CN109358132A - A kind of method of separating and assaying of the aspartic acid laumosaren glucose injection in relation to substance - Google Patents
A kind of method of separating and assaying of the aspartic acid laumosaren glucose injection in relation to substance Download PDFInfo
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Abstract
The present invention relates to Pharmaceutical Analysis fields, and in particular to a kind of method of separating and assaying of the aspartic acid laumosaren glucose injection in relation to substance.This method includes being detected using aspartic acid laumosaren glucose injection as test solution using high performance liquid chromatography, the high performance liquid chromatography uses octadecylsilane chemically bonded silica for the chromatographic column of filler, carries out linear gradient elution with mobile phase A and mobility B；The mobile phase A is phosphate solution-methanol；The Mobile phase B is acetonitrile.This method is used for detection of the aspartic acid laumosaren glucose injection in relation to substance, has the characteristics that specificity is strong, accuracy is high, reproducible, and more impurity peaks can be determined than statutory standards, the total impurities amount of detection is bigger.
Description
Technical field
The present invention relates to Pharmaceutical Analysis fields, and in particular to a kind of related object of aspartic acid laumosaren glucose injection
The method of separating and assaying of matter.
Background technique
Currently, the examination criteria of aspartic acid laumosaren glucose injection is marked through State Food and Drug Administration
It is quasi- to there is YBH30102005 (A factory), YBH15122004 (B factory) and national drug standards new drug to become a full member the 64th WS1- of standard
(X-097)-2005Z.Measuring method in above-mentioned detection method in relation to substance is all made of liquid chromatography for measuring, wherein
Measuring method regulation in YBH30102005 (A factory), YBH15122004 (B factory) in relation to substance is consistent, existing examination criteria
Related substance is all measured using isocratic elution method, retention time is usually to record to 3 times of retention times of main peak, and mobile phase is adopted
With acetonitrile-phosphate buffer (weighing potassium dihydrogen phosphate 6.7g, add phosphoric acid 2.8ml, be diluted with water to 1000ml) -0.05mol/
L tetrabutyl phosphonium bromide ammonium salt solution is mixed according to volume ratio 15:85:4, Detection wavelength: 288nm, sample volume: 20 μ l, test sample
Concentration 0.2mg/ml compares concentration: 2 μ g/ml.
Related substance detecting method has the following deficiencies: that separating degree is bad between impurity peaks in above-mentioned standard, 5- methylol
Furfural (5-HMF), Lomefloxacin main peak separate bad with other impurities peak, there is interference；Impurity peaks cannot belong to very well source (such as
Oxidation, light degradation generate)；Lomefloxacin main peak peak type is poor, there is hangover；There is ion-pairing agent (tetrabutyl phosphonium bromide in mobile phase
Ammonium), it uses for a long time larger to chromatographic column damage.Typical high-efficient liquid phase chromatogram such as Fig. 1 in YBH30102005 (according to having
Close the detection method measurement of substance) shown in, wherein peak 1 is 5 hydroxymethyl furfural impurity peaks；Peak 2 is other impurity peaks；Peak 3 is
Lomefloxacin peak.
Summary of the invention
A kind of separation survey the purpose of the present invention is to provide aspartic acid laumosaren glucose injection in relation to substance
Determine method, this method is used for detection of the aspartic acid laumosaren glucose injection in relation to substance, Lomefloxacin and impurity, with
And can be good at separating between each impurity, have the characteristics that specificity is strong, accuracy is high, reproducible, and can calibrate than method
Standard determines more impurity peaks, and the total impurities amount of detection is bigger.
Above-mentioned purpose of the invention adopts the following technical solutions to realize:
A kind of method of separating and assaying of the aspartic acid laumosaren glucose injection in relation to substance provided by the invention, packet
It includes using aspartic acid laumosaren glucose injection as test solution and is detected using high performance liquid chromatography, it is described
High performance liquid chromatography uses octadecylsilane chemically bonded silica for the chromatographic column of filler, carries out line with mobile phase A and mobility B
Property gradient elution；The mobile phase A is phosphate solution-methanol；The Mobile phase B is acetonitrile.
