Journal of pharmaceutical and biomedical analysis最新文献_第7页

Chemometric tools to comprehend a recovery process for the bioactive ingredients from purple basil (Ocimum basilicum L.): Box-Behnken design-based optimization and principal component analysis

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-17 DOI: 10.1016/j.jpba.2025.116676

İrem Toprakçı, Ebru Kurtulbaş, Selin Şahin

Medicinal and aromatic plants are alternative products to synthetics because of their antioxidant, antimicrobial, anti-inflammatory and antidiabetic effects. The objective of this study is to investigate the automated solvent extraction (ASE) process parameters for the extraction of bioactive-rich substances from purple basil (Ocimum basilicum L.). Process optimization in relation to total phenolic content (TPC), total flavonoid content (TFC), and total anthocyanin content (TAC) was performed through a chemometric approach. The ASE system was designed, modelled and optimized by 3-factor and 3-level Box-Behnken design of Response Surface method (RSM). Antioxidant activity of the samples were measured by 2 different free radical scavenging activity assays (ABTS and DPPH). By using principal component analysis (PCA) to the dataset, the impact of interactions between the parameters was also evaluated according to their antioxidant activity, TPC, TFC and TAC levels. The optimal ASE conditions (0.3 g of purple basil, 19 min of immersion time and 66 % ethanol solution) provided the highest yields of TPC (98.888 mg-GAE/g-DM), TFC (27.033 mg-CE/g-DM) and TAC (11.556 mg-C3G/g-DM), which were verified by satisfactory validation findings (the error<2 %).

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引用次数: 0

Peptide mapping analysis of synthetic semaglutide and liraglutide for generic development of drugs originating from recombinant DNA technology

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-17 DOI: 10.1016/j.jpba.2025.116682

Soo Hyun Kim , Sung Soo Kim , Hyun Jun Kim , Eun Ji Park , Dong Hee Na

Semaglutide and liraglutide are long-acting glucagon-like peptide-1 receptor agonists used to treat type-2 diabetes and obesity. Recent advances in peptide synthesis and analytical technologies have enabled the development of synthetic generic peptide for reference listed drugs (RLD) originating from recombinant DNA (rDNA) technology. Since the original semaglutide and liraglutide were produced through rDNA technology, there has been great interest in developing their synthetic peptides as generic versions of the original drugs. Therefore, this study aimed to develop a peptide mapping method to describe the primary structure of semaglutide and liraglutide using ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS), and to apply this method to demonstrate the sameness between synthetic peptides for generic drugs and rDNA peptides of the original drugs. Masses of the peptide fragments were identified using HRMS at an accurate level of mass error below 10 ppm, and their sequences were determined via MS^E sequencing using in-source fragmentation, which was also useful for identifying the fatty acid chain modification site. Full sequence coverage of each semaglutide and liraglutide was accomplished by combining peptide maps generated using Glu-C and chymotrypsin. The proposed peptide mapping method using UPLC-HRMS was useful for determining active ingredient sameness between generic synthetic peptides and previously approved peptide drug products of rDNA origin.

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引用次数: 0

Ganoderma lucidum spore oil attenuates acute liver injury by modulating lipid metabolism and gut microbiota

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-16 DOI: 10.1016/j.jpba.2025.116674

Jianying Liu , Yan Chen , Zhifeng Cen , Meiqi Hong , Binzhi Zhang , Xia Luo , Leqi Wang , Shasha Li , Xue Xiao , Qinqiang Long

