CN103499652B - Method for measuring content of hydrochlorothiazide and/or rutin in medicine and application of method - Google Patents
Method for measuring content of hydrochlorothiazide and/or rutin in medicine and application of method Download PDFInfo
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Abstract
The invention relates to a method for measuring the content of hydrochlorothiazide and/or rutin in a medicine and application of the method. The method for measuring the content of the hydrochlorothiazide and/or the rutin in the medicine comprises the following steps of preparing the medicine into a test solution, and then quantitatively detecting the content of the hydrochlorothiazide and/or the rutin in the test solution by high performance liquid chromatography by taking an anion exchange chromatographic column as an analytical column, taking a methanol-phosphate buffer system as a flowing phase and taking any wavelength being 250-280nm as detection wavelength. The method disclosed by the invention for detecting the content of the hydrochlorothiazide and/or the rutin has the advantages of ideal separating effect, high specificity, high precision, high repetitiveness and high accuracy, and a reliable technical measure is supplied to measurement of the content and the content uniformity.
Description
Technical field
The present invention relates to a kind of assay method and application thereof of medicine special component, particularly relate to content assaying method and the application thereof of Hydrochioro and/or rutin in a kind of medicine.
Background technology
Hydrochioro is diuresis type depressor, and rutin has softening blood vessel, reduces capillary permeability, increases the effects such as its tension force, be principle active component in Zhenju Jiangya Tablet side (molecular structural formula and UV spectrum are shown in Fig. 1,2).Comparatively speaking, the content of Hydrochlorothiazide In Zhenjujiangya Tablets and rutin is higher, and mother chrysanthemum Related Component is relatively little on its impact.In report about HPLC assay, except minority measures separately rutin or Hydrochioro, usually measure simultaneously.In addition, Gao Suya (Gao Suya, Li Hua, Cao Xiaoqin. capillary zone electrophresis measures the content [J] of Zhenju Jiangya Tablet effective constituent. time precious traditional Chinese medical science traditional Chinese medicines, 2008, 19 (11): 1656-2567), Yun-qiu(Su, S.-m., Yu, Y.-q.Simultaneous Determination of ClonidineHydrochloride, Hyrochlorothiazide, and Rutin in Zhenju Jiangya Tablet by CapillaryElectrophoresis [J] .Journal of Chinese Pharmaceutical Sciences, 2005, 14 (3): 173-175), fourth flies, and (fourth flies, Chen Liming. effective constituent [J] in Capillary Electrophoresis-ampere detection method Simultaneously test Chinese patent drug Zhenju Jiangya Tablet. physical and chemical inspection-chemical fascicle, 2008, 44 (5): 430-432) clonidine hydrochloride that capillary electrophoresis technique is attempted in Simultaneously test Zhenju Jiangya Tablet etc. is adopted, Hydrochioro, rutin, obtain good effect, but due to the universal problem of instrument, the method is more difficult to be promoted in routine check.Because Hydrochioro content in Zhenju Jiangya Tablet is only 5mg/ sheet, belong to small dimension preparation, uniformity of dosage units should be done and investigate, and have not yet to see relevant report.Except content assaying method research, the situation of water is insoluble in for Hydrochioro and rutin, Zhang Songming etc. adopt spectrophotometric method to adopt HPLC method to Hydrochioro and rutin to Hydrochioro, Zhang Yuehua etc., carry out the Primary Study of dissolution method respectively, had related work from now on and necessarily inspire meaning.
Summary of the invention
The object of the invention is, in order to overcome defect of the prior art, to provide assay method and the application thereof of Hydrochioro and/or rutin in a kind of medicine.
The present invention provide firstly the content assaying method of Hydrochioro and/or rutin in a kind of medicine, medicine is comprised the following steps: to be prepared into need testing solution, it is then analytical column with anion-exchange column, using methyl alcohol-phosphate buffer system as mobile phase, with wavelength optional between 250nm-280nm for determined wavelength, high performance liquid chromatography is adopted quantitatively to detect the content of Hydrochioro and/or rutin in need testing solution.
