An immunochromatographic assay (ICA) using gold nanoparticles coated with monoclonal antibody (McAb) for the detection of chromium ions (Cr) in water and serum samples was developed, optimized and validated. Gold nanoparticles coated with affinity-purified monoclonal antibodies against isothiocyanobenzyl-EDTA (iEDTA)-chelated Cr3+ were used as the detecting reagent in this completive immunoassay-based one-step test strip. The ICA was investigated to measure chromium speciation (Cr3+ and Cr6+ ions) in water samples. Chromium standard samples of 0-80 ng/mL in water were determined by the test strips. The results showed that the visual lowest detection limit (LDL) of the test strip was 50.0 ng/mL. A portable colorimetric lateral flow reader was used for the quantification of Cr. The results indicated that the linear range of the ICA with colorimetric detection was 5-80 ng/mL. The ICA was also validated for the detection of chromium ions in serum samples. The test trips showed high stability in that they could be stored at 37°C for at least 12 weeks without significant loss of activity. The test strip also showed good selectivity for Cr detection with negligible interference from other heavy metals. Because of its low cost and short testing time (within 5 min), the test strip is especially suitable for on-site large-scale screening of Cr-polluted water samples, biomonitoring of Cr exposure, and many other field applications.
Rationale: Paris polyphylla var. yunnanensis (Franch) Hand Mazz (PPY) is a traditionalChinese medicine with antitumor, antibacterial, hemostatic, and anthelmintic activities. Identification of the chemical composition in PPY is helpful to discover its active ingredients and can be used to establish its quality control protocols.Methods: The composition of PPY was identified using ultra-high-performance liquid chromatography combined with quadrupole time-of-flight mass spectrometry (UHPLC/QTOF-MS/MS) coupled with a molecular networking strategy. First, the UHPLC/QTOF-MS/MS approach was optimized for chemical compound profiling.Then, the MS data were processed using PeakView™ combined with an in-house database to quickly characterize the secondary metabolites. Finally, molecular networking excavated new molecular weights to discover unknown or trace natural products based on the characteristics of each cluster. Results:A total of 222 compounds, including 77 isospirostanols, 2 spirostanols, 19 furostanols, 10 pseudospirostanols, 6 cholesterols, 10 C 21 steroids, 5 insect metamorphosis hormones, 3 plant sterols, 6 five-ring triterpenoids, 4 flavonoids, 8 fatty acids, 2 phenylpropanoids, and 8 other compounds, were characterized in PPY by comparing their main fragmentation characteristics and pathways with the literature data, and 62 of them, 54 steroidals and 8 phenylpropanoids, were discovered or tentatively identified for the first time. Conclusions:This study extended the application of a molecular networking strategy to traditional herbal medicines and developed a molecular networking based screening approach with a significant increase in efficiency for the discovery and identification of trace novel natural products.Yumei Wang and Qian Fan contributed equally to this work as first authors. Paris polyphylla var. yunnanensis (Franch) Hand Mazz (PPY), calledChong Lou, is a well-known, valuable traditional Chinese medicine (TCM) that has been used to treat fractures, hemorrhages, snake bites, and abscess for many years. 1,2 PPY has been used as a main ingredient for the preparation of many TCMs, such as Yunnan Baiyao, Gongxuening capsules, and Jidesheng Sheyao tablets. 3 Phytochemistry studies have indicated that the dried rhizomes of Paris species contain steroidal saponins, triterpenoid saponins, flavonoids, phenylpropanoids, ecdysteroids, phenolic glycosides, and so on. 4-6 The glucosyl groups of steroidal saponins are normally attached to the steroidal aglycones through the hydroxyl groups at the C-3 and/or C-26 positions (and sometimes the C-1, C-21, C-23, and C-24 positions). Four types of steroidal sapogenins 6 (spirostanol, isospirostanol, furostanol, and pseudospirostanol) are considered major aglycones, and typical sugars are arabinose, xylose, apiose, fucose, rhamnose, glucose, and galactose. Comprehensive identification of ingredients in PPY is critical to understanding its biological mechanism and establishing quality control protocols. Because of the complexity of these components, their comprehensive...
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