CN110361474A - A kind of method for detecting purity of Allopurinol - Google Patents
A kind of method for detecting purity of Allopurinol Download PDFInfo
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- CN110361474A CN110361474A CN201910711995.5A CN201910711995A CN110361474A CN 110361474 A CN110361474 A CN 110361474A CN 201910711995 A CN201910711995 A CN 201910711995A CN 110361474 A CN110361474 A CN 110361474A
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- G01N30/02—Column chromatography
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Abstract
A kind of method for detecting purity of Allopurinol belongs to Allopurinol quality testing field, includes the following steps: step 1: configuration standard product solution；Step 2: configuration sample solution；Step 3: chromatographic column is rinsed；Step 4: the position at Allopurinol peak is determined；Step 5: sample detection.Relative complex, detection time is longer, impurity peaks and product peak fail the problems such as efficiently separating in the presence of operating for its method for detecting purity for being directed to existing Allopurinol, the technological improvement that the case where there are large errors so as to cause the purity detected, then lead to economic loss and credibility loss is made.The invention has the advantages that the present invention provides a kind of method for detecting purity of Allopurinol, its is easy to operate, the sample detection time is short, impurity peak energy is effectively separated with product peak, so that it is guaranteed that the accuracy of gained purity, avoids that buyer is caused to return goods or require economic losses and the credibility losses such as compensation because of purity error.
Description
Technical field
The invention belongs to Allopurinol quality testing field, specifically a kind of method for detecting purity of Allopurinol.
Background technique
Allopurinol also known as allopurinol are a kind of common pharmaceutical intermediates for treating gout and related disease, can
It can inhibit xanthine oxidase to play, reduce hypoxanthine and xanthine from being converted into uric acid, i.e. uric acid synthesis, in turn
Uric acid concentration in blood is reduced, reduces lithate in the calmness of bone, joint and kidney, the drug that uric acid can be inhibited to synthesize.At present
The effect of having recorded Allopurinol in detail in 2010 editions " pharmacopeia " and detection method, but wherein only give the content of Allopurinol
Liquid chromatography detecting method, but the method for detecting purity of Allopurinol is not provided, content and purity are that medicine intermediate is most heavy
Two Testing index wanted, due to ununified standardized operation, at present detect Allopurinol purity when different company all
Trial is detected using various methods, but some operations are relative complex, some have, and detection time is longer, and many method impurity
Peak fails to efficiently separate with product peak, causes the purity detected there are large error, and product purity there are errors may
Lead to a series of influences such as buyer returns goods, prestige is impaired.
Summary of the invention
In view of the above-mentioned problems existing in the prior art, this application provides a kind of method for detecting purity of Allopurinol.This hair
Bright easy to operate, the sample detection time is short, and as a result accuracy is high.
Technical scheme is as follows:
A kind of method for detecting purity of Allopurinol, includes the following steps:
Step 1: configuration standard product solution: the standard items of 40-50mg are accurately weighed, is put into the volumetric flask of 100ml and is added
After the sodium acetate solution dissolution of 50ml, with sodium acetate solution constant volume, the sodium acetate solution for accurately pipetting 1ml dissolution standard items is placed in
Mobile phase constant volume is used in 50ml volumetric flask, will be flowed the standard solution after phase dilution and is transferred in liquid phase detection bottle that tighten bottle cap standby
With；
Step 2: configuration sample solution: the standard items of 30-40mg are accurately weighed, is put into the volumetric flask of 100ml and is added
After the sodium acetate solution dissolution of 50ml, with sodium acetate solution constant volume, the sodium acetate solution for accurately pipetting 1ml sample dissolution is placed in
Mobile phase constant volume is used in 50ml volumetric flask, will be flowed the sample solution after phase dilution and is transferred in liquid phase detection bottle that tighten bottle cap standby
With；
Step 3: it rinses chromatographic column: after chromatographic column is connect with automatic sampling liquid chromatograph, being rinsed with mobile phase with 1.5-
The speed of 2.5ml/min rinses chromatographic column, and observation base, when baseline steadily after flow velocity be reduced to 1.0ml/min, baseline is steady
After prepare sample introduction；
Step 4: it determines the position at Allopurinol peak: the liquid phase for filling standard solution in step 1 detection bottle is put into
Carry out liquid phase analysis in the test board of the full-automatic sample introduction liquid phase instrument of 1200 type of Agilent, continuous sample introduction 2 times, if peak position one out
It causes, then goes out peak position for Allopurinol, if inconsistent continuation sample introduction, until peak position has result position is identical at least twice to determine out
Position Allopurinol goes out peak position；
Step 5: it is complete that the liquid phase that sample solution is filled in step 2 detection bottle sample detection: is put into Agilent1200 type
Carry out liquid phase analysis in the test board of automatic sampling liquid phase instrument, continuous sample introduction 2 times, if twice gained purity error 0.1% with
It is interior, then take the average value of purity to be twice denoted as this batch sample purity, if twice gained purity error 0.1% or more continue into
Sample, until have at least twice purity error take the average value of purity of the error in 0.1% to be denoted as this sample within 0.1%
Purity.
