Pub Date : 2024-11-26DOI: 10.1016/j.jpba.2024.116584
Jinxuan Chai , Yan Wang , Sifan Guo , Zhibo Wang , Hongwei Chen , Xian Wang , Dandan Xie , Ying Cai , Shiwei Wang , Zhencai Hu , Aihua Zhang , Shi Qiu
Diabetic nephropathy (DKD) is a diabetesrelated kidney injury with an increasing incidence every year. Metformin hydrochloride (MET), a cornerstone treatment for glucose lowering, has been widely reported for the treatment of DKD, but the specific molecular mechanisms and potential therapeutic targets still need to be further explored. We used kidney tissues from db/db mice as samples and used proteomics and bioinformatics to analyse the function, distribution and related pathways of differential proteins in DKD, focusing on the assessment of the binding energies of key proteins in the butyrate pathway and drugs at the molecular level, which showed that the expression profiles of differential proteins in kidney tissues were altered after MET treatment, involving energy metabolism. The key proteins involved in the butanoate metabolism pathway, including AACS, ACSM3, EHHADH and HMGCS2, exhibit binding energies to MET of ＜-5 kcal. It is therefore plausible that MET treatment may affect the butanoate metabolism pathway, potentially ameliorating the progression of DKD by modulating mitochondrial function and inflammatory responses.
{"title":"Proteomics exploration of metformin hydrochloride for diabetic kidney disease treatment via the butanoate metabolism pathway","authors":"Jinxuan Chai , Yan Wang , Sifan Guo , Zhibo Wang , Hongwei Chen , Xian Wang , Dandan Xie , Ying Cai , Shiwei Wang , Zhencai Hu , Aihua Zhang , Shi Qiu","doi":"10.1016/j.jpba.2024.116584","DOIUrl":"10.1016/j.jpba.2024.116584","url":null,"abstract":"<div><div>Diabetic nephropathy (DKD) is a diabetesrelated kidney injury with an increasing incidence every year. Metformin hydrochloride (MET), a cornerstone treatment for glucose lowering, has been widely reported for the treatment of DKD, but the specific molecular mechanisms and potential therapeutic targets still need to be further explored. We used kidney tissues from db/db mice as samples and used proteomics and bioinformatics to analyse the function, distribution and related pathways of differential proteins in DKD, focusing on the assessment of the binding energies of key proteins in the butyrate pathway and drugs at the molecular level, which showed that the expression profiles of differential proteins in kidney tissues were altered after MET treatment, involving energy metabolism. The key proteins involved in the butanoate metabolism pathway, including AACS, ACSM3, EHHADH and HMGCS2, exhibit binding energies to MET of ＜-5 kcal. It is therefore plausible that MET treatment may affect the butanoate metabolism pathway, potentially ameliorating the progression of DKD by modulating mitochondrial function and inflammatory responses.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"254 ","pages":"Article 116584"},"PeriodicalIF":3.1,"publicationDate":"2024-11-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142746018","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-26DOI: 10.1016/j.jpba.2024.116585
Ling Lin , Shuo Wang , Kang Wang , Zhe Zhao , Gang Li
The signal-to-noise ratio of the spectrum is a critical determinant of detection accuracy in compositional analysis utilizing spectroscopy. The spectral data acquired by the spectrometer, while intended to capture essential sample characteristics, is often interspersed with various noise interferences. This contamination can severely disrupt the integrity of measurement outcomes. Therefore, this paper proposes the "multi-band spectral data fusion" method. In order to verify the feasibility of this method, this paper takes blood detection based on dynamic spectroscopy as an example and develops two models for each of the various components in blood. The experimental results show that when compared to modeling the raw spectrum data of the samples directly, the prediction accuracy of the model constructed using the new spectra processed by the multi-band spectral data fusion method suggested in this paper is greater. The correlation coefficient of the hemoglobin prediction set has improved by 13.48 %, and the root mean square error has decreased by 21.00 %. The correlation coefficient of the blood glucose prediction set improved by 4.07 %, and the root mean square error decreased by 12.78 %. The result demonstrates that the proposed method effectively mitigates the impact of random errors without compromising the spectral information content. The approach is not limited to blood component analysis but has potential applications across diverse spectroscopic domains, providing new ideas and methods for improving the accuracy of quantitative spectroscopic analysis.
