CN103869018A - Method for measuring vincamine related substances through high performance liquid chromatography (HPLC) - Google Patents
Method for measuring vincamine related substances through high performance liquid chromatography (HPLC) Download PDFInfo
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- CN103869018A CN103869018A CN201410116092.XA CN201410116092A CN103869018A CN 103869018 A CN103869018 A CN 103869018A CN 201410116092 A CN201410116092 A CN 201410116092A CN 103869018 A CN103869018 A CN 103869018A
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- pervone
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Abstract
The invention relates to a method for measuring vincamine related substances through high performance liquid chromatography (HPLC). The method is characterized in that an octadecyl bonded silica chromatographic column is selected, an acetonitrile-0.1M ammonium carbonate solution (7:3) serves as a mobile phase for performing isocratic elution, a detector is an ultraviolet detector, and the detection wavelength is 260nm. The method has the characteristics of high separation degree, simplicity, rapidness, high specificity, high sensitivity and the like and can be used for detection and quality control of possibly generated related substances comprising intermediates, byproducts and other organic impurities in the vincamine preparation process.
Description
Technical field
The present invention relates to a kind of method of measuring pervone related substance by HPLC method, the assay method of the intermediate of especially synthetic pervone process, belongs to Pharmaceutical Analysis field.
Background technology
Pervone is the alkaloid being separated by phlox vinca (Vinca major), periwinkle (V.minor), upright catharanthus roseus (V.erecta), different in nature vinca (V.dilormis).Pervone is cerebral vasodilator, can maintain or recover cerebrovascular physiology expansion, increases the normal brain activity volume of blood flow of ischemic region, improves the utilization factor of brain to blood oxygen, improves hypoxic brain tissue metabolism.
Pervone international market demand amount is very large, at present can be semi-synthetic, its synthetic route has a lot, almost do not have about the report of its bulk drug and the control of intermediate related substance is domestic, Chinese Pharmacopoeia there is no relative quality control standard, sets up a kind of efficient, quick, accurately, its related substance is detected the efficient liquid-phase chromatography method that specificity is strong and control has very important significance.
Summary of the invention
The object of the invention is to set up a kind of method of HPLC method mensuration pervone related substance.Can be used for detection and the quality control of pervone bulk drug and pervone issuable related substance in synthetic and storage.
The invention provides a kind of method of high effective liquid chromatography for measuring pervone related substance, select octadecyl silane chromatographic column, mobile phase is acetonitrile-0.1M sal volatile (7:3), and detecting device is selected UV-detector, and detection wavelength is 260nm.
In specific embodiment of the present invention, described mobile phase is selected from 0.1M hartshorn salt and 0.2M ammonium acetate solution, preferably 0.1M sal volatile.
In specific embodiment of the present invention, acetonitrile and 0.1M sal volatile ratio are 80% ~ 55%:20% ~ 45%, and preferably acetonitrile ratio is 70%.
In specific embodiment of the present invention, flow rate of mobile phase is selected according to chromatographic column length, can select 0.8 ml/min ~ 1.5 ml/min.
In specific embodiment of the present invention, detect the preferred 260nm of wavelength.
The method of a kind of high effective liquid chromatography for measuring pervone related substance provided by the invention, its concrete implementation step comprises:
Get pervone finished product 25mg, mobile phase ultrasonic dissolution, is mixed with the need testing solution that pervone concentration is 1mg/ml.
Get pervone standard items 25mg, mobile phase ultrasonic dissolution, is mixed with the standard solution that pervone concentration is 1mg/ml, get above-mentioned standard solution appropriate, 500 times of mobile phase dilutions, are mixed with the solution that concentration is standard solution 0.2% concentration, as the reference substance solution of related substance quantitative measurement.
The end reaction liquid of getting the intermediate I of the synthetic pervone of 0.1ml, mobile phase is diluted to 10ml, as intermediate contrast solution 1.
