CN106568881A - Analysis method of preparation process of statin compound - Google Patents
Analysis method of preparation process of statin compound Download PDFInfo
- Publication number
- CN106568881A CN106568881A CN201510657055.4A CN201510657055A CN106568881A CN 106568881 A CN106568881 A CN 106568881A CN 201510657055 A CN201510657055 A CN 201510657055A CN 106568881 A CN106568881 A CN 106568881A
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- China
- Prior art keywords
- analysis method
- lovastatin
- alpha
- simvastatin
- analysis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
Abstract
The invention provides an analysis method of a preparation process of a statin compound. Reversed phase high-performance liquid chromatography is adopted in component analysis of a mixture containing Monacolin Jacid, alpha-dimethylbutyryl-S-methyl mercaptopropionate, lovastatin sodium salt, simvastatin ammonium salt, lovastatin and/or simvastatin; gradient elution is adopted; a buffer salt solution and acetonitrile are taken as a mobile phase A and a mobile phase B respectively. The analysis method can be used for analysis of related compounds in production process, influence factors are eliminated, analysis time is shortened greatly, operation is simple, reliable, and accurate, and effective means are provided for statin compound preparation process research and production process industrialized management.
Description
Technical field
The present invention relates to statin compound preparation field, in particular to a kind of statin compound preparation process
Analysis method and its in enzymatic conversion and the application of synthetically produced statin compound.
Background technology
Simvastatin is the semi-synthetic derivative of Lovastatin, and traditional production technology is the Jing with Lovastatin as raw material
Chemical synthesis obtains Simvastatin.Now can be produced by fermentation method, first pass through Lovastatin open loop in the presence of alkali and obtain
To Lovastatin sodium, then not that crin J is obtained by enzymatic hydrolysis method sour, then by acylated enzyme process simultaneously plus alpha, alpha-dimethyl
Base bytyry-S- methyl propionates (DMB-S-MMP) side chain obtains simvastatin ammonium salt, and refluxing toluene is finally passed through again
Remove ammonia and obtain Simvastatin.This is environmental friendly simple, and accessory substance is few, however it is necessary that a kind of directly perceived, quick can survey inspection
Each step reaction end is surveyed, and the scheme and means of whole detecting and tracking can be implemented to integrated artistic.
Therefore, it is necessary to seek a kind of method, so as to Accurate Determining not that crin J acid (Monacolin J are sour), α-
Dimethylbutanoyl-S- methyl propionates (DMB-S-MMP), Lovastatin sodium, simvastatin ammonium salt, Lovastatin,
The mixture of Simvastatin.
The method of phytosterol is determined in currently available technology to be had：
1.TLC TLCs, can only carry out "ball-park" estimate, using thin layer chromatography with TLC thin layer chromatography boards
Sensitivity is low, does not meet the advance of production target；
2. high performance liquid chromatography detection method, using UV detectors, and using methyl alcohol as mobile phase, Detection wavelength is
238nm (absorbing wavelength), the short interference of its wavelength is big, is easily affected by mobile phase, sample treatment, to analytic band
Carry out certain difficulty, and the sample determination time is longer, it is selective relatively poor, it is difficult to which that structure analogous product is carried out
Detection.
The content of the invention
To solve above-mentioned problems of the prior art, it is an object of the invention to provide it is a kind of it is efficient, simple to operate,
Reliable, accurate analysis method is analyzed to the preparation process of statin compound.
Specifically, technical scheme is as follows：
A kind of analysis method of statin compound preparation process, to sour comprising not that crin J in preparation process
(Monacolin J are sour), alpha-alpha-dimethyl bytyry-S- methyl propionates (DMB-S-MMP), Lovastatin sodium, pungent cut down
The mixture of statin ammonium salt, Lovastatin and/or Simvastatin carries out constituent analysis with high performance liquid chromatography, its feature
It is that the high performance liquid chromatography adopts RPLC, and adopts gradient elution, the stream of gradient elution
Dynamic phase A and B are respectively buffer salt solution and acetonitrile.
The buffer salt solution is preferably phosphate sodium dihydrogen buffer solution, and more preferably phosphate dihydrogen sodium concentration is 0.68%
(g/mL), the cushioning liquid of pH4.0.
The detector that the high performance liquid chromatography is adopted is preferably PDAD, more preferably Shimadzu
SPD-M20A PDADs.
The Detection wavelength that the high performance liquid chromatography is adopted is for 238nm.
