Pub Date : 2024-06-12DOI: 10.5584/jiomics.v14i2.232
I. Rubashvili, Mzia Tsitsagi, M. Chkhaidze, K. Ebralidze
Background: Apple pomace has garnered significant attention within the life sciences domain due to its underutilized status as a waste material from apple processing. It represents a cost-effective and abundant source of triterpene acids due to its multifunctional clinical, nutritional, and pharmaceutical benefits. Purpose: The present study aimed to develop and validate a new, selective, effective, robust and reproducible laboratory methodology based on extraction, purification and analytical procedures to obtain and determining three major triterpene acids – Ursolic acid (UA), Oleanolic acid (OA) and Betulinic acid (BA) into the dry extracted product from apple pomace. Method: A new, cost-efficient, rapid, selective and high-yield two-stage ultrasound-assisted extraction procedure was developed and the effect of critical parameters: ultrasonic power, extraction time, solvent volume, temperature, and the amount of raw material on the extraction process were investigated. The dry column vacuum chromatography technique was used for purification to remove unwanted non-polar and polar impurities from the target bioactive compounds; A new, effective, specific, sensitive, and rapid HPLC analytical procedure was developed using analytical quality by design (AQbD) approach and validated according to ICH guidelines. Conclusion: The method has a good accuracy (the mean recovery >95 %) and linearity (R2>0.999). The limit of quantitation (LOQ) is 0.0001 mg/mL for UA, 0.00005 mg/mL for OA and 0.000025 mg/mL for BA. The validation results confirm that the method is specific, precise and robust. The purity of the extracted and purified target product from apple pomace is not less than 93 %. The developed laboratory methodology is capable of being considered for industrial purposes and through the appropriate technology transfer process can be successfully transferred to the industrial scale.
背景：苹果渣作为苹果加工过程中产生的一种废料，一直未得到充分利用，因此在生命科学领域备受关注。由于三萜酸具有临床、营养和制药等多重功效，因此它是一种具有成本效益且丰富的三萜酸来源。目的：本研究旨在开发和验证一种新的、选择性强、有效、稳健且可重复的实验室方法，该方法基于提取、纯化和分析程序，用于获取和测定苹果渣干提取物中的三种主要三萜酸--熊果酸（UA）、齐墩果酸（OA）和白桦脂酸（BA）。方法：研究了超声波功率、萃取时间、溶剂用量、温度和原料用量等关键参数对萃取过程的影响。采用干柱真空层析技术进行纯化，以去除目标生物活性化合物中不需要的非极性和极性杂质；采用分析质量设计（AQbD）方法开发了一种新型、有效、特异、灵敏和快速的高效液相色谱分析程序，并根据 ICH 指南进行了验证。结果表明该方法具有良好的准确性（平均回收率>95%）和线性度（R2>0.999）。 UA 的定量限（LOQ）为 0.0001 mg/mL，OA 为 0.00005 mg/mL，BA 为 0.000025 mg/mL。 验证结果表明该方法具有特异性、准确性和稳健性。从苹果渣中提取和纯化的目标产物纯度不低于 93%。所开发的实验室方法可以考虑用于工业目的，通过适当的技术转让过程，可以成功地转移到工业规模。
{"title":"Ultrasound-Assisted Extraction- and Liquid Chromatography-Based Method Development and Validation for Obtaining and Qualitative Determination of Apple Pomace Three Triterpene Acids using Analytical Quality by Design","authors":"I. Rubashvili, Mzia Tsitsagi, M. Chkhaidze, K. Ebralidze","doi":"10.5584/jiomics.v14i2.232","DOIUrl":"https://doi.org/10.5584/jiomics.v14i2.232","url":null,"abstract":"Background: Apple pomace has garnered significant attention within the life sciences domain due to its underutilized status as a waste material from apple processing. It represents a cost-effective and abundant source of triterpene acids due to its multifunctional clinical, nutritional, and pharmaceutical benefits. \u0000Purpose: The present study aimed to develop and validate a new, selective, effective, robust and reproducible laboratory methodology based on extraction, purification and analytical procedures to obtain and determining three major triterpene acids – Ursolic acid (UA), Oleanolic acid (OA) and Betulinic acid (BA) into the dry extracted product from apple pomace. \u0000Method: A new, cost-efficient, rapid, selective and high-yield two-stage ultrasound-assisted extraction procedure was developed and the effect of critical parameters: ultrasonic power, extraction time, solvent volume, temperature, and the amount of raw material on the extraction process were investigated. The dry column vacuum chromatography technique was used for purification to remove unwanted non-polar and polar impurities from the target bioactive compounds; A new, effective, specific, sensitive, and rapid HPLC analytical procedure was developed using analytical quality by design (AQbD) approach and validated according to ICH guidelines. \u0000Conclusion: The method has a good accuracy (the mean recovery >95 %) and linearity (R2>0.999). The limit of quantitation (LOQ) is 0.0001 mg/mL for UA, 0.00005 mg/mL for OA and 0.000025 mg/mL for BA. The validation results confirm that the method is specific, precise and robust. The purity of the extracted and purified target product from apple pomace is not less than 93 %. The developed laboratory methodology is capable of being considered for industrial purposes and through the appropriate technology transfer process can be successfully transferred to the industrial scale.","PeriodicalId":37675,"journal":{"name":"Journal of Integrated OMICS","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141350932","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}
