CN105572240A - Method for detecting content of pharmaceutic adjuvant carmine by using high performance liquid chromatography - Google Patents
Method for detecting content of pharmaceutic adjuvant carmine by using high performance liquid chromatography Download PDFInfo
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- CN105572240A CN105572240A CN201410618435.2A CN201410618435A CN105572240A CN 105572240 A CN105572240 A CN 105572240A CN 201410618435 A CN201410618435 A CN 201410618435A CN 105572240 A CN105572240 A CN 105572240A
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- carmine
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- pharmaceutic adjuvant
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Abstract
The invention belongs to the field of medicine inspection, and relates to a pharmaceutic adjuvant analyzing and testing method, in particular to a method for testing the content of carmine. The method comprises the following steps: adopting an Agilent Zorbax SB-C18 (250mm*4.6mm, 5[mu]m) chromatographic column, and taking methanol and a 0.02 mol/L ammonium acetate solution (30:70) as mobile phases; testing the recycling rates of samples of 5 adjuvants, wherein the flowing speed is 1.0 mL/min, the detection wavelength is 333 nm, the column temperature is 35 DEG C, the recycling rate is 95.3-99.5%, the average recycling rate is 99.0%, and the relative standard deviation (RSD) is 1.6%. The recycling rate is greater than 80%, the linear range of the carmine is 0.050-1.000 [mu]g, r is 0.9999, and the result shows that the method is simple and accurate and can be used as a quality control method for the pharmaceutic adjuvant carmine.
Description
Technical field
The invention belongs to drug inspection field, relate to the analysis determining method veterinary inspection field of pharmaceutic adjuvant.Be specifically related to the assay method of carmine content.
Background technology
In recent years, the quality standard about medicine has obtained larger perfect, and stops on the level that relatively falls behind about the quality control standard of pharmaceutic adjuvant is also most.Quality due to pharmaceutic adjuvant quality is directly connected to pharmaceutical preparation security in vivo and validity, the research of the quality control standard of pharmaceutic adjuvant with improve extremely urgent.Famille rose is one of conventional synthetic food color, is mainly used as the colorant etc. of capsule shells, records in currency of Chinese medical industry standard YY0145-93 at pharmaceutical field.Adopt titanium trichloride titration measuring content in this standard, complex operation is time-consuming, and measurement result error is larger.This test, on the basis with reference to pertinent literature (1 ~ 3), is studied its content assaying method, establishes the high-efficient liquid phase content assay method of pharmaceutic adjuvant famille rose.
At present, the research of famille rose is mainly concentrated on it as feed coloring agent aspect both at home and abroad, and very few for the research of carmine detection method aspect, mainly contain spectrophotometer method, thin-layered chromatography and high performance liquid chromatography etc.But these method operating process are more loaded down with trivial details, be not suitable for detecting batch samples.Need badly and set up a kind of accurate, efficient, safe high performance liquid chromatography (HPLC method), residual for detecting pharmaceutic adjuvant famille rose, for ensureing that drug safety provides technical support.Therefore, study by the screening to the pre-treatment of testing sample and chromatographic condition, optimization, be intended to set up carmine residual HPLC method detection technique.
Summary of the invention
High performance liquid chromatography utilizes high pressure pump to order about the chromatographic column of mobile phase by filling Stationary liquid, because component each in potpourri is variant in character and structure, the size of having an effect with Stationary liquid is also variant, therefore under the effect of same expulsive force, the hold-up time of different component in Stationary liquid is different, thus flow out from Stationary liquid according to sequencing, reach the effect that material is separated, and carry out the method for quantitative test and qualitative analysis.
The present invention includes following steps:
(1) sample pretreatment
Accurate to draw in carmine red standard solution 1mL to 10mL volumetric flask, add water and make every 1mL and namely obtain reference substance solution containing the solution of 50 μ g.Get this product and be about 0.1g, accurately weighed, put in 100mL volumetric flask, add water appropriate dissolving, and be diluted to scale, and shake up, precision measures 1mL and puts in 20mL volumetric flask, is diluted with water to scale, shakes up, and with 0.45 μm of filtering with microporous membrane, obtains test solution.
(2) with high performance liquid chromatograph (model is Waters2695), UV-detector (model is Waters2487) and AgilentZorbaxSB-C
18(250mm × 4.6mm, 5 μm) chromatographic column detects as test apparatus, record chromatogram, and interior mark peak height (Hi) and carmine peak height (Hs), use typical curve converted score.
The assay method of carmine content according to claim 1, is characterized in that the method is AgilentZorbaxSB-C
18(250mm × 4.6mm, 5 μm) are chromatographic column, and wavelength is 333nm, and flow velocity is 1.0mL/min, and mobile phase is methyl alcohol-0.02molL
-1ammonium acetate solution (30: 70), column temperature is 35 DEG C, and sample size is 50 μ L.
The assay method of Carophyll red content according to claim 1, is characterized in that the method is SymmetryC185 μm, and 3.9 × 150mm is chromatographic column, wavelength is 475nm, and flow velocity is 1.0mL/min, and mobile phase is methyl alcohol: water=95: 5, column temperature is 30 DEG C, and sample size is 50 μ L.
The invention has the beneficial effects as follows: the present invention's application high performance liquid chromatography detects pharmaceutic adjuvant carmine content and has good veracity and precision.
