Effects of voriconazole and fluconazole on the pharmacokinetics of almonertinib in rats by UPLC–MS/MS

Almonertinib was included in the first-line treatment of non-small cell lung cancer with EGFR T790M mutations by the Chinese Society of Clinical Oncology in 2021. Considering that immunocompromised lung cancer patients are prone to opportunistic fungal infections, and most triazole antifungal drugs are moderate or strong inhibitors of CYP3A4, this study was conducted to develop and validate an accurate and rapid ultra-performance liquid chromatography tandem mass spectrometry method for quantifying almonertinib in plasma and for investigating the pharmacokinetic changes of almonertinib caused by voriconazole and fluconazole in rats. After liquid–liquid extraction with tert-butyl methyl ether, an XSelect HSS T3 column (2.1 × 100 mm, 2.5 μm, Waters) was used for the chromatographic separation of almonertinib and sorafenib-D₃ (internal standard). The analytes were detected using an AB Sciex Triple Quad 5,500 mass spectrometer in the positive ionization mode. The method exhibited great linearity (0.5–200 ng/ml, r > 0.997) and stability under the established experimental conditions. All validation experiments were in accordance with the guidelines, and the results were all within the acceptable limits. This method was successfully applied to the researches of pharmacokinetics and drug interactions for almonertinib in rats. Voriconazole and fluconazole significantly altered the pharmacokinetic profiles of almonertinib and increased the systemic exposure of almonertinib in rats to different degrees, but further human trials should be conducted to validate the results.     

Pharmacokinetic studies of a novel tubulin inhibitor SK1326 in rat plasma by UPLC–MS/MS

A sensitive method for quantitation of SK1326 in rat plasma has been established using ultra-performance liquid chromatography–electrospray ionization tandem mass spectrometry (UPLC–ESI/MS/MS). SK1326 and the internal standard (tramadol) in plasma sample were extracted using acetonitrile. A centrifuged upper layer was then evaporated and reconstituted with a mobile phase of 0.5% formic acid–acetonitrile (35:65, v/v). The reconstituted samples were injected into a C₁₈ reversed-phase column. Using MS/MS in the multiple reaction monitoring mode, SK1326 and tramadol were detected without severe interference from the rat plasma matrix. SK1326 produced a protonated precursor ion ([M + H]⁺) at m/z 432.3 and a corresponding product ion at m/z 114.4. The internal standard produced a protonated precursor ion ([M + H]⁺) at m/z 264.4 and a corresponding product ion at m/z 58.1. Detection of SK1326 in rat plasma by the UPLC–ESI/MS/MS method was accurate and precise with a quantitation limit of 1.0 ng/mL. The validation, reproducibility, stability and recovery of the method were evaluated. The method has been successfully applied to pharmacokinetic studies of SK1326 in rat plasma. The pharmacokinetic parameters of SK1326 were evaluated after intravenous (at a dose of 10 mg/kg) and oral (at a dose of 20 mg/kg) administration of SK1326 in rats. After oral administration (20 mg/kg) of SK1326, the F (fraction absorbed) value was ~77.1%.     

Study of the ESI and APCI interfaces for the UPLC–MS/MS analysis of pesticides in traditional Chinese herbal medicine

In this work, 53 selected pesticides of different chemical groups were extracted from Chinese herbal medicines and determined by ultra-high-performance liquid chromatography (UHPLC)–tandem mass spectrometry (MS/MS) using both electrospray ionization (ESI) and atmospheric-pressure chemical ionization (APCI). Extracts were obtained using the acetonitrile-based quick, easy, cheap, effective, rugged, and safe (QuEChERS) sample preparation technique. Cleanup was performed by dispersive solid-phase extraction using primary secondary amine, graphitized carbon black, and octadecylsilane. Two atmospheric-pressure interfaces, ESI and APCI, were checked and compared. The validation study, including detection limits, linearity, and matrix effects, was conducted on fritillaria, radix ginseng, folium isatidis, semen persicae, and flos lonicerae in multiple reaction monitoring mode. These matrices represent a variety of plants used in traditional Chinese medicine. Fritillaria and radix ginseng were chosen as representatives for roots, folium isatidis was chosen as a representative for leaves, semen persicae was chosen as a representative for seeds, and flos lonicerae was chosen as a representative for flowers. The limits of detection for pesticides were lower in the UHPLC–ESI-MS/MS method than in the UHPLC–APCI-MS/MS method. Matrix effects on the two ionizations were evaluated for the five matrices. Soft signal enhancement in UHPLC–APCI-MS/MS and signal suppression in UHPLC–ESI-MS/MS were observed.