The incidence of acute liver injury is increasing and poses a significant threat to human health. Ganoderma lucidum spore oil (GLSO), a lipid substance extracted from Ganoderma lucidum spore powder using supercritical CO₂ technology, has been investigated for its potential to prevent acute liver injury. However, the specific mechanism underlying the protective effects of GLSO remains incompletely understood. In this study, we investigated the preventive effect of GLSO on acute liver injury in rats, focusing on the gut microbiome and serum metabolomics. GLSO effectively alleviated liver dysfunction and reduced inflammation, leading to the prevention of acute liver injury in rats. Serum metabolomics analysis revealed that GLSO primarily modulated lipid metabolic pathways related to glycerophospholipid metabolism and sphingolipid metabolism. Specifically, GLSO decreased the levels of metabolites such as lysophosphatidylcholine (LPC), glycerophosphatidylcholine (GPC), and sphinganine 1-phosphate (SA1P), while increasing the levels of phosphatidylglycerol (PG) and digalactosylceramide (DGC). Gut microbiomics data indicated that GLSO effectively regulated the composition of the gut microbiota in rats with acute liver injury. Specifically, it increased the abundance of Firmicutes and decreased the abundance of Proteobacteria. Mantel test correlation analysis revealed a close relationship between gut microbial Burkholderiales and lipid metabolites in GLSO-mediated prevention of acute liver injury. GLSO exerts its preventive effects on acute liver injury by remodeling the gut microbiota and regulating lipid metabolism. These findings provide novel insights and potential directions for the development of new drugs targeting acute liver injury.

{"title":"Ganoderma lucidum spore oil attenuates acute liver injury by modulating lipid metabolism and gut microbiota","authors":"Jianying Liu , Yan Chen , Zhifeng Cen , Meiqi Hong , Binzhi Zhang , Xia Luo , Leqi Wang , Shasha Li , Xue Xiao , Qinqiang Long","doi":"10.1016/j.jpba.2025.116674","DOIUrl":"10.1016/j.jpba.2025.116674","url":null,"abstract":"<div><div>The incidence of acute liver injury is increasing and poses a significant threat to human health. <em>Ganoderma lucidum</em> spore oil (GLSO), a lipid substance extracted from <em>Ganoderma lucidum</em> spore powder using supercritical CO<sub>2</sub> technology, has been investigated for its potential to prevent acute liver injury. However, the specific mechanism underlying the protective effects of GLSO remains incompletely understood. In this study, we investigated the preventive effect of GLSO on acute liver injury in rats, focusing on the gut microbiome and serum metabolomics. GLSO effectively alleviated liver dysfunction and reduced inflammation, leading to the prevention of acute liver injury in rats. Serum metabolomics analysis revealed that GLSO primarily modulated lipid metabolic pathways related to glycerophospholipid metabolism and sphingolipid metabolism. Specifically, GLSO decreased the levels of metabolites such as lysophosphatidylcholine (LPC), glycerophosphatidylcholine (GPC), and sphinganine 1-phosphate (SA1P), while increasing the levels of phosphatidylglycerol (PG) and digalactosylceramide (DGC). Gut microbiomics data indicated that GLSO effectively regulated the composition of the gut microbiota in rats with acute liver injury. Specifically, it increased the abundance of <em>Firmicutes</em> and decreased the abundance of <em>Proteobacteria</em>. Mantel test correlation analysis revealed a close relationship between gut microbial <em>Burkholderiales</em> and lipid metabolites in GLSO-mediated prevention of acute liver injury. GLSO exerts its preventive effects on acute liver injury by remodeling the gut microbiota and regulating lipid metabolism. These findings provide novel insights and potential directions for the development of new drugs targeting acute liver injury.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"256 ","pages":"Article 116674"},"PeriodicalIF":3.1,"publicationDate":"2025-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143022966","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}

引用次数: 0

LC-MS/MS based analytical strategies for the detection of lipid peroxidation products in biological matrices

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-15 DOI: 10.1016/j.jpba.2025.116681

Federico Fanti , Manuel Sergi , Dario Compagnone

Oxidative stress (OS) arises mainly from exposure to reactive oxygen species (ROS) such as superoxide anion, hydroxyl radical, and hydrogen peroxide. These molecules can cause significant damage to proteins, DNA, and lipids, leading to various diseases. Cells fight ROS with detoxifying enzymes; however, an imbalance can cause damage leading to ischemic conditions, heart disease progression, and neurological disorders such as Alzheimer's disease. Accurate assessment of OS levels is then crucial and oxidized lipidic products are considered relevant OS biomarkers. In fact, lipids are particularly prone to ROS attack, leading to lipid peroxidation, cell membrane damage, and toxic by-products affecting DNA, proteins, and low-density lipoproteins. This review reports on recent advances in LC-MS/MS approaches for OS lipidic biomarkers, focusing on overcoming analytical challenges. 3 different classes of biomarkers have been reported, malondialdehyde, isoprostanes and oxidised sterols. For each class, the main analytical challenges with a particular focus on derivatisation procedure, sensitivity, matrix effect, ionisation have been described and discussed. The recent advancements of the LC-MS-MS procedures move towards simpler approaches, reducing errors and improving the reliability of the measurement thus enabling a comprehensive and robust OS assessment.

