Tong Wu, Yin Liu, Hanshuo Dong, Chunling Ai, Li Sun
Abstract Chaihu‐jia Longgu‐Muli decoction (CLMD) is a traditional Chinese medicine formula, and it has been used for more than a thousand years to treat mental and nervous system diseases. However, the active constitutions of the CLMD are unclear. Accordingly, an integrated analysis based on the ultra‐performance liquid chromatography‐quadrupole time‐of‐flight mass spectrometry method combined with the UNIFI platform was used to clarify the chemical composition of the CLMD. As a result, 102 compounds including 38 saponins, 32 flavonoids, 12 polyphenols, nine anthraquinones, and 11 others were identified or tentatively presumed. Among them, 19 compounds were confirmed unambiguously with standards. Moreover, the characteristic fragmentations and fragmentation patterns of different compounds in CLMD were summarized, and each compound was classified as an individual herb. It was demonstrated that this method is rapid and accurate and could provide a strategy for the qualitative analysis of the chemical constituents of CLMD, and these results will provide experimental evidence for the subsequent studies on the pharmacodynamic material basis and quality control of CLMD.
{"title":"Rapid identification of chemical constituents of Chaihu‐jia‐Longgu‐Muli decoction based on ultra‐performance liquid chromatography‐quadrupole time‐of‐flight mass spectrometry coupled with the UNIFI platform","authors":"Tong Wu, Yin Liu, Hanshuo Dong, Chunling Ai, Li Sun","doi":"10.1002/sscp.202300103","DOIUrl":"https://doi.org/10.1002/sscp.202300103","url":null,"abstract":"Abstract Chaihu‐jia Longgu‐Muli decoction (CLMD) is a traditional Chinese medicine formula, and it has been used for more than a thousand years to treat mental and nervous system diseases. However, the active constitutions of the CLMD are unclear. Accordingly, an integrated analysis based on the ultra‐performance liquid chromatography‐quadrupole time‐of‐flight mass spectrometry method combined with the UNIFI platform was used to clarify the chemical composition of the CLMD. As a result, 102 compounds including 38 saponins, 32 flavonoids, 12 polyphenols, nine anthraquinones, and 11 others were identified or tentatively presumed. Among them, 19 compounds were confirmed unambiguously with standards. Moreover, the characteristic fragmentations and fragmentation patterns of different compounds in CLMD were summarized, and each compound was classified as an individual herb. It was demonstrated that this method is rapid and accurate and could provide a strategy for the qualitative analysis of the chemical constituents of CLMD, and these results will provide experimental evidence for the subsequent studies on the pharmacodynamic material basis and quality control of CLMD.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":"54 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136061358","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Niroja Vadagam, Sharath Babu Haridasyam, Muvvala Venkatanarayana, Narasimha S. Lakka, Sanjeeva R. Chinnakadoori
Abstract The present research developed and validated a new stability‐indicating technique for the stereo‐selectively enantiomers of the antiviral nucleoside analog Valacyclovir hydrochloride (VAL). The chiral separation was performed using normal‐phase high‐performance liquid chromatography (HPLC) with a chiral stationary phase consisting of amylose tris(3‐chloro‐5‐methylphenylcarbamate) and a mobile phase of “ n ‐hexane, methanol, ethanol, and diethylamine”, flow rate of 0.60 mL/min, column temperature of 30°C, injection volume of 10‐μL, detection wavelength of 254‐nm, and run time of 25‐min. The enantiomers (S‐enantiomer, L‐isomer, R‐enantiomer, and D‐isomer) of Valacyclovir were separated with a resolution of 4.8 and no interference. The validation parameters verified for the proposed method, linearity in a range of 0.1002–24.3486 μg/mL (0.02–4.86%) with a regression coefficient of 0.999, and the accuracy was determined with excellent recoveries ranging from 94.38%–109.97%. The concentrations established for the detection limit and quantitation limit were 0.01% and 0.02%, respectively. The forced degradation experiments were used to assess the stability‐indicating qualities. D‐Valacyclovir impurity was successfully evaluated in release and stability samples of VAL in drug substance and tablet dosage forms using the proposed normal phase chiral HPLC approach.