{"title":"A multi-band spectral data fusion method for improving the accuracy of quantitative spectral analysis","authors":"Ling Lin , Shuo Wang , Kang Wang , Zhe Zhao , Gang Li","doi":"10.1016/j.jpba.2024.116585","DOIUrl":"10.1016/j.jpba.2024.116585","url":null,"abstract":"<div><div>The signal-to-noise ratio of the spectrum is a critical determinant of detection accuracy in compositional analysis utilizing spectroscopy. The spectral data acquired by the spectrometer, while intended to capture essential sample characteristics, is often interspersed with various noise interferences. This contamination can severely disrupt the integrity of measurement outcomes. Therefore, this paper proposes the \"multi-band spectral data fusion\" method. In order to verify the feasibility of this method, this paper takes blood detection based on dynamic spectroscopy as an example and develops two models for each of the various components in blood. The experimental results show that when compared to modeling the raw spectrum data of the samples directly, the prediction accuracy of the model constructed using the new spectra processed by the multi-band spectral data fusion method suggested in this paper is greater. The correlation coefficient of the hemoglobin prediction set has improved by 13.48 %, and the root mean square error has decreased by 21.00 %. The correlation coefficient of the blood glucose prediction set improved by 4.07 %, and the root mean square error decreased by 12.78 %. The result demonstrates that the proposed method effectively mitigates the impact of random errors without compromising the spectral information content. The approach is not limited to blood component analysis but has potential applications across diverse spectroscopic domains, providing new ideas and methods for improving the accuracy of quantitative spectroscopic analysis.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"254 ","pages":"Article 116585"},"PeriodicalIF":3.1,"publicationDate":"2024-11-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142746020","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-25DOI: 10.1016/j.jpba.2024.116582
Ruoxuan Sun, Janey Ronxhi, Xuemei Yang, Mark G. Qian, Xiaobin Zhang
The emergence of anti-drug antibodies (ADAs) poses significant impacts on the bioactivity and toxicity of biotherapeutics including proteins and peptides. Developing reliable assays to monitor the magnitudes of ADAs in blood samples is therefore considered a crucial task in animal and human studies throughout the development of biotherapeutics. Peptides represent a significant and fast-growing category of biotherapeutics for the management of a variety of indications. While peptides generally exhibit lower immunogenicity risks compared to biologics of larger sizes, drug developers are still required to conduct the risk-based immunogenicity assessment as mandated by the regulatory authorities. To address the need for efficient detection of ADAs against therapeutic peptides, we established a straightforward electrochemiluminescence immunoassay (ECLIA) based on direct binding strategy. Our assay demonstrates its applicability across various peptide therapeutics including marketed drugs and internal investigational compounds. Through stepwise tuning of the assay procedure, we identified several key factors such as buffer, detection reagent, plate type, and conjugation strategy that collectively contribute to the assay performance. Depending on the drug molecule and positive control antibody, the assay can achieve low single-digit to two-digit ng/ml sensitivity and ideal drug tolerance. In conclusion, this ECLIA platform presents a valuable and generic tool to expedite the development and validation of ADA assays for peptide-based therapeutics.
{"title":"Feasibility of a direct binding electrochemiluminescence assay to detect anti-drug antibodies against therapeutic peptides","authors":"Ruoxuan Sun, Janey Ronxhi, Xuemei Yang, Mark G. Qian, Xiaobin Zhang","doi":"10.1016/j.jpba.2024.116582","DOIUrl":"10.1016/j.jpba.2024.116582","url":null,"abstract":"<div><div>The emergence of anti-drug antibodies (ADAs) poses significant impacts on the bioactivity and toxicity of biotherapeutics including proteins and peptides. Developing reliable assays to monitor the magnitudes of ADAs in blood samples is therefore considered a crucial task in animal and human studies throughout the development of biotherapeutics. Peptides represent a significant and fast-growing category of biotherapeutics for the management of a variety of indications. While peptides generally exhibit lower immunogenicity risks compared to biologics of larger sizes, drug developers are still required to conduct the risk-based immunogenicity assessment as mandated by the regulatory authorities. To address the need for efficient detection of ADAs against therapeutic peptides, we established a straightforward electrochemiluminescence immunoassay (ECLIA) based on direct binding strategy. Our assay demonstrates its applicability across various peptide therapeutics including marketed drugs and internal investigational compounds. Through stepwise tuning of the assay procedure, we identified several key factors such as buffer, detection reagent, plate type, and conjugation strategy that collectively contribute to the assay performance. Depending on the drug molecule and positive control antibody, the assay can achieve low single-digit to two-digit ng/ml sensitivity and ideal drug tolerance. In conclusion, this ECLIA platform presents a valuable and generic tool to expedite the development and validation of ADA assays for peptide-based therapeutics.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"254 ","pages":"Article 116582"},"PeriodicalIF":3.1,"publicationDate":"2024-11-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142746019","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-22DOI: 10.1016/j.jpba.2024.116576
Isabella Sant’Anna, Rafaella Silva Arêdes, Walter Claudino P. de Souza, Renato Corrêa da Silva Lessa, Marcela Cristina de Moraes