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引用次数: 0

Nano-zirconium-silicate solid-phase extraction method for the rapid quantification of pyrrolizidine alkaloids from plant extracts by UHPLC-QTOF-MS 纳米硅酸锆固相萃取- UHPLC-QTOF-MS快速定量测定植物提取物中吡咯利西啶类生物碱。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-15 DOI: 10.1016/j.jpba.2025.116675

Benedikt Schwarz , Shah Hussain , Christian W. Huck , Thomas Jakschitz , Moritz Rubner , Günther K. Bonn

The aim of this work was to develop and validate a rapid dispersive-solid-phase extraction method for the quantification of pyrrolizidine alkaloids (PA) from plant extracts. The method was focused on the significant removal of the intricate matrix to ensure good sensitivity for the subsequent instrumental analysis of PA. This was achieved by employing nano-zirconium silicate (NZS) as a dispersive-SPE sorbent. The specific affinity of NZS for PAs allowed for the effective removal of a substantial portion of the complex matrix, thereby significantly improving the sensitivity of the method, compared to the common methods, were no specific enrichment of the PAs on the SPE sorbent is achieved. Ultra-high-performance liquid chromatography coupled to high resolution time-of-flight mass spectrometry (UHPLC/TOF MS) was used for the qualitative and quantitative analysis of PA. The procedure demonstrated high recoveries for the standard compounds spiked into a blank verbena extract at different concentrations. Recovery rates of 72–95 % for PA, and 30–70 % for their respective N-oxides (PANO) could be obtained. The method was compared to the most commonly used C18 SPE sorbent, and demonstrated a significant lower limit of quantification (LOQ) of 0.64–4.5 ng mL⁻¹ as compared to 4.98–25.7 ng mL⁻¹. The method was validated in accordance with ICH guidelines. PA standards had a linear response between 5 and 150 ng mL ⁻¹ and demonstrated a co-efficient of variance below ± 3 % with a % relative error below ± 15. The presented analytical approach was also tested for the determination of PA from contaminated Verbenae herba extract with success. The presented scheme improves the clean-up efficacy of the already used stationary phases for PA analysis and provides a great alternative analytical tool for the isolation of PAs from plant extracts.

本研究的目的是建立并验证一种快速分离-固相萃取法定量测定植物提取物中吡咯利西啶类生物碱的方法。该方法的重点是去除复杂的矩阵，以确保后续PA仪器分析的良好灵敏度。这是通过采用纳米硅酸锆（NZS）作为分散- spe吸附剂来实现的。NZS对PAs的特异性亲和力允许有效去除大部分复合基质，从而显着提高了该方法的灵敏度，与常规方法相比，SPE吸附剂上没有实现PAs的特异性富集。采用超高效液相色谱-高分辨率飞行时间质谱（UHPLC/TOF MS）对PA进行定性和定量分析。在不同浓度的空白马鞭草提取物中加入标准化合物，回收率高。PA的回收率为72 ~ 95 %，n -氧化物（PANO）的回收率为30 ~ 70 %。该方法与最常用的C18固相萃取吸附剂进行了比较，定量下限为0.64-4.5 ng mL-1，而下限为4.98-25.7 ng mL-1。该方法按照ICH指南进行验证。PA标准品在5 ~ 150 ng mL -1之间具有线性响应，方差系数低于± 3 %，相对误差低于± 15 %。用该方法对污染马鞭草提取物中PA的测定也取得了成功。该方案提高了固定相对PA分析的清洁效率，为从植物提取物中分离PA提供了一个很好的替代分析工具。