In the inventive solutions, it is preferable that the preparation method of the mobile phase A are as follows: be by mass percentage
0.6%~0.7% potassium dihydrogen phosphate and methanol is 70:30~80:20 mixing according to volume ratio, adjusts it using phosphoric acid
PH value is to 2.5~3.0.
In the inventive solutions, it is preferable that the mass percentage of the potassium dihydrogen phosphate is 0.67%,
The volume ratio of the potassium dihydrogen phosphate and the methanol is 76:24, and the pH value is 2.7.
In the inventive solutions, it is preferable that the program of the gradient elution are as follows:
In the inventive solutions, it is preferable that the model Welch Ultimate XB-C18 of the chromatographic column,
4.6 × 250mm, 5 μm.
In the inventive solutions, it is preferable that the Detection wavelength of the high performance liquid chromatography is 280~290nm；
More preferably 287nm.
In the inventive solutions, it is preferable that the flow velocity of the high performance liquid chromatography is 0.5~1.5ml/min；
More preferably 1.0ml/min.
In the inventive solutions, it is preferable that the sample volume of the high performance liquid chromatography is 5~20 μ L；It is more excellent
It is selected as 10 μ L.
In the inventive solutions, it is preferable that the chromatographic column column temperature of the high performance liquid chromatography is 35~45 DEG C；
More preferably 40 DEG C.
A kind of method of separating and assaying of the aspartic acid laumosaren glucose injection in relation to substance provided by the invention
The utility model has the advantages that
(1) method of separating and assaying provided by the invention is with Welch Ultimate XB-C18 (4.6 × 250mm, 5 μm)
Chromatographic column, using mobile phase A (phosphate solution (weighing potassium dihydrogen phosphate 6.7g, water 1000ml is added to make to dissolve)-methanol (76:
24) (with phosphorus acid for adjusting pH value to 2.7)) and Mobile phase B (acetonitrile) be mobile phase carry out linear gradient elution, can greatly improve
In relation to the separating degree between substance in existing standard, wherein 5 hydroxymethyl furfural, Lomefloxacin main peak are separated with other impurities peak
Degree is greater than 1.5, and the peak-to-peak separating degree of remaining impurity is also preferable, and Lomefloxacin main peak peak type is symmetrical, not the trailing phenomenon of original method.
This method has the characteristics that specificity is strong, accuracy is high, reproducible, and more impurity peaks can be determined than statutory standards,
The total impurities amount of detection is bigger, therefore, can be preferably applied to the quality-monitoring of aspartic acid laumosaren injection.
(2) impurity that acid, alkali, light, oxidation and high temperature generate can be detected preferably according to this method, main peak is with before
Other impurities peak energy realizes preferable separation afterwards, and separating effect is also preferable between each impurity peaks.It is broken by acid, alkali, light, oxidation
It is bad, the related material impurities in aspartic acid laumosaren glucose injection sample are belonged to, medicine is improved to enterprise
Quality provides reference.
(3) in mobile phase only have potassium dihydrogen phosphate, methanol, acetonitrile and water, be free of ion-pairing agent, to chromatographic column damage compared with
Original method is low.
Detailed description of the invention
Fig. 1 is according to the resulting high-efficient liquid phase chromatogram of existing chromatographic process；
Fig. 2 is the high-efficient liquid phase chromatogram of 1 gained test solution of embodiment；
Fig. 3 is the high-efficient liquid phase chromatogram of 1 gained contrast solution of embodiment；
Fig. 4 is the high-efficient liquid phase chromatogram of 2 gained system suitability solution of embodiment；
Fig. 5 is 3 gained Lomefloxacin linear relationship chart of embodiment；
Fig. 6 is the high-efficient liquid phase chromatogram of 7 gained specificity of embodiment test；
Fig. 7 is the high-efficient liquid phase chromatogram one of 8 gained photo damage of embodiment test；
Fig. 8 is the high-efficient liquid phase chromatogram two of 8 gained photo damage of embodiment test；
Fig. 9 is the high-efficient liquid phase chromatogram one of embodiment 8 gained high temperature and strong acid failure test；
Figure 10 is the high-efficient liquid phase chromatogram two of embodiment 8 gained high temperature and strong acid failure test；
Figure 11 is the high-efficient liquid phase chromatogram of 8 gained Oxidative demage of embodiment test；
Figure 12 is the high-efficient liquid phase chromatogram of 8 gained highly basic failure test of embodiment；
Figure 13 is the source ownership figure of the relative substance provided obtained by embodiment 8.