Preferably, described anion-exchange column is HP-SAX anion-exchange column.The filler name of described anion-exchange column is called QAE base p-propyl-toluene bonded silica gel.Its structural formula is:
Preferably, in described methyl alcohol-phosphate buffer system, described phosphate buffer is pH2.5-3.5KH
2pO
4concentration is the phosphate buffer of 10-40mmol/L, and the ratio of methyl alcohol and phosphate buffer is 5:95-20:80(volume ratio).Most preferred, described methyl alcohol-phosphate buffer system is pH3.0 and KH
2pO
4concentration is the phosphate buffer of 40mmol/L, and the volume ratio of methyl alcohol and phosphate buffer is 5:95.
Preferably, during high effective liquid chromatography for measuring, the flow velocity of mobile phase is 0.5-1.5mL/min, and optimum is 1.0mL/min.
Preferably, during high effective liquid chromatography for measuring, column temperature is 25-35 DEG C, most preferably 25 DEG C.
Preferably, during high effective liquid chromatography for measuring, determined wavelength is 268nm.
Preferably, after method medicine being prepared into need testing solution comprises the following steps: medicine and Extraction solvent to mix, ultrasonic 10-30 minute, obtains need testing solution after Separation of Solid and Liquid.Ultrasonic time most preferably 15 minutes.
As described in medicine be solid pharmaceutical preparation, before medicine and Extraction solvent are mixed, first by medicament disintegrating, or after medicine mix with Extraction solvent, medicine can be shaken fragmentation.
Preferred further, described extraction reagent is selected from methyl alcohol, percent by volume is the methanol aqueous solution of more than 50%.Most preferably be methyl alcohol.
In described medicine Hydrochioro and rutin content assaying method in, the same terms can be adopted to measure the content of Hydrochioro and/or rutin in reference substance solution, and press the content of Hydrochioro and/or rutin in one point external standard method calculating need testing solution.
After the content knowing Hydrochioro and/or rutin in need testing solution, the content of Hydrochioro and/or rutin in medicine will be easy to convert out.
In the content assaying method of Hydrochioro of the present invention and/or rutin, every optimum condition can random combination collocation.
In medicine of the present invention, the content assaying method of Hydrochioro and/or rutin can be used for the quality testing of medicine, is particularly useful for the assay of Hydrochlorothiazide In Zhenjujiangya Tablets and/or rutin or the Determination of Content Uniformity for Hydrochlorothiazide In Zhenjujiangya Tablets or the dissolution determination for chlorothiazide in Zhenju Jiangya Tablet and/or rutin.
The present invention still further provides a kind of quality determining method of Zhenju Jiangya Tablet, comprises the content adopting preceding method to detect Hydrochlorothiazide In Zhenjujiangya Tablets and rutin, and the uniformity of dosage units of Hydrochioro in Zhenju Jiangya Tablet.
Further, the quality determining method of described Zhenju Jiangya Tablet also comprises the dissolution rate detecting chlorothiazide and/or rutin in Zhenju Jiangya Tablet.
Due to Hydrochioro content only 5mg/ sheet in Zhenju Jiangya Tablet, easily affect clinical efficacy because uniformity of dosage units is defective, therefore it is considered herein that, be necessary its uniformity of dosage units to investigate to introduce in quality determining method as a quality detecting index.
Due to Hydrochioro, rutin poorly water-soluble, its stripping degree in digestive juice will affect clinical efficacy, therefore it is considered herein that, be necessary its dissolution rate to investigate to introduce in quality determining method as a quality detecting index.
Further, when detecting the uniformity of dosage units of the Hydrochioro in Zhenju Jiangya Tablet, peek sheet test sample, detects the content of Hydrochioro in every sheet test sample respectively, calculates uniformity of dosage units by Content uniformity test by preceding method.
Further, when the dissolution rate of the Hydrochioro detected in Zhenju Jiangya Tablet and/or rutin, peek sheet test sample, with the hydrochloric acid solution liquid of 0.1mol/L or simulated gastric fluid for dissolution medium, measure by Chinese Pharmacopoeia annex dissolution method and prepare need testing solution, detect the stripping quantity of Hydrochioro and/or rutin in every sheet test sample by preceding method respectively, and calculate dissolution rate.
Adopt method of the present invention to detect the content of Hydrochioro and/or rutin, there is desirable separating effect, specificity is strong, precision, repeatability and accuracy well, for the mensuration of content and uniformity of dosage units provides reliable technological means.
Accompanying drawing explanation
The molecular structural formula of Fig. 1 Hydrochioro and UV spectrum.
The molecular structural formula of Fig. 2 rutin and UV spectrum.
