An integrated strategy for the identification and screening of anti-allergy components from natural products based on calcium fluctuations and cell extraction coupled with HPLC–Q–TOF–MS

Hu, Guizhou; Li, Xin-Qi; Zhang, Jingwen; Zhang, Lijun; Qi, Jin; Yu, Boyang

doi:10.1007/s00216-021-03580-5

Cited by 15 publications

(14 citation statements)

References 46 publications

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“…QTOF‐MS/MS is a high‐resolution MS technique which allows for efficient characterization of compounds in complex natural products 17–19 . After neutral loss/diagnostic ion filtering (extraction width at ± 5 mDa), 12 polymethoxylated flavones and four polymethoxylated flavanones were fished out from the ethanol extract of lysed MCF‐7 cells.…”

Section: Resultsmentioning

confidence: 99%

“…Cell biospecific extraction procedures were performed according to previous reports with minor modifications 17,18 . MCF‐7 cells in the logarithmic growth stage were inoculated in a six‐well plate at a density of 1 × 10 6 cells/well and incubated in a 5% CO 2 incubator at 37°C for 1 h. After that, the medium was removed, 2.0 mL CRPV extract (50 mg/mL) diluted with serum‐free medium was added, and cells were incubated under the same circumstances for another 3 h. Then, the cell suspension was transferred to a centrifugal tube and centrifuged at 10,000 rpm for 10 min to remove the supernatant.…”

Section: Methodsmentioning

confidence: 99%

“…Notably, living cell‐based screening methods can identify bioactive compounds binding to different cellular targets 15,16 . More recently, Hu et al established RBL‐2H3 cell biospecific extract technology to efficiently screen antiallergy compounds in Radix Paeoniae Alba, Radix Astragali, and Radix Saposhnikoviae 17 . Zhang et al utilized a PC12 cell extraction method to specifically screen neuroprotective compounds in Huang‐Lian‐Jie‐Du decoction 18 .…”

Section: Introductionmentioning

confidence: 99%

“…In this study, potential antitumor PMFs were rapidly profiled and identified in CRPV by integrating cell biospecific extraction with neutral loss/diagnostic ion filtering‐based HPLC‐QTOF‐MS/MS. The method was established to screen and identify bioactive compounds in complex natural products rapidly, efficiently, and sensitively 14,17 …”

Section: Introductionmentioning

confidence: 99%

“…15,16 More recently, Hu et al established RBL-2H3 cell biospecific extract technology to efficiently screen antiallergy compounds in Radix Paeoniae Alba, Radix Astragali, and Radix Saposhnikoviae. 17 Zhang et al utilized a PC12 cell extraction method to specifically screen neuroprotective compounds in Huang-Lian-Jie-Du decoction. 18 Moreover, living cellbased screening methods facilitate high-throughput operation, which will accelerate the discovery of bioactive compounds in natural products.…”

Section: Introductionmentioning

confidence: 99%

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Rapid profiling of potential antitumor polymethoxylated flavonoids in natural products by integrating cell biospecific extraction with neutral loss/diagnostic ion filtering‐based high‐performance liquid chromatography–quadrupole time‐of‐flight tandem mass spectrometry

Wang

Cao

et al. 2022

Phytochemical Analysis

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Introduction Citri Reticulatae Pericarpium Viride (CRPV, Qing Pi in Chinese) has been widely used in traditional Chinese medicine. Polymethoxylated flavonoids (PMFs), which are a special group of flavonoids with strong antitumor activity, are broadly distributed in citrus peels. However, systematic investigation of antitumor PMFs in CRPV has received little attention to date. Objectives An MCF‐7 cell biospecific extraction method integrated with neutral loss/diagnostic ion filtering‐based HPLC‐QTOF‐MS/MS strategy was developed for rapid and specific profiling of antitumor PMFs and systematic identification of PMFs in CRPV. Methodology By incubating MCF‐7 cells with CRPV extract, potential antitumor PMFs specifically bound to cells and were isolated. Then, by systematic investigation of fragmentation pathways, neutral loss and diagnostic ion filtering strategies were proposed to comprehensively and accurately identify PMFs. Results Sixteen antitumor PMFs were unambiguously or tentatively identified. Among them, minor compound 15 (5‐hydroxy‐6,7,8,3′,4′‐pentamethoxyflavone with a free hydroxyl group at C‐5) exhibited excellent antitumor activity, with an IC50 value of 2.81 ± 0.76 μg/mL, which is lower than that of 5‐fluorouracil (IC50, 4.92 ± 0.83 μg/mL). Nobiletin (12) and tangeretin (16), two major PMFs, presented moderate antitumor activities with IC50 values of 13.06 ± 1.85 and 17.07 ± 1.18 μg/mL, respectively, and their contents were sensitively and precisely determined. Conclusions To the best of our knowledge, this is the first report on the systematic investigation of antitumor PMFs in CRPV. The study will lay a foundation for the quality control and clinical application of CRPV.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Rapid profiling of potential antitumor polymethoxylated flavonoids in natural products by integrating cell biospecific extraction with neutral loss/diagnostic ion filtering‐based high‐performance liquid chromatography–quadrupole time‐of‐flight tandem mass spectrometry