{"title":"Separation and quantitation of valacyclovir enantiomers using stability‐indicating chiral liquid chromatography method with a chiral stationary phase of amylose tris‐(3,5‐dimethylphenylcarbamate)","authors":"Niroja Vadagam, Sharath Babu Haridasyam, Muvvala Venkatanarayana, Narasimha S. Lakka, Sanjeeva R. Chinnakadoori","doi":"10.1002/sscp.202300145","DOIUrl":"https://doi.org/10.1002/sscp.202300145","url":null,"abstract":"Abstract The present research developed and validated a new stability‐indicating technique for the stereo‐selectively enantiomers of the antiviral nucleoside analog Valacyclovir hydrochloride (VAL). The chiral separation was performed using normal‐phase high‐performance liquid chromatography (HPLC) with a chiral stationary phase consisting of amylose tris(3‐chloro‐5‐methylphenylcarbamate) and a mobile phase of “ n ‐hexane, methanol, ethanol, and diethylamine”, flow rate of 0.60 mL/min, column temperature of 30°C, injection volume of 10‐μL, detection wavelength of 254‐nm, and run time of 25‐min. The enantiomers (S‐enantiomer, L‐isomer, R‐enantiomer, and D‐isomer) of Valacyclovir were separated with a resolution of 4.8 and no interference. The validation parameters verified for the proposed method, linearity in a range of 0.1002–24.3486 μg/mL (0.02–4.86%) with a regression coefficient of 0.999, and the accuracy was determined with excellent recoveries ranging from 94.38%–109.97%. The concentrations established for the detection limit and quantitation limit were 0.01% and 0.02%, respectively. The forced degradation experiments were used to assess the stability‐indicating qualities. D‐Valacyclovir impurity was successfully evaluated in release and stability samples of VAL in drug substance and tablet dosage forms using the proposed normal phase chiral HPLC approach.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":"2015 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136155039","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract Evogliptin (EVG) is prescribed in the treatment of type 2 diabetes. An accurate and precise high‐performance liquid chromatographic method along with forced degradation studies and design of experiments has been developed for the estimation of EVG. Agilent C 18 Column (250 × 4.6 mm, 5 μm particle size) was used as the stationary phase and methanol: 0.05% Orthophosphoric acid in water (51: 49% V/V) was used as the mobile phase. The method was linear in the concentration range of 5–30 μg/ml with a correlation coefficient (r 2 ) of 0.9996. The proposed method was validated with respect to linearity, accuracy, precision, and robustness as per the International Conference on Harmonization Q2 (R1) guideline. A forced degradation study was performed to find out the intrinsic stability of the molecules. The degradation products were well resolved from the drug peak and did not interfere with the analysis. The method was successfully applied for the analysis of EVG.
EVG是治疗2型糖尿病的处方药。通过强制降解研究和实验设计，开发了一种准确、精确的高效液相色谱法来估计EVG。以Agilent C 18色谱柱(250 × 4.6 mm, 5 μm粒径)为固定相，甲醇:0.05%正磷酸水溶液(51∶49% V/V)为流动相。该方法在5 ~ 30 μg/ml浓度范围内线性良好，相关系数(r2)为0.9996。根据国际协调会议Q2 (R1)指南，对所提出的方法进行了线性，准确度，精密度和鲁棒性验证。进行了强制降解研究，以找出分子的内在稳定性。降解产物从药峰上很好地分离出来，不干扰分析。该方法成功地应用于EVG的分析。
{"title":"Development of stability indicating liquid chromatographic method for estimation of novel anti‐diabetic drug Evogliptin","authors":"Deepanti Gajjar, Dimalkumar S. Shah","doi":"10.1002/sscp.202300082","DOIUrl":"https://doi.org/10.1002/sscp.202300082","url":null,"abstract":"Abstract Evogliptin (EVG) is prescribed in the treatment of type 2 diabetes. An accurate and precise high‐performance liquid chromatographic method along with forced degradation studies and design of experiments has been developed for the estimation of EVG. Agilent C 18 Column (250 × 4.6 mm, 5 μm particle size) was used as the stationary phase and methanol: 0.05% Orthophosphoric acid in water (51: 49% V/V) was used as the mobile phase. The method was linear in the concentration range of 5–30 μg/ml with a correlation coefficient (r 2 ) of 0.9996. The proposed method was validated with respect to linearity, accuracy, precision, and robustness as per the International Conference on Harmonization Q2 (R1) guideline. A forced degradation study was performed to find out the intrinsic stability of the molecules. The degradation products were well resolved from the drug peak and did not interfere with the analysis. The method was successfully applied for the analysis of EVG.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":"20 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135063279","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Varvara Papaioannou, Eleni V. Mikropoulou, Amalia Yanni, Vaios Karathanos, Athina Aidini, Alexandre Paccou, Maria Halabalaki