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引用次数: 0

Quantification and clinical performance of serum parathyroid hormone 1–84 via immunocapture coupled to LC–MS/MS in chronic renal failure

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-14 DOI: 10.1016/j.jpba.2025.116678

Haiwei Cao , Yuting Jin , Yingwu Wang , Hao Wang , Yijia Qin , Xinhua Guo , Suyan Tian , Jing Huang , Yanyan Li

Accurate measurement of serum parathyroid hormone (PTH) is crucial for diagnosing and managing endocrine and osteological diseases. Conventional immunoassay methods struggle with cross reactivity issues between full-length PTH and truncated fragments or post-translationally modified forms. Both the standardization of PTH assays and the peptide's stability are concerning. This study addresses these issues by establishing an immunocapture coupled with liquid chromatography–tandem mass spectrometry (LC–MS/MS) method to precisely quantify PTH1–84. PTH1–84 was isolated from one mL serum samples by immunocapture on a polystyrene bead and eluted from matrix, then quantitated by LC–MS/MS. The results from 268 serum samples were compared to an intact PTH immunoassay. The assay's linear range was 5.0–1000.0 pg/mL. The intra-assay coefficients of variation (CVs) ranged from 3.2 % to 6.8 %, and the inter-assay CVs ranged from 4.6 % to 9.5 %. The extraction efficiencies were 98.0 %–100.5 %, with no significant matrix effects observed after internal standard correction. The correlation coefficient between LC–MS/MS and immunoassay was 0.989, but the bias between the methods was substantial. Nevertheless, the immunocapture purification coupled LC–MS/MS method offers a promising approach for accurate PTH measurement.

{"title":"Quantification and clinical performance of serum parathyroid hormone 1–84 via immunocapture coupled to LC–MS/MS in chronic renal failure","authors":"Haiwei Cao , Yuting Jin , Yingwu Wang , Hao Wang , Yijia Qin , Xinhua Guo , Suyan Tian , Jing Huang , Yanyan Li","doi":"10.1016/j.jpba.2025.116678","DOIUrl":"10.1016/j.jpba.2025.116678","url":null,"abstract":"<div><div>Accurate measurement of serum parathyroid hormone (PTH) is crucial for diagnosing and managing endocrine and osteological diseases. Conventional immunoassay methods struggle with cross reactivity issues between full-length PTH and truncated fragments or post-translationally modified forms. Both the standardization of PTH assays and the peptide's stability are concerning. This study addresses these issues by establishing an immunocapture coupled with liquid chromatography–tandem mass spectrometry (LC–MS/MS) method to precisely quantify PTH1–84. PTH1–84 was isolated from one mL serum samples by immunocapture on a polystyrene bead and eluted from matrix, then quantitated by LC–MS/MS. The results from 268 serum samples were compared to an intact PTH immunoassay. The assay's linear range was 5.0–1000.0 pg/mL. The intra-assay coefficients of variation (CVs) ranged from 3.2 % to 6.8 %, and the inter-assay CVs ranged from 4.6 % to 9.5 %. The extraction efficiencies were 98.0 %–100.5 %, with no significant matrix effects observed after internal standard correction. The correlation coefficient between LC–MS/MS and immunoassay was 0.989, but the bias between the methods was substantial. Nevertheless, the immunocapture purification coupled LC–MS/MS method offers a promising approach for accurate PTH measurement.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"256 ","pages":"Article 116678"},"PeriodicalIF":3.1,"publicationDate":"2025-01-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143023505","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}

引用次数: 0

Rapid determination of etomidate and its structural analogues in e-liquid by probe electrospray ionization quadrupole time-of-flight mass spectrometry 探针电喷雾电离四极杆飞行时间质谱法快速测定电子液体中依托咪酯及其结构类似物。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-13 DOI: 10.1016/j.jpba.2025.116677

