Silvio Uhlig, Kun Cai, Krystyna Anna Liskiewicz, Maria Pain, Lene Grutle, Hanne Røberg‐Larsen, Roger Simm
The 3,3′‐linked cyclic dinucleotides (CDNs) of adenosine monophosphate (AMP) and guanosine monophosphate (GMP), cyclic di‐AMP (c‐di‐AMP), cyclic di‐GMP (c‐di‐GMP), and c‐GMP–AMP (cGAMP) are second messenger molecules in bacteria that regulate processes, such as biofilm formation, motility, virulence, stress response, and cell wall homeostasis. To analyze the profiles of the three CDNs together with their breakdown and precursor molecules, 5′‐phosphoadenylyl‐(3′ → 5′)adenine (pApA), 5′‐guanylyl‐(3′ → 5′)guanine (pGpG), 5′‐AMP, 3′‐ and 5′‐GMP, adenosine triphosphate (ATP), and GTP, we established an LC–MS/MS‐based approach for semi‐quantification and profiling. Weak anion exchange solid‐phase extraction was employed to improve selectivity and instrumental signal/noise of CDNs as well as pApA and pGpG. CDNs were analyzed using reverse‐phase UHPLC–MS/MS, whereas all other nucleotides were analyzed using hydrophilic interaction chromatography (HILIC)–MS/MS. The instrument limit of quantification ranged from 0.72 (c‐di‐AMP) to 60 nM (ATP and GTP). We applied this method to the analysis of the nine nucleotides in eight bacterial strains and found that the profiles varied widely in terms of both absolute and relative concentrations. Thus, CDN concentrations were generally <1 pmol/mg biomass, and the hydrolysis products, pApA and pGpG, were detected at lower pmol/mg concentrations. The presented method is a relatively simple and straightforward approach to profiling nucleotides with the rationale of comparing their relative levels between populations of bacterial strains.
{"title":"Profiling of cyclic di‐adenyl and ‐guanyl nucleotides and their precursors and degradation products in bacteria using LC–MS/MS","authors":"Silvio Uhlig, Kun Cai, Krystyna Anna Liskiewicz, Maria Pain, Lene Grutle, Hanne Røberg‐Larsen, Roger Simm","doi":"10.1002/sscp.202400039","DOIUrl":"https://doi.org/10.1002/sscp.202400039","url":null,"abstract":"The 3,3′‐linked cyclic dinucleotides (CDNs) of adenosine monophosphate (AMP) and guanosine monophosphate (GMP), cyclic di‐AMP (c‐di‐AMP), cyclic di‐GMP (c‐di‐GMP), and c‐GMP–AMP (cGAMP) are second messenger molecules in bacteria that regulate processes, such as biofilm formation, motility, virulence, stress response, and cell wall homeostasis. To analyze the profiles of the three CDNs together with their breakdown and precursor molecules, 5′‐phosphoadenylyl‐(3′ → 5′)adenine (pApA), 5′‐guanylyl‐(3′ → 5′)guanine (pGpG), 5′‐AMP, 3′‐ and 5′‐GMP, adenosine triphosphate (ATP), and GTP, we established an LC–MS/MS‐based approach for semi‐quantification and profiling. Weak anion exchange solid‐phase extraction was employed to improve selectivity and instrumental signal/noise of CDNs as well as pApA and pGpG. CDNs were analyzed using reverse‐phase UHPLC–MS/MS, whereas all other nucleotides were analyzed using hydrophilic interaction chromatography (HILIC)–MS/MS. The instrument limit of quantification ranged from 0.72 (c‐di‐AMP) to 60 nM (ATP and GTP). We applied this method to the analysis of the nine nucleotides in eight bacterial strains and found that the profiles varied widely in terms of both absolute and relative concentrations. Thus, CDN concentrations were generally <1 pmol/mg biomass, and the hydrolysis products, pApA and pGpG, were detected at lower pmol/mg concentrations. The presented method is a relatively simple and straightforward approach to profiling nucleotides with the rationale of comparing their relative levels between populations of bacterial strains.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2024-02-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140440848","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}
Rajesh Kanagaddi, Vaishnavi Chintala, S. Nannapaneni, N. K. Katari, Suresh Salakolusu, Jnsrc Murty, M. Ranga, Muralidharan Kaliyaperumal
The current study explains the degradation behavior of Febuxostat API, a non‐purine xanthine oxidase inhibitor used to treat hyperuricemia. A degradation study was carried out as per the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines, and the study confirms that the Febuxostat is largely stable in thermal, photolytic, oxidative, and basic hydrolytic conditions and labile in acid hydrolysis conditions. There were four different degradation products (DPs) found during acid hydrolysis; of these, DPs 2, 3, and 4 are new and have never been reported before, while DP 1 is known and has already been published. All these DPs were identified using ultra‐high‐performance liquid chromatography‐mass spectrometry (UHPLC‐MS) analysis, purified by using preparative HPLC, and characterized using high‐resolution mass spectrometry (HRMS) and nuclear magnetic resonance (NMR) spectroscopy techniques. The formed DPs are by the hydrolysis of the cyano functional group of Febuxostat and the esterification of DP‐1 under acidic conditions. All DP's structural characterization was carried out using NMR spectroscopy and HRMS. The present study describes concrete confirmation of DP structures and it explains the stability behavior of the Febuxostat. The current method is also used to identify DPs with shorter runtime in the future.