Amino acids (AAs) comprise fundamental components of the human organism since they participate in several physiological functions. Numerous methods have been developed to quantify AAs in biological fluids due to the challenges posed by matrix handling (limited sample quantities, complexity) and their physicochemical properties (protein binding, low detectability, and high polarity). This study developed and validated an LC–MS method using ion‐pairing chromatography, a sequential window acquisition of all theoretical mass spectra approach and a derivatization kit for AA analysis of physiological fluids to simultaneously quantify plasma AAs and provide data for untargeted analytical approaches. Finally, proof of concept was demonstrated through the analysis of 450 samples from a dietary intervention study with protein‐enriched wheat biscuits.
{"title":"Integration of a derivatization protocol and LC–MS sequential window acquisition of all theoretical mass spectra strategy for amino acid determination in human plasma","authors":"Varvara Papaioannou, Eleni V. Mikropoulou, Amalia Yanni, Vaios Karathanos, Athina Aidini, Alexandre Paccou, Maria Halabalaki","doi":"10.1002/sscp.202300100","DOIUrl":"https://doi.org/10.1002/sscp.202300100","url":null,"abstract":"Amino acids (AAs) comprise fundamental components of the human organism since they participate in several physiological functions. Numerous methods have been developed to quantify AAs in biological fluids due to the challenges posed by matrix handling (limited sample quantities, complexity) and their physicochemical properties (protein binding, low detectability, and high polarity). This study developed and validated an LC–MS method using ion‐pairing chromatography, a sequential window acquisition of all theoretical mass spectra approach and a derivatization kit for AA analysis of physiological fluids to simultaneously quantify plasma AAs and provide data for untargeted analytical approaches. Finally, proof of concept was demonstrated through the analysis of 450 samples from a dietary intervention study with protein‐enriched wheat biscuits.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":"58 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135736049","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jirun Jia, Rui Bao, Jiayue Hou, Mengdi Qin, Wen Li, Li Yang, Qiang Fu
Abstract Atorvastatin calcium and lisinopril are commonly used to reduce cholesterol and control blood pressure in the clinic. In this study, a sensitive and rapid method was developed and validated for the simultaneous detection of plasma concentrations of atorvastatin calcium and lisinopril using an ultra‐high performance liquid chromatography‐tandem mass spectrometry with an ACQUITY UPLC BEH C 18 column. The mobile phase of methanol and 0.1% formic acid aqueous solution was pumped at a flow rate of 0.2 mL/min. The retention times of atorvastatin calcium and lisinopril were 2.48 min and 1.99 min, respectively. The linear range of atorvastatin calcium in plasma samples was 1–2000 ng/mL, while that of lisinopril was 5–2000 ng/mL. The absolute values of intraday and inter‐day precision and accuracy were all below 15%. Furthermore, the recovery and matrix effects of atorvastatin calcium, lisinopril, and nimodipine were 80%–120% and 85%‐115%, respectively. Therefore, the developed method can be applied for the simultaneous quantification of atorvastatin calcium and lisinopril in rat plasma. The C max were about 199 ng/mL and 2059 ng/mL for atorvastatin calcium and lisinopril, respectively. In addition, the T 1/2 of atorvastatin calcium and lisinopril were 0.6 ± 0.3 and 5.4 ± 1.7 h, respectively.
