold, residue adds water 3ml, stirs evenly, and filters, filtrate adds 1 of sodium nitroprusside test solution, i.e. displaing yellow.

Chromatographic condition and system suitability test: with octadecylsilane key and silica gel is filling agent; 0.01mol/L acetonitrile: potassium dihydrogen phosphate=1: 99 is moving phase (pH=3); The detection wavelength is 276nm.Number of theoretical plate is pressed Cobastab ₁The peak calculates should be not less than 10.

The preparation of reference substance solution: precision takes by weighing Cobastab ₁, Cobastab ₆Reference substance is an amount of, adds water or methyl alcohol and makes every 1ml and contain Cobastab ₁And Cobastab ₆Be respectively the solution of 0.05mg, promptly.

The preparation of need testing solution: precision takes by weighing this product and (is equivalent to Cobastab approximately in right amount ₁5mg, Cobastab ₆5mg) put in the 50ml measuring bottle, it is an amount of to add water, ultrasonicly makes moltenly, puts coldly, adds water to scale, shakes up, and filters with the miillpore filter of 0.45 μ m, promptly.

Determination method: accurate respectively reference substance solution and each 5 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly.

Chromatographic condition and system suitability experiment: with silicone (OV-17) is stationary phase, and coating concentration is 1%; Column temperature is 200 ℃.Number of theoretical plate calculates by the vitamin E peak should be not less than 300, and the degree of separation of vitamin E peak and internal standard compound mass peak should be greater than 1.5.

Correction factor is measured: it is an amount of to get n-dotriacontane, adds n-hexane dissolution and is diluted to the solution that contains 0.25mg among every 1ml, shakes up, as inner mark solution.Other gets the about 10mg of vitamin E reference substance, and accurate the title decides, and puts in the brown tool plug conical flask, the accurate inner mark solution 10ml that adds, and close plug, jolting makes dissolving, gets 2 μ l inject gas chromatographs, the calculation correction factor.

Determination method: get the content under the content uniformity item, mixing, precision takes by weighing in right amount (being equivalent to vitamin E 10mg approximately), puts in the brown tool plug conical flask, the accurate inner mark solution 5ml that adds, close plug, jolting makes dissolving, gets 1 μ l inject gas chromatograph, measure, calculate, promptly.

Embodiments of the invention 3: the method for quality control of compound Siwei fructus ligustri lucidi capsule

Proterties: content is a chocolate brown powder.

Differentiate: (1) gets 2 of compound Siwei fructus ligustri lucidi capsules, removes capsule, porphyrize, add water 20ml, warmly make dissolving, filter, filtrate is put in the separating funnel, extract with ethyl acetate 30ml jolting, divide and get ethyl acetate liquid, filter, filtrate is put evaporate to dryness in the water-bath, residue adds ethanol 10ml makes dissolving, gets ethanol liquid 3ml and adds hydrochloric acid 3ml, the about 0.3g of magnesium powder, and solution shows orange or brownish red gradually.

(2) get discriminating (1) item remaining ethanol liquid down, be concentrated into about 2ml, drop on the filter paper, put and inspect under the ultraviolet lamp (365nm), show brown spot; Spray is with the ethanolic solution of 2% aluminium choride or smokedly then show pale brown color spot point with ammonia, and fluorescence is strengthened.

(3) get 2 of compound Siwei fructus ligustri lucidi capsules, remove capsule, porphyrize, add chloroform 30ml, jolting 10 minutes filters, and filtrate is put evaporate to dryness in the water-bath, residue adds ethanol 8ml makes dissolving, 2 of filtrate hydro-oxidation sodium test solutions promptly show glassy yellow, continue the about 2ml of dropping sodium test solution, under ultraviolet lamp (365nm), inspect, show yellow-green fluorescence.

(4) get 2 of compound Siwei fructus ligustri lucidi capsules, remove capsule, porphyrize adds water 30ml, stirs to make dissolving, filters, and gets filtrate 8ml, according to Cobastab ₁Differential method (1) down (" Chinese pharmacopoeia 2005 version two ones 784 pages) is tested apparent identical reaction from " hydro-oxidation sodium test solution 2.5ml ".

(5) get discriminating (a 4) remaining filtrate 3ml, thin up is to 15ml, according to Cobastab ₆Down differential method (1) (" Chinese pharmacopoeia 2005 version two ones 788 pages) is tested apparent identical reaction in accordance with the law from " respectively getting 1ml ".

