CN109799298A - Detection method in relation to substance in a kind of pyrrole Lun Panai bulk pharmaceutical chemicals - Google Patents
Detection method in relation to substance in a kind of pyrrole Lun Panai bulk pharmaceutical chemicals Download PDFInfo
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Abstract
The present invention relates to the detection methods in relation to substance in a kind of pyrrole Lun Panai bulk pharmaceutical chemicals, belong to Pharmaceutical Analysis technical field.Detection method of the invention qualitatively or quantitatively detects pyrrole Lun Panai and related substance using high performance liquid chromatography and/or gas chromatography.Detection method of the invention, pyrrole Lun Panai and related substance linear relationship are good, and accuracy and precision are good, and specificity is strong, and stability is high.The favorable reproducibility of this detection method is able to satisfy testing requirements of the pyrrole Lun Panai bulk pharmaceutical chemicals in relation to substance, can be used for the quality control of pyrrole Lun Panai bulk pharmaceutical chemicals.
Description
Technical field
The invention belongs to Pharmaceutical Analysis technical fields, and in particular to the detection in relation to substance in a kind of pyrrole Lun Panai bulk pharmaceutical chemicals
Method.The detection method is using high performance liquid chromatography (HPLC) and/or gas chromatography (GC) to pyrrole Lun Panai and related miscellaneous
Matter carries out qualitative or quantitative analysis.
Background technique
Pyrrole Lun Panai is the drug of Japanese Wei Cai company research and development, passes through the group of Noncompetition inhibition AMPA type glutamate receptor
Effect, for treating the partial seizures of 12 years old or more epileptic.The medicine is the first anti-with the mechanism of action of FDA approval
Epilepsy drugs.
Related substance is the starting material brought into pharmaceutical synthesis production process, intermediate, side reaction product and degradation
Impurity etc., to related substance detect can quality to drug and safety control.Current domestic Extra Pharmacopoeia Martindale is still
Detection method of the pyrrole Lun Panai in relation to substance is not included, does not have complete table for the detection method in bulk pharmaceutical chemicals in relation to substance
It states.
Summary of the invention
The purpose of the present invention is on the basis of existing technology, provide the inspection in relation to substance in a kind of pyrrole Lun Panai bulk pharmaceutical chemicals
Survey method.
Technical scheme is as follows:
Detection method in relation to substance in a kind of pyrrole Lun Panai bulk pharmaceutical chemicals, the detection method using high performance liquid chromatography and/
Or gas chromatography qualitatively or quantitatively detects pyrrole Lun Panai and related substance, high-efficient liquid phase chromatogram condition includes: to adopt
It is that mixed flow mutually carries out gradient elution with mobile phase A with Mobile phase B, mobile phase A is potassium dihydrogen phosphate aqueous solution, Mobile phase B
For acetonitrile；Its GC conditions includes: using Agilent DB-624 capillary chromatographic column.
Present invention initial proportion of mobile phase A and Mobile phase B during gradient elution is 15~25:85~75 or 80:
20；In 0-25 minutes, the volume ratio of mobile phase A and Mobile phase B is by initial proportion at the uniform velocity gradual change to 40:60；At 25-40 minutes
Interior, the volume ratio of mobile phase A and Mobile phase B keeps 40:60 constant.For example, when the initial proportion of mobile phase A and Mobile phase B is
When 80:20, specific gradient elution process is as follows:
When the present invention is detected using high performance liquid chromatography, chromatographic column selects Shimadzu Wondasil C18- WR or Dikma
Diamonsil C18, using octadecylsilane chemically bonded silica as filler.In the case where not influencing detection effect, preferred color of choice
The length for composing column is 250mm, and diameter 4.6mm, packing material size is 5 μm.Such as: Shimadzu Wondasil C18- WR column
(250x4.6mm, 5 μm) or Dikma diamonsil C18 column (250x4.6mm, 5 μm).
In a kind of scheme, mobile phase A is 9~11mmol/L potassium dihydrogen phosphate aqueous solution, is adjusted with phosphoric acid or triethylamine
PH to 3.05~6.25 adjusts pH value to 6.25 with triethylamine with phosphorus acid for adjusting pH value to 3.05.
In a preferred embodiment, mobile phase A be 9~11mmol/L potassium dihydrogen phosphate aqueous solution, pH value 4.6-5.2,
With phosphorus acid for adjusting pH value to 4.6, pH value is adjusted to 5.2 with triethylamine.
In a kind of more preferable scheme, 10mmol/L potassium dihydrogen phosphate aqueous solution is prepared, is not required to adjust, pH value 4.61.
Further, it is 200~250nm, preferably 220nm that high-efficient liquid phase chromatogram condition, which includes: Detection wavelength,；Column temperature is 25
DEG C~35 DEG C, preferably 30 DEG C.
The present invention can according to need, and select suitable sample volume sample introduction, such as: sample volume is 10 μ l, 20 μ l or 50 μ l.
The present invention carries out quantitative detection to impurity bromo propane using gas chromatography；It uses with 6% cyanogen propylbenzene-
94% dimethyl siloxane is the Agilent DB-624 capillary chromatographic column of fixer.The case where not influencing effect of the present invention
Under, it is preferable that the length of chromatographic column is 30mm, and internal diameter 0.32mm, film thickness is 1.8 μm, i.e. chromatographic column: Agilent DB-624
(30m×0.32mm×1.8μm)。
In a preferred embodiment, headspace sampling when gas chromatography detects impurity bromo propane, head space are flat
The temperature that weighs is 85~105 DEG C, preferably 95 DEG C.
Further, head space equilibration time is 20~40min, preferably 30min.
Further, initial column temperature is 35~45 DEG C, preferably 40 DEG C.
Further, gasification room temperature is 220~240 DEG C, preferably 230 DEG C.
Further, detector temperature is 240~260 DEG C, preferably 250 DEG C.
A kind of more detailed gas chromatography carries out quantitative detection to impurity bromo propane, and GC conditions include:
Chromatographic column: Agilent DB-624；Column temperature:, maintaining 5min by initial 40 DEG C, rises to 180 DEG C with 10 DEG C/min, maintains 3min；Gas
Change room temperature: 230 DEG C；Detector temperature: 250 DEG C；Carrier gas: high pure nitrogen；Split ratio: 1:5；Constant voltage mode: pressure 10psi；
Ml headspace bottle temperature: 95 DEG C；Quantitative loop temperature: 200 DEG C；Transmission line temperature: 210 DEG C；Head space equilibration time: 30min；When sample introduction
Between: 1.0min；Sampling volume: 1ml.