Fig. 3 chromatogram column temperature is investigated and is measured collection of illustrative plates.
Fig. 3 A25 DEG C of chromatogram column temperature is investigated and is measured collection of illustrative plates;
Fig. 3 B30 DEG C of chromatogram column temperature is investigated and is measured collection of illustrative plates;
Fig. 3 C35 DEG C of chromatogram column temperature is investigated and is measured collection of illustrative plates.
The Hydrochioro of reference substance solution, need testing solution and above-mentioned negative control solution and Assaying of Rutin collection of illustrative plates in Fig. 4 embodiment 1
The typical curve of Fig. 5 Hydrochioro
The typical curve of Fig. 6 rutin
Fig. 7 embodiment 2 Hydrochioro Determination of Content Uniformity collection of illustrative plates
Embodiment
The present invention once attempted using C18 chromatographic column as separating medium, adopted conventional reversed phase chromatography separation Model Establishment Hydrochlorothiazide In Zhenjujiangya Tablets and the content assaying method of rutin.List of references method, gets methanol extract liquid direct injected, has screened multiple flow phase system, finds that separating effect is all undesirable, and negative Interference Peaks is difficult to be separated.
The present invention attempts adopting SPE(Solid-Phase Extraction further) pre-service of carrying out sample eliminates interference.First be weakly acidic feature according to target compound, the anion exchange pillars such as CNW Poly-Sery MAX, CNWBOND SAX are selected to test, find to eliminate most Interference Peaks, but still have part interference component to be difficult to be separated and the recovery is on the low side, after extraction scheme having been carried out to multiple optimizations trial, have no obvious improvement.According to the deduction that interference component may be flavones ingredient in mother chrysanthemum, attempt adopting polyamide pillar Anpelclean PA to carry out removal of impurities process, find that result is very undesirable through many experiments, its impurity-eliminating effect is not even as ion-exchange pillar.
The situation analysis of major part interference impurity can be eliminated from anion exchange pillar, supposition may be because interference component and target compound belong to Acidic Compounds together, and rutin or flavones ingredient, solid phase extraction column post effect is on the low side cannot meet separation requirement, therefore attempts adopting anion-exchange column to set up analytical approach as analytical column.By series of experiments, find to adopt methyl alcohol-phosphate buffer system, HP-SAX anion-exchange column has desirable separating effect, therefore determines Hydrochioro and rutin assay method that the present invention adopts this clastotype to set up.
Below by way of specific instantiation, embodiments of the present invention are described, those skilled in the art the content disclosed by this instructions can understand other advantages of the present invention and effect easily.The present invention can also be implemented or be applied by embodiments different in addition, and the every details in this instructions also can based on different viewpoints and application, carries out various modification or change not deviating under spirit of the present invention.
Notice, in the following example, the concrete process equipment that indicates or device all adopt conventional equipment in this area or device.
In addition should be understood that the one or more method steps mentioned in the present invention do not repel and can also to there is additive method step or can also insert additive method step before and after described combination step between these steps clearly mentioned, except as otherwise noted.And, except as otherwise noted, the numbering of various method steps is only the convenient tool differentiating various method steps, but not be ordering or the enforceable scope of restriction the present invention of restriction various method steps, the change of its relativeness or adjustment, when changing technology contents without essence, when being also considered as the enforceable category of the present invention.
The assay of embodiment 1 Hydrochlorothiazide In Zhenjujiangya Tablets and rutin
1. chromatographic condition
The investigation of column temperature: by under three temperature conditions such as 25 DEG C, 30 DEG C, 35 DEG C the comparison of test sample and negative collection of illustrative plates, find the rising with column temperature, the degree of separation of rutin and impurity is on a declining curve, therefore determines that mensuration column temperature is 25 DEG C.
Chromatogram column temperature is investigated and is seen circle mark place in Fig. 3 A-3C(figure):
Measure the selection of wavelength: the DAD Spectroscopic analysis results of reference substance solution shows, rutin and Hydrochioro all have absorption maximum between 250nm-280nm, and 268nm place to obtain in chromatogram both peak height and peak area moderate, therefore preferred 268nm is determined wavelength.