{"title":"Isolation and identification of forced degradation products of Febuxostat","authors":"Rajesh Kanagaddi, Vaishnavi Chintala, S. Nannapaneni, N. K. Katari, Suresh Salakolusu, Jnsrc Murty, M. Ranga, Muralidharan Kaliyaperumal","doi":"10.1002/sscp.202300237","DOIUrl":"https://doi.org/10.1002/sscp.202300237","url":null,"abstract":"The current study explains the degradation behavior of Febuxostat API, a non‐purine xanthine oxidase inhibitor used to treat hyperuricemia. A degradation study was carried out as per the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines, and the study confirms that the Febuxostat is largely stable in thermal, photolytic, oxidative, and basic hydrolytic conditions and labile in acid hydrolysis conditions. There were four different degradation products (DPs) found during acid hydrolysis; of these, DPs 2, 3, and 4 are new and have never been reported before, while DP 1 is known and has already been published. All these DPs were identified using ultra‐high‐performance liquid chromatography‐mass spectrometry (UHPLC‐MS) analysis, purified by using preparative HPLC, and characterized using high‐resolution mass spectrometry (HRMS) and nuclear magnetic resonance (NMR) spectroscopy techniques. The formed DPs are by the hydrolysis of the cyano functional group of Febuxostat and the esterification of DP‐1 under acidic conditions. All DP's structural characterization was carried out using NMR spectroscopy and HRMS. The present study describes concrete confirmation of DP structures and it explains the stability behavior of the Febuxostat. The current method is also used to identify DPs with shorter runtime in the future.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2024-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140447812","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}
Xiaoying Lin, Xin Zhao, Bin Yang, Yinxuan Huang, Ruiwen Zhang, Manna Li, Mengjie Xiao, Hui Xie
A pinanamine‐based analog, (1R,2R,3R,5S)‐N‐((3‐cyclopropylthiophen‐2‐yl)methyl)‐2,6,6‐trimethylbicyclo[3.1.1]heptan‐3‐amine (M090) was synthesized and demonstrated with anti‐influenza activity to overcome the multi‐drug resistance. Herein, a rapid and robust liquid chromatography‐tandem mass spectrometry method was developed and validated to quantify M090 in rat plasma. With simple protein precipitation by methanol, the separation of M090 was performed on a Waters BEH‐C18 column (2.1 × 100 mm, 1.7 μm) by gradient elution at 0.4 mL/min with mobile phases consisting of water containing 0.1 % formic acid (phase A) and methanol (phase B). M090 and clenbuterol (internal standard) were detected using multiple reaction monitoring in positive electrospray ionization mode with transitions of m/z 290.3→137.0 and m/z 277.0→203.0, respectively. The method was validated over a linear range of 1.0–2400 ng/mL with a regression coefficient of 0.9974 and no endogenous interference. The intra‐ and inter‐batch precisions were within 8.29% and accuracy ranged from 100.3% to 108.1% for M090. The validated method was utilized to evaluate the in vitro metabolic stability and in vivo pharmacokinetics of M090 in rats.