Related substance in the pyrrole Lun Panai bulk pharmaceutical chemicals that the present invention refers to, including following substance: impurity 1:5- (2- pyridyl group)-
1,2- dihydropyridine -2- ketone (SMA)；Impurity 2: phenyl boric acid (SMB)；Impurity 3:2- cyanophenylboronic acid -1,3- propanediol cyclic ester
(SMC)；Impurity 4:1- phenyl -5- (2- pyridyl group) -1,2- dihydropyridine -2- ketone (intermediate 1)；Impurity 5:1- phenyl -3- is bromo-
5- (2- pyridyl group) -1,2- dihydropyridine -2- ketone (intermediate 2)；Impurity 6:1,5- diphenyl-[2,3'- bipyridyl] -6'(1'
H) -one；Impurity 7:3- (6- oxo -1- phenyl -1,6- dihydro-[2,3- bipyridyl] -5- base) benzonitrile；Impurity 8:4- (6- oxo-
1- phenyl -1,6- dihydro-[2,3- bipyridyl] -5- base) benzonitrile；Impurity 9:2- (6- oxo -1- phenyl -1,6- dihydro-[2,3-
Bipyridyl] -5- base) benzoic acid；[2,3- joins pyrrole to impurity 10:5- (2- cyano-phenyl) -6- oxo -1- phenyl -1,6- dihydro -
Pyridine] -1- oxygen；Impurity 11: bromo propane.
The present invention uses high performance liquid chromatography, is screened, is optimized from chromatographic condition etc. respectively, drafts related object
Matter method carries out methodology validation, and carries out qualitative research to impurity 1 (SMA), to impurity 2 (SMB), impurity 3 (SMC), impurity
4 (intermediates 1), impurity 5 (intermediate 2), impurity 6~10 (process impurity and degradation impurity) have carried out quantitative study.Using gas
Phase chromatography carries out quantitative study to process impurity 11 (bromo propane) issuable in this product synthesis technology, wherein miscellaneous
The correction factor of matter 2 is 1.06, and the correction factor of impurity 3 is 2.45, and the correction factor of impurity 4 is 1.24, the correction of impurity 5 because
Son is 1.89, and the correction factor of impurity 6 is 1.29, and the correction factor of impurity 7 is 0.85, and the correction factor of impurity 8 is 1.26, miscellaneous
The correction factor of matter 9 is 1.13, and the correction factor of impurity 10 is 0.65.
HPLC detection method and GC detection method the present invention provides pyrrole Lun Panai in relation to substance.Wherein, HPLC is detected
Method, including following operating procedure:
(1) solution is prepared:
Pyrrole Lun Panai is taken, 70% acetonitrile is added to dissolve and solution of every 1ml containing about pyrrole Lun Panai 0.5mg is made, as examination
Product solution.Impurity 1 plus dmso solution are taken respectively and is diluted to about 5 μ g/ml, take impurity 2, impurity 3, impurity 4, miscellaneous respectively
Matter 5, impurity 6, impurity 7, impurity 8, impurity 9, impurity 10 plus 70% acetonitrile dissolve and are diluted to about 5 μ g/ml.Take above-mentioned solution suitable
Amount mixes to obtain mixing contrast solution as solvent with 70% acetonitrile and DMSO respectively.
(2) test solution and impurity reference substance are injected separately into liquid chromatograph, chromatogram are recorded, by external standard method with peak
Areal calculation, chromatographic condition are as follows:
Chromatographic column: Shimadzu Wondasil C18- WR column (250x4.6mm, 5 μm) or Dikma diamonsil C18 column
(250x4.6mm, 5 μm), octadecylsilane chemically bonded silica is filler；
Detection wavelength: Detection wavelength is 200~250nm, preferably 220nm；
Mobile phase A: 10mmol/L potassium dihydrogen phosphate aqueous solution, pH are 3.05~6.25, preferable ph 4.6-5.2；More
Preferable ph is 4.61；Mobile phase B: acetonitrile；Mixed flow phase flow velocity is 1mL/min；Mobile phase A and stream during gradient elution
The initial proportion of dynamic phase B is 15~25:85~75 or 80:20；In 0-25 minutes, the volume ratio of mobile phase A and Mobile phase B by
Initial proportion at the uniform velocity gradual change is to 40:60；In 25-40 minutes, the volume ratio of mobile phase A and Mobile phase B keeps 40:60 constant；
Column temperature is 25 DEG C~35 DEG C, preferably 30 DEG C.
(3) measuring method: precision measures test sample and each 20 μ l of impurity reference substance solution, is injected separately into liquid chromatograph,
Record chromatogram.
Wherein, GC detection method, including following operating procedure:
(1) solution is prepared:
Take pyrrole Lun Panai sample appropriate (about 1.0g), it is accurately weighed, it sets in 10ml measuring bottle, it is dilute with n,N-dimethylacetamide
It releases to scale, shakes up, as test solution.
Bromo propane mother liquor: bromo propane (about 123mg) is set in 10ml volumetric flask, adds N, N- dimethylacetamide
Amine dissolves and is settled to scale.
The preparation of Standard Reserving Solution: precision is drawn bromo propane mother liquor 1.0ml and is placed in the measuring bottle of 100ml, and N, N- are added
Dimethyl acetamide dissolves and is settled to scale, shakes up, as Standard Reserving Solution.
Precision is drawn above-mentioned standard stock solution 10.0ml and is placed in 100ml volumetric flask, and n,N-dimethylacetamide is added to dilute
It to scale, shakes up, as reference substance solution.
(2) test solution and impurity reference substance are injected separately into gas chromatograph, chromatogram are recorded, by external standard method with peak
Areal calculation, chromatographic condition are as follows: chromatographic column: -94% dimethyl siloxane of 6% cyanogen propylbenzene is the Agilent of fixer
DB-624 (30m × 0.32mm × 1.8 μm) capillary chromatographic column；Column temperature: initial 35~45 DEG C, preferably 40 DEG C maintain 5min, with
10 DEG C/min rises to 180 DEG C, maintains 3min；Gasify room temperature: 220~240 DEG C, preferably 230 DEG C；Detector temperature: 240~
260 DEG C, preferably 250 DEG C；Carrier gas: high pure nitrogen；Split ratio: 1:5；Constant voltage mode: pressure 10psi；Ml headspace bottle temperature: 85~
105 DEG C, preferably 95 DEG C；Quantitative loop temperature: 200 DEG C；Transmission line temperature: 210 DEG C；Head space equilibration time: 20~40min, preferably
30min；Sample injection time: 1.0min.
(3) measuring method: precision measures test sample and each 1mL of impurity reference substance solution, is injected separately into gas chromatograph, remembers
Record chromatogram.
Using technical solution of the present invention, advantage is as follows:
Using detection method of the invention, pyrrole Lun Panai and related substance linear relationship are good, and accuracy and precision are equal
Well, specificity is strong, and stability is high.The favorable reproducibility of this detection method is able to satisfy inspection of the pyrrole Lun Panai bulk pharmaceutical chemicals in relation to substance
It surveys and requires, can be used for the quality control of pyrrole Lun Panai bulk pharmaceutical chemicals.