The chromatographic condition below adopted in experiment:
Chromatographic column: Sepax HP-SAX anion column (4.6mm × 150mm)
Mobile phase: the KH of methyl alcohol-40mmol/L
2pO
4solution (pH=3.0) (5:95)
Flow velocity: 1.0mL/min
Column temperature: 25 DEG C
Determined wavelength: 268nm
2. the preparation of solution
Extraction solvent is investigated: compare methyl alcohol, 50%, 70% methanol aqueous solution and sodium hydroxide solution as the extraction effect of solvent, find target compound methyl alcohol extraction effect and stability best, therefore determine that methyl alcohol is preferred Extraction solvent.
Extraction time is investigated: methyl alcohol is Extraction solvent, compares the extraction effect of ultrasonic 10min, 20min, 30min minute, and result shows that 10min just can extract completely, therefore for ensureing extraction effect, determines that extraction time is 15min.
The preparation of reference substance solution: get Hydrochioro reference substance 10mg, accurately weighed, put in the measuring bottle of 50mL, add methyl alcohol and dissolve, constant volume, shakes up, as Hydrochioro storing solution; Get control substance of Rutin 10mg, accurately weighed, put in the measuring bottle of 25mL, add methyl alcohol and dissolve, constant volume, shakes up, as rutin storing solution; Precision measures rutin storing solution 2.0mL, Hydrochioro storing solution 1.0mL, puts in 10mL measuring bottle, with 50% methanol dilution to scale, shakes up, to obtain final product.
The preparation of need testing solution: get Zhenju Jiangya Tablet fine powder 0.25g, accurately weighed, put in ground Erlenmeyer flask, precision adds methyl alcohol 25mL, jumps a queue, weighed weight, ultrasonic 15min, is chilled to room temperature, supplies the weight of less loss with methyl alcohol, shake up, filter, precision measures subsequent filtrate 1.0mL in 10mL measuring bottle, with 50% methanol dilution to scale, shake up, filter with 0.45 μm of miillpore filter, get subsequent filtrate as need testing solution.
3. determination method
Precision measures reference substance solution, each 20 μ L of need testing solution, injection liquid chromatography, measures according to high performance liquid chromatography (Chinese Pharmacopoeia 2010 editions annex VI D), and record peak area, calculates the content of Hydrochioro and rutin in test sample by one point external standard method.
4. Method validation
4.1 specificities and system suitability
Get scarce rutin negative sample (except not containing except rutin raw material, all the other supplementary materials and method for making are all identical with Zhenju Jiangya Tablet) and scarce Hydrochioro negative sample (except not containing except Hydrochioro raw material, all the other supplementary materials and method for making are all identical with Zhenju Jiangya Tablet), by need testing solution preparation method, preparation lacks rutin negative control solution and scarce Hydrochioro negative control solution respectively.Get reference substance solution, need testing solution and above-mentioned negative control solution sample introduction to measure, record DAD spectrum and chromatogram (see figure 4).With reference substance chromatographic peak same position place, negative control is noiseless; Present the chromatographic peak of retention time identical with reference substance chromatogram in test sample chromatogram, calculate with rutin, Hydrochioro chromatographic peak, theoretical cam curve is greater than equal 20000, and degree of separation is all greater than 2.0, and peak purity is greater than 995.Result shows the method specificity and system suitability well, meets assay requirement.
4.2 sample solution stability
Get with a Zhenju Jiangya Tablet need testing solution, respectively at after preparation 0,4,8,12,24h sample introduction measures, and calculates the RSD value of Hydrochioro and rutin peak area, Hydrochioro is 0.56%, rutin is 0.60%, is all not more than 2%, and interpret sample solution is stable in 24h.
4.3 linear relationships are investigated
Precision measures reference substance solution (containing Hydrochioro 21.84 μ g/mL, rutin 79.78 μ g/mL) 2 μ L, 5 μ L, 10 μ L, 20 μ L, 40 μ L sample introductions mensuration respectively, take sample size as horizontal ordinate, peak area is ordinate, drawing standard curve, and ask regression equation, related coefficient and the range of linearity.In table 1, Fig. 5,6.
Result shows, Hydrochioro regression equation is y=3.2853x+1.0712, r=0.99999, and within the scope of 42.96 ~ 859.20ng, during sample introduction, linear relationship is good; Rutin regression equation is y=1.7546x-34.038, r=0.99998, and within the scope of 159.56 ~ 3191.20ng, during sample introduction, linear dependence is good.