A kind of method for detecting purity of Allopurinol as described above, chromatographic test strip part described in step 4 and step 5
Are as follows: wavelength: 235nm；Mobile phase: using the solution of methanol-acetic acid-formic acid (73:14:13) as mobile phase；Chromatographic column: An Jie
The SB-C18 chromatographic column of the Agilent ZORBAX StableBond series of human relations company；Column temperature: 40 DEG C；Sample volume: 10 μ L；Inspection
Survey the time: 15min；Flow velocity: 1ml/min.
A kind of method for detecting purity of Allopurinol as described above, sodium acetate solution described in step 1 and step 2 are molten
Concentration be 0.1-0.3mol/L.
A kind of method for detecting purity of Allopurinol as described above, sodium acetate solution described in step 1 and step 2 are molten
Concentration be 0.2mol/L.
A kind of method for detecting purity of Allopurinol as described above, sample purity described in step 5 sample introduction six times with
On discrete distribution is still presented, then reconfigure sample solution and chromatographic column and liquid phase instrument overhauled.
A kind of method for detecting purity of Allopurinol as described above, mobile phase described in step 3 rinse the stream of chromatographic column
Speed is 2ml/min.
The invention has the advantages that the present invention provides a kind of method for detecting purity of Allopurinol, easy to operate, sample detection
Time is short, and impurity peak energy is effectively separated with product peak, so that it is guaranteed that the accuracy of gained purity, avoids because of purity error
Buyer is caused to return goods or require economic losses and the credibility losses such as compensation.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is this hair
Bright some embodiments for those of ordinary skill in the art without any creative labor, can be with
It obtains other drawings based on these drawings.
Fig. 1 is the test map of embodiment 1；
Fig. 2 is the test map of embodiment 2.
Specific embodiment
In order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below in conjunction with the embodiment of the present invention
In attached drawing, technical scheme in the embodiment of the invention is clearly and completely described, it is clear that described embodiment is
A part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, those of ordinary skill in the art
Every other embodiment obtained without creative efforts, shall fall within the protection scope of the present invention.
Embodiment 1
Step 1: configuration standard product solution: the standard items of 45mg are accurately weighed, is put into the volumetric flask of 100ml and is added
The concentration of 50ml is after 0.2mol/L sodium acetate solution dissolves, to be 0.2mol/L sodium acetate solution constant volume with concentration, accurately pipette
The 0.2mol/L sodium acetate solution of 1ml dissolution standard items, which is placed in 50ml volumetric flask, uses mobile phase constant volume, after flowing phase dilution
Standard solution be transferred in liquid phase detection bottle that tighten bottle cap spare；
Step 2: configuration sample solution: the standard items of 30-40mg are accurately weighed, is put into the volumetric flask of 100ml and is added
The concentration of 50ml is after 0.2mol/L sodium acetate solution dissolves, to be 0.2mol/L sodium acetate solution constant volume with concentration, accurately pipette
The concentration of 1ml sample dissolution is placed in 50ml volumetric flask for 0.2mol/L sodium acetate solution and uses mobile phase constant volume, and mobile phase is dilute
Sample solution after releasing is transferred in liquid phase detection bottle that tighten bottle cap spare；
Step 3: it rinses chromatographic column: after chromatographic column is connect with automatic sampling liquid chromatograph, being rinsed with mobile phase with 2ml/
The speed of min rinses chromatographic column, and observation base, when baseline steadily after flow velocity be reduced to 1.0ml/min, baseline steadily prepares afterwards
Sample introduction；
Step 4: it determines the position at Allopurinol peak: the liquid phase for filling standard solution in step 1 detection bottle is put into
Liquid phase analysis is carried out in the test board of the full-automatic sample introduction liquid phase instrument of 1200 type of Agilent, chromatographic test strip part are as follows: wavelength:
235nm；Mobile phase: using the solution of methanol-acetic acid-formic acid (73:14:13) as mobile phase；Chromatographic column: agilent company
The SB-C18 chromatographic column of Agilent ZORBAX StableBond series；Column temperature: 40 DEG C；Sample volume: 10 μ L；Detection time:
15min；Flow velocity: 1ml/min continuous sample introduction 2 times, if appearance position consistency, goes out peak position for Allopurinol, if inconsistent continuation
Sample introduction positions Allopurinol and goes out peak position until peak position has at least twice that result position is identical out；
Step 5: it is complete that the liquid phase that sample solution is filled in step 2 detection bottle sample detection: is put into Agilent1200 type
Liquid phase analysis is carried out in the test board of automatic sampling liquid phase instrument, chromatographic condition is consistent with step 4, and continuous sample introduction 2 times, if twice
Gained purity error then takes the average value of purity to be twice denoted as this batch sample purity within 0.1%, if gained purity twice
Error continues sample introduction 0.1% or more, until have at least twice purity error take error pure in 0.1% within 0.1%
The average value of degree is denoted as this sample purity, and the sample purity is still presented discrete distribution in sample introduction more than six times, then matches again
It sets sample solution and chromatographic column and liquid phase instrument is overhauled.Testing result is as shown in Figure 1.
Embodiment 2
Step 1: configuration standard product solution: the standard items of 40mg are accurately weighed, is put into the volumetric flask of 100ml and is added
The concentration of 50ml is after 0.1mol/L sodium acetate solution dissolves, to be 0.1mol/L sodium acetate solution constant volume with concentration, accurately pipette
The 0.1mol/L sodium acetate solution of 1ml dissolution standard items, which is placed in 50ml volumetric flask, uses mobile phase constant volume, after flowing phase dilution
Standard solution be transferred in liquid phase detection bottle that tighten bottle cap spare；
Step 2: configuration sample solution: the standard items of 30-40mg are accurately weighed, is put into the volumetric flask of 100ml and is added
The concentration of 50ml is after 0.1mol/L sodium acetate solution dissolves, to be 0.1mol/L sodium acetate solution constant volume with concentration, accurately pipette
The concentration of 1ml sample dissolution is placed in 50ml volumetric flask for 0.1mol/L sodium acetate solution and uses mobile phase constant volume, and mobile phase is dilute
Sample solution after releasing is transferred in liquid phase detection bottle that tighten bottle cap spare；
Step 3: rinse chromatographic column: after chromatographic column is connect with automatic sampling liquid chromatograph, with mobile phase flushing with
The speed of 1.5ml/min rinses chromatographic column, and observation base, when baseline steadily after flow velocity be reduced to 1.0ml/min, baseline is steady
After prepare sample introduction；
Step 4: it determines the position at Allopurinol peak: the liquid phase for filling standard solution in step 1 detection bottle is put into
Liquid phase analysis is carried out in the test board of the full-automatic sample introduction liquid phase instrument of 1200 type of Agilent, chromatographic test strip part are as follows: wavelength:
235nm；Mobile phase: using the solution of methanol-acetic acid-formic acid (73:14:13) as mobile phase；Chromatographic column: agilent company
The SB-C18 chromatographic column of Agilent ZORBAX StableBond series；Column temperature: 40 DEG C；Sample volume: 10 μ L；Detection time:
15min；Flow velocity: 1ml/min continuous sample introduction 2 times, if appearance position consistency, goes out peak position for Allopurinol, if inconsistent continuation
Sample introduction positions Allopurinol and goes out peak position until peak position has at least twice that result position is identical out；
Step 5: it is complete that the liquid phase that sample solution is filled in step 2 detection bottle sample detection: is put into Agilent1200 type
Liquid phase analysis is carried out in the test board of automatic sampling liquid phase instrument, chromatographic condition is consistent with step 4, and continuous sample introduction 2 times, if twice
Gained purity error then takes the average value of purity to be twice denoted as this batch sample purity within 0.1%, if gained purity twice
Error continues sample introduction 0.1% or more, until have at least twice purity error take error pure in 0.1% within 0.1%
The average value of degree is denoted as this sample purity, and the sample purity is still presented discrete distribution in sample introduction more than six times, then matches again
It sets sample solution and chromatographic column and liquid phase instrument is overhauled.Testing result is as shown in Figure 2.
