Journal of Chromatography B最新文献_第8页

Validation of a highly sensitive assay for the determination of rivastigmine in human plasma for pharmacokinetic studies. 用于药代动力学研究的人血浆中利瓦斯汀高灵敏度测定法的验证。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-26 DOI: 10.1016/j.jchromb.2025.124838

Maksim Dolov , Igor Rodin

A fast and highly sensitive LC–MS/MS method has been developed and validated for measuring rivastigmine levels in human plasma. The extraction of rivastigmine from plasma was performed using a simple and quick protein precipitation technique (PPT). The internal standard used was atazanavir-d₅. Chromatographic separation was achieved using a YMC-Triart C18 column, followed by detection with mass spectrometry. The mass transitions monitored were m/z 251.1 > 206.0 for rivastigmine, and m/z 710.5 > 168.1 for atazanavir-d₅. This method involves rapid plasma extraction, straightforward gradient chromatography, and mass spectrometric detection, allowing for the detection of rivastigmine at sub-nanogram per milliliter levels. The method was validated over a linear range of 25 to 5000 pg/ml, with a correlation coefficient of at least 0.9980. Both intra- and inter-day precision and accuracy were within 15 %. The overall recovery rates for rivastigmine and atazanavir-d₅ were close to 100 %. The total analysis time per sample was only 3 min. This method was successfully applied to determine the pharmacokinetic parameters of rivastigmine after a single oral dose of 1.5 mg (capsules) in 26 healthy volunteers.

建立了一种快速、高灵敏度的LC-MS /MS方法，用于测定人血浆中伐斯汀的含量。采用简单快速蛋白沉淀技术（PPT）从血浆中提取利瓦斯汀。内标为atazanvir -d5。采用YMC-Triart C18色谱柱进行色谱分离，然后采用质谱法进行检测。监测到的质量转移为：瑞瓦斯汀为m/z 251.1 > 206.0，阿扎那韦-d5为m/z 710.5 > 168.1。该方法包括快速血浆提取，直接梯度色谱和质谱检测，允许检测亚纳克/毫升水平的利瓦斯汀。在25 ~ 5000 pg/ml的线性范围内，相关系数至少为0.9980。日内、日间的精密度和准确度均在15%以内。瑞瓦斯汀和阿扎那韦-d5的总回收率接近100%。每个样品的总分析时间仅为3分钟。该方法成功地测定了26名健康志愿者单次口服1.5 mg（胶囊）利瓦斯汀的药动学参数。

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引用次数: 0

Development and validation of an LC-MS/MS method for Tirzepatide, a dual GIP/GLP-1 receptor agonist, in rat plasma for application to a pharmacokinetic study 大鼠血浆中双GIP/GLP-1受体激动剂tizepatide的LC-MS/MS方法的开发和验证，用于药代动力学研究。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-26 DOI: 10.1016/j.jchromb.2025.124836

Hae-In Choi , Hyeon-Cheol Jeong , Jong-Woo Jeong , Jaeyoung Lee , Da Hae Kim , Kyong-Cheol Ko , Yoon-Jee Chae , Kyeong-Ryoon Lee

A sensitive and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of tirzepatide, a dual agonist of glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) receptors in rat plasma. Tirzepatide was extracted from rat plasma by protein precipitation using methanol. Chromatographic separation was achieved using a peptide C18 column with gradient elution of water and acetonitrile containing 0.1 % formic acid. Mass spectrometric detection was performed in positive electrospray ionization mode using multiple reaction monitoring with transitions of m/z 1204.4 → 1473.6 for tirzepatide and m/z 1029.4 → 1238.4 for the internal standard, semaglutide. The developed method exhibited good linearity over a concentration range of 1–1000 ng/mL (r² > 0.99). Intra- and inter-day accuracy (−4.324–5.057 %) and precision (5.250–9.000 %) met the regulatory criteria at all quality control levels, and were stable under various plasma handling and storage conditions. The validated method was successfully applied to a pharmacokinetic study in rats following the intravenous and subcutaneous injection of tirzepatide at 0.3 mg/kg. The terminal half-lives were 10.04 h and 9.803 h after intravenous and subcutaneous administration, respectively, indicating comparable elimination profiles. The bioavailability following subcutaneous dosing was estimated to be approximately 62.38 %. These findings highlight the robustness and applicability of the developed method, suggesting its potential utility for the quantitative analysis of other peptide therapeutics with structures or mechanisms of action similar to those of tirzepatide.