Detailed description of the invention
Fig. 1 is pyrrole Lun Panai sample chromatogram figure；
Fig. 2 is pyrrole Lun Panai and related substance chromatographic peak positioning figure；
Fig. 3 is 2 linear graph of impurity；
Fig. 4 is 3 linear graph of impurity；
Fig. 5 is 4 linear graph of impurity；
Fig. 6 is 5 linear graph of impurity；
Fig. 7 is 6 linear graph of impurity；
Fig. 8 is 7 linear graph of impurity；
Fig. 9 is 8 linear graph of impurity；
Figure 10 is 9 linear graph of impurity；
Figure 11 is 10 linear graph of impurity；
Figure 12 is pyrrole Lun Panai linear graph；
Figure 13 is that head space equilibrium temperature is investigated；
Figure 14 is that head space equilibration time is investigated；
Figure 15 is the standard curve of bromo propane.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, below in conjunction with specific embodiment, to this
Invention is further elaborated.It should be appreciated that specific example described herein is only used to explain the present invention, and do not have to
It is of the invention in limiting.Unless stated otherwise, the present invention uses reagent, method and apparatus is the art conventional reagents, side
Method and equipment.
A kind of embodiment 1: HPLC detection method of the pyrrole Lun Panai in relation to substance
One, experimental material and instrument
1. drug and reagent:
Pyrrole Lun Panai (Nanjing Healthnice Medical Technology Co., Ltd.), 5- (2- pyridyl group) -1,2- dihydropyridine -2- ketone are (miscellaneous
Matter 1, Nanjing Healthnice Medical Technology Co., Ltd.), phenyl boric acid (impurity 2, Jiaxing City Ai Sen Chemical Co., Ltd.), 2- cyano benzene boron
Acid -1,3-PD cyclic ester (impurity 3, Nanjing Healthnice Medical Technology Co., Ltd.), 1- phenyl -5- (2- pyridyl group) -1,2- bis-
Pyridinium hydroxide -2- ketone (impurity 4, Nanjing Healthnice Medical Technology Co., Ltd.), the bromo- 5- of 1- phenyl -3- (2- pyridyl group) -1,2- dihydro
Pyridin-2-ones (impurity 5, Nanjing Healthnice Medical Technology Co., Ltd.), 1,5- diphenyl-[2,3'- bipyridyl] -6'(1'H) -one
(impurity 6, Nanjing Healthnice Medical Technology Co., Ltd.), 3- (6- oxo -1- phenyl -1,6- dihydro-[2,3- bipyridyl] -5- base)
Benzonitrile (impurity 7, Nanjing Healthnice Medical Technology Co., Ltd.), 4- (6- oxo -1- phenyl -1,6- dihydro-[2,3- bipyridyl] -
5- yl) benzonitrile (impurity 8, Nanjing Healthnice Medical Technology Co., Ltd.), 2- (6- oxo -1- phenyl -1,6- dihydro-[2,3- connection pyrroles
Pyridine] -5- base) benzoic acid (impurity 9, Nanjing Healthnice Medical Technology Co., Ltd.), 5- (2- cyano-phenyl) -6- oxo -1- phenyl -
1,6- dihydro-[2,3- bipyridyl] -1- oxygen (impurity 10, Nanjing Healthnice Medical Technology Co., Ltd.), bromo propane (impurity
11, Shanghai Industrial Co., Ltd. in future), acetonitrile (chromatographically pure, DIKMA), (analysis is pure, the examination of Chinese medicines group chemistry for potassium dihydrogen phosphate
Agent Co., Ltd), phosphoric acid (analyze pure, Sinopharm Chemical Reagent Co., Ltd.), (analysis is pure, Chinese medicines group chemistry for triethylamine
Reagent Co., Ltd), ammonium acetate (analyze pure, Sinopharm Chemical Reagent Co., Ltd.), n,N-dimethylacetamide (analysis
Pure, Sinopharm Chemical Reagent Co., Ltd.), ultrapure water (self-control, Millipore).
2. instrument: the title and specification of specific instrument see the table below 1.
The title and specification of the specific instrument of table 1
| AUW120D assay balance (ten a ten thousandths) | Shimadzu Corporation |
| PHS-3C digital ph | Upper Nereid section |
| LC-20A high performance liquid chromatograph | Japanese Shimadzu |
| LC-10A high performance liquid chromatograph | Japanese Shimadzu |
| LC-Solution work station | Japanese Shimadzu |
| 2695 liquid chromatograph of Waters | Waters company |
| DAD2996 detector | Waters company |
| DAD2487 detector | Waters company |
| Agilent7890A gas chromatograph (μ ECD detector) | Agilent |
| Agilent OpenLAB chromatographic work station | Agilent |
| Agilent7694E head-space sampler | Agilent |
Two, liquid phase chromatogram condition
Pyrrole Lun Panai sample is taken, 70% acetonitrile is added to dissolve and solution of every 1ml containing about pyrrole Lun Panai 0.5mg is made, as
Test solution.Chromatographic column uses Shimadzu Wondasil C18- WR column (250x4.6mm, 5 μm)；With 10mmol/L potassium dihydrogen phosphate
Aqueous solution is mobile phase A, and using acetonitrile as Mobile phase B, according to the form below carries out gradient elution；Flow velocity is 1.0ml/min.30 DEG C of column temperature,
Detection wavelength is 220nm, and precision measures 20 μ l of test solution, injects liquid chromatograph, records chromatogram 1.
Two, experimentation
1. the preparation of pyrrole Lun Panai solution
Pyrrole Lun Panai is taken, 70% acetonitrile is added to dissolve and solution of every 1ml containing about pyrrole Lun Panai 0.5mg is made, as examination
Product solution.Impurity 1 plus dmso solution are taken respectively and is diluted to about 5 μ g/ml, take impurity 2, impurity 3, impurity 4, miscellaneous respectively
Matter 5, impurity 6, impurity 7, impurity 8, impurity 9, impurity 10 plus 70% acetonitrile dissolve and are diluted to about 5 μ g/ml.Take above-mentioned solution suitable
Amount mixes to obtain mixing contrast solution as solvent with 70% acetonitrile and DMSO respectively.
2. methodology validation
2.1 specificity
Take impurity 1 plus dmso solution respectively and be diluted to about 5 μ g/ml, take respectively impurity 2, impurity 3, impurity 4,
Impurity 5, impurity 6, impurity 7, impurity 8, impurity 9, impurity 10 plus 70% acetonitrile dissolve and are diluted to about 5 μ g/ml.Take above-mentioned solution
Mixing contrast solution is mixed to obtain as solvent with 70% acetonitrile and DMSO respectively in right amount, takes above-mentioned solution and 70% acetonitrile, DMSO point
Other sample introduction, chromatogram are as shown in Figure 2.
