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JOURNAL OF CHROMATOGRAPHIC SCIENCE · 2020 ·Vol. 58 ·期 9 ·875-879 · IF 1.28

Online Liquid Microextraction Coupled with HPLC-ABTS for Rapid Screening of Natural Antioxidants: Case Study of Three Different Teas

Qian Zheng-Ming,Fang Bo-Wen,Chen Hai-Ming,Li Chun-Hong,Huang Qi,Chen Ling,Li Wen-Jia,Li De-qiang

In the present study, an online liquid extraction coupled with high-performance liquid chromatography-2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (HPLC-ABTS) system for rapid screening of antioxidants in tea samples was proposed. As an example, the tea samples were firstly extracted by online HPLC extractor with mobile phase at 70°C, then the hyphenated HPLC-ABTS was used for the chromatographic separation on a Poroshell EC C18 column by 0.3% aqueous formic acid and acetonitrile with a gradient elution at 1.5 mL·min−1, and the UV and antioxidant chromatograms with detection wavelengths at 270 nm and 750 nm were recorded, respectively. The established system integrated the processes of online HPLC sample extraction, HPLC separation and online antioxidants detection, the total analysis time of which was <20 min. The developed method was successfully applied to samples of green tea, oolong tea and black tea. As a result, 11 antioxidants were found in tea samples, including gallocatechin, epigallocatechin, catechin, chlorogenic acid, epicatechin, epigallocatechingallate, epicatechingallate, rutin, 1,4,6-trigalloylglucose, quercetin-3-glycoside and kaempferol-3-glucoside. The combined online liquid microextraction and online HPLC-ABTS method is a rapid and green approach for the quality evaluation of tea.

✓ 已验证 DOI PubMed 高影响因子
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INTERNATIONAL JOURNAL OF ENVIRONMENTAL ANALYTICAL CHEMISTRY · 2019 · IF 1.267

MnO2 nanozyme induced the chromogenic reactions of ABTS and TMB to visual detection of Fe2+ and Pb2+ ions in water

Kai Sun,Shun-Yao Li,Hui-Ling Chen,Qing-Guo Huang,Youbin Si

In this study, we used a simple and rapid colourimetric reaction for visual sensing of Fe2+ and Pb2+ ions in water by employing nano-MnO2 as a natural oxidase mimic to respectively catalyse ABTS and TMB in citrate-phosphate buffer solution (C-PBS) at 25°C and pH 3.8. It was found that nano-MnO2 possessed highly oxidase-mimicking activity with the Km values of 0.030 and 0.027 toward ABTS and TMB, respectively, indicating TMB had a stronger affinity on nano-MnO2 than ABTS. Interestingly, the presence of 0.01 mmol·L−1 Fe2+/Pb2+ ion was able to significantly down-regulate the activity of MnO2 nanozyme in nano-MnO2-mediated ABTS reaction processes (P < 0.01), which mainly due to the strong adsorption of metal ion toward nano-MnO2 surface via the electrostatic attractions, thus leading to the passivation and inactivation of MnO2 nanozyme catalytic activity. Thereinto, Fe2+ reacted with multivalent manganese by oxidation-reduction, while Pb2+ was specifically adsorbed onto the surface of MnO2 nanozyme and formed complexes. Notably, only Fe2+ ion inhibited the activity of MnO2 nanozyme-TMB with a detection limit as low as 1.0 μmol·L−1. In MnO2 nanozyme-ABTS sensing systems, Fe2+ and Pb2+ ions detection limit of 0.5 and 2.0 μmol·L−1 were, respectively, achieved with a linear response range of 0–0.02 and 0–0.8 mmol·L−1, implying the developed MnO2 nanozyme-ABTS sensor was potentially applicable for the visual determination of Fe2+ and Pb2+ ions in water. In the real water samples, MnO2 nanozyme-ABTS achieved high accuracy (relative errors: 3.4‒10.5%) and recovery (96‒110%) for respective detection of Fe2+ and Pb2+ ions. The simple and rapid MnO2 nanozyme-ABTS sensing systems might provide a practical assay for visual detection of Fe2+ and Pb2+ ions in the environmental water samples.