{"title":"Determination of a novel anti‐influenza pinanamine‐based analog in rat plasma by liquid chromatography‐tandem mass spectrometry and its application in pharmacokinetic evaluations","authors":"Xiaoying Lin, Xin Zhao, Bin Yang, Yinxuan Huang, Ruiwen Zhang, Manna Li, Mengjie Xiao, Hui Xie","doi":"10.1002/sscp.202300193","DOIUrl":"https://doi.org/10.1002/sscp.202300193","url":null,"abstract":"A pinanamine‐based analog, (1R,2R,3R,5S)‐N‐((3‐cyclopropylthiophen‐2‐yl)methyl)‐2,6,6‐trimethylbicyclo[3.1.1]heptan‐3‐amine (M090) was synthesized and demonstrated with anti‐influenza activity to overcome the multi‐drug resistance. Herein, a rapid and robust liquid chromatography‐tandem mass spectrometry method was developed and validated to quantify M090 in rat plasma. With simple protein precipitation by methanol, the separation of M090 was performed on a Waters BEH‐C18 column (2.1 × 100 mm, 1.7 μm) by gradient elution at 0.4 mL/min with mobile phases consisting of water containing 0.1 % formic acid (phase A) and methanol (phase B). M090 and clenbuterol (internal standard) were detected using multiple reaction monitoring in positive electrospray ionization mode with transitions of m/z 290.3→137.0 and m/z 277.0→203.0, respectively. The method was validated over a linear range of 1.0–2400 ng/mL with a regression coefficient of 0.9974 and no endogenous interference. The intra‐ and inter‐batch precisions were within 8.29% and accuracy ranged from 100.3% to 108.1% for M090. The validated method was utilized to evaluate the in vitro metabolic stability and in vivo pharmacokinetics of M090 in rats.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2024-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140451237","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}
Jin‐Nao‐Ning mixture (JNNm) has been clinically used to treat insomnia in China. However, the major constituents of JNNm were not revealed. In this study, an ultra‐high‐performance liquid chromatography‐tandem mass spectrometry was established and validated for simultaneous quantitation of 19 representative constituents of JNNm. The method had good specificity and adequate sensitivity. The calibration curves of the constituents displayed good linearity (r > 0.990) in appropriate ranges. The intra‐ and inter‐day precision at three quality control (QC) levels ranged from 0.68% to 5.17% and 0.42% to 5.37%, respectively. The recovery ranged from 91.76% to 110.06%, with a precision of less than 5.07%. In addition, the method had good repeatability (relative standard deviation [RSD] < 4.93%). The analytes were all stable for 24 h (RSD < 7.98%) at room temperature. The results of the quantitative analysis showed that salvianolic acid B (495.7–606.3 μg/mL) is the most abundant constituent, followed by betaine (411.3–468.3 μg/mL), salvianic acid A (10.7–34.9 μg/mL), lobetyolin (11.6–14.6 μg/mL), spinosin (4.0–7.6 μg/mL), schisandrol A (1.5–4.1 μg/mL), and schisandrol B (1.2–1.5 μg/mL). Concentrations of other constituents were below 1 μg/mL. This study would be helpful for the QC of JNNm and the revelation of its effective constituents.