Embodiment 3
Step 1: configuration standard product solution: the standard items of 40-50mg are accurately weighed, is put into the volumetric flask of 100ml and is added
The concentration of 50ml is after 0.3mol/L sodium acetate solution dissolves, to be 0.3mol/L sodium acetate solution constant volume with concentration, accurately pipette
The 0.3mol/L sodium acetate solution of 1ml dissolution standard items, which is placed in 50ml volumetric flask, uses mobile phase constant volume, after flowing phase dilution
Standard solution be transferred in liquid phase detection bottle that tighten bottle cap spare；
Step 2: configuration sample solution: the standard items of 30-40mg are accurately weighed, is put into the volumetric flask of 100ml and is added
The concentration of 50ml is after 0.3mol/L sodium acetate solution dissolves, to be 0.3mol/L sodium acetate solution constant volume with concentration, accurately pipette
The concentration of 1ml sample dissolution is placed in 50ml volumetric flask for 0.3mol/L sodium acetate solution and uses mobile phase constant volume, and mobile phase is dilute
Sample solution after releasing is transferred in liquid phase detection bottle that tighten bottle cap spare；
Step 3: rinse chromatographic column: after chromatographic column is connect with automatic sampling liquid chromatograph, with mobile phase flushing with
The speed of 2.5ml/min rinses chromatographic column, and observation base, when baseline steadily after flow velocity be reduced to 1.0ml/min, baseline is steady
After prepare sample introduction；
Step 4: it determines the position at Allopurinol peak: the liquid phase for filling standard solution in step 1 detection bottle is put into
Liquid phase analysis is carried out in the test board of the full-automatic sample introduction liquid phase instrument of 1200 type of Agilent, chromatographic test strip part are as follows: wavelength:
235nm；Mobile phase: using the solution of methanol-acetic acid-formic acid (73:14:13) as mobile phase；Chromatographic column: agilent company
The SB-C18 chromatographic column of Agilent ZORBAX StableBond series；Column temperature: 40 DEG C；Sample volume: 10 μ L；Detection time:
15min；Flow velocity: 1ml/min continuous sample introduction 2 times, if appearance position consistency, goes out peak position for Allopurinol, if inconsistent continuation
Sample introduction positions Allopurinol and goes out peak position until peak position has at least twice that result position is identical out；
Step 5: it is complete that the liquid phase that sample solution is filled in step 2 detection bottle sample detection: is put into Agilent1200 type
Liquid phase analysis is carried out in the test board of automatic sampling liquid phase instrument, chromatographic condition is consistent with step 4, and continuous sample introduction 2 times, if twice
Gained purity error then takes the average value of purity to be twice denoted as this batch sample purity within 0.1%, if gained purity twice
Error continues sample introduction 0.1% or more, until have at least twice purity error take error pure in 0.1% within 0.1%
The average value of degree is denoted as this sample purity, and the sample purity is still presented discrete distribution in sample introduction more than six times, then matches again
It sets sample solution and chromatographic column and liquid phase instrument is overhauled.
It is shown according to final test map, impurity peaks separate well with product peak, so that it is guaranteed that gained purity is accurate
Property.
Finally it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention, rather than its limitations；Although
Present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that: it still may be used
To modify the technical solutions described in the foregoing embodiments or equivalent replacement of some of the technical features；
And these are modified or replaceed, technical solution of various embodiments of the present invention that it does not separate the essence of the corresponding technical solution spirit and
Range.