(6) get 6 of compound Siwei fructus ligustri lucidi capsules, remove capsule, porphyrize, add water 30ml, stir and make dissolving, get filtrate 3ml, hydro-oxidation sodium 0.5g, make dissolving after, the heating evaporate to dryness, after the little live charcoalization, in 500 ℃ blazing 4 hours, put cold, residue adds water 8ml, stirs evenly, and filters, filtrate adds 1 of sodium nitroprusside test solution, i.e. displaing yellow.

Chromatographic condition and system suitability test: with octadecylsilane key and silica gel is filling agent; 0.05mol/L acetonitrile: potassium dihydrogen phosphate=10: 90 is moving phase (pH=3); The detection wavelength is 276nm.Number of theoretical plate is pressed Cobastab ₁The peak calculates should be not less than 3000.

The preparation of reference substance solution: precision takes by weighing Cobastab ₁, Cobastab ₆Reference substance is an amount of, adds water or methyl alcohol and makes every 1ml and contain Cobastab ₁And Cobastab ₆Be respectively the solution of 0.3mg, promptly.

The preparation of need testing solution: precision takes by weighing this product and (is equivalent to Cobastab approximately in right amount ₁20mg, Cobastab ₆20mg) put in the 100ml measuring bottle, it is an amount of to add water, ultrasonicly makes moltenly, puts coldly, adds water to scale, shakes up, and filters with the miillpore filter of 0.45 μ m, promptly.

Determination method: accurate respectively reference substance solution and each 15 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly.

Chromatographic condition and system suitability experiment: with silicone (OV-17) is stationary phase, and coating concentration is 5%; Column temperature is 300 ℃.Number of theoretical plate calculates by the vitamin E peak should be not less than 1000, and the degree of separation of vitamin E peak and internal standard compound mass peak should be greater than 3.

Correction factor is measured: it is an amount of to get n-dotriacontane, adds n-hexane dissolution and is diluted to the solution that contains 1mg among every 1ml, shakes up, as inner mark solution.Other gets the about 30mg of vitamin E reference substance, and accurate the title decides, and puts in the brown tool plug conical flask, the accurate inner mark solution 10ml that adds, and close plug, jolting makes dissolving, gets 2 μ l inject gas chromatographs, the calculation correction factor.

Determination method: get the content under the content uniformity item, mixing, precision takes by weighing in right amount (being equivalent to vitamin E 30mg approximately), puts in the brown tool plug conical flask, the accurate inner mark solution 20ml that adds, close plug, jolting makes dissolving, gets 3 μ l inject gas chromatographs, measure, calculate, promptly.

Embodiments of the invention 4: the method for quality control of compound Siwei fructus ligustri lucidi tablet

Proterties: for light brown to the sepia tablet.

Differentiate: (1) gets 2 in compound Siwei fructus ligustri lucidi tablet, and porphyrize adds water 10ml, warmly make dissolving, filter, filtrate is put in the separating funnel, extracts with ethyl acetate 15ml jolting, divide and get ethyl acetate liquid, filter, filtrate is put evaporate to dryness in the water-bath, and residue adds ethanol 5ml makes dissolving, get ethanol liquid 2ml and add hydrochloric acid 2ml, the about 0.2g of magnesium powder, solution shows orange or brownish red gradually.

(3) get 1 in compound Siwei fructus ligustri lucidi tablet, porphyrize adds chloroform 20ml, jolting 10 minutes, filter, filtrate is put evaporate to dryness in the water-bath, and residue adds ethanol 4ml makes dissolving, 2 of filtrate hydro-oxidation sodium test solutions, promptly show glassy yellow, continue the about 1ml of dropping sodium test solution, under ultraviolet lamp (365nm), inspect, show yellow-green fluorescence.

(4) get 1 in compound Siwei fructus ligustri lucidi tablet, porphyrize adds water 20ml, stirs to make dissolving, filters, and gets filtrate 5ml, according to Cobastab ₁Differential method (1) down (" Chinese pharmacopoeia 2005 version two ones 784 pages) is tested apparent identical reaction from " hydro-oxidation sodium test solution 2.5ml ".