Pub Date : 2024-04-15DOI: 10.5584/jiomics.v14i1.229
Inês F. Domingos, Luís B. Carvalho, J. Capelo, C. Lodeiro, Hugo M. Santos
Assessing total protein levels in biological samples is a common procedure in biochemistry and molecular biology. In this study, we compare tryptophan fluorescence (WF) with Bradford and BCA assays to determine total protein in serum samples. Our results indicate that tryptophan fluorescence spectrometry is an efficient, sensitive, and straightforward technique for quantifying proteins in serum. We observed minimal variation between the three methods: BCA de one with the lowers LOD and LOQ. The tryptophan method offers the possibility of reusing the intact sample that does not need colourimetric reagents for quantification. Consequently, free tryptophan serves as a reliable universal standard. This assay can be performed using a conventional fluorescence spectrometer with cuvettes or in a 96-well plate format with a plate reader. The method was successfully used as proof of concept, using serum from patients diagnosed with myeloma and serum from healthy donors.
{"title":"DTT protein equalization and Tryptophan protein quantification as a powerful tool in analytical proteomics.","authors":"Inês F. Domingos, Luís B. Carvalho, J. Capelo, C. Lodeiro, Hugo M. Santos","doi":"10.5584/jiomics.v14i1.229","DOIUrl":"https://doi.org/10.5584/jiomics.v14i1.229","url":null,"abstract":"Assessing total protein levels in biological samples is a common procedure in biochemistry and molecular biology. In this study, we compare tryptophan fluorescence (WF) with Bradford and BCA assays to determine total protein in serum samples. Our results indicate that tryptophan fluorescence spectrometry is an efficient, sensitive, and straightforward technique for quantifying proteins in serum. We observed minimal variation between the three methods: BCA de one with the lowers LOD and LOQ. The tryptophan method offers the possibility of reusing the intact sample that does not need colourimetric reagents for quantification. Consequently, free tryptophan serves as a reliable universal standard. This assay can be performed using a conventional fluorescence spectrometer with cuvettes or in a 96-well plate format with a plate reader. The method was successfully used as proof of concept, using serum from patients diagnosed with myeloma and serum from healthy donors.","PeriodicalId":37675,"journal":{"name":"Journal of Integrated OMICS","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140702926","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}
Pub Date : 2024-04-15DOI: 10.5584/jiomics.v14i1.228
Chahinez Triqui, Ilyes Zatla, Wafaa Lemerini, Nora Benmadani, Mohammed Charaf Eddine Houari, S. Gaouar, N. Soulimane
The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in December 2019 precipitated the onset of the COVID-19 pandemic, which swiftly spread across more than 214 countries and territories, posing a significant global health crisis. In response, laboratories worldwide have embarked on extensive efforts to characterize the genomic landscape of the virus, employing a myriad of sophisticated genomic analysis techniques. This study endeavors to undertake a comprehensive exploration into the genetic diversity, geographical distribution, and virulence determinants of SARS-CoV-2 clades across 11 diverse countries, employing advanced computational biology methodologies. Leveraging molecular data sourced from prominent international databases, the analysis aims to unravel the intricate phylogenetic relationships and mutational dynamics exhibited by various viral strains circulating worldwide. The findings of this investigation promise to yield invaluable insights into the evolutionary trajectory of SARS-CoV-2, shedding light on potential therapeutic targets and informing strategies for mitigating the impact of the ongoing pandemic on global public health. Results highlight significant genetic diversity among SARS-CoV-2 strains across different countries, with phylogenetic analysis revealing distinct subclass groupings within each country. A manual comparison of sequences identified numerous mutations, with certain mutations associated with increased virulence. Comparison of clade G and clade O sequences revealed differences in mutation profiles, suggesting potential links to virulence and transmissibility. These findings underscore the dynamic nature of SARS-CoV-2 evolution and the importance of monitoring genetic changes for public health interventions.