Purine nucleoside phosphorylase (PNP) from Mycobacterium tuberculosis (MtPNP) plays a crucial role in purine metabolism, making it an attractive target for developing new tuberculosis treatments. In this study, we developed a ligand screening platform using MtPNP covalently immobilized on magnetic particles (MtPNP-MPs). The immobilization process achieved a high enzyme loading and preserved the enzyme catalytic activity, enabling its use in both activity and affinity-based screening assays. The activity of MtPNP-MPs was monitored by quantifying hypoxanthine released from inosine phosphorolysis, and kinetic studies revealed Michaelis-Menten behavior for inosine and inorganic phosphate substrates, with KM values comparable to those of free MtPNP. A proof-of-concept inhibitor study using the transition state analog DI4G demonstrated the platform capability for recognizing and characterizing inhibitors, yielding an IC50 value of 91.4 nM and a competitive inhibition mechanism with a Ki of 69.2 nM. Furthermore, the MtPNP-MPs exhibited high stability, retaining over 80 % of their activity after six months of storage and more than 90 % after five consecutive reaction cycles, highlighting their potential for reuse in high-throughput assays. We optimized key parameters for ligand fishing assay, including the amount of MtPNP-MPs, incubation time, and elution conditions. While higher organic solvent concentrations and longer elution times improved ligand isolation, these conditions also reduced enzyme activity. This trade-off between ligand isolation yield and enzyme reusability suggests that elution conditions should be tailored based on the ligand binding strength. Overall, this study establishes the MtPNP-MPs platform as a versatile tool for ligand identification and inhibitor characterization, with promising applications in the screening of complex libraries, such as natural products, for bioactive compounds.
{"title":"Development of an immobilized Mycobacterium tuberculosis purine nucleoside phosphorylase platform for ligand fishing and inhibition assays","authors":"Isabella Sant’Anna, Rafaella Silva Arêdes, Walter Claudino P. de Souza, Renato Corrêa da Silva Lessa, Marcela Cristina de Moraes","doi":"10.1016/j.jpba.2024.116576","DOIUrl":"10.1016/j.jpba.2024.116576","url":null,"abstract":"<div><div>Purine nucleoside phosphorylase (PNP) from <em>Mycobacterium tuberculosis</em> (<em>Mt</em>PNP) plays a crucial role in purine metabolism, making it an attractive target for developing new tuberculosis treatments. In this study, we developed a ligand screening platform using <em>Mt</em>PNP covalently immobilized on magnetic particles (<em>Mt</em>PNP-MPs). The immobilization process achieved a high enzyme loading and preserved the enzyme catalytic activity, enabling its use in both activity and affinity-based screening assays. The activity of <em>Mt</em>PNP-MPs was monitored by quantifying hypoxanthine released from inosine phosphorolysis, and kinetic studies revealed Michaelis-Menten behavior for inosine and inorganic phosphate substrates, with <em>K</em><sub>M</sub> values comparable to those of free <em>Mt</em>PNP. A proof-of-concept inhibitor study using the transition state analog DI4G demonstrated the platform capability for recognizing and characterizing inhibitors, yielding an IC<sub>50</sub> value of 91.4 nM and a competitive inhibition mechanism with a <em>K</em><sub>i</sub> of 69.2 nM. Furthermore, the <em>Mt</em>PNP-MPs exhibited high stability, retaining over 80 % of their activity after six months of storage and more than 90 % after five consecutive reaction cycles, highlighting their potential for reuse in high-throughput assays. We optimized key parameters for ligand fishing assay, including the amount of <em>Mt</em>PNP-MPs, incubation time, and elution conditions. While higher organic solvent concentrations and longer elution times improved ligand isolation, these conditions also reduced enzyme activity. This trade-off between ligand isolation yield and enzyme reusability suggests that elution conditions should be tailored based on the ligand binding strength. Overall, this study establishes the <em>Mt</em>PNP-MPs platform as a versatile tool for ligand identification and inhibitor characterization, with promising applications in the screening of complex libraries, such as natural products, for bioactive compounds.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"254 ","pages":"Article 116576"},"PeriodicalIF":3.1,"publicationDate":"2024-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142701985","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-22DOI: 10.1016/j.jpba.2024.116580
Yanan Guo , Shuang Cao , Bin Geng , Juanjuan Ma , Bo Yao
An online solid phase extraction–ultra performance liquid chromatography–tandem mass spectrometry detection method was developed for the simultaneous determination of sacubitril and seven sartan drugs in blood serum in this study. The compounds were separated through a C18 column. Mass spectrometry of the samples was performed using a jet stream electrospray ion source (AJS, ESI+). The samples were detected via a multiple reaction monitoring (MRM) mode and quantified via a stable isotope internal standard method. 900 μL of prepared sample was injected and a run time of 12.5 minutes was obtained in this proposed method. The eight examined target compounds showed good linearity (r2＞0.994) in the corresponding mass concentration range and a lower limit of quantitation (LLOQ) was in the range of 0.05–0.1 μg/L. The recovery of the spiked serum samples ranged from 90.90 % to 106.20 % with relative standard deviations (RSDs) of 4.57 %–9.27 %. Five target compounds were detected in the actual serum samples using this method. The proposed method is simple to use, sensitive, accurate, and suitable for the trace detection of sacubitril and seven sartan drugs present in serum samples. The method can meet the needs for clinical monitoring of blood concentrations of this type of drug, while providing detection technology support for the development of compound preparations of sacubitril and other sartan drugs.