建立了一种灵敏、快速的液相色谱-串联质谱（LC-MS/MS）方法，用于定量大鼠血浆中葡萄糖依赖性胰岛素性多肽（GIP）和胰高血糖素样肽-1 （GLP-1）受体双激动剂替西肽。采用甲醇蛋白沉淀法从大鼠血浆中提取替西帕肽。色谱分离采用肽C18柱，梯度洗脱水和含有0.1%甲酸的乙腈。质谱检测采用正电喷雾电离模式，采用多重反应监测，替西肽为m/z 1204.4→1473.6，内标semaglutide为m/z 1029.4→1238.4。该方法在1 ~ 1000 ng/mL （r2 > 0.99）的浓度范围内线性良好。日内和日间准确度（-4.324- 5.057%）和精密度（5.250-9.000 %）均符合所有质量控制水平的监管标准，且在各种血浆处理和储存条件下均稳定。该方法成功应用于0.3 mg/kg替西帕肽静脉注射和皮下注射后大鼠体内的药代动力学研究。静脉和皮下给药后的终末半衰期分别为10.04 h和9.803 h，表明消除情况相当。皮下给药后的生物利用度估计约为62.38%。这些发现突出了所开发方法的稳健性和适用性，表明其对其他结构或作用机制类似于替西肽的肽疗法的定量分析的潜在效用。

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引用次数: 0

First report of borbotoxin occurrence in Brazilian Prorocentrum borbonicum isolates: LC-MS/MS detection and preliminary structural insights 巴西硼原心菌分离株中硼肉毒素的首次报道：LC-MS/MS检测和初步结构见解。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-24 DOI: 10.1016/j.jchromb.2025.124825

Cristian Rafael Kleemann , Rodrigo Barcellos Hoff , Mathias A. Schramm , Alexsandro Dallegrave , Carlos Eduardo J. de Azevedo Tibiriçá , Mauro Henrique Dartora Dutra , Mauricio Perin , Jessie Sobieski da Costa , Luiz L. Mafra Jr. , Pedro Luiz Manique Barreto

Borbotoxin is an emergent biotoxin produced by Prorocentrum borbonicum and other related dinoflagellates. This toxin is known to produce neuromuscular effects in animal models and may pose risks to human health, although its molecular structure is still unknown. Strains of Prorocentrum borbonicum isolated from the northeastern Brazilian coast were analyzed by reversed phase liquid chromatography coupled to tandem mass spectrometry. Cell pellets were extracted with methanol and water with the aid of an ultrasonic bath. Multiple reaction monitoring mode was used to search for borbotoxin A and B using mass data from literature. Putative peaks with close mass were found in two algal cell extracts. Experiments with the ion trap produced MS² spectra in agreement with previous studies. Due to the absence of analytical standards, high mass spectrometry resolution was used to confirm the masses and the occurrence of borbotoxin was thus demonstrated. To the best of our knowledge, this is the first report of borbotoxin occurrence in Brazil. Further studies will be conducted to propose a definite structure to this relatively unknown marine biotoxin.