{"title":"Navigating the molecular diversity of SARS-CoV-2: early pandemic insights from comparative phylogenetic analysis","authors":"Chahinez Triqui, Ilyes Zatla, Wafaa Lemerini, Nora Benmadani, Mohammed Charaf Eddine Houari, S. Gaouar, N. Soulimane","doi":"10.5584/jiomics.v14i1.228","DOIUrl":"https://doi.org/10.5584/jiomics.v14i1.228","url":null,"abstract":"The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in December 2019 precipitated the onset of the COVID-19 pandemic, which swiftly spread across more than 214 countries and territories, posing a significant global health crisis. In response, laboratories worldwide have embarked on extensive efforts to characterize the genomic landscape of the virus, employing a myriad of sophisticated genomic analysis techniques. This study endeavors to undertake a comprehensive exploration into the genetic diversity, geographical distribution, and virulence determinants of SARS-CoV-2 clades across 11 diverse countries, employing advanced computational biology methodologies. Leveraging molecular data sourced from prominent international databases, the analysis aims to unravel the intricate phylogenetic relationships and mutational dynamics exhibited by various viral strains circulating worldwide. The findings of this investigation promise to yield invaluable insights into the evolutionary trajectory of SARS-CoV-2, shedding light on potential therapeutic targets and informing strategies for mitigating the impact of the ongoing pandemic on global public health. Results highlight significant genetic diversity among SARS-CoV-2 strains across different countries, with phylogenetic analysis revealing distinct subclass groupings within each country. A manual comparison of sequences identified numerous mutations, with certain mutations associated with increased virulence. Comparison of clade G and clade O sequences revealed differences in mutation profiles, suggesting potential links to virulence and transmissibility. These findings underscore the dynamic nature of SARS-CoV-2 evolution and the importance of monitoring genetic changes for public health interventions.","PeriodicalId":37675,"journal":{"name":"Journal of Integrated OMICS","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140702903","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}
Pub Date : 2023-12-30DOI: 10.5584/jiomics.v13i2.224
Gilberto Igrejas, J. Capelo, Pedro Campos, Hugo Santos, C. Lodeiro, Luis Carvalho, E. Oliveira, Patricia Alexandra Curado Quintas Diniz Poeta
Letter to the editor
致编辑的信
{"title":"Antibiotic resistance and the one health approach. The importance of teaching OMICS at Universities.","authors":"Gilberto Igrejas, J. Capelo, Pedro Campos, Hugo Santos, C. Lodeiro, Luis Carvalho, E. Oliveira, Patricia Alexandra Curado Quintas Diniz Poeta","doi":"10.5584/jiomics.v13i2.224","DOIUrl":"https://doi.org/10.5584/jiomics.v13i2.224","url":null,"abstract":"Letter to the editor","PeriodicalId":37675,"journal":{"name":"Journal of Integrated OMICS","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139140678","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}
Pub Date : 2023-07-31DOI: 10.5584/jiomics.v13i1.221
Laura Mercolini, Michele Protti, Luis B. Carvalho, Pedro A.D. Teigas-Campos, Sarah Palano, Elisa Milandri, Claudia Volpi, Carlos Lodeiro, Hugo M. Santos, José L. Capelo, Roberto Mandrioli
No abstract needed for Letters to the Editor.
《致编辑信》不需要摘要。
{"title":"INSIGHTS INTO THE USE OF MICROSAMPLING FOR OMICS STUDIE","authors":"Laura Mercolini, Michele Protti, Luis B. Carvalho, Pedro A.D. Teigas-Campos, Sarah Palano, Elisa Milandri, Claudia Volpi, Carlos Lodeiro, Hugo M. Santos, José L. Capelo, Roberto Mandrioli","doi":"10.5584/jiomics.v13i1.221","DOIUrl":"https://doi.org/10.5584/jiomics.v13i1.221","url":null,"abstract":"No abstract needed for Letters to the Editor.","PeriodicalId":37675,"journal":{"name":"Journal of Integrated OMICS","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135358103","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}
Pub Date : 2022-12-31DOI: 10.5584/jiomics.v12i2.213
Thomas Webster
This is a letter to the editor so there is no astract.
这是写给编辑的信，所以没有摘要。
{"title":"Computational Nanomedicine: Helping Us Get Through COVID-19","authors":"Thomas Webster","doi":"10.5584/jiomics.v12i2.213","DOIUrl":"https://doi.org/10.5584/jiomics.v12i2.213","url":null,"abstract":"This is a letter to the editor so there is no astract.","PeriodicalId":37675,"journal":{"name":"Journal of Integrated OMICS","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44486611","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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