{"title":"Simultaneous determination of 19 constituents in the Jian‐Nao‐Ning mixture using high‐performance liquid chromatography‐tandem mass spectrometry","authors":"Ji‐Yong Fu, Ding Ding, Ru‐Mei Wang, Lyu‐Feng Zhang, Bing‐Liang Ma, Fu‐Xuan Liu, Guo‐Fu Zhu","doi":"10.1002/sscp.202300222","DOIUrl":"https://doi.org/10.1002/sscp.202300222","url":null,"abstract":"Jin‐Nao‐Ning mixture (JNNm) has been clinically used to treat insomnia in China. However, the major constituents of JNNm were not revealed. In this study, an ultra‐high‐performance liquid chromatography‐tandem mass spectrometry was established and validated for simultaneous quantitation of 19 representative constituents of JNNm. The method had good specificity and adequate sensitivity. The calibration curves of the constituents displayed good linearity (r > 0.990) in appropriate ranges. The intra‐ and inter‐day precision at three quality control (QC) levels ranged from 0.68% to 5.17% and 0.42% to 5.37%, respectively. The recovery ranged from 91.76% to 110.06%, with a precision of less than 5.07%. In addition, the method had good repeatability (relative standard deviation [RSD] < 4.93%). The analytes were all stable for 24 h (RSD < 7.98%) at room temperature. The results of the quantitative analysis showed that salvianolic acid B (495.7–606.3 μg/mL) is the most abundant constituent, followed by betaine (411.3–468.3 μg/mL), salvianic acid A (10.7–34.9 μg/mL), lobetyolin (11.6–14.6 μg/mL), spinosin (4.0–7.6 μg/mL), schisandrol A (1.5–4.1 μg/mL), and schisandrol B (1.2–1.5 μg/mL). Concentrations of other constituents were below 1 μg/mL. This study would be helpful for the QC of JNNm and the revelation of its effective constituents.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2024-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139779796","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}
Jin‐Nao‐Ning mixture (JNNm) has been clinically used to treat insomnia in China. However, the major constituents of JNNm were not revealed. In this study, an ultra‐high‐performance liquid chromatography‐tandem mass spectrometry was established and validated for simultaneous quantitation of 19 representative constituents of JNNm. The method had good specificity and adequate sensitivity. The calibration curves of the constituents displayed good linearity (r > 0.990) in appropriate ranges. The intra‐ and inter‐day precision at three quality control (QC) levels ranged from 0.68% to 5.17% and 0.42% to 5.37%, respectively. The recovery ranged from 91.76% to 110.06%, with a precision of less than 5.07%. In addition, the method had good repeatability (relative standard deviation [RSD] < 4.93%). The analytes were all stable for 24 h (RSD < 7.98%) at room temperature. The results of the quantitative analysis showed that salvianolic acid B (495.7–606.3 μg/mL) is the most abundant constituent, followed by betaine (411.3–468.3 μg/mL), salvianic acid A (10.7–34.9 μg/mL), lobetyolin (11.6–14.6 μg/mL), spinosin (4.0–7.6 μg/mL), schisandrol A (1.5–4.1 μg/mL), and schisandrol B (1.2–1.5 μg/mL). Concentrations of other constituents were below 1 μg/mL. This study would be helpful for the QC of JNNm and the revelation of its effective constituents.
{"title":"Simultaneous determination of 19 constituents in the Jian‐Nao‐Ning mixture using high‐performance liquid chromatography‐tandem mass spectrometry","authors":"Ji‐Yong Fu, Ding Ding, Ru‐Mei Wang, Lyu‐Feng Zhang, Bing‐Liang Ma, Fu‐Xuan Liu, Guo‐Fu Zhu","doi":"10.1002/sscp.202300222","DOIUrl":"https://doi.org/10.1002/sscp.202300222","url":null,"abstract":"Jin‐Nao‐Ning mixture (JNNm) has been clinically used to treat insomnia in China. However, the major constituents of JNNm were not revealed. In this study, an ultra‐high‐performance liquid chromatography‐tandem mass spectrometry was established and validated for simultaneous quantitation of 19 representative constituents of JNNm. The method had good specificity and adequate sensitivity. The calibration curves of the constituents displayed good linearity (r > 0.990) in appropriate ranges. The intra‐ and inter‐day precision at three quality control (QC) levels ranged from 0.68% to 5.17% and 0.42% to 5.37%, respectively. The recovery ranged from 91.76% to 110.06%, with a precision of less than 5.07%. In addition, the method had good repeatability (relative standard deviation [RSD] < 4.93%). The analytes were all stable for 24 h (RSD < 7.98%) at room temperature. The results of the quantitative analysis showed that salvianolic acid B (495.7–606.3 μg/mL) is the most abundant constituent, followed by betaine (411.3–468.3 μg/mL), salvianic acid A (10.7–34.9 μg/mL), lobetyolin (11.6–14.6 μg/mL), spinosin (4.0–7.6 μg/mL), schisandrol A (1.5–4.1 μg/mL), and schisandrol B (1.2–1.5 μg/mL). Concentrations of other constituents were below 1 μg/mL. This study would be helpful for the QC of JNNm and the revelation of its effective constituents.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2024-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139839823","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}
