CN112986410B - Method for detecting arylamine and aromatic hydrazine gene toxic impurities in celecoxib - Google Patents
Method for detecting arylamine and aromatic hydrazine gene toxic impurities in celecoxib Download PDFInfo
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Abstract
The invention discloses an LC-MS/MS method for detecting aniline, acetanilide, p-acetamidobenzenesulfonamide, sulfanilamide, p-hydrazinylbenzenesulfonamide hydrochloride and celecoxib impurities F in celecoxib. The method is characterized in that a reversed phase C18 column is used as a stationary phase, a mobile phase consists of an A phase and a B phase, the mobile phase is prepared by a mixture of an organic solvent and buffer salt, and gradient elution is carried out in a liquid chromatogram-mass spectrometer system. The method provided by the invention has the advantages of high efficiency, convenience, good specificity, high sensitivity and the like, and can be used for quickly and accurately qualitatively and quantitatively analyzing the residual impurities F of the aniline, the acetanilide, the p-acetamidobenzenesulfonamide, the sulfanilamide, the p-hydrazinylbenzenesulfonamide hydrochloride and the celecoxib in the celecoxib, so that the quality of the celecoxib is ensured, and the safety of clinical medication is improved.
Description
Technical Field
The invention belongs to the field of drug analysis, and particularly relates to an LC-MS/MS method for detecting aniline, acetanilide, p-acetamidobenzenesulfonamide, sulfanilamide, p-hydrazinylbenzenesulfonamide hydrochloride and celecoxib impurity F in celecoxib.
Background
Celecoxib (Celecoxib), i.e. 4- [5- (4-methylphenyl) -3- (trifluoromethyl) -1H-pyrazol-1-yl]Benzenesulfonamide of the formula C17H14F3N3O2S, molecular weight 381.38, its structural formula:
celecoxib (Celecoxib) is the first specific cyclooxygenase-2 (COX-2) inhibitor developed by sierl, usa, and is used for treating osteoarthritis and rheumatoid arthritis, and is the nonsteroidal anti-inflammatory analgesic with the largest global prescription quantity at present.
In the starting material p-aminobenzenesulfonamide for synthesizing celecoxib, various potential genotoxic impurities with warning structures are involved. By combining the production process and the aniline and acetaminophenyl structures in the literature as warning structures, genotoxic impurities containing the warning structures are analyzed.
The genotoxic impurities involved in the synthetic route of sulfanilamide are four as follows: aniline, acetanilide, p-acetamidobenzene sulfonamide and p-aminobenzene sulfonic acid. In the synthesis process of the celecoxib, hydrazine benzene sulfonamide as a synthetic intermediate and impurity F as a synthetic process are involved, and the toxicity impurities of the hydrazine benzene sulfonamide and the impurity F need to be researched.
The distribution of each gene toxic impurity in the synthetic process flow chart of the celecoxib is shown in the attached figure 1 of the specification. Aniline, acetanilide, p-acetamidobenzenesulfonamide, p-aminobenzenesulfonic acid, p-hydrazino benzenesulfonamide hydrochloride and celecoxib impurities F are six genotoxic impurities, which can be divided into three types according to the structure: anilines (aniline and sulfanilic acid), acetanilides (acetanilide and p-acetamidobenzenesulfonamide), and hydrazines (p-hydrazino benzenesulfonamide hydrochloride, celecoxib impurity F (p-tolylhydrazone benzenesulfonamide)). The maximum daily exposure of the celecoxib can reach 600mg, the European drug administration publishes that the maximum intake of genotoxic impurities (GTI) is 1.5 mu g/d, the TTC value of each genotoxic impurity is calculated according to 1.5 mu g/d, and the total content of the genotoxic impurities does not exceed 2.5 ppm.
Because of low content of genotoxic impurities and strict requirement on limit, the research adopts an HPLC-MS/MS method to carry out exclusive and sensitive inspection on the potential genotoxic impurities in celecoxib and key intermediate materials thereof. Therefore, the experimental study selects a high-sensitivity liquid chromatography-mass spectrometry detection technology to directly detect aniline, acetanilide, p-acetamidobenzenesulfonamide, sulfanilamide, p-hydrazinobenzenesulfonamide hydrochloride and celecoxib impurities F in celecoxib to a limited extent, so that the quality of celecoxib raw material medicines can be better controlled, and aniline, acetanilide, p-acetamidobenzenesulfonamide, sulfanilamide, p-hydrazinobenzenesulfonamide hydrochloride and celecoxib impurities F which may exist in the celecoxib raw material medicines can be better detected.