{"title":"Determination of sacubitril and seven sartan drugs in serum samples by online solid phase extraction-liquid chromatography-tandem mass spectrometry","authors":"Yanan Guo , Shuang Cao , Bin Geng , Juanjuan Ma , Bo Yao","doi":"10.1016/j.jpba.2024.116580","DOIUrl":"10.1016/j.jpba.2024.116580","url":null,"abstract":"<div><div>An online solid phase extraction–ultra performance liquid chromatography–tandem mass spectrometry detection method was developed for the simultaneous determination of sacubitril and seven sartan drugs in blood serum in this study. The compounds were separated through a C<sub>18</sub> column. Mass spectrometry of the samples was performed using a jet stream electrospray ion source (AJS, ESI+). The samples were detected via a multiple reaction monitoring (MRM) mode and quantified via a stable isotope internal standard method. 900 μL of prepared sample was injected and a run time of 12.5 minutes was obtained in this proposed method. The eight examined target compounds showed good linearity (<em>r</em><sup>2</sup>＞0.994) in the corresponding mass concentration range and a lower limit of quantitation (LLOQ) was in the range of 0.05–0.1 μg/L. The recovery of the spiked serum samples ranged from 90.90 % to 106.20 % with relative standard deviations (RSDs) of 4.57 %–9.27 %. Five target compounds were detected in the actual serum samples using this method. The proposed method is simple to use, sensitive, accurate, and suitable for the trace detection of sacubitril and seven sartan drugs present in serum samples. The method can meet the needs for clinical monitoring of blood concentrations of this type of drug, while providing detection technology support for the development of compound preparations of sacubitril and other sartan drugs.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"254 ","pages":"Article 116580"},"PeriodicalIF":3.1,"publicationDate":"2024-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142701983","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-22DOI: 10.1016/j.jpba.2024.116577
Zehui Wei , Wenxin Liu , Jun Zhang , Xue Dong , Shuangxian Yan , Yu Cheng , Pingyuan Wei , Suhong Wang , Mei Tian
A novel β-amyloid protein capillary microextraction column was designed and prepared using epitope molecular imprinting technology for specific recognition of trace β-amyloid proteins in complex biological matrices. Using N-terminal nonapeptide of β-amyloid protein as template molecule, choline chloride-MAA and N-hydroxymethyl acrylamide as functional monomers, ethylene glycol dimethacrylate as crosslinker, the imprinted capillary monolithic column was prepared by thermal polymerization in the acetonitrile-water system. The optimal preparation parameters were obtained with the ratio of template: functional monomer: crosslinker at 1:6:16 (mmol/mmol/mmol). The physical property evaluation through Scanning electron microscopy, Fourier transform infrared spectroscopy analysis, zeta potential analysis, particle size analysis, and Brunauer-emmett-teller showed that a porous imprinted monolithic column with a high specific surface area was successfully prepared. The static adsorption experiment showed that the theoretical maximum adsorption capacity was 0.060 μg/column and the optimal imprinting factor was 2.27. Under the optimal extraction conditions, the imprinted column exhibited good template selectivity and excellent robustness. Finally, the extraction efficiency of the capillary imprinted column in actual plasma was evaluated using ELISA method. For Aβ 40 and Aβ 42, the detection limits were 1.00 pg/mL and 0.67 pg/mL, the quantification limits were 3.00 pg/mL and 2.00 pg/mL, the detection ranges were 7.5–120.0 pg/mL and 5.0–80.0 pg/mL, respectively. After extraction of plasma samples, the measured concentrations of Aβ 40 and Aβ 42 by imprinted column were 157.31 pg/mL and 48.22 pg/mL, respectively, which were significantly higher than the measured concentrations by non-imprinted column of 89.60 pg/mL and 41.02 pg/mL. In summary, the epitope imprinted capillary monolithic column prepared in this study can effectively enrich and separate trace amounts of amyloid proteins in complex samples, and is expected to provide a new sample pretreatment method for clinical detection of amyloid proteins.