硼肉毒素是由硼原心菌和其他相关鞭毛藻产生的一种新兴生物毒素。已知这种毒素在动物模型中产生神经肌肉效应，并可能对人类健康构成风险，尽管其分子结构尚不清楚。采用反相液相色谱-串联质谱联用技术对巴西东北海岸产的硼原央菌进行了分析。在超声波浴的帮助下，用甲醇和水提取细胞颗粒。采用多反应监测模式，利用大量文献资料搜索肉毒杆菌毒素A、B。在两种藻类细胞提取物中发现了质量接近的推定峰。离子阱实验产生的MS2光谱与先前的研究一致。由于缺乏分析标准，采用高质谱分辨率对质量进行确认，从而证明了硼毒素的存在。据我们所知，这是巴西首次出现肉毒杆菌毒素的报告。将进行进一步的研究，以确定这种相对未知的海洋生物毒素的确切结构。

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引用次数: 0

Patchouli alcohol as an effective adjuvant for artemisinin-based combination therapies: Metabolic profiling, pharmacokinetics and mechanistic validation 广藿香醇作为以青蒿素为基础的联合治疗的有效佐剂：代谢谱、药代动力学和机制验证

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-24 DOI: 10.1016/j.jchromb.2025.124827

Linlin Li , Boyang Wang , Bianli Wang , Xiaolei Du , Yuanyuan Bai , Xiao Zhang , Xuesheng Yan , Ping Wang , Huimin Zhang

Malaria remains a globally fatal disease, and the emergence of drug resistance necessitates the discovery of novel adjuvant drugs for artemisinin-based combination therapies (ACTs). Patchouli alcohol (PA), the primary active component of Pogostemon cablin Benth, has demonstrated antimalarial potential. This study investigates the synergistic antimalarial effect of PA in combination with sodium artesunate (SA) and explores its pharmacokinetics and mechanism of action. Pharmacokinetic parameters were assessed using GC–MS/MS and analyzed with DAS software, while PA metabolites were identified via GC–MS and HPLC-Q-Exactive Plus Orbitrap MS. PA exhibited dose-dependent pharmacokinetics, with proportional increases in C_max and AUC across doses and a consistent T_max of 30 min. Additionally, significant gender-related differences in exposure were observed. Metabolic profiling identified 30 metabolites in plasma, urine and feces, primarily through dehydrogenation, sulfation and glucuronidation. Phase I metabolic reactions occurred more frequently in plasma, while phase II metabolic reactions more in urine and fecal samples. The rich metabolic reactions provided more possibilities for the exertion of its pharmacological effects. Network pharmacology and molecular docking predicted and validated PA's antimalarial mechanism, with strong binding interactions (M14-MMP9, SA-TNF, M14-PTGS2, SA-PTGS2, SA-CASP3, PA-MMP9). These results provide critical insights into the metabolic profile and mechanism of PA, supporting its development as a promising adjuvant for ACTs.

疟疾仍然是一种全球致命疾病，耐药性的出现要求为以青蒿素为基础的联合疗法发现新的辅助药物。广藿香醇（PA）是广藿香的主要活性成分，具有抗疟疾的潜力。本研究考察了PA与青蒿琥酯钠（SA）联用的协同抗疟作用，并探讨了其药代动力学和作用机制。使用GC-MS /MS评估药代动力学参数，并使用DAS软件进行分析，同时通过GC-MS和HPLC-Q-Exactive Plus Orbitrap MS鉴定PA代谢物，PA表现出剂量依赖的药代动力学，Cmax和AUC随剂量成比例增加，Tmax一致为30 min。此外，还观察到暴露的显著性别差异。代谢谱鉴定了血浆、尿液和粪便中的30种代谢物，主要通过脱氢、硫酸和葡萄糖醛酸化。I期代谢反应多见于血浆，II期代谢反应多见于尿液和粪便样品。丰富的代谢反应为其药理作用的发挥提供了更多的可能性。网络药理学和分子对接预测并验证了PA的抗疟机制，具有较强的结合相互作用（M14-MMP9、SA-TNF、M14-PTGS2、SA-PTGS2、SA-CASP3、PA- mmp9）。这些结果为PA的代谢特征和机制提供了重要的见解，支持其作为act的有前途的佐剂的发展。