The program of the gradient elution is preferably：Flow velocity 1.0mL/min, is temporally segmented：0～30.00min is buffered
Salting liquid is 70% to 10% (V/V, similarly hereinafter), and acetonitrile is 30% to 90%；30.00～30.01min buffer salts are molten
Liquid is 10% to 70%, and acetonitrile is 90% to 30%；30.01～35.00min buffer salt solutions are 70%, and acetonitrile is
30%.
Preferably, the high performance liquid chromatography adopts octadecylsilane chemically bonded silica post (C18 posts), and size is
250 × 4.6mm, column temperature is 40 DEG C, and sample size is 3～20 μ L.
The analysis method further includes to be measured the content of each composition in the mixture, prepares respectively first
Not that crin J acid of variable concentrations, alpha-alpha-dimethyl bytyry-S- methyl propionates, Lovastatin sodium, simvastatin ammonium salt,
The standard liquid of Lovastatin, Simvastatin, using the high performance liquid chromatography peak of each standard liquid is determined respectively
Area, and their peak area-concentration standard curve is drawn respectively；Then the mixture is configured to into sample solution,
Using the peak area of sample solution described in the high effective liquid chromatography for measuring；According to the calibration curve is calculated
The concentration of each composition in sample solution, and thus calculate the content of each composition in the mixture.
The analysis method of the statin compound preparation process that the present invention is provided can be applicable to the preparation life of statin compound
In product, including：Constituent analysis is carried out to the zymotic fluid in the microbial conversion process.According to the result of constituent analysis,
Determine in the zymotic fluid not that crin J acid, alpha-alpha-dimethyl bytyry-S- methyl propionates, Lovastatin sodium, pungent cut down him
The concentration of spit of fland ammonium salt, Lovastatin, to microorganism conversion and synthesis Simvastatin selection analysis are carried out；And thereby determine that
The process of production and/or the terminal of production.
The present invention's has the advantages that compared with prior art：
The invention discloses a kind of analysis method of statin compound preparation process, it can be used for statin compound system
The middle control analysis of standby production process and production terminal, can simultaneously to not that crin J acid (Monacolin J are sour), alpha, alpha-dimethyl
It is base bytyry-S- methyl propionates (DMB-S-MMP), Lovastatin sodium, simvastatin ammonium salt, Lovastatin, pungent
The mixture for cutting down statin is analyzed, and eliminates disturbing factor, substantially reduces analysis time, simple to operate, reliable,
Accurately, the factors such as many conventional operating procedures, analysis time length, disturbing factor be more are changed, therefore is to provide him
The effective means that spit of fland class compound preparation research work is managed with production process industrialized production.
Specific embodiment
Below by way of the description of specific embodiment, the invention will be further described, but this is not the limit to the present invention
System, those skilled in the art's basic thought of the invention, various modifications may be made or improves, but as long as not
Depart from the basic thought of the present invention, within the scope of the present invention.
It is an object of the invention to provide a kind of a kind of efficient, simple to operate, reliable, accurately statin compound preparation
The analysis method of process.
It is high to the mixture of statin compound preparation process in a kind of embodiment of the analysis method of the present invention
Effect liquid phase chromatogram method carries out constituent analysis, and high performance liquid chromatography is RPLC, and is washed using gradient
De-, high-efficient liquid phase chromatogram condition is：
Chromatographic column：C18Post, 250 × 4.6nm (ID)；
Mobile phase：A：Buffer salt solution (weighs 6.8g sodium dihydrogen phosphates, adds water to 1000mL, adjust pH to 4.0, shake
, filter and obtain final product)；B：Acetonitrile.
The gradient elution program of table 1
| Time (minute) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 70 | 30 |
| 30.00 | 10 | 90 |
| 30.01 | 70 | 30 |
| 35.00 | 70 | 30 |
Column temperature：40℃；
Wavelength：238nm (DAD detectors)；
Sample size：3～20 μ L (preferably 10 μ L).
Reagent and medicine：
Not that crin J acid (Monacolin J are sour) reference substance (content >=99%)；
Alpha-alpha-dimethyl bytyry-S- methyl propionates (DMB-S-MMP) reference substance (content >=99%)；
Lovastatin sodium reference substance (content >=99%)；
Simvastatin ammonium salt reference substance (content >=99%)；
Lovastatin reference substance (content >=99%)；
Simvastatin reference substance (content >=99%)；
Acetonitrile (HPLC levels)；
Sodium dihydrogen phosphate (AR levels).
Mobile phase and solution water are redistilled water in test.