(6) get 4 in compound Siwei fructus ligustri lucidi tablet, porphyrize adds water 20ml, stirring makes dissolving, gets filtrate 2ml, hydro-oxidation sodium 0.3g, after making dissolving, the heating evaporate to dryness is after the little live charcoalization, in 500 ℃ blazing 2 hours, put coldly, residue adds water 5ml, stir evenly, filter, filtrate adds 1 of sodium nitroprusside test solution, i.e. displaing yellow.

Check: should meet every regulation relevant under the tablet item (" two appendix I of Chinese pharmacopoeia version in 2005 E).

The preparation of need testing solution: precision takes by weighing the compound Siwei fructus ligustri lucidi tablet and (is equivalent to Cobastab approximately in right amount ₁10mg, Cobastab ₆10mg) put in the 100ml measuring bottle, it is an amount of to add water, ultrasonicly makes moltenly, puts coldly, adds water to scale, shakes up, and filters with the miillpore filter of 0.45 μ m, promptly.

Embodiments of the invention 5: the method for quality control of compound Siwei fructus ligustri lucidi tablet

Proterties: for light brown to the sepia tablet.

Differentiate: (1) gets 1 in compound Siwei fructus ligustri lucidi tablet, and porphyrize adds water 5ml, warmly make dissolving, filter, filtrate is put in the separating funnel, extracts with ethyl acetate 10ml jolting, divide and get ethyl acetate liquid, filter, filtrate is put evaporate to dryness in the water-bath, and residue adds ethanol 2ml makes dissolving, get ethanol liquid 1ml and add hydrochloric acid 1ml, the about 0.1g of magnesium powder, solution shows orange or brownish red gradually.

(3) get 1 in compound Siwei fructus ligustri lucidi tablet, porphyrize adds chloroform 10ml, jolting 10 minutes, filter, filtrate is put evaporate to dryness in the water-bath, and residue adds ethanol 2ml makes dissolving, 2 of filtrate hydro-oxidation sodium test solutions, promptly show glassy yellow, continue the about 0.5ml of dropping sodium test solution, under ultraviolet lamp (365nm), inspect, show yellow-green fluorescence.

(4) get 1 in compound Siwei fructus ligustri lucidi tablet, porphyrize adds water 10ml, stirs to make dissolving, filters, and gets filtrate 2ml, according to Cobastab ₁Differential method (1) down (" Chinese pharmacopoeia 2005 version two ones 784 pages) is tested apparent identical reaction from " hydro-oxidation sodium test solution 2.5ml ".

(6) get 2 in compound Siwei fructus ligustri lucidi tablet, porphyrize adds water 10ml, stirring makes dissolving, gets filtrate 1ml, hydro-oxidation sodium 0.1g, after making dissolving, the heating evaporate to dryness is after the little live charcoalization, in 500 ℃ blazing 2 hours, put coldly, residue adds water 3ml, stir evenly, filter, filtrate adds 1 of sodium nitroprusside test solution, i.e. displaing yellow.

Embodiments of the invention 6: the method for quality control of compound Siwei fructus ligustri lucidi tablet

Proterties: for light brown to the sepia tablet.

Differentiate: (1) gets 2 in compound Siwei fructus ligustri lucidi tablet, and porphyrize adds water 20ml, warmly make dissolving, filter, filtrate is put in the separating funnel, extracts with ethyl acetate 30ml jolting, divide and get ethyl acetate liquid, filter, filtrate is put evaporate to dryness in the water-bath, and residue adds ethanol 10ml makes dissolving, get ethanol liquid 3ml and add hydrochloric acid 3ml, the about 0.3g of magnesium powder, solution shows orange or brownish red gradually.

(3) get 2 in compound Siwei fructus ligustri lucidi tablet, porphyrize adds chloroform 30ml, jolting 10 minutes, filter, filtrate is put evaporate to dryness in the water-bath, and residue adds ethanol 8ml makes dissolving, 2 of filtrate hydro-oxidation sodium test solutions, promptly show glassy yellow, continue the about 2ml of dropping sodium test solution, under ultraviolet lamp (365nm), inspect, show yellow-green fluorescence.

(4) get 2 in compound Siwei fructus ligustri lucidi tablet, porphyrize adds water 30ml, stirs to make dissolving, filters, and gets filtrate 8ml, according to Cobastab ₁Differential method (1) down (" Chinese pharmacopoeia 2005 version two ones 784 pages) is tested apparent identical reaction from " hydro-oxidation sodium test solution 2.5ml ".

