THERAPEUTIC DRUG MONITORING
·
2025
·
IF 2.4
Dong Zhonghua,Wang Peng,Cui Xueyan,Liu Jinjuan,Shi Haiyan,Yang Yilei,Xu Jingya,Yi Qiaoyan
Background: Perampanel (PER), a novel antiepileptic drug, exhibits high protein binding. Given the drug's susceptibility to various factors, including drug interactions, and the considerable variability in blood concentrations among individuals, solely measuring the total plasma concentration of PER may not provide comprehensive insights. This study aimed to establish an HPLC-FLD method to quantify both total and free PER in clinical samples. Methods: Analysis of total PER involved a straightforward sample preparation process involving plasma protein precipitation. Plasma samples were ultrafiltered to isolate the free portion of PER. Chromatographic separation was achieved on an InertSustain C18 column at a 1-mL/min flow rate using a gradient of acetonitrile and aqueous sodium acetate buffer at pH 4.4. Results: Calibration curves for total and free PER in plasma exhibited excellent linearity over the concentration ranges of 10–3000 and 0.5–100 ng/mL, respectively. The method was applied to analyze blood samples from patients with epilepsy quantitatively; the total concentration of perampanel in the plasma of the patients was in the range of 90.45–563.39 ng/mL, whereas the free concentration was in the range of 1.82–15.29 ng/mL. The plasma protein binding ratio of perampanel was normally between 97.09% and 99.29% and decreased in cases of hypoproteinemia. Conclusions: The method was rigorously validated for selectivity, accuracy, precision, and stability in accordance with established FDA and EMA guidelines. The developed method enables the rapid and accurate quantification of both total and free PER concentrations in the plasma of patients with epilepsy, offering technical support for its subsequent clinical application.
研究领域:
临床药理学
·
分析化学
·
神经病学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
THERAPEUTIC DRUG MONITORING
·
2015
·Vol. 37
·期 6
·776
·
IF 2.376
Zhang Zhi-qing,Dong Wei-chong,Yang Xiu-ling,Zhang Jin-feng,Jiang Xin-Hui,Jing Shao-jun,Yang Hong-le,Jiang Ye
Background: Free drug analysis is increasingly becoming popular in therapeutic drug monitoring (TDM). Centrifugal ultrafiltration (CF-UF) is the primary method to separate free drug from that of bound drug. However, the volume ratio of ultrafiltrate to sample solution ( V u / V s ) affects the accuracy of CF-UF, which highly depends on the different plasma conditions. Plasma protein concentrations in patients are different from those observed in healthy subjects, and there are also significant differences among patients with different diseases. Only very few studies have reported on the effect of protein concentration on the analysis methodology of free drug by CF-UF. Methods: In this study, valproic acid was used as the representative drug, and plasma samples with different albumin concentrations were analyzed by CF-UF and hollow fiber centrifugal ultrafiltration (HFCF-UF). Results: There was no significant difference of free drug concentrations by HFCF-UF and CF-UF when plasma albumin concentrations ranged 40–60 g/L. However, at low albumin concentrations (<40 g/L), a considerable difference was detected, and the difference was increased with the decrease of plasma albumin concentration. When the albumin concentration was as low as 10 g/L, the free drug concentration was 17.3 mcg/mL by CF-UF, whereas it was 10.2 mcg/mL by HFCF-UF. Conclusions: The accuracy of free drug measurement by CF-UF was albumin concentration dependent. However, such an effect was not observed when samples were prepared by HFCF-UF, which was more suitable for TDM of plasma samples from different patients. Therefore, this method could be readily applied to the measurement of free valproic acid plasma concentrations for TDM in patients.
研究领域:
药理学
·
临床医学
✓ 已验证
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Binghong Gao,Chuanyan Rao,Xingmeng Lei,Siqi Li,Xiaochuan Huang,Yanlin Liu,Dongqing Ye
This study investigated the effects of molecular weight regulation on mannoproteins (MPs) in encapsulating both monomeric and oligomeric proanthocyanidins (MOPC). To achieve this, two different conformations of MPs were fractionated by ultrafiltration into two main molecular weight components. The results indicated that regulating molecular weight through ultrafiltration altered the conformation of MPs chains, which in turn affected the intermolecular forces with MOPC. Specifically, the interactions among the different molecular weight components of MPs formed a cross-linking network. High molecular weight components provided external support to this network, while low molecular weight components contributed to internal stabilization. The dense, flexible, and irregular coil structure of MPs facilitated the entry of MOPC into the internal spaces of the cross-linking network. Additionally, protein unfolding into a more stable β-sheet structure further stabilized MPs' cross-linking network, enhancing its capacity to encapsulate MOPC. The presence of more hydrophobic regions and positive charges within the proteins increased the number of binding sites for MOPC. This study aimed to identify the primary structural forms of MPs with potent initial bitter inhibition capabilities on MOPC, and a proposed ultrafiltration method was introduced to modify the triple helix structure of polysaccharide, offering a foundational theoretical framework for enhancing the functional attributes of current or prospective MPs. Additionally, the mixed regulation of polysaccharides with varying conformations was also beneficial for guiding the production of high-efficiency polysaccharide products.
