Mass Spectrometry-Sensitive Probes Coupled with Direct Analysis in Real Time for Simultaneous Sensing of Chemical and Biological Properties of Botanical Drugs

Abstract

The development of botanical materials as therapeutic agents involves the meticulous assessment of safety, efficacy, and quality. Compared with small-molecule drugs, quality control of botanical drugs confronts with more significant challenges due to their inherent complexity. Current quality control methods for botanical drugs, either prevailing chemical tests or emerging biological assays, are not able to meet recent demands of multiplexing, sensitivity, and speed.Here, we propose an on-demand strategy based on a direct analysis in real time-mass spectrometry (DART-MS) platform, which is capable of simultaneously analyzing multiple constituents and bioactivities of botanical drugs. Notably, the bioactivities are assessed by a multiple-enzyme assay that adopts cleavable mass spectrometry probes as enzymatic substrates: these probes labeled with a piperazine tag make possible sensitive, multiplexed, and quantitative enzyme activity measurements. The concept is successfully demonstrated via a case study of Danshen (Salvia miltiorrhiza) Injection where simultaneous detection of 34 constituents and inhibitory activities on two target enzymes can be achieved in just minutes. This proof-of-concept application also gives evidence that combining MSsensitive probes with DART-MS can provide an environmentally friendly, highly sensitive analytical approach for botanical quality control.

Screening Immunoactive Compounds of Ganoderma lucidum Spores by Mass Spectrometry Molecular Networking Combined With in vivo Zebrafish Assays

“…These herbal constituents generate thousands of molecular ions and fragment ions in MS analysis, rendering it challenging to annotate the detected chemical signatures. To address this issue, many MS data processing strategies have been developed to accelerate the dereplication and discovery process (Wang et al, 2016b(Wang et al, , 2019Li et al, 2017Li et al, , 2019. Among these approaches, molecular networking (Watrous et al, 2012) is an emerging tool well suited to this task.…”

Emerging biotechnology applications in natural product and synthetic pharmaceutical analyses

“… Category Technique Response Application Ref. Biological-chemical-coupling technology DART-MS-enzyme Chemical components and biological activities Danshen injection 61 ABTS ± CE Major antioxidants in injection Shuxuening injection 62 Metabolomics-chemometrics Bioactive-chemical quality marker combination Pollen of T. orientalis 63 Biosensors for PAT Enzyme based SPR Acceptable therapeutic monitoring Bromocriptine 64 HF-AS Lipase-binding ligands Lotus leaf extracts 65 TPE-fluorescent probe Monitor ACE activity Tongmaiyangxin pill 66 FRET fluorescent acceptor Patulin and zearalenone Extracts of TCM 67 Aptamer biosensor Rapid detection of pathogenic E. coli Licorice extracts 68 Protein chip for target enzyme PAD Hypoglycemic and xanthine oxidase inhibitory activity Mulberry, lotus and Salvia miltiorrhiza extracts 69 , 70 , 71 Dual-channel microfluidic chip Multiple biomarker assay Qishen Yiqi Pill 72 Biological soft sensing technology HSI-AI-CNN Physical, chemical and biological properties Shuxuening injection 73 NIR-machine learning Chemical compounds and anti-inflammatory activity Chrysanthemum extracts 74 …”

“…Li et al. 61 proposed an on-demand strategy based on direct analysis in real time-mass spectrometry (DARTMS), which uses an MS probe as the substrate of the enzyme to determine the biological activity of selected thrombin and angiotensin-converting enzyme (ACE). Each probe consists of a specific peptide sequence and 1-(2-pyrimidyl) piperazine as a label with high ionization efficiency.…”

ASAP‐MS and DART‐MS as ancillary tools for direct analysis of the lichen metabolome

“…DART‐MS is especially popular for the direct analysis of complex biological samples (e.g., botanical drugs, enzymatic substrates, 14 or crude propolis 15 ). This method has even been previously used directly on lichen thalli: DART‐MS has notably been used for the chemical profiling of the lichen Ophioparma ventosa and for the thermochemical study of mycosporine serinol from Lichina pygmaea 16,17 …”