Test result is shown: under the chromatographic condition, baseline is steady, and solvent measures noiseless, pyrrole Lun Panai master to this product
Peak theoretical cam curve 105666, mixing each impurity separating degree in control sample, well (1.5 or more), each impurity can be detected
Out, specificity is preferable.Impurity 7 and impurity 8 are the position isomer of this product, under this condition same time appearance, when reservation
Between it is consistent, retention time is in 27min or so.
In the case where this is in relation to material conditions, select solvent can effectively to detect SMA when DMSO, since SMA is through four-step reaction
To final finished, a possibility that residual, is smaller, and is controlled SMA in intermediate detection, therefore in finished product
Quantitative analysis is not carried out to SMA, selects 70% acetonitrile as detection solvent.
2.2 failure test
The issuable catabolite of pyrrole Lun Panai can be detected under selected chromatographic condition to investigate, respectively with height
The drastic conditions such as temperature, acid, alkali, oxidation, illumination destroy this product, and the sample after destruction is dissolved with 70% acetonitrile and is prepared into
Test solution, it is accurate respectively to measure above-mentioned each 20 μ l of solution, liquid chromatograph is injected, chromatogram is recorded, specific method is shown in Table 2.
2 failure test result of table
This product is aoxidized in alkali, acid, has different degrees of degradation under illumination condition, this product is by soda acid, oxidation and illumination item
Part is affected, and various conditions are destroyed generated catabolite and can be detected, the separating degree of each catabolite and main peak
Well, related substance-measuring yield is good, and principal component peak purity is good, and therefore, which can be used for the related object of this product
Matter measurement.
The determination of 2.3 quantitative limits, detection limit
It takes each impurity to compare respectively and certain density sample is prepared in pyrrole Lun Panai control, 20 μ l of sample introduction after gradually diluting,
With signal-to-noise ratio S/N=3, S/N=10 measurement, it the results are shown in Table 3.
The detection of table 3 limit and quantitative limit result
| Title | Quantitative limit (ng) | Detection limit (ng) |
| Pyrrole Lun Panai | 1.0 | 0.3 |
| Impurity 2 | 2.9 | 0.9 |
| Impurity 3 | 1.6 | 0.5 |
| Impurity 4 | 1.3 | 0.4 |
| Impurity 5 | 1.6 | 0.6 |
| Impurity 6 | 1.7 | 0.6 |
| Impurity 7 | 1.5 | 0.5 |
| Impurity 8 | 1.5 | 0.4 |
| Impurity 9 | 1.6 | 0.4 |
| Impurity 10 | 1.0 | 0.3 |
2.4 sample solution stability tests
Take pyrrole Lun Panai sample, add the dissolution of 70% acetonitrile that the sample solution of about 0.5mg/ml is made, respectively after preparation 0h,
2h, 4h, 6h, 8h, 12h sample introduction, to investigate the content of each impurity and principal component in sample solution, in terms of area normalization method.Knot
Fruit is shown in Table 4.
4 sample solution stability test of table
The result shows that: sample solution is being placed in 12 hours in relation to substance 0.055% or so, and impurity 6 is 0.009%
Left and right, main peak peak area RSD are 0.68%, and sample solution is good in 12 hours internal stabilities.
2.5 linear
2.5.1 impurity stock solution is prepared
It takes impurity 2, impurity 3, impurity 4, impurity 5, impurity 6, impurity 7, impurity 8, impurity 9 appropriate respectively, acetonitrile is added to prepare
At the solution of about 5 μ g/ml of concentration, storeed as impurity 2, impurity 3, impurity 4, impurity 5, impurity 6, impurity 7, impurity 8, impurity 9
Liquid takes impurity 10 appropriate, adds acetonitrile to be configured to the solution of about 0.625 μ g/ml of concentration, as 10 stock solution of impurity, in Leng Chu
It places.
2.5.2 linear
Each impurity control stock solution of accurate absorption is set in right amount in 10ml measuring bottle respectively, with 70% acetonitrile constant volume, is shaken up, is obtained
A series of solution of concentration.Precision draws above-mentioned each 20 μ l of graded series strength solution, and successively sample introduction divides from low concentration to high concentration
Analysis records chromatogram, and with impurity reference substance solution concentration C (μ g/ml) for abscissa, impurity reference substance peak area is ordinate,
It carries out linear regression and finds out regression equation, the results are shown in Table 5 and Fig. 3~Figure 12.
Table 5 linearly investigates result
2.6 sample introduction precision tests
4 solution of linear term lower linear is taken, continuous sample introduction measures 6 times, investigates the situation of change of peak area and retention time.Knot
Fruit is shown in Table 6.
6 sample introduction Precision test result of table
The result shows that sample introduction precision is good.
2.7 mixing contrast solution stability
Mixing contrast solution is taken, 0,2,4,6,8,10 after preparation hour measurement.It the results are shown in Table 7.
7 impurity contrast solution stability test result of table
| Time (hour) | 0 | 2 | 4 | 6 | 8 | RSD (%) |
| 2 peak area of impurity | 22153 | 22560 | 22542 | 22445 | 22484 | 0.7 |
| 3 peak area of impurity | 40264 | 40332 | 39724 | 39314 | 41294 | 1.9 |
| 4 peak area of impurity | 33932 | 33907 | 33760 | 32982 | 33528 | 1.2 |
| 5 peak area of impurity | 29905 | 29684 | 28552 | 29207 | 29589 | 1.8 |
| 6 peak area of impurity | 31529 | 31486 | 31467 | 31248 | 31868 | 0.7 |
| Impurity 7,8 peak areas | 83845 | 84895 | 84330 | 83001 | 85048 | 1.0 |
| 9 peak area of impurity | 38663 | 38933 | 38554 | 37667 | 38296 | 1.2 |
| 10 peak area of impurity | 45065 | 43890 | 45185 | 43275 | 45210 | 2.0 |
| Pyrrole Lun Panai peak area | 530837 | 531695 | 520769 | 525269 | 524644 | 0.9 |
The result shows that being placed at room temperature for after the preparation of impurity contrast solution more stable in 10 hours.
2.8 repetitive test
Take pyrrole Lun Panai sample appropriate, precision weighs 6 parts, adds 70% acetonitrile to dissolve and is respectively prepared in every 1ml containing 0.5mg
Solution as test solution.Precision measures above-mentioned each 20 μ l of solution, liquid chromatograph is injected separately into, by external standard method with peak face
Product calculates the content of each impurity in this product.Measurement result is shown in Table 8.