N. V. V. D. P. Boppy, S. Haridasyam, Niroja Vadagam, M. Venkatanarayana, Sanjeeva R. Chinnakadoori, Dr. Narasimha S. Lakka, PhD
A simple and robust high‐performance liquid chromatography (HPLC) method was developed for organic impurities of hydroxyzine hydrochloride in pharmaceuticals. The developed method was designed to estimate all organic impurities of hydroxyzine. The HPLC separation was achieved using C18 column (150 × 3.9 mm, 5 μm) along with a binary gradient consisting of mobile phases A (0.1%, trifluoroacetic acid in purified water) and B (0.05%, trifluoroacetic acid in acetonitrile), a flow rate of 0.7–mL/min, a column temperature of 30°C and a sample temperature of 25°C. The detection wavelength used was 230 nm for the estimation of impurity‐A, impurity‐B, and all unspecified impurities and degradation products, whereas impurity‐C was quantitated using 254 nm. The stability‐indicating property of the developed HPLC technique was assessed using stress testing conditions of hydrolysis, oxidation, thermal, photo‐light, and humidity. The validation study was performed for the limit of detection and limit of quantification, linearity, and recoveries were 0.03%, 0.05%, and 0.1132–2.9920 μg/mL (R2 > 0.999), and 84.09%–109.74%, respectively. The proposed method is highly suitable for the determination of assay, organic impurities, and degradation products of the hydroxyzine. The chemical structure of degradation product 1 (hydroxyzine N‐Oxide) and degradation product 2 (O‐Acetyl hydroxyzine) were identified with the supporting data of LC‐mass spectrometry (LC‐MS) and high‐resolution MS.
{"title":"Separation and quantification of organic‐related impurities of anti‐histamine drug hydroxyzine in pharmaceutical dosage forms using stability‐indicating high‐performance liquid chromatography, liquid chromatography‐mass spectrometry, and high‐resolution mass spectrometry techniques","authors":"N. V. V. D. P. Boppy, S. Haridasyam, Niroja Vadagam, M. Venkatanarayana, Sanjeeva R. Chinnakadoori, Dr. Narasimha S. Lakka, PhD","doi":"10.1002/sscp.202300157","DOIUrl":"https://doi.org/10.1002/sscp.202300157","url":null,"abstract":"A simple and robust high‐performance liquid chromatography (HPLC) method was developed for organic impurities of hydroxyzine hydrochloride in pharmaceuticals. The developed method was designed to estimate all organic impurities of hydroxyzine. The HPLC separation was achieved using C18 column (150 × 3.9 mm, 5 μm) along with a binary gradient consisting of mobile phases A (0.1%, trifluoroacetic acid in purified water) and B (0.05%, trifluoroacetic acid in acetonitrile), a flow rate of 0.7–mL/min, a column temperature of 30°C and a sample temperature of 25°C. The detection wavelength used was 230 nm for the estimation of impurity‐A, impurity‐B, and all unspecified impurities and degradation products, whereas impurity‐C was quantitated using 254 nm. The stability‐indicating property of the developed HPLC technique was assessed using stress testing conditions of hydrolysis, oxidation, thermal, photo‐light, and humidity. The validation study was performed for the limit of detection and limit of quantification, linearity, and recoveries were 0.03%, 0.05%, and 0.1132–2.9920 μg/mL (R2 > 0.999), and 84.09%–109.74%, respectively. The proposed method is highly suitable for the determination of assay, organic impurities, and degradation products of the hydroxyzine. The chemical structure of degradation product 1 (hydroxyzine N‐Oxide) and degradation product 2 (O‐Acetyl hydroxyzine) were identified with the supporting data of LC‐mass spectrometry (LC‐MS) and high‐resolution MS.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2024-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139780059","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}
N. V. V. D. P. Boppy, S. Haridasyam, Niroja Vadagam, M. Venkatanarayana, Sanjeeva R. Chinnakadoori, Dr. Narasimha S. Lakka, PhD