Specific embodiment
It in order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below will be in the embodiment of the present invention
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, according to normal conditions or manufacturer builds
The condition of view carries out.Reagents or instruments used without specified manufacturer is the conventional production that can be obtained by commercially available purchase
Product.
Unless otherwise specified, the reference substance Lomefloxacin used in the following embodiments is examined from Chinese food drug
Determine research institute's (lot number: 130452-200902, content: 90.3%)；The present embodiments relate to test sample L-aminobutanedioic acid Lome
Husky star glucose injection, specification are 100ml, wherein the aspartic acid laumosaren contained is with Lomefloxacin
(C17H19F2N3O3) meter 0.2g, glucose 5g；Aspartic acid laumosaren raw material involved in the embodiment of the present invention refers to a winter
The solid material medicine of propylhomoserin Lomefloxacin；High performance liquid chromatograph model LC-20AT (the band PDA that the embodiment of the present invention uses
Detector), it is purchased from Shimadzu Corporation；Electronic balance model MS205DU is purchased from METTLER company.
Feature and performance of the invention are described in further detail with reference to embodiments.
Embodiment 1: the foundation of chromatographic condition
Chromatographic column: Welch Ultimate XB-C18 (4.6 × 250mm, 5 μm)
Mobile phase A: phosphate solution (weighing potassium dihydrogen phosphate 6.7g, water 1000ml is added to make to dissolve)-methanol (76:24)
(with phosphorus acid for adjusting pH value to 2.7)
Mobile phase B: acetonitrile
Detection wavelength: 287nm
Flow velocity: 1ml/min
Column temperature: 40 DEG C
Sample volume: 10 μ l.
Elution program according to the form below 1 carries out linear gradient elution:
1 elution program of table
Solution is prepared: taking aspartic acid laumosaren glucose injection is test solution (2.0mg/ containing Lomefloxacin
Ml), accurate to measure test solution 1ml, add mobile phase A to be diluted to 500ml, as contrast solution (the 40 μ g/ containing Lomefloxacin
ml)。
Measuring method: test solution and each 10 μ l injection liquid chromatograph of contrast solution are taken, according to above-mentioned chromatographic condition
It is measured respectively, records chromatogram, the high-efficient liquid phase chromatogram of test solution is shown in Fig. 2, the high-efficient liquid phase color of contrast solution
Spectrogram is shown in Fig. 3.
From Fig. 2~3 it is found that related substance in existing standard can be greatly improved by carrying out gradient elution using above-mentioned mobile phase
Between separating degree, wherein 5 hydroxymethyl furfural (retention time 4.915min), Lomefloxacin main peak and other impurities peak point
It is greater than 1.5 from degree, the peak-to-peak separating degree of remaining impurity is also preferable, and Lomefloxacin main peak peak type is symmetrical, and the hangover of original method is not existing
As.
Embodiment 2: system suitability
Lomefloxacin reference substance about 22mg is taken, adds 30% hydrogenperoxide steam generator 1ml, heating water bath 2 after adding water 1ml to dissolve again
It is hour (Oxidative demage), cooling, add mobile phase A to be diluted to 10ml, shake up, precision measures 10 μ l of this solution and injects liquid chromatogram
Instrument is measured according to the chromatographic condition in embodiment 1, records chromatogram, as a result as shown in Figure 4.
From fig. 4, it can be seen that Lomefloxacin peak retention time is about 22.0min, Lomefloxacin peak and other impurities peak point
1.5 (5.7 and 2.1) are all larger than from degree.