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引用次数: 0

Study on the chemical composition and quality evaluation of Fufangxueteng medicinal wine based on UHPLC-Q orbitrap-HRMS and HPLC combined with fingerprinting and content determination analysis 基于UHPLC-Q轨道谱- hrms和HPLC结合指纹图谱和含量测定分析的复方雪藤药酒化学成分及质量评价研究。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-24 DOI: 10.1016/j.jchromb.2025.124835

Wenshuang Tang , Zengdan Ran , Banghui Zhu , Xin Hu , Yang Wang , Yueting Li , Jia Sun , Yongjun Li , Xue Ma

Objective

This study aims to rapidly identify the chemical components of Fufangxueteng Medicinal Wine (FFXTMW) and establish a quality control method based on fingerprinting and quantitative analysis.

Methods

Qualitative analysis was carried out using UHPLC-Q Orbitrap-HRMS; high-performance liquid chromatography (HPLC) was employed for fingerprint analysis and content determination.

Results

A total of 152 compounds were identified in FFXTMW, with 20 confirmed by reference standards; the identified components mainly included flavonoids, organic acids, alkaloids, and phenylpropanoids. For 13 batches of FFXTMW, their fingerprints showed 72 common peaks with a similarity of 0.926–0.993, and 12 components were characterized. Additionally, six key components (isofraxidin, toddalolactone, periplocin, syringin, naringin, hesperidin) exhibited good linearity with peak areas in their respective concentration ranges (r² ≥ 0.9997), with average spiked recoveries of 98.71%–103.08% and relative standard deviations (RSD) of 1.12%–2.49%.

Conclusion

UHPLC-Q Orbitrap-HRMS enables rapid and accurate identification of the main chemical components in FFXTMW. The established HPLC-based fingerprinting and quantitative methods are efficient and precise, and applicable for FFXTMW quality control. These results lay a foundation for upgrading FFXTMW's quality standards.

目的：快速鉴定复方雪藤药酒（FFXTMW）的化学成分，建立基于指纹图谱和定量分析的质量控制方法。方法：采用UHPLC-Q Orbitrap-HRMS进行定性分析；采用高效液相色谱法进行指纹分析和含量测定。结果：共鉴定出152个化合物，其中20个经参比标准品确认；鉴定出的主要成分有黄酮类化合物、有机酸类化合物、生物碱类化合物和苯丙素类化合物。13个批次的FFXTMW指纹图谱有72个共同峰，相似度为0.926 ~ 0.993，共鉴定了12个组分。6种关键成分（异黄皮苷、牙陀内酯、洋槐苷、紫丁香苷、柚皮苷、橙皮苷）在各自浓度范围内与峰面积呈良好的线性关系（r2≥0.9997），平均加标回收率为98.71% ~ 103.08%，相对标准偏差（RSD）为1.12% ~ 2.49%。结论：UHPLC-Q Orbitrap-HRMS技术能够快速、准确地鉴定出药材中的主要化学成分。所建立的hplc指纹图谱和定量方法高效、准确，适用于FFXTMW的质量控制。这些结果为提高FFXTMW的质量标准奠定了基础。