Table 1 linear relationship investigates result
4.4 instrument precisions are investigated
Precision measures reference substance solution 20 μ L, injection liquid chromatography, replication 6 times, calculates each pair of index components peak area RSD value.The peak area RSD value of Hydrochioro is 0.16%, the peak area RSD value of rutin is 1.10%, shows that the method instrument precision is good.In table 2.
Table 2 instrument precision investigates result
4.5 repeated experiment
Get same batch of Zhenju Jiangya Tablet (lot number is 101122), by the parallel preparation of need testing solution preparation method 6 parts of need testing solutions, measure in accordance with the law, with each index components cubage RSD value recorded.The RSD value of Hydrochioro content is 0.60%, the RSD value of rutin content is 0.46%, shows that the method repeatability is good.In table 3.
Table 3 repeated experiment result
4.6 Intermediate precision experiments
Get with a collection of Zhenju Jiangya Tablet test sample, investigate the Intermediate precision under different time, different personnel, different instrument three variables respectively.Result shows, the RSD value under three variables is all less than 2.0%, and therefore the method Intermediate precision is good.In table 4.
Table 4 Intermediate precision experimental result
4.7 recovery experiments
Get 6 parts, the Zhenju Jiangya Tablet powder with a collection of known content, every part of 0.125g, precision adds reference substance solution (containing Hydrochioro 0.1031mg/mL, rutin 0.3371mg/mL) 25mL, by legal system available test sample solution below " 1.3.2 " item, measure, the calculation sample recovery and RSD value thereof in accordance with the law.The Hydrochioro recovery is 101.99%, RSD value is 0.33%; The rutin recovery is 101.67%, RSD value is 0.82%, shows that the method accuracy is good.In table 5.
Table 5 recovery experimental result
5. sample determination
Get 3 batches of large production samples of Zhenju Jiangya Tablet, measure content in accordance with the law.Result shows, Hydrochioro average content is 4.86mg/ sheet, is equivalent to 97.27% of labelled amount, conforms with the regulations; Rutin average content is 15.74mg/ sheet, and be equivalent to 91.67% of labelled amount (be 85.85% calculating according to rutin bulk drug HPLC method content), content is on the low side.In table 6.
Table 6 three batches of Zhenju Jiangya Tablet assay results
The Determination of Content Uniformity of embodiment 2 Hydrochlorothiazide In Zhenjujiangya Tablets
1 chromatographic condition
With embodiment 1.
The preparation of 2 solution
Hydrochioro reference substance 10mg is got in the preparation of reference substance solution, accurately weighed, puts in the measuring bottle of 50mL, and add methyl alcohol and dissolve, constant volume, shakes up, as Hydrochioro storing solution; Precision measures Hydrochioro storing solution 1.0mL, puts in 10mL measuring bottle, with 50% methanol dilution to scale, shakes up, to obtain final product.
Zhenju Jiangya Tablet 1 is got in the preparation of need testing solution, puts in ground Erlenmeyer flask, adds 15g beaded glass (Φ 4-6mm), precision adds methyl alcohol 25mL again, jumps a queue, weighed weight, put oscillator vibration completely broken to label, take out, ultrasonic 15min, be chilled to room temperature, supply the weight of less loss with methyl alcohol, shake up, filter, precision measures subsequent filtrate 1.0mL in 10mL measuring bottle, with 50% methanol dilution to scale, shake up, filter with 0.45 μm of miillpore filter, get subsequent filtrate as need testing solution.
3 determination methods
Get test sample 10, prepare need testing solution by " need testing solution preparation " respectively; Precision measures reference substance solution and each 20 μ L of need testing solution, injection liquid chromatography, measure according to high performance liquid chromatography (" Chinese Pharmacopoeia " 2010 editions one annex VI D), record peak area, calculates the content of Hydrochioro in every sheet test sample by one point external standard method; Uniformity of dosage units is calculated by " Chinese Pharmacopoeia " 2010 editions two annex X E Content uniformity tests.
4 Method validation
4.1 specificities and system suitability
Get scarce Hydrochioro negative sample, by need testing solution preparation method, lack Hydrochioro negative control solution.Get reference substance solution, need testing solution and above-mentioned negative control solution sample introduction to measure, record DAD spectrum and chromatogram (see figure 7).With reference substance chromatographic peak same position place, negative control is noiseless; Present the chromatographic peak of retention time identical with reference substance chromatogram in test sample chromatogram, calculate with Hydrochioro chromatographic peak, theoretical cam curve is greater than 20000, and degree of separation is greater than 2.0, and peak purity is greater than 995.Result shows the method specificity and system suitability well, meets assay requirement.