研究领域:
食品科学
·
生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SCIENCE OF THE TOTAL ENVIRONMENT
·
2019
·
IF 5.589
Yuxi Lu,Xuelu Gao,Jinming Song,Chen-Tung Arthur Chen,Jinling Chu
The environmental characteristics of colloidal toxic trace metals Cd, Cu and Pb in riverine and estuarine waters collected from two urban rivers of Yantai City in eastern China, the Guangdang and Xin'an Rivers, were investigated using a modified centrifugal ultrafiltration (CUF) method in conjunction with acid extraction and inductively coupled plasma mass spectrometry. The target metals in dissolved pool were divided into four CUF fractions, i.e. <1 kDa, 1–3 kDa, 3–10 kDa and 10 kDa–0.2 μm, and the results showed that colloidal Cd, Cu and Pb were dominated by 1–10 kDa (1–3 and 3–10 kDa), 1–3 kDa and 10 kDa–0.2 μm fractions, respectively. The coagulation/flocculation of low-molecular-weight (1–10 kDa) colloidal Cd and Cu in the estuaries was obvious and strong, while the enrichment of dissolved Pb in the 10 kDa–0.2 μm fraction may be mainly related to its biogeochemical interactions with Fe-oxides, which is easy to occur in macromolecular colloids. In addition, the actual molecular weight cutoffs (MWCOs) of the three used CUF units with nominal MWCOs of 1, 3 and 10 kDa were determined to be 4.9, 8.5 and 33.9 kDa, respectively, indicating that membrane calibration is essential for explaining the actual fraction of dissolved trace metals and verifying the integrity of ultrafiltration membrane. Overall, the results in this study provide a further understanding of the heterogeneity in biogeochemical features, migration and fate of toxic trace metals in aquatic ecosystems, especially that of the river-sea mixing zone.
研究领域:
环境科学
·
海洋学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Frontiers in Plant Science
·
2017
·
IF 4.291
Chen Guilin,Guo Mingquan
Topoisomerase I (Topo I) catalyzes topological interconversion of duplex DNA during DNA replication and transcription, and has been deemed as important antineoplastic targets. In this study, the fraction R.d-60 from ethyl acetate extracts of Rhamnus davurica showed higher inhibitory rates against SGC-7901 and HT-29 compared with the R.d-30 fraction in vitro. However, the specific active components of R.d-60 fraction remain elusive. To this end, a method based on bio-affinity ultrafiltration and high performance liquid chromatography/electrospray mass spectrometry (HPLC- ESI-MS/MS) was developed to rapidly screen and identify the Topo I inhibitors in this fraction. The enrichment factors (EFs) were calculated to evaluate the binding affinities between the bioactive constituents and Topo I. As a result, eight ligands were identified and six of which with higher EFs showed more potential antitumor activity. Furthermore, antiproliferative assays in vitro (IC50 values) with two representative candidates (apigenin, quercetin) against SGC-7901, HT-29 and Hep G2 cells were conducted and further validated. Finally, the structure-activity relationships revealed that flavones contain a C2-C3 double bond of C ring exhibited higher bio-affinities to Topo I than those without it. This integrated method combining Topo I ultrafiltration with HPLC-MS/MS proved to be very efficient in rapid screening and identification of potential Topo I inhibitors from the complex extracts of medicinal plants, and could be further explored as a valuable high-throughput screening platform in the early drug discovery stage.
研究领域:
药理学
·
分析化学
·
肿瘤学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2018
·Vol. 410
·期 18
·4311-4322
·
IF 3.307
Wu Si-Qi,Song Hui-Peng,Li Bin,Liu Run-Zhou,Yang Hua,He Ling,Li Ping
Development of fast and accurate methods to discover lead compounds for drug candidates is highly important. In this study, a reliable and effective post-column on-flow biochemical assay (POBA) was established to screen potent peroxidase inhibitors from complex chemical mixtures (e.g., natural product extracts). Multiple factors such as flow rate, organic phase, detection wavelength, and reaction coil were carefully investigated. To better understand the features of POBA, another emerging technology of ultrafiltration LC-MS was used for comparison. The result showed that POBA had advantages in saving time, avoiding false positives, and improving the accuracy. To illustrate the practicality of the method, Radix Salvia Miltiorrhizae, a traditional herb for cardiovascular disease treatment, was applied as the research objective. Finally, six compounds including tanshinol, protocatechuic aldehyde, salvianolic acid D, rosmarinic acid, lithospermic acid, and salvianolic acid B were determined as novel peroxidase inhibitors. Their bioactivities were validated by microplate-based assay, molecular docking, and pharmacophore modeling. This study demonstrates a great potential of POBA in the efficient and accurate discovery of drug candidates.