{"title":"Study on the chemical composition and quality evaluation of Fufangxueteng medicinal wine based on UHPLC-Q orbitrap-HRMS and HPLC combined with fingerprinting and content determination analysis","authors":"Wenshuang Tang , Zengdan Ran , Banghui Zhu , Xin Hu , Yang Wang , Yueting Li , Jia Sun , Yongjun Li , Xue Ma","doi":"10.1016/j.jchromb.2025.124835","DOIUrl":"10.1016/j.jchromb.2025.124835","url":null,"abstract":"<div><h3>Objective</h3><div>This study aims to rapidly identify the chemical components of Fufangxueteng Medicinal Wine (FFXTMW) and establish a quality control method based on fingerprinting and quantitative analysis.</div></div><div><h3>Methods</h3><div>Qualitative analysis was carried out using UHPLC-Q Orbitrap-HRMS; high-performance liquid chromatography (HPLC) was employed for fingerprint analysis and content determination.</div></div><div><h3>Results</h3><div>A total of 152 compounds were identified in FFXTMW, with 20 confirmed by reference standards; the identified components mainly included flavonoids, organic acids, alkaloids, and phenylpropanoids. For 13 batches of FFXTMW, their fingerprints showed 72 common peaks with a similarity of 0.926–0.993, and 12 components were characterized. Additionally, six key components (isofraxidin, toddalolactone, periplocin, syringin, naringin, hesperidin) exhibited good linearity with peak areas in their respective concentration ranges (r<sup>2</sup> ≥ 0.9997), with average spiked recoveries of 98.71%–103.08% and relative standard deviations (RSD) of 1.12%–2.49%.</div></div><div><h3>Conclusion</h3><div>UHPLC-Q Orbitrap-HRMS enables rapid and accurate identification of the main chemical components in FFXTMW. The established HPLC-based fingerprinting and quantitative methods are efficient and precise, and applicable for FFXTMW quality control. These results lay a foundation for upgrading FFXTMW's quality standards.</div></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":"1267 ","pages":"Article 124835"},"PeriodicalIF":2.8,"publicationDate":"2025-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145395831","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

A paradigm for sustainable analysis: QbD-driven HPLC-DAD for triple antiallergics in pharmaceutical, biological, and environmental samples 可持续分析的范例：qbd驱动的HPLC-DAD用于药物，生物和环境样品中的三重抗过敏药。

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-24 DOI: 10.1016/j.jchromb.2025.124815

Aya Magdy Saad , Jenny Jeehan Mohamed Nasr , Asmaa Kamal El-Deen

This study introduces an innovative approach combining Quality by Design (QbD) principles with high-performance liquid chromatography and diode array detection (HPLC-DAD) for the simultaneous analysis of tranilast (TNL), pranoprofen (PPF), and chlorpheniramine maleate (CPM), a novel antiallergic combination therapy. The QbD framework was systematically employed to optimize critical method parameters, including mobile phase composition, flow rate, and column temperature, to achieve robust separation of the target analytes within a short analysis time. Method validation was conducted following ICH guidelines, demonstrating excellent linearity within the concentration range of 4–40, 2–50 and 1–140 μg/mL, with detection limits of 1.034, 0.661, and 0.328 μg/mL and quantitation limits of 3.133, 2.0, and 0.994 μg/mL for CPM, PPF, and TNL, respectively. The method was further applied to triple combination eye drops, aqueous humor, and environmental water samples, with a recovery of > 97 %. The application of QbD enhanced the method's reliability and confirmed consistent performance. A holistic sustainability assessment confirmed the method's environmental and economic viability, featuring low solvent use, minimal sample preparation, and long-term durability, in line with green analytical chemistry principles. This integrated strategy provides an efficient solution for the quality control of complex ophthalmic formulations, facilitating the development of safer and more effective antiallergic therapies.

本研究提出了一种结合设计质量（QbD）原理、高效液相色谱和二极管阵列检测（HPLC-DAD）的创新方法，用于同时分析曲尼拉斯特（TNL）、普萘洛芬（PPF）和马来酸氯苯那敏（CPM）这一新型抗过敏联合疗法。系统地利用QbD框架优化关键方法参数，包括流动相组成、流速和柱温，以在较短的分析时间内实现目标分析物的稳定分离。方法按照ICH标准进行验证，在4-40、2-50和1-140 μg/mL的浓度范围内线性良好，CPM、PPF和TNL的检出限分别为1.034、0.661和0.328 μg/mL，定量限分别为3.133、2.0和0.994 μg/mL。该方法进一步应用于三联滴眼液、体液和环境水样，回收率为0.97%。QbD的应用提高了方法的可靠性，保证了方法的一致性。全面的可持续性评估证实了该方法的环境和经济可行性，其特点是溶剂使用量低，样品制备最少，长期耐用，符合绿色分析化学原则。这种综合策略为复杂眼科配方的质量控制提供了有效的解决方案，促进了更安全、更有效的抗过敏疗法的发展。