Embodiment 3: linear relationship
Precision weighs Lomefloxacin reference substance 21.84mg, sets in 100ml measuring bottle, and the mobile phase A for adding embodiment 1 to provide is molten
Scale is solved and be diluted to, is shaken up, the reference substance solution that concentration containing Lomefloxacin is about 0.2mg/ml is made.Accurate measurement is above-mentioned again
Appropriate reference substance solution, concentration, which is made, with mobile phase A dilution respectively may be about the solution of 40 μ g/ml, 20 μ g/ml and 2 μ g/ml.Point
Reference substance solution 10 the μ l, 20 μ g/ml of inaccurate 20 μ l of reference substance solution, the 10 μ l for measuring 0.2mg/ml and 5 μ l, 40 μ g/ml
10 μ l of reference substance solution and 5 μ l, 2 μ g/ml 10 μ l of reference substance solution inject liquid chromatograph, by the chromatography in embodiment 1
Condition measures peak area, and using integrating peak areas value as ordinate, control quality is abscissa, draws standard curve, recurrence side
Journey is y=5791.4x+69480, R2=0.9999, gained standard curve is shown in Fig. 5.
As seen from Figure 5, Lomefloxacin is in good linear between 19.72~3944ng in quality.
Embodiment 4: detection limit, quantitative limit
Precision measures Lomefloxacin the reference substance solution 660 μ l and 220 μ l that concentration is about 2 μ g/ml, sets in 10ml measuring bottle,
Add mobile phase A to be diluted to scale, shake up, difference is accurate to measure 10 μ l, injects in liquid chromatograph, as detection limit and quantitatively
Limit, chromatogram show that its signal-to-noise ratio is respectively 5.09 and 17.72, and the detection limit and quantitative limit of this method Lomefloxacin are respectively
0.43ng and 1.30ng.
Embodiment 5: precision test
Precision measure concentration be 40 μ g/ml 10 μ l of Lomefloxacin reference substance solution, inject liquid chromatograph in, continuously into
Sample 6 times, chromatogram is recorded, peak area is respectively 2366997,2365606,2366583,2365493,2365539,2366008,
RSD is 0.03%, shows that this method precision is good.
Embodiment 6: repetitive test
Take 6 parts of aspartic acid laumosaren glucose injection (B factory, lot number 161103, specification are as follows: 100ml, Lome are husky
Star 0.2g and glucose 5g), it is measured according to the chromatographic condition that embodiment 1 provides, as a result see Table 2 for details:
2 repetitive test data of table
As shown in Table 2, for RSD less than 2%, single impurity RSD is respectively less than 6% to total impurities (average 0.3%), shows this method
It is reproducible.
Embodiment 7: specificity test
Blank test: take 5% glucose solution as blank auxiliary solution.Precision measures blank auxiliary solution and mobile phase
Each 10 μ l of solution A, is measured according to the chromatographic condition in embodiment 1 respectively, records chromatogram, as a result sees Fig. 6, wherein bent
Line 4 is the test result of 5% glucose solution；Curve 5 is the test result of mobile phase A.
As seen from Figure 6, mobile phase A is noiseless to this law, in blank auxiliary the peak 5-HMF in about 5min or so appearance,
And other impurity peaks are not detected in this position in sample, it is seen then that mobile phase A and glucose to the detection in relation to substance substantially not
There are interference, test mode provided by the invention has good for the measurement of aspartic acid laumosaren glucose injection
Specificity.In addition, can also find out that 5-HMF is able to achieve with impurity in sample and is kept completely separate by Fig. 6, illustrate that this method is also applicable in
The detection of 5-HMF in aspartic acid laumosaren glucose injection.
Embodiment 8: destructive testing
Forced degradation test is the strong degradation condition in simulation strong acid, highly basic, oxidation, illumination and high temperature etc., acceleration pair
Aspartic acid laumosaren is destroyed, it is therefore an objective to by the separation for investigating the catabolite and main peak and known impurities of sample
Situation, the efficiency and applicability of analysis and assessment method.
In order to preferably investigate the specificity and stability of this method, the strong of acid, alkali, oxidation, illumination and high temperature is devised
Degrading experiment processed.