Compared with a classical method of ultrafiltration LC-MS, the newly developed method of on-flow bioassay shows advantages in saving time, avoiding false positives and improving the accuracy
研究领域:
药理学
·
分析化学
·
天然产物研究
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Central South University
·
2014
·Vol. 21
·期 7
·2645-2651
·
IF 0.464
Zhao Jin-yao,Li Li,Jiao Fei-peng,Ren Feng-lian
The human plasma protein binding of water soluble flavonoids in the peels of five spices of citrus fruits was studied by ultrafiltration combined with HPLC. The flavonoids were extracted separately by hot and cold water, and higher total flavonoid contents were detected in the former extracts than the latter ones. All the extracts show significant scavenging abilities to both ABTS and DPPH free radicals, which indicates the health benefits of the water extracts of citrus fruits peels. For DPPH radical, the IC 50 values of hot extract follow as Navel orange (NO)≈Mandarin orange (MO)< Lemon (LE)< Lo tangerine (LO)< Pomelo (PO), while the rank is NO< PO
研究领域:
食品科学
·
药理学
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
Frontiers in Plant Science
·
2022
·
IF 6.627
Zhu Lian,Ma Sheng-jun,Liu Ming-juan,Li Kai-lin,E Shuai,Wang Zi-ming,Li Sha-ni,Zhang Sheng-lan,Cai Wei
Arnebiae Radix (dried root of Arnebia euchroma (Royle) Johnst.) is a traditional Chinese medicine (TCM) used to treat macular eruptions, measles, sore throat, carbuncles, burns, skin ulcers, and inflammations. The Arnebiae Radix extract can exert anti-breast cancer effects through various mechanisms of action. This study aimed to rapidly screen potential estrogen receptor (estrogen receptor α and estrogen receptor β) ligands from the Arnebiae Radix extract. In this study, an analytical method based on affinity ultrafiltration coupled with UHPLC-Q-Exactive Orbitrap mass spectrometry was established for rapidly screening and identifying estrogen receptor ligands. Then, bindings of the components to the active site of estrogen receptor (estrogen receptor α and estrogen receptor β) were investigated via molecular docking. Moreover, surface plasmon resonance (SPR) experiments with six compounds were performed to verify the affinity. As a result, a total of 21 ligands were screened from Arnebiae Radix using affinity ultrafiltration. Among them, 14 and 10 compounds from Arnebiae Radix showed affinity with estrogen receptor α and estrogen receptor β, respectively. All of those ligands could have a good affinity for the multiple amino acid residues of the estrogen receptor based on molecular docking. In addition, six compounds display the great affinity by SPR. The method established in the study could be used to rapidly screen estrogen receptor ligands in Traditional Chinese medicine. The results demonstrated that the affinity ultrafiltration–UHPLC-Q-Exactive Orbitrap mass spectrometry method not only aids in the interpretation of the potential bioactive components and possible mechanisms of action of Arnebiae Radix but also provides a further effective basis for the quality control of this valuable herb medicine.