{"title":"A rapid and sensitive ultra‐high‐performance liquid chromatography‐tandem mass spectrometry method for the simultaneous determination of atorvastatin calcium and lisinopril in rat plasma and its application in a pharmacokinetic study","authors":"Jirun Jia, Rui Bao, Jiayue Hou, Mengdi Qin, Wen Li, Li Yang, Qiang Fu","doi":"10.1002/sscp.202300129","DOIUrl":"https://doi.org/10.1002/sscp.202300129","url":null,"abstract":"Abstract Atorvastatin calcium and lisinopril are commonly used to reduce cholesterol and control blood pressure in the clinic. In this study, a sensitive and rapid method was developed and validated for the simultaneous detection of plasma concentrations of atorvastatin calcium and lisinopril using an ultra‐high performance liquid chromatography‐tandem mass spectrometry with an ACQUITY UPLC BEH C 18 column. The mobile phase of methanol and 0.1% formic acid aqueous solution was pumped at a flow rate of 0.2 mL/min. The retention times of atorvastatin calcium and lisinopril were 2.48 min and 1.99 min, respectively. The linear range of atorvastatin calcium in plasma samples was 1–2000 ng/mL, while that of lisinopril was 5–2000 ng/mL. The absolute values of intraday and inter‐day precision and accuracy were all below 15%. Furthermore, the recovery and matrix effects of atorvastatin calcium, lisinopril, and nimodipine were 80%–120% and 85%‐115%, respectively. Therefore, the developed method can be applied for the simultaneous quantification of atorvastatin calcium and lisinopril in rat plasma. The C max were about 199 ng/mL and 2059 ng/mL for atorvastatin calcium and lisinopril, respectively. In addition, the T 1/2 of atorvastatin calcium and lisinopril were 0.6 ± 0.3 and 5.4 ± 1.7 h, respectively.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":"72 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136024312","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Muhammad Wasim, Haq Nawaz Khan, Abdul Tawab, Fazal e Habib, Mazhar Iqbal, Fazli Rabbi Awan
Abstract Plasma amino acids are generally analyzed through ion exchange chromatography, a reproducible but time‐consuming method. Here, we report the optimization of a reverse‐phase‐high‐performance liquid chromatography with fluorescence detector (RP‐HPLC‐FLD) assay for the detection and quantification of plasma amino acids for potential applications in metabolic disorders (e.g., aminoacidopathies, a rare group of Inborn Errors of Metabolism). For assay development, initially standard amino acids were derivatized with ortho‐phthalaldehyde‐3‐mercaptopropionic acid (OPA‐3‐MPA) and filtered through a 0.20 μm syringe filter. The excitation and emission wavelengths of 240–450 nm (λex—λem) were used for the detection of amino acids. Chromatographic separation was achieved by gradient RP‐HPLC‐FLD through C18 symmetry column (150 × 4.6 mm, particle size 3.5 μm). HPLC assay was successfully optimized and was able to detect amino acids in the range of 10–400 ng/mL and good linearity (R 2 > 0.98) was achieved in the mixture for each standard amino acid. Moreover, the current assay showed great efficiency with two additional advantages: the use of low‐cost mobile phases, and the detection and quantification of amino acids at low level (ng/mL) concentration in biofluids. This assay could be applied for the analysis of human plasma to identify aminoacidopathies in newborn screening programs, and other metabolic disorders.