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CHEMICAL ENGINEERING JOURNAL · 2021 · IF 13.273

Rapid determination of monopersulfate with bromide ion-catalyzed oxidation of 2, 2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS)

Xin Liu,Qi Fu,Peng Xu,Pengfei Zhu,Zhuoyu Yang,John C. Crittenden

A sensitive spectrophotometric method (the Br − -ABTS method) for measuring monopersulfate (PMS) of 0–20 μM in water was established. The Br − -ABTS method was based on the oxidation coloration reaction where Br − as catalyst promoted PMS for oxidation of ABTS forming the green-colored ABTS radical (ABTS • + ) under pH 3.0 (50 mM phosphate buffer). ABTS • + could be quantitatively measured at 415 nm, 650 nm, 732 nm and 820 nm using UV–vis spectrophotometer. There was a well linear relationship between the absorbance of ABTS • + and PMS concentration with the value of R 2 being 0.999. The proposed Br − -ABTS method was highly sensitive, and the absorbance generated by Br − -ABTS at 415 nm was 6.59 × 10 4 cm −1 per M (mol L –1 ) of PMS. The changes of pH from 3.0 to 9.0 had an insignificant effect on the calibration curve of Br − -ABTS method. The reaction of PMS with ABTS had a stoichiometric coefficient of 1:1.96 at the pH 3.0. The residual absorbance of ABTS •+ in ultrapure water, Xiangjiang water and Hou-lake water was stable within 30 min. Moreover, the trace PMS concentration of 0.03–20 μM in ultrapure water, 0.05–20 μM in natural water (e. g. Xiangjiang water and Hou-lake water) and 1–20 μM in wastewater was accurately measured with the proposed Br − -ABTS method. Additionally, the interference of natural water matrix (e. g. humic acid), metal ions (e. g. Co(II), Fe(III) and Cu(II)) and hydroxylamine had barely interference for the proposed Br − -ABTS method. Furthermore, the Br − -ABTS method was more accurate and flexible to determine PMS concentration than DPD method during the decolorization of rhodamine B in the Co(II)/PMS system, Cu(II)/PMS system and hydroxylamine/Fe(III)/PMS process.

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BIOMEDICAL CHROMATOGRAPHY · 2026 ·Vol. 40 ·期 7 ·e70526 · IF 1.7

Combining Online HPLC-ABTS Screening With Machine Learning to Predict the Bioactivity of Antioxidant Components in Pandani Tectorii Radix et Rhizoma

Shu-Zhen Chen,Yong-Lin Ma,Dong-Xiao Li,Lian-Lin Yang,Ji-Wen Wang,Qian-Qian Hu,Hong-Liang Ma

This study developed an online HPLC-ABTS method to screen antioxidant phenolic acids in Pandani tectorii radix et rhizoma (PT) from 20 geographical origins. Six common constituents were screened and assigned by standards: neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, and isochlorogenic acid C. Using these six components as variables, four regression models were constructed to predict ABTS radical scavenging rate: support vector regression (SVR), decision tree regression, K-nearest neighbors (KNN) regression, and extreme gradient boosting (XGBoost) regression. From 80 samples, 64 were used for model development, and 16 were reserved for hold-out validation. Model selection employed 5-fold, nested, and leave-one-out cross-validations, complemented by permutation tests and learning curve analysis. The SVR model performed best, with nested CV R 2 = 0.9298, RMSE = 4.1207, MAE = 2.2313, and an optimism gap of 0.0137. After final tuning (C = 100, gamma = 0.1, and epsilon = 0.5), it achieved R 2 = 0.9000, RMSE = 4.0602, and MAE = 2.2986 on the hold-out set. This efficient and accurate method enables simultaneous screening of antioxidant constituents and prediction of radical scavenging activity, providing a new technical tool and scientific basis for PT quality evaluation.