Disclosure of Invention
The invention aims to establish an LC-MS/MS method for detecting residual aniline, acetanilide, p-acetamido benzene sulfonamide, sulfanilamide, p-hydrazino benzene sulfonamide hydrochloride and celecoxib impurity F in celecoxib, and the method provides an accurate and efficient detection method for the content of potential genotoxic impurities in celecoxib.
In order to achieve the purpose, the invention provides the following technical scheme:
the invention provides an LC-MS/MS method for detecting aniline, acetanilide, p-acetamido benzene sulfonamide, sulfanilamide, p-hydrazino benzene sulfonamide hydrochloride and celecoxib impurity F in celecoxib, which comprises the following steps:
1. solution preparation
(1) Preparing a carbamazepine internal standard solution: weighing about 10mg of carbamazepine, precisely weighing, placing in a volumetric flask with the capacity of 100mL, dissolving by using methanol as a solvent, diluting to a scale, shaking uniformly to be used as a carbamazepine stock solution (100 mu g/mL), and storing at the temperature of minus 20 ℃;
(2) preparing a test solution: taking about 80mg of celecoxib sample, precisely weighing, placing in a 2.0mL poly-plastic centrifuge tube, precisely adding 500 muL of methanol, properly shaking to completely dissolve the sample, sequentially and precisely adding 20 muL of 1% ammonium acetate (pH is adjusted to 8-10 by ammonia water) aqueous solution of 5% Dithiothreitol (DTT), 20 muL of internal standard carbamazepine working solution (500 ng/mL), vortex and uniformly mixing for 30s, then adding 500 muL of purified water, shaking to precipitate celecoxib, freezing and centrifuging at 15800rpm/min for 10min, sucking supernatant, filtering through a 0.22μm filter membrane, discarding a proper amount of primary filtrate, placing the secondary filtrate in a 2mL glass sample injection vial provided with a plastic lining tube, taking 20 muL, and performing LC-MS/MS analysis;
(3) preparing an impurity stock solution: weighing aniline, acetanilide, p-acetamidobenzenesulfonamide, p-hydrazinobenzenesulfonamide hydrochloride, p-aminobenzenesulfonamide and celecoxib impurity F reference substances, wherein the amount of each reference substance is about 10mg, precisely weighing, and placing in a 100mL brown volumetric flask. Adding a proper amount of methanol, ultrasonically dissolving, diluting to a scale, shaking uniformly to serve as a stock solution (100 mu g/mL) of a mixed reference substance of aniline, acetanilide, p-acetamidobenzenesulfonamide, p-hydrazinobenzenesulfonamide hydrochloride, p-aminobenzenesulfonamide and celecoxib impurity F, and storing at the temperature of minus 20 ℃;
(4) preparing an impurity control solution: precisely sucking appropriate amounts of impurity stock solution and carbamazepine internal standard solution to prepare mixed standard working solution containing 5 mu g/mL of impurity concentration and 500ng/mL of carbamazepine internal standard solution; precisely sucking 500 mu L of methanol, placing the methanol into a 2.0mL poly-plastic centrifuge tube, precisely adding 20 mu L of 1% ammonium acetate (pH is adjusted to be 8-10 by ammonia water) aqueous solution of 5% Dithiothreitol (DTT) in sequence, adding 20 mu L of mixed standard working solution, uniformly mixing by vortex for 30s, adding 500 mu L of purified water, shaking up to prepare a mixed control solution with 50% limit concentration (aniline, acetanilide, p-acetamidobenzene sulfonamide, p-aminobenzenesulfonamide, p-hydrazinobenzene sulfonamide hydrochloride and celecoxib impurities F are all 100ng/mL, and internal standard carbamazepine is 10 ng/mL), taking 20 mu L, and performing LC-MS/MS analysis;
(5) preparation of blank solution: 1mL of 50% methanol is precisely added with 20 mu L of 1% ammonium acetate (pH is adjusted to 8-10 by ammonia water) aqueous solution of 5% Dithiothreitol (DTT).