Meiting Lin , Zhen Zhang , Qun He , Hongyuan Hao , Ping Xiang , Junbo Zhao

Etomidate and its structural analogues, which have anesthetic effects, are classified as controlled psychotropic drugs. Electronic cigarettes (e-cigarettes) have become more and more popular. With the increase of adding etomidate and its analogues to electronic liquids (e-liquids), there is a trend of abuse, which is a tough problem urgently need to be solved. This seriously affects the health and security of the public and the development of society. A simple, rapid and effective screening method is very crucial for their identification. In this study, we applied a newly developed method, probe electrospray ionization quadrupole time-of-flight mass spectrometry (PESI-QTOF-MS) with DPiMS QT ion source to analyze etomidate and its analogues in e-liquids. It allowed identification in 0.3 min with lower sample usage. Isomers can be distinguished by ion abundance ratios at collision energy (CE) 15 eV, which provided possibility for distinguishing more isomers by in-situ mass spectrometry. Limit of detection (LOD) and limit of quantitation (LOQ) of four substances were 20 ng/mL and 50 ng/mL, respectively. Good linear relationships were obtained in the concentration range of 50–5000 ng/mL with little matrix effect. The accuracy, precision, dilution effect and carryover of the method were also validated. Positive specimens (n = 38) were analyzed by both PESI-QTOF-MS and gas chromatography-mass spectrometry (GC-MS). There were five impurities including nicotine, cooling agent and flavorings were investigated by PESI-QTOF-MS, which provided the possibility for tracing the origin of illegal e-liquids. This study will help solve the backlog of cases and improve work efficiency effectively by reducing analysis time. Furthermore, it meets the need of addressing current situation of drug control and can assist forensic laboratories in investigating cases. It also demonstrates the application prospects of rapid screening in new drugs.

依托咪酯及其结构类似物具有麻醉作用，被列为管制精神药物。电子烟（e-cigarette）已经变得越来越流行。随着依托咪酯及其类似物在电子液体中添加量的增加，有滥用的趋势，这是一个亟待解决的棘手问题。这严重影响了公众的健康安全和社会的发展。一种简单、快速、有效的筛选方法对其鉴别至关重要。在这项研究中，我们采用了一种新的方法，探针电喷雾电离四极杆飞行时间质谱（PESI-QTOF-MS）与DPiMS QT离子源分析电子液体中的依托咪酯及其类似物。它可以在0.3 min内以较低的样本使用量进行鉴定。用碰撞能（CE） 15 eV的离子丰度比可以区分异构体，这为原位质谱法区分更多异构体提供了可能性。4种物质的检出限和定量限分别为20 ng/mL和50 ng/mL。在50 ~ 5000 ng/mL浓度范围内呈良好的线性关系，基质效应较小。验证了该方法的准确度、精密度、稀释效果和延展性。阳性标本（n = 38）采用PESI-QTOF-MS和气相色谱-质谱（GC-MS）分析。利用PESI-QTOF-MS分析了电子烟中尼古丁、冷却剂和调味剂等5种杂质，为非法电子烟的来源溯源提供了可能。通过减少分析时间，有助于解决积压的案例，有效提高工作效率。此外，它满足了解决药物管制现状的需要，并可协助法医实验室调查案件。同时也展示了快速筛选技术在新药中的应用前景。

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引用次数: 0

Chromatographic analysis and pKa evaluation of active pharmaceutical ingredients in anti-metastatic breast cancer: Green vs. conventional RPLC 抗转移性乳腺癌有效药物成分的色谱分析和pKa评价：绿色与传统RPLC。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-11 DOI: 10.1016/j.jpba.2025.116671

Fatmanur Kalkir , Ebru Çubuk Demi̇ralay , Yaşar Doğan Daldal , Hülya Yilmaz

This study aimed to determine the chromatographic retention and dissociation/protonation constant (pK_a) values of lapatinib and tamoxifen, key drugs used in metastatic breast cancer treatment, at 37°C using both conventional and green high-performance liquid chromatography (HPLC) methods. Qualitative analysis was conducted on an XTerra C18 column (250 ×4.6 mm I.D., 5 μm particle size) at a flow rate of 1 mL/min. Hydroorganic mixtures with 45 %, 50 %, 55 %, and 60 % (v/v) organic modifiers were used to evaluate the retention times of the compounds. The compatibility of pK_a values