(6) get 6 in compound Siwei fructus ligustri lucidi tablet, porphyrize adds water 30ml, stirring makes dissolving, gets filtrate 3ml, hydro-oxidation sodium 0.5g, after making dissolving, the heating evaporate to dryness is after the little live charcoalization, in 500 ℃ blazing 4 hours, put coldly, residue adds water 8ml, stir evenly, filter, filtrate adds 1 of sodium nitroprusside test solution, i.e. displaing yellow.

Embodiments of the invention 7: the method for quality control of compound Siwei fructus ligustri lucidi granule

Proterties: content is a chocolate brown powder.

Differentiate: (1) gets 2 bags of compound Siwei fructus ligustri lucidi granules, removes unlap, porphyrize, add water 10ml, warmly make dissolving, filter, filtrate is put in the separating funnel, extract with ethyl acetate 15ml jolting, divide and get ethyl acetate liquid, filter, filtrate is put evaporate to dryness in the water-bath, residue adds ethanol 5ml makes dissolving, gets ethanol liquid 2ml and adds hydrochloric acid 2ml, the about 0.2g of magnesium powder, and solution shows orange or brownish red gradually.

(3) get 1 bag of compound Siwei fructus ligustri lucidi granule, remove unlap, porphyrize, add chloroform 20ml, jolting 10 minutes filters, and filtrate is put evaporate to dryness in the water-bath, residue adds ethanol 4ml makes dissolving, 2 of filtrate hydro-oxidation sodium test solutions promptly show glassy yellow, continue the about 1ml of dropping sodium test solution, under ultraviolet lamp (365nm), inspect, show yellow-green fluorescence.

(4) get 1 bag of compound Siwei fructus ligustri lucidi granule, remove unlap, porphyrize adds water 20ml, stirs and makes dissolving, filters, and gets filtrate 5ml, according to Cobastab ₁Differential method (1) down (" Chinese pharmacopoeia 2005 version two ones 784 pages) is tested apparent identical reaction from " hydro-oxidation sodium test solution 2.5ml ".

(6) get 4 bags of compound Siwei fructus ligustri lucidi granules, remove unlap, porphyrize, add water 20ml, stir and make dissolving, get filtrate 2ml, hydro-oxidation sodium 0.3g, make dissolving after, the heating evaporate to dryness, after the little live charcoalization, in 500 ℃ blazing 2 hours, put cold, residue adds water 5ml, stirs evenly, and filters, filtrate adds 1 of sodium nitroprusside test solution, i.e. displaing yellow.

Check: should meet every regulation relevant under the granule item (" two appendix IE of Chinese pharmacopoeia version in 2005).

The preparation of need testing solution: precision takes by weighing the compound Siwei fructus ligustri lucidi granule and (is equivalent to Cobastab approximately in right amount ₁10mg, Cobastab ₆10mg) put in the 100ml measuring bottle, it is an amount of to add water, ultrasonicly makes moltenly, puts coldly, adds water to scale, shakes up, and filters with the miillpore filter of 0.45 μ m, promptly.

Embodiments of the invention 8: the method for quality control of compound Siwei fructus ligustri lucidi granule

Proterties: content is a chocolate brown powder.

Differentiate: (1) gets 1 bag of compound Siwei fructus ligustri lucidi granule, removes unlap, porphyrize, add water 5ml, warmly make dissolving, filter, filtrate is put in the separating funnel, extract with ethyl acetate 10ml jolting, divide and get ethyl acetate liquid, filter, filtrate is put evaporate to dryness in the water-bath, residue adds ethanol 2ml makes dissolving, gets ethanol liquid 1ml and adds hydrochloric acid 1ml, the about 0.1g of magnesium powder, and solution shows orange or brownish red gradually.

(3) get 1 bag of compound Siwei fructus ligustri lucidi granule, remove unlap, porphyrize, add chloroform 10ml, jolting 10 minutes filters, and filtrate is put evaporate to dryness in the water-bath, residue adds ethanol 2ml makes dissolving, 2 of filtrate hydro-oxidation sodium test solutions promptly show glassy yellow, continue the about 0.5ml of dropping sodium test solution, under ultraviolet lamp (365nm), inspect, show yellow-green fluorescence.