Preparing standard solution：Accurately weigh each Intermediate control product (not that crin J acid (Monacolin J are sour), alpha-alpha-dimethyl
Bytyry-S- methyl propionates (DMB-S-MMP), Lovastatin sodium, simvastatin ammonium salt, Lovastatin, Simvastatin)
About 20mg is in 25mL measuring bottles, plus methyl alcohol makes to dissolve and be diluted to scale in right amount, shakes up, and obtains final product.
The analysis method of the present invention can be applied in statin compound preparation process, can be in the microbial conversion process
The zymotic fluid in each stage is acquired sample, then carries out constituent analysis using analysis method of the present invention.The present invention should
One embodiment can be：According to the result of the constituent analysis of the sample of reaction, not that crin is determined in reactant liquor
J acid (Monacolin J are sour), alpha-alpha-dimethyl bytyry-S- methyl propionates (DMB-S-MMP), Lovastatin sodium, pungent cut down
Statin ammonium salt, Lovastatin, the concentration of Simvastatin, and thus being controlled, tracking, giving birth to reaction substrate conversion
The endpoint of product.Such as, the concentration of Lovastatin sodium reaches peak value in certain stage sample and the concentration of Lovastatin reaches
To minimum, then can determine that now the stage production terminates.
And for example, the concentration of that crin J acid (Monacolin J are sour) does not reach peak value and Lovastatin in certain stage sample
The concentration of sodium reaches minimum, then can determine that now the stage production terminates.
Further explain and describe present invention below by way of the mode of example, but these examples be understood not to it is right
The restriction of the scope of the present invention.
Laboratory apparatus in example below is：
Shimadzu LC-2010CHT type high performance liquid chromatograph, Shimadzu SPD-M20A detector, chromatography record
Completed by Shimadzu work station.
Chromatographic condition is as described above, sample size is 10 μ L.
Embodiment 1：The selection of chromatographic condition
The selection of 1.1 mobile phases
Why the present invention is mobile phase from acetonitrile and water, is in order at the consideration of following factor：Eliminate and flowed when determining
The disturbing factor of relative detection, to improve reliability, accuracy；Improve each peak-to-peak separating degree；And washed using gradient
It is de- to make not that crin J acid (Monacolin J are sour), alpha-alpha-dimethyl bytyry-S- methyl propionates (DMB-S-MMP), Lip river
Fluvastatin sodium, simvastatin ammonium salt, Lovastatin, Simvastatin reach baseline and efficiently separate.
The selection of 1.2 Detection wavelengths
Detected in 238nm wavelength.
1.3 sample solutions are prepared
Precision weighs test sample in right amount, plus methyl alcohol makes to dissolve and be diluted to scale in right amount, shakes up, and obtains the confession of about 800 μ g/mL
Test sample solution.
Hybrid standard liquid：
Precision weighs reference substance not that crin J acid (Monacolin J are sour), alpha-alpha-dimethyl bytyry-S- methyl propionates
(DMB-S-MMP), Lovastatin sodium, simvastatin ammonium salt, Lovastatin, Simvastatin respectively about 20mg in same
In 25mL measuring bottles, plus methyl alcohol makes to dissolve and be diluted to scale in right amount, shakes up, and obtains final product.
Embodiment 2：The measure of standard repeatability
The μ L of hybrid standard liquid sample introduction 10 are taken, continuously enters 6 pins, with calculated by peak area RDS%, the results are shown in Table 2.
Table 2 is repeated
Embodiment 3
Not that crin J acid timing sampling detection is obtained by the conversion of enzymatic hydrolysis method to Lovastatin sodium.
Test sample：The measure of the 80h tunnings of microbe conversion production phase：
Precision weighs test sample about 0.1g and dissolves in 10mL volumetric flasks methyl alcohol, and ultrasonic 7min, shakes up constant volume, has
0.22 μm of membrane filtration of machine is obtained final product, and is need testing solution, sample size：10 μ L, conversion obtains not that crin J acid and is
More than 99%.
Claims (10)
1. a kind of analysis method of statin compound preparation process, in preparation process comprising not that crin J acid, alpha, alpha-dimethyl
Base bytyry-S- methyl propionates, Lovastatin sodium, simvastatin ammonium salt, Lovastatin and/or Simvastatin it is mixed
Compound carries out constituent analysis with high performance liquid chromatography, it is characterised in that the high performance liquid chromatography adopts anti-phase height
Effect liquid phase chromatogram, and gradient elution is adopted, the mobile phase A and B of gradient elution are respectively buffer salt solution and acetonitrile.