Quantitation by GC–MS of Methylglyoxal as a Marker in Anxiety-Related Studies

Methylglyoxal (MG) is present in the human system. In the cell, MG is degraded by glyoxalase 1 (Glo 1), an enzyme differentially expressed in several mouse models of anxiety-related behavior. It has also been associated with the formation of advanced glycation and lipoxidation end products, contributing to diabetes-related complications. Approaches developed for accurate quantitation of MG in this study include the synthesis of a deuterated MG analog to serve as an internal standard; skillful application of the standard addition protocol (with “reiteration”) to quantitate MG in “blank” samples, allowing the preparation of calibration standards using the same test specimen matrix system; and various pretreatment methods, including extraction and derivatization, to prepare complex brain tissue specimens for gas chromatography–mass spectrometry analysis. Using our method, brain MG levels in high- and low-anxiety-related behavior (HAB and LAB) mice were determined as 5.11 ± 0.89 and 13.05 ± 0.89 μg g^?1, respectively. These data supported the conclusion that MG level in brain is negatively correlated with anxiety in mouse models of anxiety-related studies, contributing to better understanding of the molecular mechanisms underlying reported observations.     

Identification of the absorptive constituents and their metabolites in vivo of Ziziphi Spinosae Semen by UPLC–ESI–Q-TOF–MS/MS

In this research, ultra-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC–ESI–Q-TOF–MS/MS) was used for detection and identification of the absorptive constituents and their metabolites in rat plasma, urine and feces following oral administration of Ziziphi Spinosae Semen alcohol extract. After structure elucidation, a total of 12 compounds in rat plasma, comprising seven prototypes and five metabolites, 28 compounds in urine, comprising 17 prototypes and 11 metabolites, and 23 compounds in feces, comrpising 17 prototypes and six metabolites, have been tentatively identified by comparison with standard compounds and reference literature information. To the best of our knowledge, this is the first comprehensive and systematical metabolic study on the seed. Mostly importantly, we propose that gastric acid could convert jujubosides into an absorbable form of ebelin lactone oligosaccharides, which may be responsible for the low bioavailability and specific bioactivities of these compounds. Additionally, we deduced that the absorption site of ebelin lactone oligosaccharides is located in the stomach, and that the ebelin lactone form of jujubosides may be more suitable for absorption than its hydrolysis product. Our investigation will be helpful to narrow the scope for potentially active ingredients of the seed, and pave the way for determination of the pharmacological mechanism of the seed.     

超声辅助分散液液微萃取/超高效液相色谱-串联质谱法检测水中多种磺胺类抗生素残留

建立了水中13种磺胺类抗生素的超声辅助分散液液微萃取/超高效液相色谱-串联质谱(UA-DLLME/UPLC-MS/MS)测定方法。以乙腈为分散剂,四氯乙烷为萃取剂,通过超声分散方式协同萃取水样中的目标化合物,采用C18色谱柱(100mm×2.1mm,1.7μm)分离,以乙腈-0.1%甲酸溶液为流动相,采用UPLC-MS/MS多反应监测模式(MRM)测定。优化了萃取剂和分散剂的类型和用量、萃取时间、氯化钠浓度和pH值等条件。在优化条件下,13种磺胺类抗生素在一定质量浓度范围内线性关系良好,相关系数均大于0.998,方法检出限(S/N＝3)为0.6~2.4ng/L;3个加标水平的平均回收率为80.3%~101.8%,相对标准偏差(RSD,n＝5)为0.7%~4.5%。该方法前处理简单、环保、灵敏,适用于水样中多种磺胺类抗生素的测定。     

超高效液相色谱-电喷雾串联质谱法测定饲料中残留的三聚氰胺

饲料样品经1%三氯乙酸-二甲基亚砜提取,Waters Oasis MCX柱净化,超高效液相色谱分离,最终采用电喷雾串联四极杆质谱进行检测。结果表明,三聚氰胺在饲料中的含量范围为10～5000 μg/kg时,线性关系良好(r＞0.99)。在10～100 μg/kg 的添加水平范围内的平均回收率为83%～94%,相对标准偏差为4.2%～6.5%。该方法的检出限为10 μg/kg。     