8 repetitive test result of table
| Serial number | 1 | 2 | 3 | 4 | 5 | 6 | It is average | RSD (%) |
| Impurity 6 (%) | 0.010 | 0.011 | 0.010 | 0.011 | 0.011 | 0.010 | 0.010 | 6.8 |
| Total miscellaneous (%) | 0.042 | 0.042 | 0.042 | 0.042 | 0.043 | 0.041 | 0.042 | 1.5 |
2.9 recovery test
Precision weighs nine parts of pyrrole Lun Panai samples, is separately added into 80%, 100%, 120% impurity of Light absorbing impurty
Control adds 70% dilution in acetonitrile to scale, and accurate respectively to measure 20 μ l, injection liquid chromatograph carries out determination of recovery rates.As a result
It is shown in Table 9~table 17.
9 impurity of table, 2 recovery test result
10 impurity of table, 3 recovery test result
11 impurity of table, 4 recovery test result
12 impurity of table, 5 recovery test result
13 impurity of table, 6 recovery test result
14 impurity of table, 7 recovery test result
15 impurity of table, 8 recovery test result
16 impurity of table, 9 recovery test result
17 impurity of table, 10 recovery test result
2.10 Intermediate precision is tested
Take pyrrole Lun Panai sample appropriate, precision weighs 6 parts, adds 70% acetonitrile to dissolve and is respectively prepared in every 1ml containing 0.5mg
Solution as test solution.Precision measures above-mentioned each 20 μ l of solution, liquid chromatograph is injected separately into, by external standard method with peak face
Product calculates the content of each impurity in this product, the results are shown in Table 18.
18 impurity of table, 6 Intermediate precision test result
Instrument 1: Shimadzu LC-20A high performance liquid chromatograph number: 100106
Instrument 2: Shimadzu LC-10A high performance liquid chromatograph number: 100107
Instrument 3: Shimadzu LC-10A high performance liquid chromatograph number: 100108
Test result shows: this method Intermediate precision is good.
2.11 correction factor measures
Impurity 2,3,4,5,6,8,9, using 3 root chromatogram columns, amounts to 6 times on 2 liquid chromatographs, in quantitative limit to rule
In the 250% of threshold quantity concentration, 6 parts of samples are prepared altogether, each peak area are returned with each constituent concentration, and calculate impurity phase
To the correction factor of pyrrole Lun Panai；Impurity 7,10 calculates impurity phase using the ratio of senior middle school's low concentration and pyrrole Lun Panai peak area
To the correction factor of pyrrole Lun Panai, 19~table 28 the results are shown in Table.
The measurement of 19 impurity of table, 2 correction factor
20 impurity of table, 3 correction factor calculated result
21 impurity of table, 4 correction factor calculated result
22 impurity of table, 5 correction factor calculated result
23 impurity of table, 6 correction factor calculated result
24 impurity of table, 8 correction factor calculated result
25 impurity of table, 9 correction factor calculated result
26 pyrrole Lun Panai correction factor calculated result of table
27 impurity of table, 7 correction factor calculated result
| Impurity 7 | Correction factor |
| 1 | 0.85 |
| 2 | 0.86 |
| 3 | 0.85 |
| 4 | 0.84 |
| 5 | 0.85 |
| 6 | 0.83 |
| Average value | 0.85 |
28 impurity of table, 10 correction factor calculated result
| Impurity 10 | Correction factor |
| 1 | 0.65 |
| 2 | 0.65 |
| 3 | 0.59 |
| 4 | 0.69 |
| 5 | 0.63 |
| 6 | 0.68 |
| Average value | 0.65 |
Test result shows: the correction factor of impurity 2 is 1.06, and the correction factor of impurity 3 is 2.45, the correction of impurity 4
The factor is 1.24, and the correction factor of impurity 5 is 1.89, and the correction factor of impurity 6 is 1.29, and the correction factor of impurity 7 is 0.85,
The correction factor of impurity 8 is 1.26, and the correction factor of impurity 9 is 1.13, and the correction factor of impurity 10 is 0.65.
2.12 durability is investigated
The present invention further inquires into the tolerance degree of the detection method when minor change occurs for chromatographic condition, and has carried out resistance to
It is tested with property, is mixing contrast solution and sample solution with test solution, project of investigating includes starting mobile phase ratio, column
Temperature, different flowing phase pH values and different chromatographic columns, to mix contrast solution peak number, main peak and separating degree, the main peak of adjacent peak
The number of theoretical plate of principal component, principal component retention time and normalization purity are inspection target in tailing factor, sample solution, to this
The durability of method is investigated.Measurement result is shown in Table 29~table 32.
2.12.1 the variation of starting aqueous phase ratio
As hereinbefore, starting aqueous phase ratio is 15% to other high-efficient liquid phase chromatogram conditions during gradient elution,
20%, separating degree, the main peak tailing factor, sample of mixing contrast solution peak number, main peak and adjacent peak are investigated under conditions of 25%
The number of theoretical plate of principal component, principal component retention time and the situation of change for normalizing purity in product solution.The results showed that just
When beginning watr-proportion changes in 15%~25% range, shadow suffered by principal component number of theoretical plate, separating degree and related substance-measuring
Sound is smaller, and retention time difference is larger.Measurement result is shown in Table 29.
29 durability of table investigates (initial watr-proportion)
2.12.2 the variation of flowing phase pH value
Other high-efficient liquid phase chromatogram conditions in aqueous pH values are respectively 4.6 during gradient elution as hereinbefore,
Separating degree, the main peak tailing factor, sample of mixing contrast solution peak number, main peak and adjacent peak are investigated under conditions of 4.9 and 5.2
The number of theoretical plate of principal component, principal component retention time and the situation of change for normalizing purity in solution.The results showed that water phase
When pH value changes in 4.6~5.2 ranges, each parameter is influenced little.Measurement result is shown in Table 30.
30 durability of table investigates (aqueous pH values)
2.12.3 the variation of column temperature
Other high-efficient liquid phase chromatogram conditions as hereinbefore, mixing control are investigated when column temperature is 25 DEG C, 30 DEG C and 35 DEG C
Solution peak number, the separating degree of main peak and adjacent peak, main peak tailing factor, the number of theoretical plate of principal component in sample solution, it is main at
Divide retention time and normalizes the situation of change of purity.The results showed that when column temperature changes within the scope of 25 DEG C~35 DEG C, it is right
Each parameter influences little.Measurement result is shown in Table 31.
31 durability of table investigates (column temperature)
2.12.4 the variation of chromatographic column
Other high-efficient liquid phase chromatogram conditions as hereinbefore, investigate different chromatographic columns to mixing contrast solution peak number, master
The separating degree of peak and adjacent peak, main peak tailing factor the number of theoretical plate of principal component, principal component retention time and are returned in sample solution
One changes the situation of change of purity etc..The results showed that being influenced on each parameter little using different chromatographic columns.Measurement result is shown in
Table 32.