{"title":"Preparation of amyloid N-terminal nonapeptide imprinted monolithic column and evaluation of adsorption properties","authors":"Zehui Wei , Wenxin Liu , Jun Zhang , Xue Dong , Shuangxian Yan , Yu Cheng , Pingyuan Wei , Suhong Wang , Mei Tian","doi":"10.1016/j.jpba.2024.116577","DOIUrl":"10.1016/j.jpba.2024.116577","url":null,"abstract":"<div><div>A novel β-amyloid protein capillary microextraction column was designed and prepared using epitope molecular imprinting technology for specific recognition of trace β-amyloid proteins in complex biological matrices. Using N-terminal nonapeptide of β-amyloid protein as template molecule, choline chloride-MAA and N-hydroxymethyl acrylamide as functional monomers, ethylene glycol dimethacrylate as crosslinker, the imprinted capillary monolithic column was prepared by thermal polymerization in the acetonitrile-water system. The optimal preparation parameters were obtained with the ratio of template: functional monomer: crosslinker at 1:6:16 (mmol/mmol/mmol). The physical property evaluation through Scanning electron microscopy, Fourier transform infrared spectroscopy analysis, zeta potential analysis, particle size analysis, and Brunauer-emmett-teller showed that a porous imprinted monolithic column with a high specific surface area was successfully prepared. The static adsorption experiment showed that the theoretical maximum adsorption capacity was 0.060 μg/column and the optimal imprinting factor was 2.27. Under the optimal extraction conditions, the imprinted column exhibited good template selectivity and excellent robustness. Finally, the extraction efficiency of the capillary imprinted column in actual plasma was evaluated using ELISA method. For Aβ 40 and Aβ 42, the detection limits were 1.00 pg/mL and 0.67 pg/mL, the quantification limits were 3.00 pg/mL and 2.00 pg/mL, the detection ranges were 7.5–120.0 pg/mL and 5.0–80.0 pg/mL, respectively. After extraction of plasma samples, the measured concentrations of Aβ 40 and Aβ 42 by imprinted column were 157.31 pg/mL and 48.22 pg/mL, respectively, which were significantly higher than the measured concentrations by non-imprinted column of 89.60 pg/mL and 41.02 pg/mL. In summary, the epitope imprinted capillary monolithic column prepared in this study can effectively enrich and separate trace amounts of amyloid proteins in complex samples, and is expected to provide a new sample pretreatment method for clinical detection of amyloid proteins.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"254 ","pages":"Article 116577"},"PeriodicalIF":3.1,"publicationDate":"2024-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142746116","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-22DOI: 10.1016/j.jpba.2024.116578
Elias Vanneste , Quinten Speleers , Anke Meyers , Karyna Krupianskaya , Annick Gillet , Bart Croonenborghs , Aaron DeMent , Erik Haghedooren , Ann Van Schepdael
Literature about sterilization of pharmaceutical substances is limited. The aim of this study was to evaluate the effect of nitrogen dioxide (NO2) sterilization, a new emerging technology, on five different ophthalmic active pharmaceutical ingredients, i.e., tetracycline hydrochloride, aciclovir, dexamethasone, methylprednisolone, and triamcinolone. The NO2 process concentrations tested were 5, 10, and 20 mg/L. The applied temperature was 21 °C and the relative humidity 30 %. The process cycle consisted of two pulses with a dwell time of 10 min each. Non-processed samples were used as a blank. The effect of the sterilization method was assessed by high performance liquid chromatography coupled to an ultraviolet/visible detector, used for the quantitative analysis of the degradation products and the relative content of the evaluated ophthalmic medicines. For tetracycline hydrochloride and aciclovir, an increase of impurities was observed by increasing the NO2 concentration. The maximum permissible NO2 concentrations were estimated to be 10 mg/L and 2.5 mg/L, respectively, considering the requirement for the impurities to be within the limit stated in the European Pharmacopoeia (Ph. Eur.). For both compounds, samples subjected to 20 mg/L NO2 demonstrated a significant difference in content compared to the non-processed sample. For methylprednisolone, dexamethasone, and triamcinolone, impurities complied with the limits of the Ph. Eur. for each NO2 concentration and relative contents were not significantly affected. Sterilization of tetracycline hydrochloride and aciclovir with NO2 is not recommended due to extensive degradation. NO2 sterilization of methylprednisolone, dexamethasone, and triamcinolone could find its application within the aseptic processing procedure of related pharmaceuticals.