Embodiment
To the specific embodiment of the present invention be described in detail below:
1. instrument and reagent:
Instrument:
Waters2695 high performance liquid chromatograph (band DAD detecting device)
Reagent:
Edible synthesized coloring matter carmine red standard solution (0.500mgmL
-1, lot number: GBW (E) 100004a09001, by metering scientific research, institute provides); Carmine (lot number: 20100901,20100902,20100903 is provided by certain manufacturing enterprise); Methyl alcohol is chromatographically pure, and water is ultrapure water, and it is pure that other reagent is analysis.
2. method and result
Chromatographic condition and system suitability
Adopt AAgilentZorbaxSB-C
18(250mm × 4.6mm, 5 μm) are chromatographic column, and wavelength is 333nm, and flow velocity is 1.0mL/min, and mobile phase is methyl alcohol-0.02molL
-1ammonium acetate solution (30: 70), column temperature is 35 DEG C, and sample size is 50 μ L, and number of theoretical plate should be not less than 4000 in carmine peak.Under this chromatographic condition, accurate draw blank solvent, reference substance solution and need testing solution 10 μ L, injection liquid chromatography.
The making of typical curve
The carmine reference substance solution of accurate absorption (50 μ gmL
-1) 1,5,10,15,20 μ L, inject
High performance liquid chromatograph, measures, obtains peak area.Take peak area as ordinate, sample size (μ g) is horizontal ordinate, drawing standard curve, obtains regression equation and is: Y=3536.9X+21.753, r=0.9999 (n=5).Result shows, carmine good in 0.050 ~ 1.000 μ g scope internal linear relation.
Precision test
Accurate absorption need testing solution 10 μ L, difference sample introduction 6 times under above-mentioned chromatographic condition, the RSD of carmine integrating peak areas value is 0.9% (n=6).Result shows: instrument precision is good.
Stability test
Get same need testing solution, respectively at 0,1,5,10,15,20,24h carries out measuring its peak area, its RSD is 1.1% (n=6).Result shows that need testing solution is stable in 24h.
Replica test
Get carmine sample, prepare 6 increment product by the compound method of above-mentioned need testing solution, under above-mentioned chromatographic condition, sample introduction measures, and average content is 93.12%, RSD is 0.8% (n=6).Result shows: the method is reproducible.
Recovery test
The carmine sample getting known content (93.12%) is about 0.05g totally 6 parts, accurately weighed, is placed in 100mL volumetric flask, is settled to scale, shakes up after appropriate dissolving that add water.Precision measures 1mL and puts in 10mL volumetric flask, and precision adds 100 μ gmL respectively
-1carmine red standard solution is (in accurate absorption carmine red standard solution 5mL to 25mL volumetric flask, add water and make the solution of every 1mL containing 100 μ g) 5mL, add water and be settled to scale, shake up, as need testing solution, measure carmine content by chromatographic condition, calculate the recovery, result average recovery rate is 99.0%, RSD is 1.6% (n=6).
Sample size measures
Sample thief 3 batches, by need testing solution preparation method preparation, measures in accordance with the law and calculates, in table 1.
Table 1
Claims (2)
1. the assay method of carmine content, is characterized in that after testing sample pre-service, detects, comprise the following steps with high performance liquid chromatography:
Sample pretreatment
(1) accurately to draw in carmine red standard solution 1mL to 10mL volumetric flask, add water and make every 1mL and namely obtain reference substance solution containing the solution of 50 μ g; Get this product and be about 0.1g, accurately weighed, put in 100mL volumetric flask, add water appropriate dissolving, and be diluted to scale, and shake up, precision measures 1mL and puts in 20mL volumetric flask, is diluted with water to scale, shakes up, and with 0.45 μm of filtering with microporous membrane, obtains test solution;
(2) with high performance liquid chromatograph (model is Waters2695), UV-detector (model is Waters2487) and AgilentZorbaxSB-C
18(250mm × 4.6mm, 5 μm) chromatographic column detects as test apparatus, record chromatogram, and interior mark peak height (Hi) and carmine peak height (Hs), use typical curve converted score.
2. the assay method of carmine content according to claim 1, is characterized in that the method is AgilentZorbaxSB-C
18(250mm × 4.6mm, 5 μm) are chromatographic column, and wavelength is 333nm, and flow velocity is 1.0mL/min, and mobile phase is methyl alcohol-0.02molL
-1ammonium acetate solution (30: 70), column temperature is 35 DEG C, and sample size is 50 μ L.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106483205A (en) * | 2015-08-31 | 2017-03-08 | 江苏维赛科技生物发展有限公司 | A kind of method that employing high performance liquid chromatography detects pharmaceutic adjuvant carmine content |
| CN107037152A (en) * | 2017-04-14 | 2017-08-11 | 汤臣倍健股份有限公司 | The detection method of carminic acid content in a kind of gelatine capsule |
| CN108387657A (en) * | 2018-03-07 | 2018-08-10 | 吉林师范大学 | The detection method of colorant content in Waterelon Frost Lozenges |
-
2014
- 2014-11-06 CN CN201410618435.2A patent/CN105572240A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106483205A (en) * | 2015-08-31 | 2017-03-08 | 江苏维赛科技生物发展有限公司 | A kind of method that employing high performance liquid chromatography detects pharmaceutic adjuvant carmine content |
| CN107037152A (en) * | 2017-04-14 | 2017-08-11 | 汤臣倍健股份有限公司 | The detection method of carminic acid content in a kind of gelatine capsule |
| CN107037152B (en) * | 2017-04-14 | 2019-04-09 | 汤臣倍健股份有限公司 | The detection method of carminic acid content in a kind of gelatine capsule |
| CN108387657A (en) * | 2018-03-07 | 2018-08-10 | 吉林师范大学 | The detection method of colorant content in Waterelon Frost Lozenges |
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