4.2 sample solution stability
Get with a Zhenju Jiangya Tablet need testing solution, respectively at after preparation 0,4,8,12,24h sample introduction measures, calculate the RSD value of Hydrochioro and rutin peak area, Hydrochioro is 0.52%, and interpret sample solution is stablized in 24h.
4.3 linear relationships are investigated
With embodiment 1.
4.4 instrument precision experiments
With embodiment 1.
4.5 repeated experiment
Get with a collection of Zhenju Jiangya Tablet test sample, by the parallel preparation of need testing solution preparation method 6 parts of test liquids, measure, with Hydrochioro content meter in the test sample recorded, RSD is 1.20% in accordance with the law, shows that the method repeatability is good.In table 7.
Table 7 repeated experiment result
4.6 Intermediate precision experiments
Get with a collection of Zhenju Jiangya Tablet test sample, investigate the Intermediate precision under different time, different personnel, different instrument three variables respectively.With Hydrochioro content meter in the test sample recorded, different time Intermediate precision RSD value is 1.05%, and different personnel's Intermediate precision RSD value is 1.59%, and different instrument Intermediate precision RSD value is 0.60%, shows that the method Intermediate precision is good.In table 8.
Table 8 Intermediate precision test findings
4.7 recovery experiments
Get 6 parts, the Zhenju Jiangya Tablet powder with a collection of known content, every part of 0.125g, put in ground Erlenmeyer flask, add 15g beaded glass (Φ 4-6mm), then precision adds Hydrochioro reference substance solution (0.1025mg/mL) 25mL, jump a queue, weighed weight, puts oscillator vibration 30min, by legal system available test sample solution below " 2.1.2 " item, measure in accordance with the law, calculate the recovery and the RSD value thereof of Hydrochioro.In table 9.
5 sample determinations
Get 3 batches of large production samples of Zhenju Jiangya Tablet, measure uniformity of dosage units in accordance with the law.Result shows, three batches of large production sample uniformity of dosage units are all defective, and the preparation technology of prompting product or process control method existing problems, need be improved the uniformity of dosage units requirement meeting product.In table 10.
Table 9 recovery test result
Table 10 three batches of Zhenju Jiangya Tablet Determination of Content Uniformity results
The dissolution determination of embodiment 3 Hydrochlorothiazide In Zhenjujiangya Tablets and rutin
1 chromatographic condition
With embodiment 1.
2 determination methods
Hydrochioro reference substance 10mg is got in the preparation of reference substance solution, accurately weighed, puts in the measuring bottle of 50mL, and add methyl alcohol and dissolve, constant volume, shakes up, as Hydrochioro storing solution; Get control substance of Rutin 10mg, accurately weighed, put in the measuring bottle of 25mL, add methyl alcohol and dissolve, constant volume, shakes up, as rutin storing solution; Precision measures rutin storing solution 5.0mL, Hydrochioro storing solution 1.0mL, puts in 25mL measuring bottle, with 0.1mol/L(9 → 1000) hydrochloric acid solution be diluted to scale, shake up, to obtain final product.
The preparation of need testing solution is according to dissolution method (" Chinese Pharmacopoeia " version in 2010 two annex X C first methods), with 0.1mol/L(9 → 1000) hydrochloric acid solution 1000ml be dissolution medium, rotating speed is 75 turns per minute, operate in accordance with the law, through 60 minutes time, get liquid, filter with miillpore filter, get subsequent filtrate as need testing solution.
Precision measures reference substance solution, each 20 μ l injection liquid chromatographies of need testing solution, measures by chromatographic condition in embodiment 1, and record peak area, calculates the stripping quantity of Hydrochioro and rutin in every sheet by external standard method.The dissolution determination of three batch samples the results are shown in Table 11:
Table 11 three batches of Zhenju Jiangya Tablet dissolution determination results
The above; be only preferred embodiment of the present invention; not to any formal and substantial restriction of the present invention; should be understood that; for those skilled in the art; under the prerequisite not departing from the inventive method, also can make some improvement and supplement, these improve and supplement and also should be considered as protection scope of the present invention.All those skilled in the art, without departing from the spirit and scope of the present invention, a little change made when utilizing disclosed above technology contents, the equivalent variations of modifying and developing, be Equivalent embodiments of the present invention; Meanwhile, all according to substantial technological of the present invention to the change of any equivalent variations that above-described embodiment is done, modify and differentiation, all still belong in the scope of technical scheme of the present invention.