{"title":"Nitrogen dioxide sterilization of a set of five ophthalmic active pharmaceutical ingredients: Impact on impurity profile and content","authors":"Elias Vanneste , Quinten Speleers , Anke Meyers , Karyna Krupianskaya , Annick Gillet , Bart Croonenborghs , Aaron DeMent , Erik Haghedooren , Ann Van Schepdael","doi":"10.1016/j.jpba.2024.116578","DOIUrl":"10.1016/j.jpba.2024.116578","url":null,"abstract":"<div><div>Literature about sterilization of pharmaceutical substances is limited. The aim of this study was to evaluate the effect of nitrogen dioxide (NO<sub>2</sub>) sterilization, a new emerging technology, on five different ophthalmic active pharmaceutical ingredients, i.e., tetracycline hydrochloride, aciclovir, dexamethasone, methylprednisolone, and triamcinolone. The NO<sub>2</sub> process concentrations tested were 5, 10, and 20 mg/L. The applied temperature was 21 °C and the relative humidity 30 %. The process cycle consisted of two pulses with a dwell time of 10 min each. Non-processed samples were used as a blank. The effect of the sterilization method was assessed by high performance liquid chromatography coupled to an ultraviolet/visible detector, used for the quantitative analysis of the degradation products and the relative content of the evaluated ophthalmic medicines. For tetracycline hydrochloride and aciclovir, an increase of impurities was observed by increasing the NO<sub>2</sub> concentration. The maximum permissible NO<sub>2</sub> concentrations were estimated to be 10 mg/L and 2.5 mg/L, respectively, considering the requirement for the impurities to be within the limit stated in the European Pharmacopoeia (Ph. Eur.). For both compounds, samples subjected to 20 mg/L NO<sub>2</sub> demonstrated a significant difference in content compared to the non-processed sample. For methylprednisolone, dexamethasone, and triamcinolone, impurities complied with the limits of the Ph. Eur. for each NO<sub>2</sub> concentration and relative contents were not significantly affected. Sterilization of tetracycline hydrochloride and aciclovir with NO<sub>2</sub> is not recommended due to extensive degradation. NO<sub>2</sub> sterilization of methylprednisolone, dexamethasone, and triamcinolone could find its application within the aseptic processing procedure of related pharmaceuticals.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"254 ","pages":"Article 116578"},"PeriodicalIF":3.1,"publicationDate":"2024-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142701982","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-19DOI: 10.1016/j.jpba.2024.116572
Yi Wang , Dongcao Xu , Xinxin Liu , Mengchun Cheng , Jingsong Huang , Dan Liu , Xiaozhe Zhang , Lihua Zhang
The prevalence of major depressive disorder (MDD) is higher in females than males, emphasizing the need to identify gender-specific biomarkers to improve diagnosis accuracy. In this study, a cross-sectional investigation with 258 samples was conducted to evaluate the discriminative power of potential gender-specific biomarkers for MDD. Eighteen MDD-related differential metabolites have been identified, involving pathways of phospholipids, glycerolipids, fatty acids, sphingolipids, cholesterol, vitamin E, and heme. A potential biomarker combination consisting of palmitelaidic acid, gamma carboxyethyl hydroxychroman (gamma-CEHC), and lysoPE(16:0) was confirmed for predicting depression in women using binary logistic regression analysis. To evaluate the panel's specificity, nine generalized anxiety disorder (GAD) samples, which share highly similar clinical symptoms with MDD, were included in the validation set. The discovery and validation sets yielded an area under the receiver operating characteristic curve of 0.86 and 0.83, respectively. All nine female GAD samples were correctly predicted as non-MDD, demonstrating the panel's specificity in diagnosing female MDD. Remarkably, this composite panel achieved a 75 % prediction accuracy in female samples in both the discovery and validation sets, but it did not reach 60 % prediction accuracy in male samples in either set. Our findings highlight the importance of gender-specific molecular diagnostics in developing practical and accurate diagnostic methods for MDD.