Wang

Cao

et al. 2022

Phytochemical Analysis

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Identification and quantification of cucurbitacin in watermelon frost using molecular networking integrated with ultra‐high‐performance liquid chromatography‐tandem mass spectrometry

Liu

2023

J of Separation Science

Self Cite

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Watermelon frost, a traditional Chinese medicine produced using watermelon and Glauber's salt, has been widely used for the therapy of oral and throat disorders. Watermelon contains various phytochemical compounds including cucurbitacins and their glycoside derivatives, which have attracted considerable attention because of their medicinal values. However, whether the composition of cucurbitacins existed in watermelon frost was rarely reported. In this study, three cucurbitacins including cucurbitacin B, isocucurbitacin B, and cucurbitacin E were found from watermelon frost extract assisted by ultra-highperformance liquid chromatography-tandem mass spectrometry and molecular networking guided strategy, and the compounds were verified using standard solutions. Furthermore, a quantification method for simultaneously targeted analysis of cucurbitacins was established using ultra-high-performance liquid chromatography-tandem mass spectrometry operating in the multiple reaction monitoring mode. Among them, cucurbitacin B and cucurbitacin E in watermelon frost samples were determined, and the concentrations were 3.78 ± 0.18 and 0.86 ± 0.19 ng/ml, respectively. While isocucurbitacin B was not detected due to the lower content possibly. In conclusion, ultra-high-performance liquid chromatography-tandem mass spectrometry combined with molecular networking is a very useful technique for the rapid identification of unknown cucurbitacin components in watermelon frost.

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A novel method for evaluating pseudoallergy based on β‐hexosaminidase and its application for traditional Chinese medicine injections

Yang,

Zhang,

et al. 2024

J of Separation Science

Self Cite

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Pseudoallergy is a typical and common adverse drug reaction to injections, especially in traditional Chinese medicine injections (TCMIs). At present, the evaluation methods for pseudoallergy include cell methods in vitro and animal methods in vivo. The mast cell evaluation method based on the β‐hexosaminidase (β‐Hex)‐catalyzed substrate, 4‐nitrophenyl‐β‐N‐acetyl‐D‐glucosaminide (4‐NPG), is an important method for the evaluation of drug‐induced pseudoallergy, but it is prone to false positive results and has insufficient sensitivity. In this study, a novel β‐Hex evaluation system with rat basophilic leukemia‐2H3 cells based on high‐performance liquid chromatography‐fluorescence detection (HPLC‐FLD) was established, which effectively increased the sensitivity and avoided false positive results. Cell viabilities were measured by 3‐(4,5‐dimethyl‐2‐thiazolyl)‐2,5‐diphenyl tetrazolium bromide assay. In addition, a method for the determination of histamine, which is another indicator in the development of pseudoallergy, was established to validate the above method. The results of this novel method indicated that two TCMIs (Shuxuening injection and Shenqi Fuzheng injection), which were considered to be pseudoallergenic using 4‐NPG, were not pseudoallergenic. Overall, the novel β‐Hex/HPLC‐FLD evaluation system using Rat basophilic leukemia‐2H3 cells established was effective and precise. It could be used for the evaluation of pseudoallergic reactions caused by TCMIs and other injections.

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An integrated strategy for the identification and screening of anti-allergy components from natural products based on calcium fluctuations and cell extraction coupled with HPLC–Q–TOF–MS

Cited by 15 publications

References 46 publications

Rapid profiling of potential antitumor polymethoxylated flavonoids in natural products by integrating cell biospecific extraction with neutral loss/diagnostic ion filtering‐based high‐performance liquid chromatography–quadrupole time‐of‐flight tandem mass spectrometry

Rapid profiling of potential antitumor polymethoxylated flavonoids in natural products by integrating cell biospecific extraction with neutral loss/diagnostic ion filtering‐based high‐performance liquid chromatography–quadrupole time‐of‐flight tandem mass spectrometry

Identification and quantification of cucurbitacin in watermelon frost using molecular networking integrated with ultra‐high‐performance liquid chromatography‐tandem mass spectrometry

A novel method for evaluating pseudoallergy based on β‐hexosaminidase and its application for traditional Chinese medicine injections

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