(1) photo damage is tested
Day photo damage: aspartic acid laumosaren glucose injection (B factory, lot number 161103, specification: 100ml: Lip river are taken
U.S. sand star 0.2g and glucose 5g) in intensity of illumination 4000lx, it is surveyed after placing 24 hours according to the chromatographic condition in embodiment 1
It is fixed.
1 (ultraviolet destruction) of ultraviolet destruction: aspartic acid laumosaren glucose injection (B factory, lot number 161103, rule are taken
Lattice: 100ml: Lomefloxacin 0.2g sets in glass evaporating dish with glucose 5g) liquid, and ultraviolet light irradiates after 2h according to embodiment 1
In chromatographic condition measurement.
Control: aspartic acid laumosaren raw material (without destroying) (D factory provides, lot number 20151201) plus water is taken to be made
The solution of the about 2.0mg/ml containing Lomefloxacin is measured according to the chromatographic process in comparative examples 1.
2 (ultraviolet destructions) of ultraviolet destruction: taking aspartic acid laumosaren raw material (D factory provides, lot number 20151201), ultraviolet
Line measures after irradiating 6h according to the chromatographic condition in embodiment 1.
The chromatogram of above-mentioned photo damage test result is shown in Fig. 7~8, and in Fig. 7, curve 6 is the test knot of ultraviolet destruction 1
Fruit, curve 7 are the test result of day photo damage, and curve 8 is the test result of control；Fig. 8 is the test result of ultraviolet destruction 2.
(2) high temperature and strong acid failure test
High temperature 1: aspartic acid laumosaren raw material (D factory provides, lot number 20151201) about 27mg is taken, water 10ml is added
Dissolution, is made the solution of the about 2mg/ml containing Lomefloxacin, and boiling water bath measures after heating 4h according to the chromatographic condition in embodiment 1.
High temperature 2: take aspartic acid laumosaren raw material (D factory provide, lot number 20151201) respectively at 130 DEG C and
After heating 2h under the conditions of 160 DEG C, raw material about 27mg is taken, water 10ml is added to dissolve, is measured according to the chromatographic condition in embodiment 1.
Strong acid destroys: taking aspartic acid laumosaren raw material (D factory provides, lot number 20151201) about 27mg, adds water 1ml molten
The solution of the about 20mg/ml containing Lomefloxacin is made in solution, and 10mol/L hydrochloric acid solution 1ml is added, after boiling water bath heating 2 as a child,
With 10mol/L sodium hydroxide solution 1ml neutralize, then plus mobile phase A be diluted to 10ml, according in embodiment 1 chromatographic condition survey
It is fixed.
High temperature 1 and strong acid failure test result are shown in Fig. 9, wherein curve 9 is the test result of high temperature 1, curve
10 test results destroyed for strong acid；The test result of high temperature 2 is shown in Figure 10, wherein curve 11 is the test knot at 160 DEG C
Fruit, curve 12 are the test result at 130 DEG C.
(3) Oxidative demage is tested
Oxidative demage: aspartic acid laumosaren raw material (D factory provides, lot number 20151201) about 27mg is taken, adds water 1ml molten
Solution, is made the solution of the about 20mg/ml containing Lomefloxacin, adds 30% hydrogenperoxide steam generator 1ml, after boiling water bath 2 hours, then plus
Mobile phase A is diluted to 10ml, measures according to above-mentioned chromatogram condition.
Control 1: taking aspartic acid laumosaren glucose injection, (B factory, lot number 161103, specification: 100ml: Lome is husky
Star 0.2g and glucose 5g) without destruction, it is measured according to the chromatographic process in embodiment 1.
Control 2: aspartic acid laumosaren glucose injection (A factory, lot number 21612201-1, specification: 100ml: Lip river are taken
U.S. sand star 0.2g and glucose 5g) without destruction, it is measured according to the chromatographic process in embodiment 1.
Test result is shown in Figure 11, and wherein curve 13 is the test result of Oxidative demage, and curve 14 is the measurement knot for compareing 1
Fruit, curve 15 are the test result for compareing 2.