{"title":"Discovery of potential female-specific biomarkers for major depressive disorder by LC–MS-based metabolomics","authors":"Yi Wang , Dongcao Xu , Xinxin Liu , Mengchun Cheng , Jingsong Huang , Dan Liu , Xiaozhe Zhang , Lihua Zhang","doi":"10.1016/j.jpba.2024.116572","DOIUrl":"10.1016/j.jpba.2024.116572","url":null,"abstract":"<div><div>The prevalence of major depressive disorder (MDD) is higher in females than males, emphasizing the need to identify gender-specific biomarkers to improve diagnosis accuracy. In this study, a cross-sectional investigation with 258 samples was conducted to evaluate the discriminative power of potential gender-specific biomarkers for MDD. Eighteen MDD-related differential metabolites have been identified, involving pathways of phospholipids, glycerolipids, fatty acids, sphingolipids, cholesterol, vitamin E, and heme. A potential biomarker combination consisting of palmitelaidic acid, gamma carboxyethyl hydroxychroman (gamma-CEHC), and lysoPE(16:0) was confirmed for predicting depression in women using binary logistic regression analysis. To evaluate the panel's specificity, nine generalized anxiety disorder (GAD) samples, which share highly similar clinical symptoms with MDD, were included in the validation set. The discovery and validation sets yielded an area under the receiver operating characteristic curve of 0.86 and 0.83, respectively. All nine female GAD samples were correctly predicted as non-MDD, demonstrating the panel's specificity in diagnosing female MDD. Remarkably, this composite panel achieved a 75 % prediction accuracy in female samples in both the discovery and validation sets, but it did not reach 60 % prediction accuracy in male samples in either set. Our findings highlight the importance of gender-specific molecular diagnostics in developing practical and accurate diagnostic methods for MDD.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"254 ","pages":"Article 116572"},"PeriodicalIF":3.1,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142701984","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Senescence in renal cells has attracted wide attention as the critical factor promoting renal fibrosis and chronic kidney disease. Establishing a reliable cellular model is essential to study mechanisms underlying renal cell senescence. Herein, we compared various inducers to define the most suitable senescence inducer for HK-2 proximal tubular cells. These inducers included hydrogen peroxide (H2O2), high-temperature (HT), glucose, mannitol and hydroxyurea (HU). To screen for optimal concentration/level, the highest concentration/level of each inducer that did not increase cell death (to avoid severe toxicity) was selected for senescence induction and comparative analysis using the two most appropriate markers for HK-2 cell senescence as recently established. The data revealed that 0.4 mM, 43 °C, 80 mM, 80 mM and 100 μM were the optimal concentrations/levels of H2O2, HT, glucose, mannitol and HU, respectively. Comparative analysis using optimal concentration/level of each marker revealed that 0.4 mM H2O2, HT at 43 °C, 80 mM glucose and 80 mM mannitol were the weak senescence inducers. The most effective inducer for HK-2 senescence was 100 μM HU, which provided the greatest fold-changes of cell area and granularity when compared with other stimuli in a time-dependent manner. Based on these data comparing H2O2, HT, glucose, mannitol and HU at their optimal concentrations/levels, 100 μM HU seems to be most effective for senescence induction in HK-2 cells for in vitro study of proximal renal tubular cells.
肾脏细胞的衰老作为促进肾脏纤维化和慢性肾脏疾病的关键因素已引起广泛关注。建立可靠的细胞模型对研究肾细胞衰老的机制至关重要。在此，我们比较了各种诱导剂，以确定最适合 HK-2 近端肾小管细胞的衰老诱导剂。这些诱导剂包括过氧化氢（H2O2）、高温（HT）、葡萄糖、甘露醇和羟基脲（HU）。为了筛选最佳浓度/水平，我们选择了不增加细胞死亡（以避免严重毒性）的最高浓度/水平的诱导剂进行衰老诱导，并使用最近确定的两种最合适的 HK-2 细胞衰老标记进行比较分析。数据显示，0.4 mM、43 ℃、80 mM、80 mM 和 100 μM 分别是 H2O2、HT、葡萄糖、甘露醇和 HU 的最佳浓度/水平。利用各标记物的最佳浓度/水平进行的比较分析表明，0.4 mM H2O2、43 ℃下的HT、80 mM葡萄糖和80 mM甘露醇是弱衰老诱导剂。对 HK-2 衰老最有效的诱导剂是 100 μM HU，与其他刺激相比，它能以时间依赖的方式使细胞面积和颗粒度发生最大的倍数变化。基于这些数据，比较了 H2O2、HT、葡萄糖、甘露醇和 HU 的最佳浓度/水平，100 μM HU 似乎是体外研究近端肾小管细胞时诱导 HK-2 细胞衰老的最有效方法。