{"title":"High‐performance liquid chromatography‐based assay optimization for the detection of plasma amino acids for applications in metabolic disorders in developing countries","authors":"Muhammad Wasim, Haq Nawaz Khan, Abdul Tawab, Fazal e Habib, Mazhar Iqbal, Fazli Rabbi Awan","doi":"10.1002/sscp.202300119","DOIUrl":"https://doi.org/10.1002/sscp.202300119","url":null,"abstract":"Abstract Plasma amino acids are generally analyzed through ion exchange chromatography, a reproducible but time‐consuming method. Here, we report the optimization of a reverse‐phase‐high‐performance liquid chromatography with fluorescence detector (RP‐HPLC‐FLD) assay for the detection and quantification of plasma amino acids for potential applications in metabolic disorders (e.g., aminoacidopathies, a rare group of Inborn Errors of Metabolism). For assay development, initially standard amino acids were derivatized with ortho‐phthalaldehyde‐3‐mercaptopropionic acid (OPA‐3‐MPA) and filtered through a 0.20 μm syringe filter. The excitation and emission wavelengths of 240–450 nm (λex—λem) were used for the detection of amino acids. Chromatographic separation was achieved by gradient RP‐HPLC‐FLD through C18 symmetry column (150 × 4.6 mm, particle size 3.5 μm). HPLC assay was successfully optimized and was able to detect amino acids in the range of 10–400 ng/mL and good linearity (R 2 > 0.98) was achieved in the mixture for each standard amino acid. Moreover, the current assay showed great efficiency with two additional advantages: the use of low‐cost mobile phases, and the detection and quantification of amino acids at low level (ng/mL) concentration in biofluids. This assay could be applied for the analysis of human plasma to identify aminoacidopathies in newborn screening programs, and other metabolic disorders.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":"2019 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-09-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136365143","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pintu B. Prajapati, Abhinandan Shahi, Aneri Acharya, V. Pulusu, Shailesh Shah
According to the concept of green analytical chemistry, the analytical method development should be carried out by avoiding or minimizing the usage of toxic organic solvents for the safety of human and aquatic animal life and the protection of the environment. Hence, the green analytical chemistry (GAC)‐assisted robust liquid chromatographic method has been developed for chromatographic analysis of azilsartan medoxomil and cilnidipine (CIL) using safe organic solvents. The chromatographic method was developed by the implementation of analytical quality by design using chemometrics and response surface modeling. The chromatographic separation was carried out using Shim‐Pack C18 (250 × 4.6 mm, 5.0 μm) column as stationary phase and ethanol: 0.1% V/V ammonia solution (45:55, %V/V) as mobile phase. The chromatographic peak of AZL and CIL was found to be at the retention time of 3.5 and 4.5 min, respectively. The flow rate was kept at 1.0 mL/min and the column oven temperature was set to 40˚C. The developed method was found to be validated as per the International Council for Harmonization Q2 (R1) guideline. The method was applied for the assay of fixed‐dose combination and results were found in compliance with the labeled claim. The greenness of the method was evaluated using GAC tools.
{"title":"Robust method operable design region for economical and eco‐friendly chromatographic analysis of azilsartan medoxomil and cilnidipine by incorporating a hybrid approach of green analytical chemistry and analytical quality by design","authors":"Pintu B. Prajapati, Abhinandan Shahi, Aneri Acharya, V. Pulusu, Shailesh Shah","doi":"10.1002/sscp.202300111","DOIUrl":"https://doi.org/10.1002/sscp.202300111","url":null,"abstract":"According to the concept of green analytical chemistry, the analytical method development should be carried out by avoiding or minimizing the usage of toxic organic solvents for the safety of human and aquatic animal life and the protection of the environment. Hence, the green analytical chemistry (GAC)‐assisted robust liquid chromatographic method has been developed for chromatographic analysis of azilsartan medoxomil and cilnidipine (CIL) using safe organic solvents. The chromatographic method was developed by the implementation of analytical quality by design using chemometrics and response surface modeling. The chromatographic separation was carried out using Shim‐Pack C18 (250 × 4.6 mm, 5.0 μm) column as stationary phase and ethanol: 0.1% V/V ammonia solution (45:55, %V/V) as mobile phase. The chromatographic peak of AZL and CIL was found to be at the retention time of 3.5 and 4.5 min, respectively. The flow rate was kept at 1.0 mL/min and the column oven temperature was set to 40˚C. The developed method was found to be validated as per the International Council for Harmonization Q2 (R1) guideline. The method was applied for the assay of fixed‐dose combination and results were found in compliance with the labeled claim. The greenness of the method was evaluated using GAC tools.