(4) highly basic failure test
Highly basic destroys 1: taking aspartic acid laumosaren raw material (D factory provides, lot number 20151201) about 27mg, adds water 1ml
The solution of the about 20mg/ml containing Lomefloxacin is made in dissolution, is added 10mol/L sodium hydroxide solution 1ml (can generate precipitating), boiling
Heating water bath 2 as a child after, with 10mol/L hydrochloric acid solution 1ml neutralize, then plus mobile phase A be diluted to 10ml, according to embodiment 1
In chromatographic condition measurement.
Highly basic destroys 2: taking aspartic acid laumosaren raw material (D factory provides, lot number 20151201) about 27mg, adds water 1ml
The solution of the about 20mg/ml containing Lomefloxacin is made in dissolution, and 10mol/L sodium hydroxide solution about 0.4ml is added, and (it is heavy not generate
Form sediment), boiling water bath heating 2 as a child after, with 10mol/L hydrochloric acid solution 0.4ml neutralize, then plus mobile phase A be diluted to 10ml, according to
Chromatographic condition measurement in embodiment 1.
Control: aspartic acid laumosaren raw material (without destroying) (D factory provides, lot number 20151201) plus water is taken to be made
The solution of the about 2.0mg/ml containing Lomefloxacin is measured according to the chromatographic process in comparative examples 1.
Test result is shown in Figure 12, wherein curve 16 is the test result that highly basic destroys 1, and curve 17 is the survey that highly basic destroys 2
Test result, while curve 8 is set as control.
(5) destructive testing conclusion
By Fig. 7~12 it is found that can preferably be examined according to this method to the impurity that acid, alkali, light, oxidation and high temperature generate
It surveys, main peak is preferably separated with the realization of front and back other impurities peak energy, and separating effect is also preferable between each impurity peaks.Secondly, discovery
Aspartic acid laumosaren aqueous solution is stablized in boiling water bath heating 2h, and 130 DEG C of heating 2h of aspartic acid laumosaren raw material are generated
A small amount of impurity, 160 DEG C of heating 2h generate more impurity and degradation impurity amount also increases very greatly；Impurity is generated in the case where acid, alkali destroy
It is less；It degrades and accelerates under illumination condition, generation impurity is more, and 2h irradiation solution colour reforms into depth especially under ultraviolet light
Yellow；It is most using hydrogen peroxide oxidation destruction generation impurity, it degrades most severe.The source of each impurity is belonged to, is as a result seen
Figure 13, wherein wherein curve 18 is the survey of aspartic acid laumosaren raw material (A factory provides, lot number 20151201) in relation to substance
Test result (without destroying)；The meaning of remaining curve is as described above；It is by destructive testing as a result, provided by the invention following
The source of each impurity belongs to, and the results are shown in Table 3:
3 impurity source of table ownership
Remarks: Lomefloxacin main peak retention time is about 22min
Embodiment 9: sample test
In order to verify the accuracy of this method test, the chromatographic condition provided using the embodiment of the present invention 1 is to A factory and B factory
Aspartic acid laumosaren glucose injection (specification is 100ml: Lomefloxacin 0.2g and glucose 5g) is measured respectively,
Simultaneously using statutory standards [YBH30102005 (A factory), YBH15122004 (B factory)] respectively to the sample of above-mentioned A factory and B factory into
Row test, test result are as shown in table 4:
Wherein, the calculation method of relative substance content: (5- methylol chaff is removed if any impurity peaks in test solution chromatogram
Outside aldehyde), impurity level in test solution is calculated by principal component Self-control method.
Maximum single impurity peak area/(contrast solution main peak area × right in maximum single impurity %=test sample map
According to solution extension rate) × 100%
Impurity peak area and/(contrast solution main peak area × contrast solution dilution times in total impurities %=test sample map
Number) × 100%
4 test result of table
From table 4, it can be seen that the product quality of Liang Jia enterprise can be better discriminated between using new method, and in terms of result, A factory
Product quality is better than B factory, and the related substance-measuring method that the present invention establishes is able to detect that more miscellaneous than statutory standards method
Matter, average total impurities amount (0.23%) is bigger than statutory standards result by 0.1%, and method specificity is stronger.