A simple and robust high‐performance liquid chromatography (HPLC) method was developed for organic impurities of hydroxyzine hydrochloride in pharmaceuticals. The developed method was designed to estimate all organic impurities of hydroxyzine. The HPLC separation was achieved using C18 column (150 × 3.9 mm, 5 μm) along with a binary gradient consisting of mobile phases A (0.1%, trifluoroacetic acid in purified water) and B (0.05%, trifluoroacetic acid in acetonitrile), a flow rate of 0.7–mL/min, a column temperature of 30°C and a sample temperature of 25°C. The detection wavelength used was 230 nm for the estimation of impurity‐A, impurity‐B, and all unspecified impurities and degradation products, whereas impurity‐C was quantitated using 254 nm. The stability‐indicating property of the developed HPLC technique was assessed using stress testing conditions of hydrolysis, oxidation, thermal, photo‐light, and humidity. The validation study was performed for the limit of detection and limit of quantification, linearity, and recoveries were 0.03%, 0.05%, and 0.1132–2.9920 μg/mL (R2 > 0.999), and 84.09%–109.74%, respectively. The proposed method is highly suitable for the determination of assay, organic impurities, and degradation products of the hydroxyzine. The chemical structure of degradation product 1 (hydroxyzine N‐Oxide) and degradation product 2 (O‐Acetyl hydroxyzine) were identified with the supporting data of LC‐mass spectrometry (LC‐MS) and high‐resolution MS.
{"title":"Separation and quantification of organic‐related impurities of anti‐histamine drug hydroxyzine in pharmaceutical dosage forms using stability‐indicating high‐performance liquid chromatography, liquid chromatography‐mass spectrometry, and high‐resolution mass spectrometry techniques","authors":"N. V. V. D. P. Boppy, S. Haridasyam, Niroja Vadagam, M. Venkatanarayana, Sanjeeva R. Chinnakadoori, Dr. Narasimha S. Lakka, PhD","doi":"10.1002/sscp.202300157","DOIUrl":"https://doi.org/10.1002/sscp.202300157","url":null,"abstract":"A simple and robust high‐performance liquid chromatography (HPLC) method was developed for organic impurities of hydroxyzine hydrochloride in pharmaceuticals. The developed method was designed to estimate all organic impurities of hydroxyzine. The HPLC separation was achieved using C18 column (150 × 3.9 mm, 5 μm) along with a binary gradient consisting of mobile phases A (0.1%, trifluoroacetic acid in purified water) and B (0.05%, trifluoroacetic acid in acetonitrile), a flow rate of 0.7–mL/min, a column temperature of 30°C and a sample temperature of 25°C. The detection wavelength used was 230 nm for the estimation of impurity‐A, impurity‐B, and all unspecified impurities and degradation products, whereas impurity‐C was quantitated using 254 nm. The stability‐indicating property of the developed HPLC technique was assessed using stress testing conditions of hydrolysis, oxidation, thermal, photo‐light, and humidity. The validation study was performed for the limit of detection and limit of quantification, linearity, and recoveries were 0.03%, 0.05%, and 0.1132–2.9920 μg/mL (R2 > 0.999), and 84.09%–109.74%, respectively. The proposed method is highly suitable for the determination of assay, organic impurities, and degradation products of the hydroxyzine. The chemical structure of degradation product 1 (hydroxyzine N‐Oxide) and degradation product 2 (O‐Acetyl hydroxyzine) were identified with the supporting data of LC‐mass spectrometry (LC‐MS) and high‐resolution MS.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2024-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139840116","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}
A sensitive ultra‐high‐performance liquid chromatography (UHPLC) method was developed for the quantification of trace levels of nilotinib and its related substances (RSs) in the nilotinib capsule dosage form. Critical method parameters were optimized using the design of experiments, employing the ACQUITY UHPLC BEH Phenyl column (2.1 × 100 mm, 1.7 μm) at a constant flow rate of 0.6 mL/min. The isocratic mobile phase, consisting of aqueous and organic phases, achieved efficient chromatographic separation within 8 min at 261 nm. The mean retention time for nilotinib was 6.1339, and for RSs, it ranged from 2.10 to 6.91 min, with a resolution exceeding 2 for all peaks. The method demonstrated robustness, separating known impurities and degradation products. The linearity was assessed over a concentration range of 0.02–80 ppm for nilotinib and 0.015–0.12 ppm for impurities. The limit of detection and limit of quantitation (LOQ) for nilotinib were 0.01 and 0.02 μg/mL, respectively, and 0.01 and 0.015 μg/mL for individual impurities. Recovery studies at LOQ, 100%, and 150% of the specification limit yielded percent recoveries ranging from 92.27% to 102.45%. Precision results showed low relative standard deviations, below 2% for nilotinib and below 8% for impurities. This method is deemed suitable for pharmaceutical formulation quantification.