2. And respectively injecting the blank solvent, the test sample solution and the impurity control solution into a liquid chromatography-mass spectrometry combination instrument, recording a chromatogram, and calculating by peak area according to an internal standard method. The specific conditions are as follows:
setting chromatographic conditions:
and (3) chromatographic column: thermo Scientific BDS HYPERSIL C18（100×4.6 mm, 2.4 μm）
Mobile phase: phase A: 95% methanol 0.1% formic acid 0.1% ammonium acetate solution
Phase B: 5% methanol 0.1% formic acid 0.1% ammonium acetate solution
The elution gradient is shown in table 1:
TABLE 1 elution gradiometer
Flow rate: 0.65 mL/min, and performing LC-MS/MS measurement by splitting flow 6:4 after the column;
column temperature: 40 ℃;
setting mass spectrum conditions:
electrospray positive ionization (ESI)+) The nozzle voltage was 4000V, the atomization temperature was 100 deg.C, the purge gas pressure was 0.5psi, the sheath gas pressure was 35psi, the assist gas pressure was 5psi, the capillary temperature was 350 deg.C, and the collision gas argon pressure was 1.2 mTorr. [ M + H ] for quantitation]+ The ion reaction was monitored as shown in table 2:
TABLE 2 impurities for quantification [ M + H]+ Ion reaction monitoring meter
3. Formula for calculating impurity content
The internal standard method has the following calculation formula:
correction factors:
the concentration of the test sample:
in the formula: as-peak area of internal standard substance in control solution; a. theR-peak area of the substance to be measured in the control solution; a. thex-peak area of the substance to be measured in the test sample solution; as' — peak area of internal standard in test solution; cs-concentration of internal standard substance in control solution (ng/mL); cR-concentration of test substance in control solution (ng/mL); cs' — concentration of internal standard substance in the test solution (ng/mL); cx-concentration of analyte in test solution (ng/mL).
The invention provides a method for detecting residual contents of aniline, acetanilide, p-acetamido benzene sulfonamide, sulfanilamide, p-hydrazino benzene sulfonamide hydrochloride and celecoxib impurities F in celecoxib.
Description of the drawings:
FIG. 1: the production route of each genotoxic impurity of celecoxib is explained.
The specific implementation mode is as follows:
the invention provides a method for detecting residual contents of aniline, acetanilide, p-acetamido-benzene sulfonamide, sulfanilamide, p-hydrazino-benzene sulfonamide hydrochloride and celecoxib impurity F in celecoxib, and a person skilled in the art can use the contents for reference and appropriately improve process parameters to realize the detection. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
The raw material medicines, reagents or instruments or reference substances used in the method for detecting the aniline, acetanilide, p-acetamido benzene sulfonamide, sulfanilamide, p-hydrazino benzene sulfonamide hydrochloride and celecoxib impurity F in the celecoxib can be purchased from the market.
The invention is further illustrated below with reference to comparative examples and examples:
example 1 determination of blank solvent
Blank solvent: 1mL of 50% methanol is precisely added with 20 mu L of 1% ammonium acetate (pH is adjusted to 8-10 by ammonia water) aqueous solution of 5% Dithiothreitol (DTT).
Impurity control solution: precisely sucking 500 mu L of methanol, placing the methanol into a 2.0mL poly-plastic centrifuge tube, precisely adding 20 mu L of 1% ammonium acetate (pH is adjusted to be 8-10 by ammonia water) aqueous solution of 5% Dithiothreitol (DTT), adding 20 mu L of mixed standard working solution (containing 5 mu g/mL of impurities and 500ng/mL of internal standard carbamazepine), uniformly mixing for 30s by vortex, adding 500 mu L of purified water, and uniformly shaking to obtain the product.
And accurately and respectively taking the impurity control solution for continuous sampling for 6 times, recording the spectrum, and calculating the RSD.
As a result: the method continuously samples for 6 times, and the ratios RSD of six impurity peak areas of aniline, acetanilide, p-acetamidobenzene sulfonamide, p-aminobenzenesulfonamide, p-hydrazino benzenesulfonamide hydrochloride and celecoxib impurity F to the internal standard peak area are all less than 5%. Indicating that the precision of the established genotoxic impurity inspection method meets the requirement. The RSD of the retention time is less than 1 percent, and the experimental result shows that the precision of the instrument is good.