" independent samples t test " is carried out with SPSS software to the total impurities data of A factory and B factory, as a result, it has been found that, use is legal
When standard method is examined, for two factory's total impurities mean values without significant difference, p value is 0.456 (p > 0.05), is illustrated using statutory standards
Method analyzes the quality difference that cannot distinguish between A factory and B factory；When being examined using related substance method provided by the invention, two factories are total
Impurity mean value is 0.001 (p < 0.01) there are significant difference, p value, illustrates that there are extremely significant differences for Liang Jia enterprise sample detection result
It is different, and A factory product quality is significantly better than B factory product.It can be seen that the present invention provides compared with existing statutory standards method
Method it is more accurate, can be preferably applied to monitoring aspartic acid laumosaren glucose injection quality.
Embodiments described above is a part of the embodiment of the present invention, instead of all the embodiments.Reality of the invention
The detailed description for applying example is not intended to limit the range of claimed invention, but is merely representative of selected implementation of the invention
Example.Based on the embodiments of the present invention, obtained by those of ordinary skill in the art without making creative efforts
Every other embodiment, shall fall within the protection scope of the present invention.
Claims (10)
1. a kind of method of separating and assaying of the aspartic acid laumosaren glucose injection in relation to substance, which is characterized in that including
It is detected using aspartic acid laumosaren glucose injection as test solution using high performance liquid chromatography, the height
Effect liquid phase chromatogram method uses octadecylsilane chemically bonded silica for the chromatographic column of filler, is carried out with mobile phase A and mobility B linear
Gradient elution；The mobile phase A is phosphate solution-methanol；The Mobile phase B is acetonitrile.
2. the method according to claim 1, wherein the preparation method of the mobile phase A are as follows: contain quality percentage
The potassium dihydrogen phosphate and methanol that amount is 0.6%~0.7% are that 70:30~80:20 is mixed according to volume ratio, using phosphoric acid tune
Its pH value is saved to 2.5~3.0.
3. according to the method described in claim 2, it is characterized in that, the mass percentage of the potassium dihydrogen phosphate is
0.67%, the volume ratio of the potassium dihydrogen phosphate and the methanol is 76:24, and the pH value is 2.7.
4. the method according to claim 1, wherein the program of the linear gradient elution are as follows:
5. the method according to claim 1, wherein the model Welch Ultimate XB- of the chromatographic column
C18,4.6 × 250mm, 5 μm.
6. the method according to claim 1, wherein the Detection wavelength of the high performance liquid chromatography be 280~
290nm；Preferably 287nm.
7. the method according to claim 1, wherein the flow velocity of the high performance liquid chromatography be 0.5~
1.5ml/min；Preferably 1.0ml/min.
8. the method according to claim 1, wherein the sample volume of the high performance liquid chromatography is 5~20 μ L；
Preferably 10 μ L.
9. the method according to claim 1, wherein the chromatographic column column temperature of the high performance liquid chromatography be 35~
45℃。
10. the method according to claim 1, wherein the chromatographic column column temperature of the high performance liquid chromatography is 40
℃。
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| CN111208221A (en) * | 2020-01-09 | 2020-05-29 | 宜宾市南溪区红光制药有限公司 | Method for detecting lomefloxacin hydrochloride related substances |
| CN112034060A (en) * | 2020-08-25 | 2020-12-04 | 开封康诺药业有限公司 | Analysis method of pantoprazole sodium related substance for injection |
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| CN101315351B (en) * | 2008-06-26 | 2011-01-05 | 江南大学 | HPLC-ESI-MS/MS measuring method for simultaneously detecting 19 kinds of carbostyril medicaments |
| CN106093236A (en) * | 2016-06-05 | 2016-11-09 | 浙江海洋大学 | A kind of detect the method for lomefloxacin enantiomer in aquatic products |
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2018
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111208221A (en) * | 2020-01-09 | 2020-05-29 | 宜宾市南溪区红光制药有限公司 | Method for detecting lomefloxacin hydrochloride related substances |
| CN112034060A (en) * | 2020-08-25 | 2020-12-04 | 开封康诺药业有限公司 | Analysis method of pantoprazole sodium related substance for injection |
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