{"title":"Systematic optimization of reverse phase ultra‐high‐performance liquid chromatography method for quantification of nilotinib and its related substances in bulk drug and pharmaceutical formulation using quality by design approach","authors":"Shishir Kumar Prasad, Divekar Kalpana","doi":"10.1002/sscp.202300185","DOIUrl":"https://doi.org/10.1002/sscp.202300185","url":null,"abstract":"A sensitive ultra‐high‐performance liquid chromatography (UHPLC) method was developed for the quantification of trace levels of nilotinib and its related substances (RSs) in the nilotinib capsule dosage form. Critical method parameters were optimized using the design of experiments, employing the ACQUITY UHPLC BEH Phenyl column (2.1 × 100 mm, 1.7 μm) at a constant flow rate of 0.6 mL/min. The isocratic mobile phase, consisting of aqueous and organic phases, achieved efficient chromatographic separation within 8 min at 261 nm. The mean retention time for nilotinib was 6.1339, and for RSs, it ranged from 2.10 to 6.91 min, with a resolution exceeding 2 for all peaks. The method demonstrated robustness, separating known impurities and degradation products. The linearity was assessed over a concentration range of 0.02–80 ppm for nilotinib and 0.015–0.12 ppm for impurities. The limit of detection and limit of quantitation (LOQ) for nilotinib were 0.01 and 0.02 μg/mL, respectively, and 0.01 and 0.015 μg/mL for individual impurities. Recovery studies at LOQ, 100%, and 150% of the specification limit yielded percent recoveries ranging from 92.27% to 102.45%. Precision results showed low relative standard deviations, below 2% for nilotinib and below 8% for impurities. This method is deemed suitable for pharmaceutical formulation quantification.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2024-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139856360","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}
A sensitive ultra‐high‐performance liquid chromatography (UHPLC) method was developed for the quantification of trace levels of nilotinib and its related substances (RSs) in the nilotinib capsule dosage form. Critical method parameters were optimized using the design of experiments, employing the ACQUITY UHPLC BEH Phenyl column (2.1 × 100 mm, 1.7 μm) at a constant flow rate of 0.6 mL/min. The isocratic mobile phase, consisting of aqueous and organic phases, achieved efficient chromatographic separation within 8 min at 261 nm. The mean retention time for nilotinib was 6.1339, and for RSs, it ranged from 2.10 to 6.91 min, with a resolution exceeding 2 for all peaks. The method demonstrated robustness, separating known impurities and degradation products. The linearity was assessed over a concentration range of 0.02–80 ppm for nilotinib and 0.015–0.12 ppm for impurities. The limit of detection and limit of quantitation (LOQ) for nilotinib were 0.01 and 0.02 μg/mL, respectively, and 0.01 and 0.015 μg/mL for individual impurities. Recovery studies at LOQ, 100%, and 150% of the specification limit yielded percent recoveries ranging from 92.27% to 102.45%. Precision results showed low relative standard deviations, below 2% for nilotinib and below 8% for impurities. This method is deemed suitable for pharmaceutical formulation quantification.