研究领域:
中药学
·
生物化学
·
药物化学
✓ 已验证
DOI
PubMed
高影响因子
MOLECULES
·
2021
·Vol. 26
·期 9
·2467
·
IF 4.412
Yueqi Wang,Liping Guo,Chunming Liu,Yuchi Zhang,Sainan Li
Extracts of the fungusInonotus obliquusexhibit cytotoxic properties against different cancers; hence, this fungal species has been extensively studied. This study aimed to extract total triterpenoids fromInonotus obliquususing ionic liquids (ILs) and separate potential lactate dehydrogenase (LDH) inhibitors via ultrafiltration (UF)-high-speed countercurrent chromatography (HSCCC). Total triterpenoids fromInonotus obliquuswere extracted by performing a single-factor experiment and employing a central composite design via ultrasonic-assisted extraction (UAE) and heat-assisted extraction (HAE). The extract was composed of 1-butyl-3-methylimidazolium bromide as the IL and methanol as the dispersant. Ultrafiltration-liquid chromatography (UF-LC) was used to rapidly scan the LDH inhibitors and betulin and lanosterol were identified as potential inhibitors. To obtain these target compounds, betulin and lanosterol with the purities of 95.9% and 97.8% were isolated from HSCCC within 120 min. Their structures were identified using several techniques, among which IL-HAE was fast and effective. This study reports the extraction of triterpenoids fromInonotus obliquusby IL for the first time. Collectively, the findings demonstrate that UF-LC is an effective tool for screening potential LDH inhibitors from crude extracts ofI. obliquusand may help to identify bioactive substances against myocardial infarction, whereas high-purity compounds can be separated via UF-HSCCC.Keywords:Inonotus obliquus;ultrafiltration;liquid chromatography;ultrasonic-assisted extraction;lactate dehydrogenase inhibitor;myocardial infarction
研究领域:
药理学
·
分析化学
·
中药学
✓ 已验证
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2021
·
IF 7.514
Songjie Li,Ruimin Wang,Xiaoping Hu,Congfa Li,Lu Wang
Cerasus humilis (Bge.) Sok. leaf-tea (CLT) has a potential anti- α -glucosidase effect. However, its anti- α -glucosidase functional compositions remain unclear. Results showed that 70% methanol extract of CLT (IC 50 = 36.57 μg/mL) with the highest total phenolic/flavonoid contents exhibited significantly higher α -glucosidase inhibitory activity ( α -GIA) than acarbose (IC 50 = 189.57 μg/mL). Additionally, phenolic constituents of the CLT extract were analyzed for the first time in this work. Ten major potential α -glucosidase inhibitors ( α -GIs) with high bio-affinity degree in the CLT extract were recognized using a bio-affinity ultra-filtration and HPLC-ESI-qTOF-MS/MS method. In vitro α -GIA assay confirmed that myricetin (IC 50 = 36.17 μg/mL), avicularin (IC 50 = 69.84 μg/mL), quercitrin , isoquercitrin, prunin and guajavarin were responsible for the α -GIA of the CLT extract. More importantly, the interaction mechanism between α -GIs and α -glucosidase was investigated via in silico analysis. This study provides a high-throughput screening platform for identification of the potential α -GIs from natural products.
研究领域:
植物化学
·
药理学
·
分析化学
✓ 已验证
DOI
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2011
·
IF 2.733
Shu Liu,Jun Yan,Junpeng Xing,Fengrui Song,Zhiqiang Liu,Shuying Liu
The liquid chromatography–tandem mass spectrometry (LC–MS/MS) methods were used to identify the pharmacologically active n-butanol extract from Compound Indigowoad Root Granule . As a result, eighteen compounds belonging to various structural classes such as nucleosides, purines, flavonoids and amino acid were unambiguously identified. Then an in vitro neuraminidase (NA) inhibition assay was carried out to examine the inhibitory activity of the standard samples and extracts on NA. After which, ultrafiltration liquid chromatography with photodiode array detection coupled to electrospray ionization tandem mass spectrometry (ultrafiltration LC–MS/MS) was used to study NA inhibitory activity of standard flavones and n-butanol extract of Compound Indigowoad Root Granule. This method is highly selective and sensitive, and it could be used for characterization of bioactive compounds and botanical extracts. The result provides some enlightenment for the explanation of the antiviral activity of Compound Indigowoad Root Granule and some guidance for natural anti-influenza medicine development.
研究领域:
药理学
·
分析化学
·
病毒学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF SEPARATION SCIENCE
·
2017
·Vol. 40
·期 12
·2565-2574
·
IF 2.557
Senlin Li,Sainan Li,Yu Huang,Chunming Liu,Lina Chen,Yuchi Zhang
The separation of a compound of interest from its structurally similar homologues to produce high-purity natural products is a challenging problem. This work proposes a novel method for the separation of iristectorigenin A from its structurally similar homologues by ionic-liquid-based ultrasound-assisted extraction and the subsequent screening and isolation of potential α-glucosidase inhibitors via ultrafiltration and semipreparative high-performance liquid chromatography. Ionic-liquid-based ultrasound-assisted extraction was successfully applied to the extraction of tectorigenin, iristectorigenin A, irigenin, and irisflorentin from Belamcanda chinensis . The optimum conditions for the efficient extraction of isoflavones were determined as 1.0 M 1-ethyl-3-methylimidazolium tetrafluoroborate with extraction time of 30 min and a solvent to solid ratio of 30 mL/g. Ultrafiltration with liquid chromatography and mass spectrometry was applied to screen and identify α-glucosidase inhibitors from B. chinensis , followed by the application of semipreparative high-performance liquid chromatography to separate and isolate the active constituents. Four major compounds including tectorigenin, iristectorigenin A, irigenin, and irisflorentin were screened and identified as α-glucosidase inhibitors, and then the four active compounds abovementioned were subsequently isolated by semipreparative high-performance liquid chromatography (99.89, 88.97, 99.79, and 99.97% purity, respectively). The results demonstrate that ionic liquid extraction can be successfully applied to the extraction of isoflavones from B. chinensis .
研究领域:
天然产物化学
·
药理学
·
分析化学
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）