The end reaction liquid of getting the intermediate II of the synthetic pervone of 0.1ml, mobile phase is diluted to 10ml, as intermediate contrast solution 2.
Accurately draw respectively need testing solution, reference substance solution, intermediate contrast solution 1 and intermediate contrast solution (2) 5 μ l, injection liquid chromatography, pervone main peak should be greater than 1.5 with the peak-to-peak degree of separation of phase related substance impurity, meet the requirement of Chinese Pharmacopoeia to degree of separation, press main peak pervone peak and calculate, theoretical cam curve is 11549, is greater than 5000, the symmetrical factor at main peak and other contrast solution peaks is respectively 0.98 and 0.96, all meets the standard of Chinese Pharmacopoeia.
What the inventive method was applicable to pervone mensuration can be pervone sterling, crude product, the product take pervone as major component such as bulk drug.Assay method of the present invention is also applicable to mensuration and the quality control of intermediate I, intermediate II and other unknown intermediate products that produce in building-up process in building-up process.
Accompanying drawing explanation
Fig. 1 is pervone need testing solution chromatogram.
Fig. 2 is reference substance solution chromatogram.
Fig. 3 is the chromatogram of intermediate I and intermediate II in pervone building-up process.
Embodiment
Embodiment 1
Pervone sterling, purchased from European sigma company, purity is 99.83%.High performance liquid chromatograph is selected U.S. Agilent1260 model instrument, detecting device is DAD detecting device, chromatographic column is thermo C18,4.6 × 250mm, 5 μ m, detect wavelength 260nm, sample size: 5 μ l (G1315D automatic sampler), flow velocity 1ml/min, column temperature: 35 ℃, mobile phase: acetonitrile-0.1M sal volatile (7:3) isocratic elution.
Experimental procedure:
Get pervone sterling 25mg, mobile phase ultrasonic dissolution, is mixed with the need testing solution that pervone concentration is 1mg/ml.
Get pervone standard items 25mg, mobile phase ultrasonic dissolution, is mixed with the standard solution that pervone concentration is 1mg/ml, get above-mentioned standard solution appropriate, 500 times of mobile phase dilutions, are mixed with the solution that concentration is standard solution 0.2% concentration, as the reference substance solution of related substance quantitative measurement.
Get respectively need testing solution and reference substance solution, according to chromatographic condition of the present invention, respectively injection liquid chromatography, according to analysis condition analysis of the present invention and record chromatogram.
Fig. 1 is pervone need testing solution gained chromatogram, and pervone retention time is 8.8min, 5.4min, and 11.4min is respectively related impurities in pervone sterling.Result shows, under chromatographic condition of the present invention, pervone and other impurity peaks degree of separation are all greater than 1.5, and main peak and related substance peak symmetrical factor are respectively 0.98,0.95 and 1.02, meet Chinese Pharmacopoeia relevant criterion.Fig. 2 is reference substance solution collection of illustrative plates, and gained main peak area is 7.41437, and related substance percentage composition (C%) is calculated by following formula: c=(A*W1*25*1*100)/(B*W2*500*25)
A is the peak area of impurity peaks in need testing solution
B is the molten product solution reference substance peak area of contrast
W1 is reference substance weight
W2 is test sample weight
As calculated, obtain two its related substances in pervone sterling and be respectively 0.1% and 0.05%, pervone purity is 99.85%, and accurately, reliably, this detection method can be used for related substance in ability pervone sterling and detects and quality control testing result.
Embodiment 2
In employing and embodiment 1, similar instrument and equipment and operation steps are tested, and the intermediate in our company's pervone building-up process is measured and quality control.
High performance liquid chromatograph is selected U.S. Agilent1260 model instrument, detecting device is VWD detecting device, chromatographic column is thermo C18,4.6 × 150mm, 5 μ m, detect wavelength 260nm, sample size: 5 μ l (G1315D automatic sampler), flow velocity 1ml/min, column temperature: 35 ℃, mobile phase: acetonitrile-0.1M sal volatile (7:3) isocratic elution.