{"title":"Comparative analysis of various senescence inducers in proximal renal tubular cells","authors":"Piyaporn Rattananinsruang, Chadanat Noonin, Visith Thongboonkerd","doi":"10.1016/j.jpba.2024.116571","DOIUrl":"10.1016/j.jpba.2024.116571","url":null,"abstract":"<div><div>Senescence in renal cells has attracted wide attention as the critical factor promoting renal fibrosis and chronic kidney disease. Establishing a reliable cellular model is essential to study mechanisms underlying renal cell senescence. Herein, we compared various inducers to define the most suitable senescence inducer for HK-2 proximal tubular cells. These inducers included hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>), high-temperature (HT), glucose, mannitol and hydroxyurea (HU). To screen for optimal concentration/level, the highest concentration/level of each inducer that did not increase cell death (to avoid severe toxicity) was selected for senescence induction and comparative analysis using the two most appropriate markers for HK-2 cell senescence as recently established. The data revealed that 0.4 mM, 43 °C, 80 mM, 80 mM and 100 μM were the optimal concentrations/levels of H<sub>2</sub>O<sub>2</sub>, HT, glucose, mannitol and HU, respectively. Comparative analysis using optimal concentration/level of each marker revealed that 0.4 mM H<sub>2</sub>O<sub>2</sub>, HT at 43 °C, 80 mM glucose and 80 mM mannitol were the weak senescence inducers. The most effective inducer for HK-2 senescence was 100 μM HU, which provided the greatest fold-changes of cell area and granularity when compared with other stimuli in a time-dependent manner. Based on these data comparing H<sub>2</sub>O<sub>2</sub>, HT, glucose, mannitol and HU at their optimal concentrations/levels, 100 μM HU seems to be most effective for senescence induction in HK-2 cells for <em>in vitro</em> study of proximal renal tubular cells.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"254 ","pages":"Article 116571"},"PeriodicalIF":3.1,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142695529","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-16DOI: 10.1016/j.jpba.2024.116570
Pengwei Zhao , Zhixuan Song , Yunhan Li , Xiaorui Liu , Zhengjin Jiang , Qing Zhu , Jia-Huan Qu
Chlorogenic acid (CGA) is a key component in Aidi injection, known for its anti-cancer properties and ability to reduce toxicity. Therefore, accurate detection of CGA levels in Aidi injection is essential for monitoring therapeutic efficacy and minimizing adverse effects. This study presents a rapid and simple fluorescent method for detecting CGA in Aidi injection using aggregation-induced emission (AIE) nanoclusters, i.e. D(-)-penicillamine (DPA)-capped bimetallic gold/copper nanoclusters (DPA-Au/CuNCs). Upon the addition of CGA, the aggregation state of DPA-Au/CuNCs was disrupted through hydrogen bond formation and ligand exchange, leading to fluorescence quenching. The prepared DPA-Au/CuNCs exhibited a rapid response time of 0.5 min, and demonstrated good sensitivity for CGA, with a limit of detection of 3.75 μg/mL, and a linear detection range of 12.5–200 μg/mL. This method was successfully applied for the analysis of CGA in Aidi injection and plasma with good recovery rates and minimal matrix effect, highlighting its potential for the applications in pharmaceutical products and clinical samples.
{"title":"Rapid and simple fluorescent detection of chlorogenic acid in Aidi injection using aggregation-induced emission (AIE) nanoclusters","authors":"Pengwei Zhao , Zhixuan Song , Yunhan Li , Xiaorui Liu , Zhengjin Jiang , Qing Zhu , Jia-Huan Qu","doi":"10.1016/j.jpba.2024.116570","DOIUrl":"10.1016/j.jpba.2024.116570","url":null,"abstract":"<div><div>Chlorogenic acid (CGA) is a key component in Aidi injection, known for its anti-cancer properties and ability to reduce toxicity. Therefore, accurate detection of CGA levels in Aidi injection is essential for monitoring therapeutic efficacy and minimizing adverse effects. This study presents a rapid and simple fluorescent method for detecting CGA in Aidi injection using aggregation-induced emission (AIE) nanoclusters, i.e. D(-)-penicillamine (DPA)-capped bimetallic gold/copper nanoclusters (DPA-Au/CuNCs). Upon the addition of CGA, the aggregation state of DPA-Au/CuNCs was disrupted through hydrogen bond formation and ligand exchange, leading to fluorescence quenching. The prepared DPA-Au/CuNCs exhibited a rapid response time of 0.5 min, and demonstrated good sensitivity for CGA, with a limit of detection of 3.75 μg/mL, and a linear detection range of 12.5–200 μg/mL. This method was successfully applied for the analysis of CGA in Aidi injection and plasma with good recovery rates and minimal matrix effect, highlighting its potential for the applications in pharmaceutical products and clinical samples.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"254 ","pages":"Article 116570"},"PeriodicalIF":3.1,"publicationDate":"2024-11-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142682021","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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