UPLC-MS/MS法测定中药材中齐墩果酸与熊果酸的含量

建立了一种同时检测中药材中齐墩果酸（OA）和熊果酸（UA）含量的超高效液相色谱串联质谱方法。采用超高效液相色谱-三级四极杆质谱(UPLC-TQMS)法对样品进行测定,WatersAcquityUPLCBEHC18色谱柱(50mm×2.1mm,1.7μm)进行分离,以乙腈-5mmol/L乙酸铵水溶液(氨水调至pH9.24)为流动相,梯度洗脱;负离子模式下检测。结果表明,齐墩果酸和熊果酸在0.5~50.0ng/mL浓度范围内线性关系良好,相关系数(r2)分别为0.9998和0.9997;检出限(S/N=3)分别为0.0066,0.0128ng/mL,定量下限(S/N=10)分别为0.0020,0.0038ng/mL;对OA和UA进行加标回收实验,平均回收率分别为101.1%和100.8%,相对标准偏差(RSD,n=9)分别为1.8%和0.04%。对10种不同中药材中齐墩果酸和熊果酸含量进行检测,结果表明该方法快速简便、准确度高、重现性好,可用于含有齐墩果酸和熊果酸的中药材含量测定。     

超高效液相色谱-串联质谱法测定鸡血浆中利巴韦林及其主要代谢物

建立了同时检测血浆中利巴韦林(Ribavirin)及其主要代谢物1H-1,2,4-三氮-3-甲酰胺(TCONH2)和1-β-D-呋喃基核糖三氮唑-3-羧酸(RTCOOH)的超高效液相色谱-串联质谱(UPLC-MS/MS)分析方法。鸡血浆样品加入13C5利巴韦林内标,以甲醇沉淀蛋白,经ZORBAXSB-Aq(100mm×3mm,1.8μm)色谱柱分离,以0.1%甲酸水-甲醇为流动相,梯度洗脱,采用正离子多反应监测模式(MRM),内标法定量。结果表明：利巴韦林、TCONH2与RTCOOH的线性范围分别为5~5000,5~5000,50~5000μg/L,相关系数(r2)分别为0.9947,0.9992,0.9976。质控样品的加标回收率为84.1%~108.2%,批内相对标准偏差(RSD,n=6)为1.6%~13.4%,批间RSD(n=12)为3.8%~16.9%。样品的检出限(S/N=3)为2~10μg/L。本方法快速简便,有效排除了内源性干扰,方法的线性范围宽,重复性好,适用于血浆样品中利巴韦林及主要代谢产物的测定。     

分散固相萃取净化超高效液相色谱串联质谱法研究茶叶与茶汤中茚虫威残留降解规律

利用分散固相萃取净化茶叶、固相萃取富集净化茶汤,超高效液相色谱串联质谱法测定,建立了茶叶和茶汤中茚虫威残留的分析方法,并研究了150g/L茚虫威乳油在茶园茶叶中的残留降解规律,对茚虫威在茶叶-茶汤过程中的浸出率和饮用安全性进行风险评估。结果表明：茚虫威在0.01～20.0mg/L浓度范围内呈线性,其相关系数(r)为0.9937,仪器检出限(LOD)为0.025ng,在高、中、低3种水平下的平均加标回收率为96%～107%,RSD(n=6)为1.3%～7.8%,茚虫威在茶鲜叶、成茶中的定量下限(S/N=10)为5μg/kg,茶汤中的定量下限为0.10μg/L;150g/L茚虫威乳油在茶园鲜叶上的半衰期为1.42～2.47d;茚虫威在成茶-茶汤过程中的3次总浸出率小于10%,茶汤中茚虫威残留量及浸出率随冲泡次数的增加逐渐下降,按照风险评估制定茶叶中茚虫威残留量MRL值为3mg/kg是安全的,人体摄入的茚虫威残留量仅占每日允许摄入量(ADI)的0.619%。     

高效液相色谱-串联质谱法测定动物组织中的16种喹诺酮类药物残留

建立了动物组织样品中萘啶酸、恶喹酸、氟甲喹、诺氟沙星、依诺沙星、环丙沙星、洛美沙星、丹诺沙星、恩诺沙星、氧氟沙星、沙拉沙星、二氟沙星、麻保沙星、培氟沙星、司帕沙星、奥比沙星等16种喹诺酮类兽药多残留量的高效液相色谱-串联质谱测定方法。用酸性乙腈萃取样品中的16种喹诺酮类药物残留,然后用正己烷脱脂,旋转蒸发浓缩,以Inertsil C8-3色谱柱分离,在正离子模式下以电喷雾电离串联质谱进行测定。在10,50,100 μg/kg 3个加标水平下进行了验证试验,方法的线性范围为10~100 μg/kg,平均回收率为62.4%~102%,相对标准偏差为1.4%~11.9%。该方法简便、快速、准确,各项技术指标满足国内外法规的要求,可用于鸡肉、鸡肝和鱼肉等动物组织样品中喹诺酮类药物多残留的确证检测。     