Claims (6)
1. a kind of method for detecting purity of Allopurinol, characterized by the following steps:
Step 1: configuration standard product solution: the standard items of 40-50mg are accurately weighed, is put into the volumetric flask of 100ml and 50ml is added
Sodium acetate solution dissolution after, with sodium acetate solution constant volume, the sodium acetate solution for accurately pipetting 1ml dissolution standard items is placed in 50ml
Mobile phase constant volume is used in volumetric flask, will be flowed the standard solution after phase dilution and is transferred in liquid phase detection bottle that tighten bottle cap spare；
Step 2: configuration sample solution: accurately weighing the standard items of 30-40mg, is put into the volumetric flask of 100ml and is added 50ml's
After sodium acetate solution dissolution, with sodium acetate solution constant volume, the sodium acetate solution for accurately pipetting 1ml sample dissolution is placed in 50ml capacity
Mobile phase constant volume is used in bottle, will be flowed the sample solution after phase dilution and is transferred in liquid phase detection bottle that tighten bottle cap spare；
Step 3: it rinses chromatographic column: after chromatographic column is connect with automatic sampling liquid chromatograph, being rinsed with mobile phase with 1.5-
The speed of 2.5ml/min rinses chromatographic column, and observation base, when baseline steadily after flow velocity be reduced to 1.0ml/min, baseline is steady
After prepare sample introduction；
Step 4: it determines the position at Allopurinol peak: the liquid phase for filling standard solution in step 1 detection bottle is put into Agilent
Carry out liquid phase analysis in the test board of the full-automatic sample introduction liquid phase instrument of 1200 types, continuous sample introduction 2 times, if appearance position consistency, for
Allopurinol goes out peak position, if inconsistent continuation sample introduction, until out peak position have at least twice result position it is identical position it is not fast
Alcohol goes out peak position；
Step 5: it is full-automatic that the liquid phase that sample solution is filled in step 2 detection bottle sample detection: is put into Agilent1200 type
Carry out liquid phase analysis in the test board of sample introduction liquid phase instrument, continuous sample introduction 2 times, if gained purity error is within 0.1% twice,
The average value of purity twice is taken to be denoted as this batch sample purity, if purity error obtained by twice continues sample introduction 0.1% or more, directly
To have at least twice purity error take the average value of purity of the error in 0.1% to be denoted as this sample purity within 0.1%.
2. a kind of method for detecting purity of Allopurinol according to claim 1, it is characterised in that: in step 4 and step 5
The chromatographic test strip part are as follows: wavelength: 235nm；Mobile phase: the solution conduct of methanol-acetic acid-formic acid (73:14:13) is used
Mobile phase；Chromatographic column: the SB-C18 chromatographic column of the Agilent ZORBAX StableBond series of agilent company；Column temperature: 40
℃；Sample volume: 10 μ L；Detection time: 15min；Flow velocity: 1ml/min.
3. a kind of method for detecting purity of Allopurinol according to claim 1, it is characterised in that: in step 1 and step 2
The molten concentration of the sodium acetate solution is 0.1-0.3mol/L.
4. a kind of method for detecting purity of Allopurinol according to claim 3, it is characterised in that: in step 1 and step 2
The molten concentration of the sodium acetate solution is 0.2mol/L.
5. a kind of method for detecting purity of Allopurinol according to claim 1, it is characterised in that: sample described in step 5
Discrete distribution is still presented in sample introduction in product purity more than six times, then reconfigure sample solution and to chromatographic column and liquid phase instrument into
Row maintenance.
6. a kind of method for detecting purity of Allopurinol according to claim 1, it is characterised in that: stream described in step 3
The dynamic flow velocity for mutually rinsing chromatographic column is 2ml/min.
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| JP2000290188A (en) * | 1999-04-05 | 2000-10-17 | Ito En Ltd | Xanthine oxydase inhibotor |
| US20060263840A1 (en) * | 2005-05-20 | 2006-11-23 | Mayo Foundation For Medical Education And Research , A Minnesota Corporation | Method for rapid determination of thiopurine methyltransferase activity |
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2019
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2000290188A (en) * | 1999-04-05 | 2000-10-17 | Ito En Ltd | Xanthine oxydase inhibotor |
| US20060263840A1 (en) * | 2005-05-20 | 2006-11-23 | Mayo Foundation For Medical Education And Research , A Minnesota Corporation | Method for rapid determination of thiopurine methyltransferase activity |
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| Title |
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| M.K. REINDERS ET AL.: "A simple method for quantification of allopurinol and oxipurinol in human serum by high-performance liquid chromatography with UV-detection", 《JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS》 * |
| X. LIU ET AL.: "Determination of allopurinol and oxypurinol in human plasma and urine by liquid chromatography-tandem mass spectrometry", 《JOURNAL OF CHROMATOGRAPHY B》 * |
| 国家药典委员会编: "《中华人民共和国药典 2010年版 2部》", 31 January 2010, 中国医药科技出版社 * |
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