(4) get 1 bag of compound Siwei fructus ligustri lucidi granule, remove unlap, porphyrize adds water 10ml, stirs and makes dissolving, filters, and gets filtrate 2ml, according to Cobastab ₁Differential method (1) down (" Chinese pharmacopoeia 2005 version two ones 784 pages) is tested apparent identical reaction from " hydro-oxidation sodium test solution 2.5ml ".

(6) get 2 bags of compound Siwei fructus ligustri lucidi granules, remove unlap, porphyrize, add water 10ml, stir and make dissolving, get filtrate 1ml, hydro-oxidation sodium 0.1g, make dissolving after, the heating evaporate to dryness, after the little live charcoalization, in 500 ℃ blazing 2 hours, put cold, residue adds water 3ml, stirs evenly, and filters, filtrate adds 1 of sodium nitroprusside test solution, i.e. displaing yellow.

Embodiments of the invention 9: the method for quality control of compound Siwei fructus ligustri lucidi granule

Proterties: content is a chocolate brown powder.

Differentiate: (1) gets 2 bags of compound Siwei fructus ligustri lucidi granules, removes unlap, porphyrize, add water 20ml, warmly make dissolving, filter, filtrate is put in the separating funnel, extract with ethyl acetate 30ml jolting, divide and get ethyl acetate liquid, filter, filtrate is put evaporate to dryness in the water-bath, residue adds ethanol 10ml makes dissolving, gets ethanol liquid 3ml and adds hydrochloric acid 3ml, the about 0.3g of magnesium powder, and solution shows orange or brownish red gradually.

(3) get 2 bags of compound Siwei fructus ligustri lucidi granules, remove unlap, porphyrize, add chloroform 30ml, jolting 10 minutes filters, and filtrate is put evaporate to dryness in the water-bath, residue adds ethanol 8ml makes dissolving, 2 of filtrate hydro-oxidation sodium test solutions promptly show glassy yellow, continue the about 2ml of dropping sodium test solution, under ultraviolet lamp (365nm), inspect, show yellow-green fluorescence.

(4) get 2 bags of compound Siwei fructus ligustri lucidi granules, remove unlap, porphyrize adds water 30ml, stirs and makes dissolving, filters, and gets filtrate 8ml, according to Cobastab ₁Differential method (1) down (" Chinese pharmacopoeia 2005 version two ones 784 pages) is tested apparent identical reaction from " hydro-oxidation sodium test solution 2.5ml ".

(6) get 6 bags of compound Siwei fructus ligustri lucidi granules, remove unlap, porphyrize, add water 30ml, stir and make dissolving, get filtrate 3ml, hydro-oxidation sodium 0.5g, make dissolving after, the heating evaporate to dryness, after the little live charcoalization, in 500 ℃ blazing 4 hours, put cold, residue adds water 8ml, stirs evenly, and filters, filtrate adds 1 of sodium nitroprusside test solution, i.e. displaing yellow.

Claims

1. the quality determining method of a compound Siwei fructus ligustri lucidi preparation, described preparation is made capsule by the medicine of following weight, tablet, granule: oryzanol 10～20g, vitamin B1 5～15g, vitamin B6 5～15g, vitamin E 5～15g, biotin 1500～2005 μ g, taurine 10～50g, Herba Epimedii extractum 50～100g, Fructus Ligustri Lucidi extractum 80～150g, cultivated land medicinal extract 40～80g, puncturevine caltrop fruit medicinal extract 40～80g, it is characterized in that: described quality determining method comprises proterties, differentiate, check and assay, described proterties comprises: for capsule, the product content thing is that light brown is to brown granular or powder; For tablet, medicine shows light brown to sepia; For granule, product is that light brown is to tan particle;