({}_{s}^{s}{pK}_{a})

of the compounds in water-organic solvent mixtures obtained from these studies, which were carried out without any significant change in liquid chromatography performance, with the values obtained by the conventional method is remarkable. The

{}_{s}^{s}{pK}_{a}

values determined in this study were correlated with the macroscopic parameters of acetonitrile, methanol, ethanol and the pK_a (

{}_{w}^{w}{pK}_{a})

values of lapatinib and tamoxifen in water were calculated. The

{}_{w}^{w}{pK}_{a}

values calculated from these studies are compatible with each other and with the literature values. The environmental impact of the study, which was carried out using the green method and the conventional RPLC method, was evaluated using the Green Solvent Selection Tool (GSST), Green Analytical Procedures Index (GAPI), and Analytical Greenness Metric Approach (AGREE).

本研究旨在采用传统高效液相色谱法和绿色高效液相色谱法测定转移性乳腺癌治疗的关键药物拉帕替尼和他莫昔芬在37℃下的色谱保留和解离/质子化常数（pKa）值。定性分析采用XTerra C18色谱柱（250 ×4.6 mm内径，5 μm粒度），流速为1 mL/min。采用含45 %、50 %、55 %和60 % （v/v）有机改性剂的水有机混合物评价化合物的保留时间。从这些研究中得到的化合物在水-有机溶剂混合物中的pKa值pKass与传统方法得到的值的相容性是显著的，这些研究在液相色谱性能没有任何明显变化的情况下进行。本研究测定的pKass值与乙腈、甲醇、乙醇的宏观参数相关，并计算拉帕替尼和他莫昔芬在水中的pKa （pKaww）值。这些研究计算的pKaww值相互吻合，也与文献值吻合。本研究采用绿色方法和常规RPLC方法进行，并使用绿色溶剂选择工具（GSST）、绿色分析程序指数（GAPI）和分析绿色度度量方法（AGREE）对研究的环境影响进行评估。

{"title":"Chromatographic analysis and pKa evaluation of active pharmaceutical ingredients in anti-metastatic breast cancer: Green vs. conventional RPLC","authors":"Fatmanur Kalkir , Ebru Çubuk Demi̇ralay , Yaşar Doğan Daldal , Hülya Yilmaz","doi":"10.1016/j.jpba.2025.116671","DOIUrl":"10.1016/j.jpba.2025.116671","url":null,"abstract":"<div><div>This study aimed to determine the chromatographic retention and dissociation/protonation constant (pK<sub>a</sub>) values of lapatinib and tamoxifen, key drugs used in metastatic breast cancer treatment, at 37°C using both conventional and green high-performance liquid chromatography (HPLC) methods. Qualitative analysis was conducted on an XTerra C18 column (250 ×4.6 mm I.D., 5 μm particle size) at a flow rate of 1 mL/min. Hydroorganic mixtures with 45 %, 50 %, 55 %, and 60 % (v/v) organic modifiers were used to evaluate the retention times of the compounds. The compatibility of pK<sub>a</sub> values <span><math><mrow><mfenced><mrow><mmultiscripts><mrow><mi>pK</mi></mrow><mrow><mi>a</mi></mrow><none></none><mprescripts></mprescripts><mrow><mi>s</mi></mrow><mrow><mi>s</mi></mrow></mmultiscripts></mrow></mfenced></mrow></math></span> of the compounds in water-organic solvent mixtures obtained from these studies, which were carried out without any significant change in liquid chromatography performance, with the values obtained by the conventional method is remarkable. The <span><math><mmultiscripts><mrow><mi>pK</mi></mrow><mrow><mi>a</mi></mrow><none></none><mprescripts></mprescripts><mrow><mi>s</mi></mrow><mrow><mi>s</mi></mrow></mmultiscripts></math></span> values determined in this study were correlated with the macroscopic parameters of acetonitrile, methanol, ethanol and the pK<sub>a</sub> (<span><math><mrow><mmultiscripts><mrow><mi>pK</mi></mrow><mrow><mi>a</mi></mrow><none></none><mprescripts></mprescripts><mrow><mi>w</mi></mrow><mrow><mi>w</mi></mrow></mmultiscripts><mrow><mo>)</mo><mspace></mspace></mrow></mrow></math></span>values of lapatinib and tamoxifen in water were calculated. The <span><math><mmultiscripts><mrow><mi>pK</mi></mrow><mrow><mi>a</mi></mrow><none></none><mprescripts></mprescripts><mrow><mi>w</mi></mrow><mrow><mi>w</mi></mrow></mmultiscripts></math></span> values calculated from these studies are compatible with each other and with the literature values. The environmental impact of the study, which was carried out using the green method and the conventional RPLC method, was evaluated using the Green Solvent Selection Tool (GSST), Green Analytical Procedures Index (GAPI), and Analytical Greenness Metric Approach (AGREE).</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"256 ","pages":"Article 116671"},"PeriodicalIF":3.1,"publicationDate":"2025-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143007221","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}

