Screening of thrombin inhibitors from phenolic acids using enzyme-immobilized magnetic beads through direct covalent binding by ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry

Cao, Jun; Xu, Jingjing; Liu, Xun-Gao; Wang, Shuling; Peng, Li-Qing

doi:10.1016/j.chroma.2016.09.022

Cited by 36 publications

(18 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Among the identified constituents, danshensu, salvianolic acid B, salvianolic acid C, rosmarinic acid produced good thrombin inhibitions, while ononin and danshensu showed ACE inhibitions. Some activities to the enzymes are reported for the first time, and the present finding indicates that direct enzyme inhibition is possibly one of the mechanisms responsible for their pharmacological effects, such as platelet aggregation inhibition and thrombin time reduction 42 – 44 . Especially, salvianolic acid B and rosmarinic acid exhibit potent thrombin inhibitory activities with IC 50 of 81.9 and 92.0 μM, the latter of which is also similar to the value reported by a previous study 45 .…”

Section: Discussionmentioning

confidence: 57%

A multiple biomarker assay for quality assessment of botanical drugs using a versatile microfluidic chip

et al. 2017

Sci Rep

View full text Add to dashboard Cite

Quality control is critical for ensuring the safety and effectiveness of drugs. Current quality control method for botanical drugs is mainly based on chemical testing. However, chemical testing alone may not be sufficient as it may not capture all constituents of botanical drugs. Therefore, it is necessary to establish a bioassay correlating with the drug’s known mechanism of action to ensure its potency and activity. Herein we developed a multiple biomarker assay to assess the quality of botanicals using microfluidics, where enzyme inhibition was employed to indicate the drug’s activity and thereby evaluate biological consistency. This approach was exemplified on QiShenYiQi Pills using thrombin and angiotensin converting enzyme as “quality biomarkers”. Our results demonstrated that there existed variations in potency across different batches of the intermediates and preparations. Compared with chromatographic fingerprinting, the bioassay provided better discrimination ability for some abnormal samples. Moreover, the chip could function as “affinity chromatography” to identify bioactive phytochemicals bound to the enzymes. This work proposed a multiple-biomarker strategy for quality assessment of botanical drugs, while demonstrating for the first time the feasibility of microfluidics in this field.

show abstract

Section: Discussionmentioning

confidence: 57%

A multiple biomarker assay for quality assessment of botanical drugs using a versatile microfluidic chip

et al. 2017

Sci Rep

View full text Add to dashboard Cite

show abstract

“…However, ultrafiltration and centrifugation can disrupt the weak binding between small molecule compounds and their macromolecular binding partners. In recent years, enzyme immobilization has been used for protein purification and enzyme inhibitor screening . Magnetic beads (MBs) are considered ideal solid support for enzyme immobilization due to their high operational stability , low nonspecific binding, and convenience of magnetic separation .…”

Section: Introductionmentioning

confidence: 99%

Screening inhibitors of xanthine oxidase from natural products using enzyme immobilized magnetic beads by high‐performance liquid chromatography coupled with tandem mass spectrometry

Wang

et al. 2017

J of Separation Science

View full text Add to dashboard Cite

In this study, high-performance liquid chromatography coupled with tandem mass spectrometry was used to assess the results of bioactive compound screening from natural products using immobilized enzyme magnetic beads. We compared three commercial magnetic beads with modified amino, carboxy, and N-hydroxysuccinimide groups, respectively. Amino magnetic beads performed best for immobilization and were selected for further experiments. Xanthine oxidase was immobilized on amino magnetic beads and applied to screen potential inhibitors in fresh Zingiber officinale Roscoe, extracts of Scutellaria baicalensis Georgi, and Pueraria lobata Ohwi. In total, 12 potential xanthine oxidase ligands were identified from fresh Zingiber root and Scutellaria root extracts, of which eight were characterized and the concentration required for 50% inhibition was determined. Preliminary structure-function relationships were discussed based on these results. A convenient and effective method was therefore developed for the identification of active compounds from complex natural product mixtures.

show abstract

“…In order to obtain the best screening performance for active compounds in the complex matrix, some important parameters of the method including incubation time, TYR concentration, and sample concentration should be optimized. According to previous research, the incubation time might be controlled at the range of 30–120 min because a short incubation time (less than 30 min) might prevent the identification of target molecules which are not firmly bound to TYR, while a long incubation time (about 120 minutes) would not make significant influence on the screening result [ 24 , 25 ]. After investigation, a proper TYR concentration (800 U/mL) increased the sensitivity and number of bioactive constituents detected in the sample; meanwhile, the inhibition effect of tyrosinase is most stable by the incubation time of 40 min (data not shown).…”

Section: Resultsmentioning

confidence: 99%

Screening and Characterizing Tyrosinase Inhibitors fromSalvia miltiorrhizaandCarthamus tinctoriusby Spectrum-Effect Relationship Analysis and Molecular Docking

Wang

Zhang

et al. 2018

Journal of Analytical Methods in Chemistry

View full text Add to dashboard Cite

Tyrosinase (TYR) is a rate-limiting enzyme in the synthesis of melanin, while direct TYR inhibitors are a class of important clinical antimelanoma drugs. This study established a spectrum-effect relationship analysis method and high-performance liquid chromatography-mass spectrometry (LC-MS) analysis method to screen and identify the active ingredients that inhibited TYR in Salvia miltiorrhiza–Carthamus tinctorius (Danshen–Honghua, DH) herbal pair. Seventeen potential active compounds (peaks) in the extract of DH herbal pair were predicted, and thirteen of them were tentatively identified by LC-MS analysis. Furthermore, TYR inhibitory activities of five pure compounds obtained from the DH herbal pair were validated in the test in which kojic acid served as a positive control drug. Among them, three compounds including protocatechuic aldehyde, hydroxysafflor yellow A, and tanshinone IIA were verified to have high TYR inhibitory activity (IC50 value of 455, 498, and 1214 μM, resp.) and bind to the same amino acid residues in TYR catalytic pocket according to the results of the molecular docking test. However, the other two compounds lithospermic acid and salvianolic acid A had a weak effect on TYR, as they do not combine with the active amino acid residues or act on the active center of TYR. Therefore, the developed methods (spectrum-effect relationship analysis and molecular docking) could be used to effectively screen TYR inhibitors in complex mixtures such as natural products.

show abstract

Screening of thrombin inhibitors from phenolic acids using enzyme-immobilized magnetic beads through direct covalent binding by ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry

Cited by 36 publications

References 28 publications

A multiple biomarker assay for quality assessment of botanical drugs using a versatile microfluidic chip

A multiple biomarker assay for quality assessment of botanical drugs using a versatile microfluidic chip

Screening inhibitors of xanthine oxidase from natural products using enzyme immobilized magnetic beads by high‐performance liquid chromatography coupled with tandem mass spectrometry

Screening and Characterizing Tyrosinase Inhibitors fromSalvia miltiorrhizaandCarthamus tinctoriusby Spectrum-Effect Relationship Analysis and Molecular Docking

Contact Info

Product

Resources

About