UPLC-MS/MS法对动物源性食品中12种大环内酯类抗生素残留的测定

建立了超高效液相色谱-串联质谱(UPLC-MS/MS)法测定动物源性食品中12种大环内酯类抗生素(林可霉素、阿奇霉素、螺旋霉素、替米考星、竹桃霉素、红霉素、泰乐霉素、吉他霉素、罗红霉素、克拉霉素、麦迪霉素、交沙霉素)的方法.样品均质后,用乙腈提取,正己烷净化,无水硫酸钠脱水.乙腈提取液减压浓缩后,氮气流吹干,甲醇溶解定容;采用UPLC-MS/MS电喷雾多反应监测模式检测,基质匹配标准曲线定量.实验结果表明,12种大环内酯化合物在5 ～100 μg/kg范围内线性关系良好,检出限均为5.0 μg/kg,定量下限为10 μg/kg.5种空白基质样品中,10、25、50 μg/kg加标水平的平均回收率为60% ～117%,相对标准偏差均在20%以内.该方法灵敏度高、重复性好,各项技术指标均满足国内外相关法规要求,可用于动物源性食品中12种大环内酯类抗生素残留的检测.     

基于UPLC-MS/MS的2型糖尿病患者血清胆汁酸代谢产物分析

应用基于超高效液相色谱-串联质谱(UPLC-MS/MS)技术的代谢组学方法检测2型糖尿病患者血清胆汁酸代谢谱,获得代谢标志物,发掘可预测糖尿病的潜在预防治疗靶点。采用WatersACQUITYUPLCBEHC18色谱柱(150mm×2.1mm,1.7μm)分离,0.05%乙酸水溶液-乙腈为流动相,以0.40mL/min流速进行梯度洗脱,用此方法测定了30例糖尿病患者和50例正常人血清胆汁酸代谢谱。41种胆汁酸标准品在30min内均得到良好分离,各成分在一定浓度范围内呈良好线性关系,检出限(LOD)为0.24~7.81nmol/L,定量下限(LOQ)为0.49~15.63nmol/L,相关系数(r2)为0.9867~0.9997。血清中胆汁酸代谢谱与两组间存在相关性,通过正交偏最小二乘法-判别分析(OPLS-DA)发现,有12种胆汁酸(包括甘氨熊脱氧胆酸、甘氨胆酸、牛磺鹅脱氧胆酸、胆酸、熊去氧胆酸、甘氨鹅脱氧胆酸、甘氨脱氧胆酸、12酮基石胆酸、鹅脱氧胆酸、脱氧胆酸、异石胆酸、石胆酸)承载了分组的重要信息。结合受试者工作特征(ROC)分析,最终获得牛磺鹅脱氧胆酸(AUC=0.68)、甘氨鹅脱氧胆酸(AUC=0.66)、甘氨熊脱氧胆酸(AUC=0.63)、胆酸(AUC=0.63)、脱氧胆酸(AUC=0.63)、甘氨脱氧胆酸(AUC=0.58)6种胆汁酸代谢标志物,对于糖尿病的早期诊断及深入研究具有潜在的科研及临床价值。     

抗阿尔兹海默症藏药三味豆蔻汤的化学成分分析研究

三味豆蔻汤作为藏药的经典方剂,由药材豆蔻、香旱芹、荜茇和牦牛奶熬煮制成,其对阿尔兹海默症(Alzheimer′sdisease,AD)具有一定的治疗作用。该方剂化学成分复杂且极性、挥发性差异大,相关的物质基础研究较少。该研究分别采用气相色谱-质谱(GC-MS)、超高效液相色谱-高分辨串联质谱(UPLC-HRMS/MS)技术针对三味豆蔻汤中不同极性组分进行检测分析,以了解其物质组成。结果显示：从三味豆蔻汤中共发现并鉴定出99个化学成分,分别为豆蔻和香旱芹中含有的含氧单萜和单萜烯类成分(如桉油精)、荜茇的生物碱类成分(如胡椒碱)以及牦牛奶含有的脂类成分(如甘油三酯等)。该结果可为了解三味豆蔻汤的药效物质基础提供重要依据。