32 durability of table investigates (different chromatographic columns)
2.13 determinations in relation to substance-measuring method
It is verified by the above method, shows that the method tentatively drafted is suitable for this product Related substances separation.
Each impurity correction factor (impurity 2:1.06, impurity 3:2.45, impurity 4:1.24, impurity 5:1.89, impurity 6:
1.29, impurity 7:0.85, impurity 8:1.26, impurity 9:1.13, impurity 10:0.65), according to impurity correction factor size, determine
Impurity calculation.Because impurity 10 is genotoxicity impurity, limit is lower, is calculated using external standard method, other known impurities
It is calculated using own control correction up factorization method, remaining impurity is calculated using Self-control method.
It is temporarily that the determination of related substance is as follows:
This product is taken, solubilizer (70% acetonitrile) dissolves and solution of every 1ml containing about pyrrole Lun Panai 0.5mg is made, as confession
Test sample solution；Precision measurement test solution is appropriate, and solubilization dilution agent is made in every 1ml containing about the solution of 5 μ g, molten as compareing
Liquid；Separately take impurity 6,10 reference substance of impurity appropriate, solubilizer (70% acetonitrile), which dissolves and quantifies dilution, to be made in every 1ml containing about miscellaneous
Matter 6 is 0.5 μ g, the mixed solution that impurity 10 is 0.06 μ g, as reference substance solution.According to high performance liquid chromatography (V D of annex)
Measurement.It is filler with octadecylsilane chemically bonded silica；Using 10mmol/L potassium dihydrogen phosphate aqueous solution as mobile phase A, acetonitrile
For Mobile phase B, according to the form below carries out gradient elution；Detection wavelength is 220nm；30 DEG C of column temperature.Number of theoretical plate is based on the peak pyrrole Lun Panai
It calculates and is not less than 3000.20 μ l of contrast solution is taken, liquid chromatograph is injected, detection sensitivity is adjusted, makes the peak of principal component chromatographic peak
Height is about the 20% of full scale；It is accurate again to measure reference substance solution and each 20 μ l of test solution, it is injected separately into liquid chromatograph,
Record chromatogram.Peak sequence is followed successively by impurity 6, impurity 10 in reference substance solution, and impurity 10 is measured using external standard method, miscellaneous
Matter 6 is measured using own control correction up factorization method, remaining impurity is measured using Self-control method, remaining always collects widely
It is measured with Self-control method.
Impurity content is calculated by following formula:
10 calculation formula of impurity is as follows:
Impurity (%)=(rU/rS) × (CS/CU) × 100
The peak area of impurity 10 in rU=sample solution
The peak area of impurity 10 in rS=reference substance solution
The concentration (mg/ml) that impurity 10 compares in CS=standard solution
Pyrrole Lun Panai concentration (mg/ml) in CU=sample solution
6 calculation formula of impurity is as follows:
Impurity (%)=(rU/rS) × F
6 peak area of rU=sample solution impurity
1% own control peak area of rS=pyrrole Lun Panai
The correction factor of F=impurity 6
Other single impurity calculation formula are as follows:
Impurity (%)=rU/rS
Each impurity peak area of rU=sample solution
1% own control peak area of rS=pyrrole Lun Panai
Other total miscellaneous calculation formula are as follows:
Other total miscellaneous (%)=rU/rS
RU=sample solution all impurity summations in addition to impurity 6,10
1% own control peak area of rS=pyrrole Lun Panai
According to the above method, the related substance of three batches of samples is calculated, the results are shown in Table 33.
33 pilot scale Related substances separation result of table
A kind of embodiment 2: GC detection method of the pyrrole Lun Panai in relation to substance
One, detection method is tentatively established
The solvent isopropanol that this product uses during the preparation process may be reacted with NBS generates bromo propane, the chemical combination
There is object genotoxicity impurity to have similar structures, judge that the compound has genotoxicity.It is 1.5 μ g/ days, this product day according to TTC
Dose maximum be 12mg calculate the limit of bromo propane is 125ppm.
Since bromo propane boiling point to be detected is lower (59 degrees Celsius), capillary column head space temperature programming can be used
Method measures its content.Bromo propane belongs to halogenated hydrocarbons, detects on FID unobvious, there is higher response, and energy on ECD
Reach the requirement of detection, therefore select μ ECD (electron capture detector) as detector, and uses headspace sampling.
Two, solution is prepared
1 Standard Reserving Solution is prepared
According to sample dissolubility, sample solution concentration is set to 0.1g/ml, so the standard limitation solution of bromo propane is dense
Degree should be 12.5 μ g/ml.
Bromo propane mother liquor: bromo propane 124.89mg is set in 10ml volumetric flask, adds N, N- dimethylacetamide
Amine dissolves and is settled to scale.
The preparation of Standard Reserving Solution: precision is drawn bromo propane mother liquor 1.0ml and is placed in the measuring bottle of 100ml, and N, N- are added
Dimethyl acetamide dissolves and is settled to scale, shakes up, as Standard Reserving Solution.(CBromo propane=124.89 μ g/ml).
2 reference substances (standard limitation) solution is prepared
Precision is drawn above-mentioned standard stock solution 10.0ml and is placed in 100ml volumetric flask, and n,N-dimethylacetamide is added to dilute
It to scale, shakes up, as reference substance solution.(CBromo propane=12.49 μ g/ml)
3 test solutions are prepared
Take pyrrole Lun Panai sample appropriate (about 1.0g), it is accurately weighed, it sets in 10ml measuring bottle, it is dilute with n,N-dimethylacetamide
It releases to scale, shakes up, as test solution.
Three, GC conditions
The selection of 1 chromatographic column
Using -94% dimethyl siloxane of 6% cyanogen propylbenzene as the DB-624 capillary chromatographic column of fixer it is available compared with
Good separating effect.
2 chromatographic conditions are drafted
Chromatographic column: Agilent DB-624 (30m × 0.32mm × 1.8 μm)；
Column temperature:, maintaining 5min by initial 40 DEG C, rises to 180 DEG C with 10 DEG C/min, maintains 3min；Gasify room temperature: 230 DEG C；
Detector temperature: 250 DEG C；Carrier gas: high pure nitrogen；Split ratio: 1:5；Constant voltage mode: pressure 10psi；
Ml headspace bottle temperature: 95 DEG C；Quantitative loop temperature: 200 DEG C；Transmission line temperature: 210 DEG C；Head space equilibration time: 30min
Sample injection time: 1.0min；Sampling volume: 1ml.
3 head space temperature and equilibration time are investigated
The investigation of 3.1 equilibrium temperatures
It takes reference substance solution 5ml to be placed in 20ml headspace sampling bottle, seals, automatic sampling.When setting head space balance
Between be 30min, investigating equilibrium temperature is respectively 85 DEG C, 95 DEG C, 105 DEG C, is influenced on sensitivity, the result is shown in Figure 13.