TABLE 3 Selenxib raw material medicine added with 50% limit concentration aniline, acetanilide, p-acetamidobenzenesulfonamide, p-aminobenzenesulfonamide, p-hydrazino benzenesulfonamide hydrochloride and celecoxib impurity F (both 100 ng/mL), internal standard carbamazepine (10 ng/mL) peak area of sample introduction precision
Note: a represents aniline; b represents acetanilide; c represents p-acetamido benzene sulfonamide; d represents sulfanilamide; e represents p-hydrazinylbenzenesulfonamide hydrochloride; f represents celecoxib impurity F, the same as below.
TABLE 4 celecoxib raw material drug is added with 50% limit concentration aniline, acetanilide, p-acetamidobenzenesulfonamide, p-aminobenzenesulfonamide, p-hydrazino benzenesulfonamide hydrochloride and celecoxib impurity F (both 100 ng/mL), internal standard carbamazepine (10 ng/mL) sample injection retention time (min) precision
Comparative example 1
Blank solvent: 50% methanol.
Impurity control solution: precisely sucking 500 mu L of methanol, placing the methanol into a 2.0mL poly-plastic centrifuge tube, adding 20 mu L of mixed standard working solution (containing 5 mu g/mL of impurities and 500ng/mL of internal standard carbamazepine), uniformly mixing the solution for 30s by vortex, adding 500 mu L of purified water, and uniformly shaking to obtain the product.
And precisely and respectively taking the impurity control solution, continuously injecting the sample for 6 times, recording the spectrum, and calculating the RSD.
As a result: the reason is presumed that the hydrazine-benzenesulfonamide (SHH) hydrochloride in the impurity control solution is extremely unstable in aqueous solution, the chromatographic peak difference of two adjacent samples is large, the RSD of six consecutive analyses is over 50%, and the response is obviously reduced gradually.
Analyzing and solving the problems: the apparent pH of the sample solution is measured to be acidic by 50 percent methanol, and the phenylhydrazine compound is supposed to be easy to generate oxidation reaction in an acidic aqueous solution, so that a proper amount of antioxidant is considered to be added. Dithiothreitol is a common antioxidant, can play a good role in antioxidation when the pH is between 8 and 10, but cannot play a role in antioxidation under an acidic condition. Therefore, the pH of the sample solution 50% methanol is adjusted to 8-9 by using concentrated ammonia water. Preparing 1% ammonium acetate (pH is adjusted to 8-10 by ammonia water) aqueous solution of 5% Dithiothreitol (DTT), and adding 20 mu L of dithiothreitol solution into each sample. After the test that the stabilizer dithiothreitol is added and the pH value of the sample dissolving solution is adjusted, the stability of the SHH aqueous solution is obviously improved.
Example 2 Linear test
Chromatographic condition setting:
and (3) chromatographic column: thermo Scientific BDS HYPERSIL C18（100×4.6 mm, 2.4 μm）；
Mobile phase: phase A: 95% methanol 0.1% formic acid 0.1% ammonium acetate solution; phase B: 5% methanol 0.1% formic acid 0.1% ammonium acetate solution;
the elution gradient is shown in table 1;
flow rate: 0.65 mL/min, and performing LC-MS/MS determination by shunting 6:4 behind the column; column temperature: at 40 ℃;
setting mass spectrum conditions:
electrospray positive ionization (ESI)+) The nozzle voltage was 4000V, the atomization temperature was 100 deg.C, the purge gas pressure was 0.5psi, the sheath gas pressure was 35psi, the assist gas pressure was 5psi, the capillary temperature was 350 deg.C, and the collision gas argon pressure was 1.2 mTorr. For quantification of [ M + H]+ The monitoring of the ionic reaction is shown in table 2;
blank solvent: 1mL of 50% methanol is precisely added with 20 mu L of 1% ammonium acetate (pH is adjusted to 8-10 by ammonia water) aqueous solution of 5% Dithiothreitol (DTT).