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引用次数: 0

Novel isotope-coded derivatization of carboxylic acids provides accurate quantification of Azaspiracids in shellfish by liquid chromatography–tandem mass spectrometry 新型的同位素编码衍生化羧酸提供了液相色谱-串联质谱法对贝类中氮杂螺酸的准确定量

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-22 DOI: 10.1016/j.jchromb.2025.124826

Shimba Kawasue, Kyoko Kuniyoshi, Takashi Kurohara, Koji Fujihara, Yutaka Abe, Naoki Sugimoto, Masashi Uema, Naomasa Oshiro

The combination of isotope-coded derivatization (ICD) and liquid chromatography–tandem mass spectrometry (LC–MS/MS) is an excellent analytical method that could be applied to various compounds. However, the ICD reagents are expensive, which limits the use of this method for general testing including the evaluation of food safety and quality. Therefore, we synthesized reasonable ICD reagents, isopropyl piperidine carboxylic acid hydrazide (IPPAH) and isopropyl-piperidine carboxylic acid hydrazide-d₆ (IPPAH-d₆) using readily available acetone‑d₆, which reacts with the carboxy groups in the presence of a condensing agent to give a mass difference of six. It is applicable to carboxylic acids with a wide range of molecular weights. The application of the combination of ICD and LC/MS method to the analysis of the shellfish toxin azaspiracid in shellfish showed a significant improvement in the matrix effects (93.5–99.6 %). The method quality was evaluated by analyzing the certified reference material. The established method achieved an accurate quantification (95.7–103 %) of AZAs in shellfish specimens.

同位素编码衍生化（ICD）与液相色谱-串联质谱（LC-MS /MS）相结合是一种适用于多种化合物的优良分析方法。然而，ICD试剂价格昂贵，这限制了该方法用于一般检测，包括食品安全和质量评价。因此，我们利用丙酮-d6合成了合理的ICD试剂，异丙基哌啶羧酸肼（IPPAH）和异丙基哌啶羧酸肼-d6 (IPPAH-d6)，丙酮-d6在缩合剂的存在下与羧基反应，得到6的质量差。适用于分子量范围广的羧酸。将ICD与LC/MS相结合的方法应用于贝类中贝类毒素氮氮唑酸的分析，基质效应明显改善（93.5 ~ 99.6%）。通过对标准物质的分析，评价了方法的质量。建立的方法对贝类标本中AZAs的定量准确度为95.7 ~ 103%。

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引用次数: 0

Establishment of a LC-MS/MS method for the simultaneous quantitative determination of trimethylamine N-oxide and four precursors in human saliva 建立同时定量测定人唾液中三甲胺n -氧化物及四种前体的LC-MS/MS方法

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-22 DOI: 10.1016/j.jchromb.2025.124829

Jianyue Li , Bin Hu , Jie Wu , Dazhen Wang , Junyan Xia , Yanan Li , Yonghong Gao , Baorong Chen , Jianfeng Zhou , Zenghe Li