The result shows that: raising of each solvent with equilibrium temperature, sensitivity increase.But head space equilibrium temperature increases, ml headspace bottle
In pressure increase it is difficult to will lead to sample introduction needle sample introduction when hypertonia.In summary situation preferably selects 95 DEG C to be used as top
Empty equilibrium temperature.
The selection of 3.2 equilibration times
It takes reference substance solution 5ml to be placed in 20ml headspace sampling bottle, seals, automatic sampling, head space equilibrium temperature is
At 95 DEG C, when investigation equilibration time is respectively 20min, 30min, 40min, the influence to sensitivity, the result is shown in Figure 14.
The result shows that: when equilibration time is 30min, bromo propane peak area is maximum, and vapor-liquid two phases concentration basically reaches
It balances, regulation residual solvent head space equilibration time is generally 30~45min in pharmacopeia, and overlong time is unfavorable for the guarantor of air-tightness
It holds, therefore now preferably using 30min as head space equilibration time.
4 methodology validations
4.1 system suitability
According under above-mentioned chromatographic condition, precision draws the above-mentioned reference substance solution 5ml prepared and is put into 20ml headspace sampling
Bottle, headspace sampling record chromatogram.The result shows that: theoretical cam curve: nBromo propane=17634.
4.2 specificity
It draws bromo propane and is interpreted into a certain concentration, sample introduction with n,N-dimethylacetamide.The result shows that: bromo propane
Retention time be 5.114min；Separately taking n,N-dimethylacetamide and sample solution, sample introduction is analyzed, as the result is shown N, N- diformazan
Yl acetamide is noiseless to measuring.
4.3 detection limits and quantitative limit determine
Standard Reserving Solution is successively diluted, respectively using signal-to-noise ratio S:N=10:1 and 3:1 when concentration as quantitative limit and inspection
Limit is surveyed, is as a result measured: the quantitative limit of bromo propane are as follows: 0.0624 μ g/ml, detection are limited to: 0.0156 μ g/ml.
4.4 linear test
Precision draws Standard Reserving Solution 0.5ml, 0.8ml, 1.0ml, 1.2ml and 1.5ml into 10ml volumetric flask, with N, N-
Dimethyl acetamide is diluted to scale, shakes up.Under above-mentioned chromatographic condition, successively sample introduction is analyzed from low concentration to high concentration, note
Chromatogram is recorded, with reference substance solution concentration C (μ g/ml) for abscissa, the peak area A of sampleSFor ordinate, linear regression is carried out
And regression equation is found out, test result is shown in Table 34 and Figure 15.
34 linear relationship of table investigates result
Above-mentioned test result shows: in 6.25~18.74 μ g/ml concentration ranges, the peak area of bromo propane with it is dense
Preferable linear relationship is presented between degree.
4.5 sample introduction precision tests
Above system applicability contrast solution is taken, 6 needle of continuous sample introduction investigates this method as index using the peak area of sample
Sample introduction precision, the results showed that the sample introduction precision of this method is good.It the results are shown in Table 35.
35 sample introduction Precision test result of table
4.6 repetitive test
6 parts of 140101 batches of samples are taken, every part of about 1.0g is accurately weighed, sets in 10ml measuring bottle and uses n,N-dimethylacetamide
It is diluted to scale, is shaken up, as solution to be measured.6 parts of solution to be measured are prepared in parallel.Precision measures test solution and reference substance is molten
Each 5ml of liquid, headspace sampling calculate the bromo propane content in sample by external standard method.The result shows that: it is not detected as in sample
Bromo propane.It the results are shown in Table 36.
36 repetitive test result of table
| Number | 1 | 2 | 3 | 4 | 5 | 6 | Mean value | RSD (%) |
| Bromo propane (%) | - | - | - | - | - | - | / | / |
4.7 recovery test
9 parts of 140101 batches of samples are taken, every part of about 1.0g is accurately weighed, is respectively placed in 10ml measuring bottle, accurate respectively to be added
Standard Reserving Solution 0.8ml, 1.0ml, 1.2ml are diluted to scale with n,N-dimethylacetamide, be configured to 80%, 100%,
Each three parts of 120% recycling solution.Accurate respectively to measure 5ml, headspace sampling carries out determination of recovery rates, the results are shown in Table 37.As a result
Show: the rate of recovery of this method is good.
37 recovery test result of table
4.8 serviceability test
As hereinbefore, when minor change occurs for GC conditions, this detection method is resistance to for other GC conditions
By degree, serviceability test is carried out, investigation factor includes column temperature, gasification room temperature, and detector temperature, inspection target is bromo
Number of theoretical plate, retention time, symmetrical factor and the separating degree of isopropyl alkane.Test result is shown in Table 38~table 40.
The durability of 38 column temperature of table investigates result
| Column temperature (DEG C) | Number of theoretical plate | Retention time (min) | Symmetrical factor |
| 35 | 21564 | 5.638 | 0.94 |
| 40 | 17040 | 5.116 | 0.93 |
| 45 | 14519 | 4.640 | 0.92 |
The durability of the gasification room temperature of table 39 investigates result
| Gasify room temperature (DEG C) | Number of theoretical plate | Retention time (min) | Symmetrical factor |
| 220 | 17256 | 5.116 | 0.96 |
| 230 | 17040 | 5.116 | 0.93 |
| 240 | 17062 | 5.116 | 0.96 |
The durability of 40 detector temperature of table investigates result
Conclusion: when minor change occurs for column temperature, gasification room temperature or detector temperature, the chromatographic behavior variation of solvent is not
Greatly, within the acceptable range；The good tolerance of this method.
Every experiment investigations such as this method is linear, precision, repeatability, the rate of recovery, durability, as a result meet correlation
Regulation, can be used for the detection of bromo propane content in this product.
The measurement of 5 samples
According to the above method to sample, bromo propane detection in pyrrole Lun Panai sample is carried out, the results are shown in Table 41.
Impurity bromo propane testing result in 41 pyrrole Lun Panai of table
| Lot number | Bromo propane (ppm) |
| 140101 | It is not detected |
| 140201 | It is not detected |
| 140202 | It is not detected |
| 2mg lists product 108647 | It is not detected |
| 4mg lists product 107299 | It is not detected |
Bromo propane is not detected as in three batches of samples and listing product.Therefore related substance detection meets rule in sample
It is fixed.
The present invention is respectively adopted high performance liquid chromatography and gas chromatography and studies this product impurity that may be present.