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":" ","pages":""},"PeriodicalIF":1.1,"publicationDate":"2023-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46419719","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract For the simultaneous estimation of Benidipine hydrochloride (BEN) and Telmisartan (TEL) in combined pharmaceutical dosage form, a stability‐indicating high‐performance thin‐layer chromatography (HPTLC) method has been developed and validated. It involves the use of HPTLC plates pre‐coated with silica gel 60F 254 and a mobile phase comprising of Methanol: Acetonitrile (1:9 v/v). The measurements of both drugs were performed at 237 nm. The drugs were subjected to acid‐alkali hydrolysis, photolytic, thermal, and oxidation degradation. The linearity of BEN was found to be between 200–1000 ng/band (R 2 = 0.9917), while for TEL, linearity was found to be between 2000 and 10000 ng/band (R 2 = 0.9979). The limit of detection and limit of quantitation were found to be 3.28 and 9.94 ng/spot for BEN and 158.79 and 481.17 ng/spot for TEL. The % relative standard deviation of the precision study was determined to be less than 2%. Various system suitability parameters were determined. The R f values of BEN and TEL were found to be 0.890 ± 0.0105 and 0.173 ± 0.0151, respectively. The resolution between BEN and TEL peak were found to be 4.48 and capacity factors were found to be 0.21 and 0.74 for BEN and TEL, respectively.
{"title":"Development and validation of stability indicating thin‐layer chromatography method for simultaneous estimation of Benidipine hydrochloride and Telmisartan","authors":"Laxmibharti B. P. Mandal, Ketan Shah","doi":"10.1002/sscp.202300075","DOIUrl":"https://doi.org/10.1002/sscp.202300075","url":null,"abstract":"Abstract For the simultaneous estimation of Benidipine hydrochloride (BEN) and Telmisartan (TEL) in combined pharmaceutical dosage form, a stability‐indicating high‐performance thin‐layer chromatography (HPTLC) method has been developed and validated. It involves the use of HPTLC plates pre‐coated with silica gel 60F 254 and a mobile phase comprising of Methanol: Acetonitrile (1:9 v/v). The measurements of both drugs were performed at 237 nm. The drugs were subjected to acid‐alkali hydrolysis, photolytic, thermal, and oxidation degradation. The linearity of BEN was found to be between 200–1000 ng/band (R 2 = 0.9917), while for TEL, linearity was found to be between 2000 and 10000 ng/band (R 2 = 0.9979). The limit of detection and limit of quantitation were found to be 3.28 and 9.94 ng/spot for BEN and 158.79 and 481.17 ng/spot for TEL. The % relative standard deviation of the precision study was determined to be less than 2%. Various system suitability parameters were determined. The R f values of BEN and TEL were found to be 0.890 ± 0.0105 and 0.173 ± 0.0151, respectively. The resolution between BEN and TEL peak were found to be 4.48 and capacity factors were found to be 0.21 and 0.74 for BEN and TEL, respectively.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":"106 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135891007","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pooja Dhurjad, Kajal Gupta, Akash P. Sakla, N. Shankaraiah, R. Sonti
Brassinin, a phytochemical in cruciferous vegetables, is an indoleamine 2,3‐dioxygenase inhibitor and possesses a therapeutic opportunity in cancer immunotherapy. Here, we have developed and validated a novel, specific, and rapid bioanalytical method in rat plasma that is unavailable in the literature for preclinical studies. The method development involved single‐step protein precipitation as an extraction method using acetonitrile with an absolute and relative mean recovery of 79.32% and 93.59%, respectively. The method is accurate and precise over the linearity range of 1.0–50.0 μg/mL with a retention time of 5.3 and 3.5 min of brassinin and telmisartan, an internal standard. The intra‐day and inter‐day precision of the method expressed as a %coefficient of variation, ranging from 0.03% to 2.82% and 0.39% to 5.82%, whereas intra‐day and inter‐day accuracy ranged from 99.75% to 103.40% and 99.76% to 104.25%, respectively. The high‐performance liquid chromatography bioanalytical method was successfully validated as per Food and Drug Administration guidelines, and it can be applied in routine bioanalysis and pharmacokinetic studies.