2. analysis method as claimed in claim 1, it is characterised in that the buffer salt solution is phosphate sodium dihydrogen buffer solution.
3. analysis method as claimed in claim 2, it is characterised in that the buffer salt solution be phosphate dihydrogen sodium concentration be 0.68%,
The cushioning liquid of pH4.0.
4. analysis method as claimed in claim 1, it is characterised in that the high performance liquid chromatography adopts PDAD.
5. analysis method as claimed in claim 4, it is characterised in that the PDAD is Shimadzu SPD-M20A.
6. analysis method as claimed in claim 1, it is characterised in that the Detection wavelength of the high performance liquid chromatography is 238nm.
7. analysis method as claimed in claim 1, it is characterised in that the program of gradient elution is：Flow velocity 1.0mL/min, on time
Between be segmented：0～30.00min buffer salt solutions are 70% to 10%, and acetonitrile is 30% to 90%；30.00～30.01min
Buffer salt solution is 10% to 70%, and acetonitrile is 90% to 30%；30.01～35.00min buffer salt solutions are 70%,
Acetonitrile is 30%.
8. analysis method as claimed in claim 1, it is characterised in that the high performance liquid chromatography is bonded using octadecylsilane
Silicagel column, size is 250 × 4.6mm, and column temperature is 40 DEG C, and sample size is 3～20 μ L.
9. analysis method as claimed in claim 1, it is characterised in that the analysis method is included to each composition in the mixture
Content is measured：Prepare respectively first not that crin J acid of variable concentrations, alpha-alpha-dimethyl bytyry-S- methyl propionates,
Lovastatin sodium, simvastatin ammonium salt, Lovastatin, the standard liquid of Simvastatin, using the high-efficient liquid phase color
Spectrometry determines respectively the peak area of each standard liquid, and draws their peak area-concentration standard curve respectively；Then by institute
State mixture and be configured to sample solution, using the peak area of sample solution described in the high effective liquid chromatography for measuring；According to institute
State the concentration that calibration curve calculates each composition in the sample solution, and thus calculate containing for each composition in the mixture
Amount.
10. analysis method as claimed in claim 1, it is characterised in that described comprising not that crin J acid, alpha-alpha-dimethyl butyryl
The mixture of base-S- methyl propionates, Lovastatin sodium, simvastatin ammonium salt, Lovastatin and/or Simvastatin is
Refer to microbial conversion process in zymotic fluid, the composition of zymotic fluid is analyzed, by analysis result determines production process with
/ or production terminal.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510657055.4A CN106568881A (en) | 2015-10-12 | 2015-10-12 | Analysis method of preparation process of statin compound |
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|---|---|---|---|
| CN201510657055.4A CN106568881A (en) | 2015-10-12 | 2015-10-12 | Analysis method of preparation process of statin compound |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1987449A (en) * | 2005-12-19 | 2007-06-27 | 北京维信学知科技发展有限公司 | Method for detecting quality of blood fat recovery capsule |
| US20120258123A1 (en) * | 2011-04-08 | 2012-10-11 | Zora Biosciences Oy | Biomarkers for sensitive detection of statin-induced muscle toxicity |
| CN103063779A (en) * | 2013-01-06 | 2013-04-24 | 江苏长泰药业有限公司 | Detection method of simvastatin nicotinate tablet related impurities |
-
2015
- 2015-10-12 CN CN201510657055.4A patent/CN106568881A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1987449A (en) * | 2005-12-19 | 2007-06-27 | 北京维信学知科技发展有限公司 | Method for detecting quality of blood fat recovery capsule |
| US20120258123A1 (en) * | 2011-04-08 | 2012-10-11 | Zora Biosciences Oy | Biomarkers for sensitive detection of statin-induced muscle toxicity |
| CN103063779A (en) * | 2013-01-06 | 2013-04-24 | 江苏长泰药业有限公司 | Detection method of simvastatin nicotinate tablet related impurities |
Non-Patent Citations (3)
| Title |
|---|
| 刘放等: "HPLC法测定辛伐他汀胶囊的含量和有关物质", 《西北药学杂志》 * |
| 王学军等: "反相高液相色谱法测定辛伐他汀及其有关物质洛伐他汀的含量", 《中国现代应用药学杂志》 * |
| 陈吉汉等: "超高效液相色谱-串联质谱法测定鸡肉与鸡蛋中6种他汀类药物残留", 《分析化学（FENXI HUAXUE）研究报告》 * |
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Application publication date: 20170419 |
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