Mixing standard curve working solution: accurately sucking a proper amount of mixed reference substance stock solutions (each 100 mu g/mL) containing aniline, acetanilide, p-acetamidobenzenesulfonamide, p-aminobenzenesulfonamide, p-hydrazinobenzenesulfonamide hydrochloride and celecoxib impurity F and a proper amount of carbamazepine stock solutions (each 100 mu g/mL) in a 10mL volumetric flask, diluting the mixed reference substance stock solutions to a scale mark by using methanol, and preparing the mixture into the mixture containing aniline, acetanilide, p-acetamidobenzenesulfonamide, p-aminobenzenesulfonamide, p-hydrazinobenzenesulfonamide hydrochloride and celecoxib impurity F with the concentrations of 1.0 mu g/mL, 1.5 mu g/mL, 2.5 mu g/mL, 5 mu g/mL, 7.5 mu g/mL and 10 mu g/mL respectively; the series concentration mixed standard solution with the internal standard carbamazepine of 500ng/mL is used as a mixed standard curve working solution (which is used for preparation now) for measuring aniline, acetanilide, p-acetamidobenzenesulfonamide, p-aminobenzenesulfonamide, p-hydrazinobenzenesulfonamide hydrochloride and celecoxib impurity F.
Standard curve test solutions: precisely sucking 500 mu L of methanol, placing the methanol into a 2mL poly-plastic centrifuge tube, precisely adding 20 mu L of a series of concentration standard curve working solutions of 1% ammonium acetate (the pH value is adjusted to be 8-10 by ammonia water) of 5% Dithiothreitol (DTT), precisely adding 20 mu L of aniline, acetanilide, p-acetamidobenzene sulfonamide, p-aminobenzenesulfonamide, p-hydrazinobenzene sulfonamide hydrochloride and celecoxib impurity F, after uniformly mixing the solution for 30s by vortex, precisely adding 500 mu L of purified water, shaking the solution uniformly to prepare the mixture containing aniline, acetanilide, p-acetamidobenzene sulfonamide, p-aminobenzenesulfonamide, p-hydrazinobenzene sulfonamide hydrochloride and celecoxib impurity F, wherein the concentrations of the impurity F are respectively 20.00ng/mL, 30.00ng/mL, 50.00ng/mL, 100.0ng/mL, 150.0ng/mL and 200.0 ng/mL; the internal standard carbamazepine is a series of solutions to be tested with concentration standard curves of 10.00 ng/mL.
Respectively and precisely measuring 20 mu L of sample introduction of the standard curve solution to be detected and the blank solution to be detected, recording chromatograms, and performing linear regression on the concentration C (ng/mL) of each genotoxic impurity component according to the ratio (Ax/Ai) of the peak area of the impurity F of aniline, acetanilide, p-acetamidobenzenesulfonamide, p-aminobenzenesulfonamide, p-hydrazinobenzenesulfonamide hydrochloride and celecoxib to the peak area of the internal standard carbamazepine according to the least square method.
The result shows that under the measuring conditions, aniline, acetanilide, p-acetamido benzene sulfonamide, p-aminobenzene sulfonamide, p-hydrazino benzene sulfonamide hydrochloride and celecoxib impurity F have good linear relation within the concentration range of 20-200 ng/mL.
TABLE 5 Standard Curve data for Aniline
TABLE 6 acetanilide Standard Curve data
TABLE 7 Standard Curve data for Paracetanilide
TABLE 8 Standard Curve data for celecoxib impurity F
TABLE 9 Standard Curve data for sulfanilamide
TABLE 10 Standard Curve data for hydrazinobenzenesulfonamide hydrochloride
Example 3 recovery test
The celecoxib bulk drug test sample is added with three levels of low (50%), medium (100%) and high (150%) concentration as samples of an accuracy experiment.
Recovery control solution: precisely absorbing 500 mu L of methanol, placing the methanol into a 2.0mL poly-plastic centrifuge tube, sequentially and precisely adding 20 mu L of 1% ammonium acetate (pH is adjusted to 8-10 by ammonia water) aqueous solution containing 5% Dithiothreitol (DTT), adding aniline, acetanilide and p-acetamidobenzene sulfonamide, 20 mu L of mixed standard working solution of sulfanilamide, p-hydrazino benzenesulfonamide hydrochloride, celecoxib impurity F (the concentration is 5 mu g/mL) and internal standard carbamazepine (500 ng/mL), uniformly mixing by vortex for 30s, adding 500 mu L of purified water to prepare a mixed control solution with 50% limit concentration (aniline, acetanilide, p-acetamidobenzenesulfonamide, p-sulfanilamide, p-hydrazino benzenesulfonamide hydrochloride and celecoxib impurity F are 100ng/mL, and the internal standard carbamazepine is 10 ng/mL) as a recovery rate control solution.