It using high performance liquid chromatography (HPLC), screened, optimized from chromatographic condition etc. respectively, draft related substance
Method carries out methodology validation, and carries out qualitative research to impurity 1 (SMA), to impurity 2 (SMB), impurity 3 (SMC), impurity 4
(intermediate 1), impurity 5 (intermediate 2), impurity 6~10 (process impurity and degradation impurity) have carried out quantitative study.Using gas phase
Chromatography (GC) carries out quantitative study to process impurity 11 (bromo propane) issuable in this product synthesis technology.This hair
Bright detection method, pyrrole Lun Panai and related substance linear relationship are good, and accuracy and precision are good, and specificity is strong, surely
Qualitative height.The favorable reproducibility of this detection method is able to satisfy the testing requirements in relation to substance in pyrrole Lun Panai bulk pharmaceutical chemicals, can be used for pyrrole
The quality of Lun Panai bulk pharmaceutical chemicals controls.
Claims (10)
1. the detection method in relation to substance in a kind of pyrrole Lun Panai bulk pharmaceutical chemicals, which is characterized in that the detection method is using efficient
Liquid chromatogram and/or gas chromatography qualitatively or quantitatively detect pyrrole Lun Panai and related substance,
Its high-efficient liquid phase chromatogram condition includes: that mobile phase A is used mutually to carry out gradient elution with Mobile phase B for mixed flow, described
Mobile phase A is potassium dihydrogen phosphate aqueous solution, and the Mobile phase B is acetonitrile；
Its GC conditions includes: using Agilent DB-624 capillary chromatographic column.
2. the detection method in relation to substance in pyrrole Lun Panai bulk pharmaceutical chemicals according to claim 1, which is characterized in that the ladder
Spending the initial proportion of mobile phase A and Mobile phase B in elution process is 15~25:85~75 or 80:20；In 0-25 minutes, stream
The volume ratio of dynamic phase A and Mobile phase B is by initial proportion at the uniform velocity gradual change to 40:60；In 25-40 minutes, mobile phase A and mobile phase
The volume ratio of B keeps 40:60 constant.
3. the detection method in relation to substance in pyrrole Lun Panai bulk pharmaceutical chemicals according to claim 1 or 2, which is characterized in that institute
State high-efficient liquid phase chromatogram condition include: chromatographic column be Shimadzu Wondasil C18- WR or Dikma diamonsil C18；It is preferred that
The length of chromatographic column is 250mm, and diameter 4.6mm, packing material size is 5 μm.
4. the detection method in relation to substance in pyrrole Lun Panai bulk pharmaceutical chemicals according to claim 3, which is characterized in that the stream
Dynamic phase A is 9~11mmol/L potassium dihydrogen phosphate aqueous solution, adjusts pH to 3.05~6.25 with phosphoric acid or triethylamine；Preferable ph
For 4.6-5.2；More preferable pH value is 4.61.
5. the detection method in relation to substance in pyrrole Lun Panai bulk pharmaceutical chemicals according to claim 3, which is characterized in that the height
Effect liquid phase chromatogram condition includes: that Detection wavelength is 200~250nm, preferably 220nm；Column temperature be 25 DEG C~35 DEG C, preferably 30
℃；Sample volume is 20 μ l.
6. the detection method in relation to substance in pyrrole Lun Panai bulk pharmaceutical chemicals according to claim 1, which is characterized in that use gas
Phase chromatography carries out quantitative detection to impurity bromo propane；It uses with -94% dimethyl siloxane of 6% cyanogen propylbenzene as fixation
The Agilent DB-624 capillary chromatographic column of liquid；It is preferred that the length of chromatographic column is 30mm, internal diameter 0.32mm, film thickness 1.8
μm。
7. the detection method in relation to substance in pyrrole Lun Panai bulk pharmaceutical chemicals according to claim 6, which is characterized in that the gas
Headspace sampling when phase chromatography detects impurity bromo propane, head space equilibrium temperature be 85~105 DEG C, preferably 95
℃；Head space equilibration time is 20~40min, preferably 30min；Initial column temperature is 35~45 DEG C, preferably 40 DEG C；Gasify room temperature
Degree is 220~240 DEG C, preferably 230 DEG C；Detector temperature is 240~260 DEG C, preferably 250 DEG C.
8. the detection method in relation to substance in pyrrole Lun Panai bulk pharmaceutical chemicals according to claim 7, which is characterized in that the gas
Phase chromatographic condition includes: chromatographic column: Agilent DB-624；Column temperature: initial 40 DEG C, 5min is maintained, rises to 180 with 10 DEG C/min
DEG C, maintain 3min；Gasify room temperature: 230 DEG C；Detector temperature: 250 DEG C；Carrier gas: high pure nitrogen；Split ratio: 1:5；Constant pressure mould
Formula: pressure 10psi；Ml headspace bottle temperature: 95 DEG C；Quantitative loop temperature: 200 DEG C；Transmission line temperature: 210 DEG C；Head space equilibration time:
30min；Sample injection time: 1.0min；Sampling volume: 1ml.
9. the detection method in relation to substance in pyrrole Lun Panai bulk pharmaceutical chemicals according to claim 1, which is characterized in that described to have
Closing substance includes following substance: impurity 1:5- (2- pyridyl group) -1,2- dihydropyridine -2- ketone；Impurity 2: phenyl boric acid；Impurity 3:2-
Cyanophenylboronic acid -1,3- propanediol cyclic ester；Impurity 4:1- phenyl -5- (2- pyridyl group) -1,2- dihydropyridine -2- ketone；Impurity 5:
The bromo- 5- of 1- phenyl -3- (2- pyridyl group) -1,2- dihydropyridine -2- ketone；Impurity 6:1,5- diphenyl-[2,3'- bipyridyl] -6'
(1'H) -one；Impurity 7:3- (6- oxo -1- phenyl -1,6- dihydro-[2,3- bipyridyl] -5- base) benzonitrile；Impurity 8:4- (6- oxygen
Generation -1- phenyl -1,6- dihydro-[2,3- bipyridyl] -5- base) benzonitrile；Impurity 9:2- (6- oxo -1- phenyl -1,6- dihydro-[2,
3- bipyridyl] -5- base) benzoic acid；[2,3- joins pyrrole to impurity 10:5- (2- cyano-phenyl) -6- oxo -1- phenyl -1,6- dihydro -
Pyridine] -1- oxygen；Impurity 11: bromo propane.
10. the detection method in relation to substance in pyrrole Lun Panai bulk pharmaceutical chemicals according to claim 1, which is characterized in that impurity 2
Correction factor be 1.06, the correction factor of impurity 3 is 2.45, and the correction factor of impurity 4 is 1.24, the correction factor of impurity 5
It is 1.89, the correction factor of impurity 6 is 1.29, and the correction factor of impurity 7 is 0.85, and the correction factor of impurity 8 is 1.26, impurity
9 correction factor is 1.13, and the correction factor of impurity 10 is 0.65.
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