{"title":"Systematic optimization of reverse phase ultra‐high‐performance liquid chromatography method for quantification of nilotinib and its related substances in bulk drug and pharmaceutical formulation using quality by design approach","authors":"Shishir Kumar Prasad, Divekar Kalpana","doi":"10.1002/sscp.202300185","DOIUrl":"https://doi.org/10.1002/sscp.202300185","url":null,"abstract":"A sensitive ultra‐high‐performance liquid chromatography (UHPLC) method was developed for the quantification of trace levels of nilotinib and its related substances (RSs) in the nilotinib capsule dosage form. Critical method parameters were optimized using the design of experiments, employing the ACQUITY UHPLC BEH Phenyl column (2.1 × 100 mm, 1.7 μm) at a constant flow rate of 0.6 mL/min. The isocratic mobile phase, consisting of aqueous and organic phases, achieved efficient chromatographic separation within 8 min at 261 nm. The mean retention time for nilotinib was 6.1339, and for RSs, it ranged from 2.10 to 6.91 min, with a resolution exceeding 2 for all peaks. The method demonstrated robustness, separating known impurities and degradation products. The linearity was assessed over a concentration range of 0.02–80 ppm for nilotinib and 0.015–0.12 ppm for impurities. The limit of detection and limit of quantitation (LOQ) for nilotinib were 0.01 and 0.02 μg/mL, respectively, and 0.01 and 0.015 μg/mL for individual impurities. Recovery studies at LOQ, 100%, and 150% of the specification limit yielded percent recoveries ranging from 92.27% to 102.45%. Precision results showed low relative standard deviations, below 2% for nilotinib and below 8% for impurities. This method is deemed suitable for pharmaceutical formulation quantification.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2024-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139796377","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}
S. Katna, Arvind Kumar, T. Banshtu, N. Devi, Shubhra Singh, Hema Prasad
The study was carried out to determine the dissipation kinetics, persistence, and dietary risk assessment of tetraniliprole. Three applications of tetraniliprole were given at 50.00 g a.i./hac (X) and 62.50 g a.i/hac (1.25X) on chilli and brinjal at the fruiting stage. The recoveries in all the matrices were within the acceptable range of 70%–120%. The initial residues of tetraniliprole on chilli fruits were 0.379 and 0.593 μg/g and on brinjal fruits, the residues were 0.559 and 0.916 μg/g at 50.00 and 62.50 g a.i./ha, respectively. In red chilli, chilli field soil, and brinjal field soil the residues were below the limit of quantitation. The tetraniliprole follows first‐order dissipation kinetics with the half‐life (RL50) 1.58 and 1.99 days in chilli and 1.42 and 1.79 days in brinjal at X and 1.25X doses. In chilli, 9.89 and 12.11 days whereas, in brinjal, 10.75 and 13.25 days were recommended for harvesting the crops after the last application of tetraniliprole at X and 1.25 X doses, respectively. The hazard quotient values in chilli and brinjal for both males and females were below 1 indicating that the use of tetraniliprole does not cause any health risk to the consumers.
{"title":"Dissipation kinetics, persistence, and dietary risk assessment of tetraniliprole in two solanaceous vegetables, chilli and brinjal","authors":"S. Katna, Arvind Kumar, T. Banshtu, N. Devi, Shubhra Singh, Hema Prasad","doi":"10.1002/sscp.202300121","DOIUrl":"https://doi.org/10.1002/sscp.202300121","url":null,"abstract":"The study was carried out to determine the dissipation kinetics, persistence, and dietary risk assessment of tetraniliprole. Three applications of tetraniliprole were given at 50.00 g a.i./hac (X) and 62.50 g a.i/hac (1.25X) on chilli and brinjal at the fruiting stage. The recoveries in all the matrices were within the acceptable range of 70%–120%. The initial residues of tetraniliprole on chilli fruits were 0.379 and 0.593 μg/g and on brinjal fruits, the residues were 0.559 and 0.916 μg/g at 50.00 and 62.50 g a.i./ha, respectively. In red chilli, chilli field soil, and brinjal field soil the residues were below the limit of quantitation. The tetraniliprole follows first‐order dissipation kinetics with the half‐life (RL50) 1.58 and 1.99 days in chilli and 1.42 and 1.79 days in brinjal at X and 1.25X doses. In chilli, 9.89 and 12.11 days whereas, in brinjal, 10.75 and 13.25 days were recommended for harvesting the crops after the last application of tetraniliprole at X and 1.25 X doses, respectively. The hazard quotient values in chilli and brinjal for both males and females were below 1 indicating that the use of tetraniliprole does not cause any health risk to the consumers.","PeriodicalId":21639,"journal":{"name":"SEPARATION SCIENCE PLUS","volume":null,"pages":null},"PeriodicalIF":1.1,"publicationDate":"2024-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139870353","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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