Recovery of sample solution: taking about 80mg of a celecoxib raw material sample, precisely weighing, placing the sample in a 2.0mL poly-plastic centrifuge tube, precisely adding 500 muL of methanol, properly shaking to completely dissolve the sample, sequentially and precisely adding 20 muL of 1% ammonium acetate (pH is adjusted to 8-10 by ammonia water) aqueous solution of 5% Dithiothreitol (DTT), adding 20 muL of aniline, acetanilide, p-acetamidobenzene sulfonamide, p-aminobenzenesulfonamide, p-hydrazino benzene sulfonamide hydrochloride and celecoxib impurity F (the concentrations are respectively 2.5 mug/mL, 5 mug/mL or 7.5 mug/mL), adding 20 muL of standard working solution containing 500ng/mL of carbamazepine, adding 500 muL of purified water, shaking to separate out celecoxib, freezing and centrifuging at a high speed of 15800rpm/min for 10min, sucking supernatant fluid to pass 0.22μm, discarding a proper amount of filtrate, taking a subsequent filtrate as a low (50%), the test article at medium (100%) or high (150%) concentration levels is added to the standard control solution.
3 parts of the mixture are prepared in parallel at each addition level and are respectively injected for determination. According to the peak area ratio, calculating aniline, acetanilide, p-acetamidobenzene sulfonamide, p-aminobenzenesulfonamide, p-hydrazino benzenesulfonamide hydrochloride and celecoxib impurities F according to an internal standard method, measuring the concentration of the six impurities, and deducting the background content in the sample to calculate the recovery rate.
And the results show that the calculated recovery rates of low, medium and high concentrations are all between 80 and 120 percent, and the accuracy is good.
TABLE 11 test of aniline recovery in celecoxib drug substance
TABLE 12 experiment of acetanilide recovery in celecoxib drug substance
TABLE 13 Paracetamol/Aminobenzene/sulfonamide recovery test in celecoxib drug substance
TABLE 14 test of the recovery of hydrochloride salt of hydrazino benzenesulfonylamine from celecoxib drug substance
TABLE 15 test of recoveries of sulfanilamide from celecoxib drug substance
TABLE 16 celecoxib impurity F recovery test in celecoxib bulk drug
Claims (1)
1. An LC-MS/MS method for detecting aniline, acetanilide, p-acetamido benzene sulfonamide, sulfanilamide, p-hydrazino benzene sulfonamide hydrochloride and celecoxib impurities F in celecoxib, wherein chromatographic conditions are set as follows: a chromatographic column: thermo Scientific BDS HYPERSIL C18And chromatographic column parameters: 100X 4.6 mm, 2.4 μm;
mobile phase: phase a, 95% methanol 0.1% formic acid 0.1% ammonium acetate solution; phase B, 5% methanol 0.1% formic acid 0.1% ammonium acetate solution;
elution gradient:
flow rate: 0.65 mL/min, and performing LC-MS/MS determination by shunting 6:4 behind the column; column temperature: at 40 ℃;
setting mass spectrum conditions: electrospray positive ionization ESI+The nozzle voltage 4000V, the atomization temperature 100 ℃, the purge gas pressure 0.5psi, the sheath gas pressure 35psi, the auxiliary gas pressure 5psi, the capillary temperature 350 ℃, and the collision gas argon pressure 1.2mTorr were measured and used for quantitative hydrogenation positive ion reaction monitoring as follows:
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| Isolation, synthesis and characterization of impurities in Celecoxib a cox-2 inhibitor;U.Satyanarayana et al.;《Journal of Pharmaceutical and Biomedical Analysis》;20041231;第35卷;全文 * |
| Stability-Indicating HPLC Method for Quantification of Celecoxib and Diacerein Along With Its Impurities in Capsule Dosage Form;Hanimi Reddy Bapatu et al.;《Journal of Chromatographic Science》;20151231;第53卷;摘要，第145页 * |
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