In this work, a liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed to simultaneously quantitatively detect the choline (CHL), trimethylamine N-oxide (TMAO), γ-butyrobetaine (GBB), betaine (BET), and ʟ-carnitine (CAR) in human saliva for the first time. Saliva samples collected from healthy volunteers were treated with acetonitrile for protein precipitation, followed by direct injection into the LC-MS/MS system for analysis. The method validation was performed in accordance with the CLSI C62-A standard guidelines. The linearity was in the range of 0.300–300.000 μmol/L for CHL, 0.005–5.000 μmol/L for TMAO, 0.050–50.000 μmol/L for GBB, 0.020–20.000 μmol/L for BET, and 0.100–100.000 μmol/L for CAR. The intra-day and inter-day precision ranged from 1.13 % to 8.34 % and from 1.63 % to 10.65 %, respectively. The recoveries and matrix effects of these analytes ranged from 94.31 % to 108.00 % and from 90.59 % to 111.37 %, respectively. The performance of the method met the requirements of the guidelines. The validated method was subsequently applied to analyze test saliva samples from 43 healthy volunteers and 4 coronary heart disease (CHD) inpatients, to evaluate the distributions of five analytes in healthy people and CHD inpatients. This study provides a reliable and accurate analytical approach for the clinical detection of five analytes in saliva.

本文首次建立了液相色谱-串联质谱（LC-MS/MS）同时定量检测人唾液中胆碱（CHL）、三甲胺n -氧化物（TMAO）、γ-丁甜菜碱（GBB）、甜菜碱（BET）和肉碱（CAR）的方法。健康志愿者唾液样品经乙腈沉淀蛋白后，直接注入LC-MS/MS系统分析。方法验证按照CLSI C62-A标准指南进行。CHL的线性范围为0.300 ~ 300.000 μmol/L， TMAO为0.005 ~ 5.000 μmol/L， GBB为0.050 ~ 50.000 μmol/L， BET为0.020 ~ 20.000 μmol/L， CAR为0.100 ~ 100.000 μmol/L。日内精度为1.13% ~ 8.34%，日内精度为1.63% ~ 10.65%。加样回收率为94.31% ~ 108.00%，基质效应为90.59% ~ 111.37%。该方法的性能满足指南的要求。随后将验证的方法应用于43名健康志愿者和4名冠心病住院患者的唾液样本分析，评估5种分析物在健康人群和冠心病住院患者中的分布。本研究为临床检测唾液中5种分析物提供了可靠、准确的分析方法。

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引用次数: 0

Pharmacokinetic profile and bioavailability assessment of rubusoside in mouse plasma utilizing UPLC-MS/MS analysis 利用UPLC-MS/MS分析评价鲁布苏苷在小鼠血浆中的药动学特征及生物利用度

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-22 DOI: 10.1016/j.jchromb.2025.124832

Meiling Zhang , Siyu Zhuo , Fangmei Zhang , Jianshe Ma , Lufeng Hu , Xianqin Wang

The ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) technique was utilized to detect rubusoside in mouse plasma and evaluate its pharmacokinetic behavior following oral administration at doses of 10 mg/kg and 20 mg/kg, as well as intravenous administration at 10 mg/kg. Plasma samples were prepared using a protein precipitation method employing acetonitrile as the precipitating agent and isoscoparin as the internal standard (IS). Chromatographic separation was performed on a UPLC high strength silica (HSS) T3 column under gradient elution conditions, using acetonitrile and 0.1 % formic acid in water as the mobile phase system. The calibration curve for rubusoside demonstrated excellent linearity across a concentration range of 3.5 to 1120 ng/mL. Both intra-day and inter-day precision values remained below 7 %, with accuracy ranging from 92.3 % to 108.1 %. Matrix effects were consistently within the range of 107.1 % to 110.0 %, and the recovery rate exceeded 74.2 %. Following oral administration, the absolute bioavailability of rubusoside was determined to be 1.00 % and 0.98 % at doses of 10 mg/kg and 20 mg/kg, respectively. These findings confirm that the developed UPLC-MS/MS method effectively supports the pharmacokinetic evaluation of rubusoside