Experimental procedure:
Get respectively the synthetic end reaction liquid of intermediate I of pervone of 0.1ml and the end reaction liquid of the intermediate II of synthetic pervone, mobile phase is diluted to 10ml, as the qualitative contrast liquid of pervone related substance.Synthetic pervone finished product 25mg between separately picking up the car, mobile phase ultrasonic dissolution, is mixed with the need testing solution that pervone concentration is 1mg/ml.
Get respectively vincamine intermediate I, intermediate II contrast liquid and need testing solution, measure and analytic record chromatogram by chromatographic condition of the present invention.
In Fig. 3, compound 1 and 2 is respectively as intermediate I and intermediate II, peak degree of separation between intermediate I and intermediate II is 1.53, peak symmetrical factor is respectively 1.03 and 0.95, all meet Chinese Pharmacopoeia standard, this detection method can be used for detection and the quality control of the intermediate in pervone building-up process.In Fig. 3, intermediate I and intermediate II relative retention time are 2.1min and 3.8min, in need testing solution, related substance retention time is respectively 5.4min and 11.4min, the related substance that our company's production pervone comprises is other unknown impurities that produce in building-up process, non-intermediate I and intermediate II.Result shows, the inventive method can separate the intermediate in pervone and pervone building-up process preferably, can be for detection and the quality control of intermediate in pervone building-up process.
Claims (4)
1. HPLC method is measured a method for pervone related substance, it is characterized in that: select octadecyl silane chromatographic column, take acetonitrile-0.1M sal volatile (7:3) as mobile phase isocratic elution, detecting device is UV-detector, and detection wavelength is 260nm.
2. method according to claim 1, is characterized in that, the grain diameter of described octadecyl silane chromatographic column filler is 3-5 μ m.
3. method according to claim 1, is characterized in that, described mobile phase ratio is that acetonitrile volume fraction is that 70%, 0.1M sal volatile volume fraction is 30%.
4. method according to claim 1, is characterized in that, detecting device is UV-detector, and DAD detecting device and VWD detecting device are all suitable for, and detection wavelength is 260nm.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112034058A (en) * | 2020-08-21 | 2020-12-04 | 开封康诺药业有限公司 | Method for detecting isomer impurities in vincamine |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0045918A1 (en) * | 1980-08-04 | 1982-02-17 | Paolo Corvi Mora | A process for the synthesis of vincamine and related indole alkaloids |
| CN101721361A (en) * | 2009-12-08 | 2010-06-09 | 李荣立 | Pervone vincamine injection and preparation process thereof |
-
2014
- 2014-03-27 CN CN201410116092.XA patent/CN103869018A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0045918A1 (en) * | 1980-08-04 | 1982-02-17 | Paolo Corvi Mora | A process for the synthesis of vincamine and related indole alkaloids |
| CN101721361A (en) * | 2009-12-08 | 2010-06-09 | 李荣立 | Pervone vincamine injection and preparation process thereof |
Non-Patent Citations (4)
| Title |
|---|
| MOSTAFA A.M. SHEHATA ET AL.: "Stability-indicating methods for determination of vincamine in presence of its degradation product", 《JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS》 * |
| PIERGIORFIO PIETTA: "High-performance liquid chromatographic determination of vincamine", 《JOURNAL OF CHROMATOGRAPHY》 * |
| 张超: "长春胺全合成研究进展", 《2008年中国药学会学术年会暨第八届中国药师周论文集》 * |
| 郑璐侠 等: "高效液相色谱电化学法测定长春胺的有关物质表长春胺", 《药物分析杂志》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112034058A (en) * | 2020-08-21 | 2020-12-04 | 开封康诺药业有限公司 | Method for detecting isomer impurities in vincamine |
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Application publication date: 20140618 |