引用次数: 0

Hydrophilic interaction liquid chromatography-tandem mass spectrometry analysis of oligosaccharides and iridoid glycosides in rat plasma: Pharmacokinetic characterization of previously overlooked oligosaccharides from Radix Rehmanniae 大鼠血浆中低聚糖和环烯醚萜苷的亲水相互作用-液相色谱-串联质谱分析：以前被忽视的地黄低聚糖的药代动力学表征。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-09 DOI: 10.1016/j.jpba.2025.116670

Hanyang Wang , Yue Du , Chunwei Zhou , Yan Wang , Xin Wang

Radix Rehmanniae (RR) is a widely used herb in traditional Chinese Medicine with properties of tonifying the kidneys and nourishing the blood. Both raw and processed RR are effective for the treatment of diabetes in clinical practice. Oligosaccharides and iridoid glycosides are the primary active components responsible for the anti-diabetic effects of RR. In this study, a rapid and sensitive hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC−MS/MS) method was developed for simultaneous determination of oligosaccharides (raffinose, manninotriose and stachyose) and iridoid glycosides (catalpol and ajugol) in rat plasma. Significant analytical challenges were encountered during method development, including distinct retention behaviors of oligosaccharides and iridoid glycosides, low ionization and extraction efficiency of oligosaccharides, thermal instability of catalpol and reduced column performance. The strategies to overcome these challenges were presented by optimizing chromatographic separation, mass spectrometric detection and sample preparation. The best separation was achieved using an Accucore-150-Amide-HILIC column (100 mm × 2.1 mm, 2.6 μm) at 50 °C with mobile phase consisted of acetonitrile and ammonium acetate (2.5 mM) under gradient elution. Ammonium adduct ions produced by positive electrospray ionization were chosen as precursor ions for multiple reaction monitoring transitions. The established HILIC−MS/MS method exhibited good linearity (r > 0.9937) with the lower limits of quantification of 0.01–0.2 μg/mL using only 50 µL of plasma sample. The method was successfully applied to pharmacokinetic characterization of oligosaccharides and iridoid glycosides in normal and type 2 diabetic rats following intragastric administration of raw and processed RR extracts.

地黄（RR）是一种广泛使用的中药，具有补肾养血的特性。在临床实践中，生的和加工的红霉素对糖尿病的治疗都是有效的。低聚糖和环烯醚萜苷是RR抗糖尿病作用的主要活性成分。本研究建立了快速、灵敏的亲水相互作用液相色谱-串联质谱（HILIC-MS/MS）同时测定大鼠血浆中低聚糖（棉子糖、甘露糖和水苏糖）和环烯醚萜苷（catalpol和ajugol）的方法。在方法开发过程中遇到了重大的分析挑战，包括低聚糖和环烯醚萜苷的独特保留行为，低聚糖的低电离和提取效率，catalpol的热不稳定性以及色谱柱性能降低。通过优化色谱分离、质谱检测和样品制备，提出了克服这些挑战的策略。Accucore-150-Amide-HILIC色谱柱（100 mm × 2.1 mm, 2.6 μm），温度为50 °C，流动相为乙腈和乙酸铵（2.5 mm），梯度洗脱，分离效果最佳。采用正电喷雾电离产生的铵加合物离子作为前体离子，用于多反应监测跃迁。所建立的HILIC-MS/MS方法具有良好的线性关系（r > 0.9937），定量下限为0.01 ~ 0.2 μg/mL，仅需50 µL血浆样品。该方法成功地应用于正常和2型糖尿病大鼠灌胃RR提取物后低聚糖和环烯醚萜苷的药动学表征。