Claims (10)
1. the content assaying method of a Hydrochlorothiazide In Zhenjujiangya Tablets and/or rutin, medicine is comprised the following steps: to be prepared into need testing solution, it is then analytical column with anion-exchange column, using methyl alcohol-phosphate buffer system as mobile phase, with wavelength optional between 250nm-280nm for determined wavelength, high performance liquid chromatography is adopted quantitatively to detect the content of Hydrochioro and/or rutin in need testing solution; Described anion-exchange column is HP-SAX anion-exchange column; In described methyl alcohol-phosphate buffer system, described phosphate buffer is pH2.5-3.5KH
2pO
4concentration is the phosphate buffer of 10-40mmol/L, and the volume ratio of methyl alcohol and phosphate buffer is 5:95-20:80;
During high effective liquid chromatography for measuring, the flow velocity of mobile phase is 0.5-1.5mL/min, and column temperature is 25-35 DEG C;
After method medicine being prepared into need testing solution comprises the following steps: medicine and Extraction solvent to mix, ultrasonic 10-30 minute, obtains need testing solution after Separation of Solid and Liquid; Described extraction reagent is methyl alcohol or percent by volume is the methanol aqueous solution of more than 50%.
2. the content assaying method of Hydrochlorothiazide In Zhenjujiangya Tablets and/or rutin as claimed in claim 1, it is characterized in that, described methyl alcohol-phosphate buffer system is pH3.0 and KH
2pO
4concentration is the phosphate buffer of 40mmol/L, and the volume ratio of methyl alcohol and phosphate buffer is 5:95.
3. the content assaying method of Hydrochlorothiazide In Zhenjujiangya Tablets and/or rutin as claimed in claim 1, it is characterized in that, during high effective liquid chromatography for measuring, the flow velocity of mobile phase is 1.0mL/min.
4. the content assaying method of Hydrochlorothiazide In Zhenjujiangya Tablets and/or rutin as claimed in claim 1, it is characterized in that, during high effective liquid chromatography for measuring, column temperature is 25 DEG C.
5. the content assaying method of Hydrochlorothiazide In Zhenjujiangya Tablets and/or rutin as claimed in claim 1, it is characterized in that, during high effective liquid chromatography for measuring, determined wavelength is 268nm.
6. the content assaying method of Hydrochlorothiazide In Zhenjujiangya Tablets and/or rutin as claimed in claim 1, it is characterized in that, after method medicine being prepared into need testing solution comprises the following steps: medicine and Extraction solvent to mix, ultrasonic 15 minutes, after Separation of Solid and Liquid, obtain need testing solution.
7. the content assaying method of Hydrochlorothiazide In Zhenjujiangya Tablets and/or rutin as described in as arbitrary in claim 1-6, it is characterized in that, in the content assaying method of described Hydrochlorothiazide In Zhenjujiangya Tablets and rutin, adopt the same terms to measure the content of Hydrochioro and/or rutin in reference substance solution, and press the content that one point external standard method calculates Hydrochioro and/or rutin in need testing solution.
8. a purposes for the content assaying method of Hydrochlorothiazide In Zhenjujiangya Tablets and/or rutin as described in claim as arbitrary in claim 1-7 is the assay for Hydrochlorothiazide In Zhenjujiangya Tablets and/or rutin or the Determination of Content Uniformity for Hydrochlorothiazide In Zhenjujiangya Tablets or the dissolution determination for chlorothiazide and/or rutin in Zhenju Jiangya Tablet.
9. a quality determining method for Zhenju Jiangya Tablet, comprises and adopts method described in claim 1-7 arbitrary claim to detect the content of Hydrochlorothiazide In Zhenjujiangya Tablets and rutin, and the uniformity of dosage units of Hydrochioro in Zhenju Jiangya Tablet.
10. quality determining method as claimed in claim 9, also comprises and utilizes method described in arbitrary claim in claim 1-5 and 7 to detect the dissolution rate of chlorothiazide and/or rutin in Zhenju Jiangya Tablet.
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