Described discriminating comprises: it is an amount of that (1) gets compound Siwei fructus ligustri lucidi capsule, tablet or granule, removes unlap, porphyrize, add water 10m1, warmly make dissolving, filter, filtrate is put in the separating funnel, extract with ethyl acetate 15ml jolting, divide and get ethyl acetate liquid, filter, filtrate is put evaporate to dryness in the water-bath, residue adds ethanol 5ml makes dissolving, gets ethanol liquid 2ml and adds hydrochloric acid 2ml, magnesium powder approximately O.2g, and solution shows orange or brownish red gradually; (2) get above-mentioned remaining ethanol liquid, be concentrated into about 1ml, drop on the filter paper, put under the ultraviolet lamp that wavelength is 365nm and inspect, show brown spot; Spray is with the ethanolic solution of 1% aluminium choride or smokedly then show pale brown color spot point with ammonia, and fluorescence is strengthened; (3) get 1 part of compound Siwei fructus ligustri lucidi capsule, tablet or granule, porphyrize adds chloroform 20ml, jolting 10 minutes, filter, filtrate is put evaporate to dryness in the water-bath, and residue adds ethanol 4m1 makes dissolving, 2 of filtrate hydro-oxidation sodium test solutions, promptly show glassy yellow, continuing dropping sodium test solution 1ml, is to inspect under the ultraviolet lamp of 365nm at wavelength, shows yellow-green fluorescence; (4) get 1 part of compound Siwei fructus ligustri lucidi capsule, tablet or granule, porphyrize, add water 20ml, stirring makes dissolving, filter, get filtrate 5m1, according to " differential method (1) item under two 784 pages of vitamin B1 items of Chinese pharmacopoeia version in 2005 shows identical reaction from " hydro-oxidation sodium test solution 2.5ml " test; (5) get above-mentioned remaining filtrate 2m1, thin up is to 10ml, according to " differential method (1) item is tested from " respectively getting 1m1 " under two 788 pages of vitamin B6 items of Chinese pharmacopoeia version in 2005, shows identical reaction in accordance with the law; (6) get 4 parts of compound Siwei fructus ligustri lucidi capsule, tablet or granules, porphyrize adds water 20ml, stirring makes dissolving, gets filtrate 2ml, and hydro-oxidation sodium O.3g, after making dissolving, the heating evaporate to dryness is after the little live charcoalization, in 500 ℃ blazing 2 hours, put coldly, residue adds water 5m1, stir evenly, filter, filtrate adds sodium nitroprusside test solution l to be dripped, i.e. displaing yellow;

Described content assaying method comprises:

The assay of vitamin B1, vitamin B6: (1) chromatographic condition and system suitability test: with octadecylsilane key and silica gel is filling agent; O.025mol/L acetonitrile: potassium dihydrogen phosphate=3:97 is a moving phase; The detection wavelength is 276nm, and number of theoretical plate calculates by the vitamin B1 peak should be not less than 1500; (2) system of reference substance solution each: precision takes by weighing vitamin B1, the vitamin B6 reference substance is an amount of, adds water or methyl alcohol and makes every 1m1 and contain vitamin B1 and vitamin B6 and be respectively O.1mg solution, promptly; (3) system of need testing solution respectively: it is an amount of that precision takes by weighing compound Siwei fructus ligustri lucidi capsule, tablet or granule, be equivalent to VB11 0mg, vitamin B6 lOmg puts in the lOOml measuring bottle, and it is an amount of to add water, ultrasonic make molten, put coldly, add water to scale, shake up, with the O.45 miillpore filter filtration of μ m, promptly; (4) determination method: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject liquid chromatograph, measure, promptly;

The assay of vitamin E comprises: (1) chromatographic condition and system suitability experiment: the silicone with 0V-17 is a stationary phase, and coating concentration is 2%; Column temperature is 265 ℃, and number of theoretical plate calculates by the vitamin E peak should be not less than 500, and the degree of separation of vitamin E peak and internal standard compound mass peak should be greater than more than 2; (2) correction factor is measured: it is an amount of to get n-dotriacontane, add n-hexane dissolution and be diluted to the solution that contains among every 1m1 O.5mg, shake up, as inner mark solution, other gets vitamin E reference substance 20mg, the accurate title, decide, and puts in the brown tool plug conical flask, the accurate inner mark solution 10ml that adds, close plug, jolting makes dissolving, gets 2 μ, 1 inject gas chromatograph, the calculation correction factor; (3) determination method: the content of getting compound Siwei fructus ligustri lucidi capsule, tablet or granule, mixing, precision take by weighing in right amount, are equivalent to vitamin E 20mg, put in the brown tool plug conical flask, the accurate inner mark solution 10ml that adds, close plug, jolting makes dissolving, get 2 μ, 1 inject gas chromatograph, measure, calculate, promptly.