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引用次数: 0

Cellulose filter paper immobilized acetylcholinesterase for rapid screening of enzyme inhibitors in Phyllanthus emblica L. 纤维素滤纸固定化余甘子乙酰胆碱酯酶抑制剂的快速筛选。

IF 3.1 3区医学 Q2 CHEMISTRY, ANALYTICAL

Journal of pharmaceutical and biomedical analysis

Pub Date : 2025-01-08 DOI: 10.1016/j.jpba.2025.116669

Yangzom Dawa , Yong-Chen Hua , Fang-Di Hu , Juan Chen

Acetylcholinesterase (AChE) is widely recognized as a promising therapeutic target enzyme for Alzheimer's disease (AD). The screening of AChE inhibitors (AChEIs) holds great significance for the treatment of AD. In this study, cellulose ﬁlter paper (CFP) -immobilized AChE was prepared and firstly applied to screening AChEIs from 30 % ethanol extract of Phyllanthus emblica L. fruits combined with ultra-high performance liquid chromatography quadrupole time-of-fight mass spectrometry (UHPLC-Q-TOF-MS/MS). Using CFP-immobilized AChE as the bait, AChEIs were harvested and the instantaneous separation characteristics of CFP were utilized to further facilitate the separation of the complex from the inactive components. Ultimately, 27 compounds specifically bound with AChE were screened and identified using UHPLC-Q-TOF-MS/MS. Additionally, molecular docking was employed to explore the binding mechanisms between screened potential inhibitors and AChE. The results show that, most of the screened compounds were found to exhibit higher affinity that of the positive control (huperzine A), and all the compounds expect mucic acid to be well embedded into the active pocket of AChE. To verify the reliability of the screening method and molecular docking, two commercial standards geraniin and ellagic acid were experimented with an AChE inhibition assay in vitro. The results showed that both compounds were found to effectively inhibit AChE with IC₅₀ values of 42.42 ± 7.10 μM, 172.43 ± 9.22 μM. The developed method exhibits the advantages of rapidness and effectiveness in screening of AChEIs from complex herbal extracts.

乙酰胆碱酯酶（AChE）被广泛认为是治疗阿尔茨海默病（AD）的一种有前景的靶酶。AChE抑制剂（AChEIs）的筛选对AD的治疗具有重要意义。本研究制备了纤维素滤纸（CFP）固定化乙酰氨基甲酸乙酯（AChE），并首次应用超高效液相色谱-四极杆对抗时间质谱（UHPLC-Q-TOF-MS/MS）技术从30 %乙醇的余甘子果实提取物中筛选乙酰氨基甲酸乙酯。以CFP固定的AChE为诱饵，收获AChEIs，利用CFP的瞬时分离特性进一步促进配合物与非活性组分的分离。最终，通过UHPLC-Q-TOF-MS/MS筛选和鉴定了27个与AChE特异性结合的化合物。此外，采用分子对接的方法探索筛选出的潜在抑制剂与AChE的结合机制。结果表明，筛选的大部分化合物都比阳性对照石杉碱A具有更高的亲和力，并且所有化合物都期望乙酸能很好地嵌入AChE的活性口袋中。为了验证筛选方法和分子对接的可靠性，我们对两种商业标准天竺葵苷和鞣花酸进行了体外AChE抑制实验。结果表明，两种化合物均能有效抑制AChE， IC50值分别为42.42 ± 7.10 μM和172.43 ± 9.22 μM。该方法对复方草药提取物中乙酰胆碱酯类化合物的筛选具有快速、有效的优点。

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引用次数: 0