采用超高效液相色谱-串联质谱（UPLC-MS/MS）技术检测小鼠血浆中鲁布苏苷的药代动力学行为，并对口服剂量为10 mg/kg、口服剂量为20 mg/kg、静脉给药剂量为10 mg/kg的鲁布苏苷进行了评价。血浆样品采用蛋白质沉淀法制备，以乙腈为沉淀剂，异莨菪碱为内标。以乙腈和0.1%甲酸水溶液为流动相，在UPLC高强度二氧化硅（HSS） T3柱上梯度洗脱。在3.5 ~ 1120 ng/mL的浓度范围内，冬冬苷的校准曲线具有良好的线性关系。日内和日间精度值均低于7%，精度范围为92.3% ~ 108.1%。基质效应在107.1% ~ 110.0%范围内，回收率超过74.2%。口服给药后，在10 mg/kg和20 mg/kg剂量下，鲁布苏苷的绝对生物利用度分别为1.00%和0.98%。这些结果证实了所建立的UPLC-MS/MS方法可以有效地支持鲁布冬苷的药动学评价

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引用次数: 0

Determination of the pyrrolo[2,3-d] pyrimidine antifolate AGF94 from a preclinical murine study using a simple UHPLC-MS/MS assay 用简单的UHPLC-MS/MS法测定临床前小鼠研究中的吡罗[2,3-d]嘧啶抗叶酸盐AGF94

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B

Pub Date : 2025-10-21 DOI: 10.1016/j.jchromb.2025.124833

Tryston T. Metz , Rachel C. Ashe , Alexis C. Duncan , Ethan C. Wilner , Keith T. Schmidt , Kristbjorn O. Gudmundsson , Zhanjun Hou , Aleem Gangjee , Larry H. Matherly , Serguei V. Kozlov , William D. Figg

A UHPLC-MS/MS method for the detection and quantification of the 6-substituted pyrrolo[2,3-d] pyrimidine AGF94, a novel antifolate, in mouse plasma and tissue was developed and validated. The developed method relies on a simple protein precipitation with methanol, followed by separation on a C18 column using a gradient solvent system of acetonitrile and water with 0.1 % formic acid in both. Detection and quantification of AGF94 were achieved by multiple reaction monitoring using a Sciex QTRAP 5500 mass spectrometer operated in positive electrospray ionization mode. The transitions for AGF94 and the internal standard were m/z 448 to 137 and 442 to 295, respectively. The calibration curve ranged from 5 to 500 ng/mL in mouse plasma with a linearity of R² = 0.99611 ± 0.00280 across multiple days. Accuracy of the assay ranged from −6.22 to 5.56 % and precision was less than 11.58 % off from nominal concentrations. Benchtop, freeze/thaw cycling, and autosampler stabilities did not indicate any substantial changes in concentrations during processing. The overall process efficiency was greater than 96 % for both the analyte and internal standard. The precision and accuracy of the assay were established, and the assay was utilized to analyze preclinical samples from a pharmacokinetic study using AGF94 in a murine pancreatic cancer model. Pharmacokinetic parameters from a noncompartmental analysis of AGF94 in multiple matrices were reported utilizing the validated method

建立了一种检测和定量小鼠血浆和组织中6-取代吡咯[2,3-d]嘧啶AGF94的UHPLC-MS/MS方法。所开发的方法依赖于用甲醇进行简单的蛋白质沉淀，然后在C18柱上使用乙腈和水的梯度溶剂系统（两者都含有0.1%甲酸）进行分离。AGF94的检测和定量采用Sciex QTRAP 5500质谱仪进行多反应监测，采用正电喷雾电离模式。AGF94和内标的转捩分别为m/z 448 ~ 137和442 ~ 295。小鼠血浆的校准曲线范围为5 ~ 500 ng/mL，在多天内线性R2 = 0.99611±0.00280。测定的准确度范围为- 6.22%至5.56%，准确度低于标称浓度11.58%。台式、冷冻/解冻循环和自动进样器稳定性未显示处理过程中浓度有任何实质性变化。分析物和内标品的总体工艺效率均大于96%。建立了该方法的精密度和准确性，并利用该方法分析了AGF94在小鼠胰腺癌模型中的药代动力学研究的临床前样品。利用验证的方法报告了AGF94在多种基质中的非区隔分析的药代动力学参数

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