{"title":"A validated high‐performance liquid chromatography method for the determination of brassinin, an indoleamine 2,3‐dioxygenase inhibitor in rat plasma","authors":"Pooja Dhurjad, Kajal Gupta, Akash P. Sakla, N. Shankaraiah, R. Sonti","doi":"10.1002/sscp.202300073","DOIUrl":"https://doi.org/10.1002/sscp.202300073","url":null,"abstract":"Brassinin, a phytochemical in cruciferous vegetables, is an indoleamine 2,3‐dioxygenase inhibitor and possesses a therapeutic opportunity in cancer immunotherapy. Here, we have developed and validated a novel, specific, and rapid bioanalytical method in rat plasma that is unavailable in the literature for preclinical studies. The method development involved single‐step protein precipitation as an extraction method using acetonitrile with an absolute and relative mean recovery of 79.32% and 93.59%, respectively. The method is accurate and precise over the linearity range of 1.0–50.0 μg/mL with a retention time of 5.3 and 3.5 min of brassinin and telmisartan, an internal standard. The intra‐day and inter‐day precision of the method expressed as a %coefficient of variation, ranging from 0.03% to 2.82% and 0.39% to 5.82%, whereas intra‐day and inter‐day accuracy ranged from 99.75% to 103.40% and 99.76% to 104.25%, respectively. The high‐performance liquid chromatography bioanalytical method was successfully validated as per Food and Drug Administration guidelines, and it can be applied in routine bioanalysis and pharmacokinetic studies.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":" ","pages":""},"PeriodicalIF":1.1,"publicationDate":"2023-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46907907","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Joan Bergman, Lainey Harvill, Joe S. Smith, Ellen Haynes, Christopher A. Cleveland, M. Yabsley, S. Coker, Wided Najahi-Missaoui, D. Elder, S. Cox
The purpose of this study was to establish a reliable method for the quantification of flubendazole and its metabolite, 2‐aminoflubendazole, in small‐volume canine plasma samples. Following liquid extraction with chloroform, samples were separated by reverse‐phase high‐performance liquid chromatography on an XBridge C18 4.6 × 250 mm column (5 μm). Quantification was performed using ultraviolet detection at 246 nm. A mixture of 5 mM potassium phosphate monobasic and acetonitrile (72:28) was used as the mobile phase. The standard curve ranged from 2.5 to 1000 ng/mL. Intra‐ and interassay variance for flubendazole and 2‐aminoflubendazole was less than 6%, while the recovery ranged from 91 to 101%. The lower limit of quantification was 2.5 ng/mL. This method was successfully validated and applied to the analysis of flubendazole and 2‐aminoflubendazole samples.
{"title":"Validation of a high‐performance liquid chromatography method for detecting flubendazole and 2‐aminoflubendazole in canine plasma","authors":"Joan Bergman, Lainey Harvill, Joe S. Smith, Ellen Haynes, Christopher A. Cleveland, M. Yabsley, S. Coker, Wided Najahi-Missaoui, D. Elder, S. Cox","doi":"10.1002/sscp.202300065","DOIUrl":"https://doi.org/10.1002/sscp.202300065","url":null,"abstract":"The purpose of this study was to establish a reliable method for the quantification of flubendazole and its metabolite, 2‐aminoflubendazole, in small‐volume canine plasma samples. Following liquid extraction with chloroform, samples were separated by reverse‐phase high‐performance liquid chromatography on an XBridge C18 4.6 × 250 mm column (5 μm). Quantification was performed using ultraviolet detection at 246 nm. A mixture of 5 mM potassium phosphate monobasic and acetonitrile (72:28) was used as the mobile phase. The standard curve ranged from 2.5 to 1000 ng/mL. Intra‐ and interassay variance for flubendazole and 2‐aminoflubendazole was less than 6%, while the recovery ranged from 91 to 101%. The lower limit of quantification was 2.5 ng/mL. This method was successfully validated and applied to the analysis of flubendazole and 2‐aminoflubendazole samples.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":" ","pages":""},"PeriodicalIF":1.1,"publicationDate":"2023-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41763337","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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