✓ 已验证 DOI PubMed 高影响因子
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CHEMICAL ENGINEERING JOURNAL · 2021 · IF 13.273

Enhanced diclofenac elimination in Fe(II)/peracetic acid process by promoting Fe(III)/Fe(II) cycle with ABTS as electron shuttle

Jinbin Lin,Yuye Hu,Junyang Xiao,Yixin Huang,Mengyun Wang,Haoyu Yang,Jing Zou,Baoling Yuan,Jun Ma

Fe(II)/peracetic acid (Fe(II)/PAA) process exhibits a great potential in abatement of micropollutants because of the fast reaction between Fe(II) and PAA. Similar to Fenton system, the slow reduction from Fe(III) to Fe(II) leads to the high dosage of Fe(II) in Fe(II)/PAA process. This study introduced 2,2′-azino-bis(3-ethylbenz- -othiazoline-6-sulfonate) (ABTS) as electron shuttle to improve the oxidation capacity of Fe(II)/PAA process by accelerating the redox cycle of Fe(III)/Fe(II). ABTS/Fe(II)/PAA process possessed high elimination efficiency of diclofenac in acidic conditions. Alcohols quenching experiments confirmed that ABTS •+ was the primary reactive species responsible for diclofenac elimination, although Fe IV O 2+ and carbon-centered radicals (R-O • ) also existed in ABTS/Fe(II)/PAA process. ABTS •+ was generated via various pathways, including the reactions of ABTS with Fe(III), Fe IV O 2+ and R-O • . ABTS was not an ideal electron shuttle because R-O • could decompose partial ABTS •+ to sulfoxides and sulfones. Two possible degradation pathways of diclofenac were proposed based on the detected transformation products. ABTS/Fe(II)/PAA process was highly-efficient for diclofenac elimination in underground water and reservoir water, while high concentration of humic acid had inhibition effect on diclofenac degradation. The present study provides a promising way to alleviate the intrinsic drawbacks of Fe(II)/PAA process and might advance its potential application in water treatment.

CAS / MDL: 15307-86-5 28752-68-3
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FOOD CHEMISTRY · 2025 · IF 9.8

Identification of 13 natural antioxidants in green calyx plum using AAPH, ABTS, and FRAP-coupled HPLC-DAD via QTOF-MS/MS

Jing-Wen Hao,Wei-Ting Wang,Nai-Dong Chen,Yan Shen

The novel application of a combined screening method, utilizing AAPH, ABTS, and FRAP assays with HPLC, is first used for screening natural antioxidants in Green Calyx plums (GCP). This method was assumed that the peak areas of compounds with antioxidant activity in HPLC chromatograms would be significantly reduced or disappeared after incubating with the AAPH, ABTS, and FRAP. The thirteen screened antioxidants can be identified by Q-TOF-MS/MS. kaempferol-3-O-rutinoside, narcissoside, and astragalin were first reported in GCP. respectively. The thirteen screened antioxidants were quantified simultaneously at their maximum absorption wavelengths, with concentrations ranging from 0.61 to 79.30 mg/g across three distinct flowering stages. Furthermore, the in vitro activity and molecular docking of oxidative stress-related enzyme xanthine oxidase (XOD) were utilized to evaluate the antioxidant capacity of the screened compounds. Consequently, our method effectively screens for antioxidants within complex mixtures, positioning GCP as a promising reservoir of natural bioactive compounds.

CAS / MDL: 64-18-6
✓ 已验证 产品关联 DOI PubMed 高影响因子
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Journal of Food Measurement and Characterization · 2021 ·Vol. 15 ·期 5 ·3873-3883 · IF 2.431

Comparison of the chemical-antioxidant profiles of different parts of Citrus reticulata Blanco (Rutaceae) based on OLE-HPLC-DAD-MS/MS-ABTS assay

Qian Zheng-ming,Huang Qi,Feng Xiao-long,Li De-qiang

Given the safety of synthetic antioxidants, people are increasingly inclined to develop and use natural antioxidants. However, rapid screening and identification of active ingredients from the complex systems of natural products is one of the major challenges for the modernization of traditional Chinese medicine. In this study, online extraction-high performance liquid chromatography-diode array detection-quadrupole time-of-flight mass spectrometry coupled with ABTS antioxidant assay (OLE-HPLC-DAD-QTOF-MS/MS-ABTS), in which sample preparation, separation, detection and antioxidant assay were integrated in a single step, was used to rapidly identify antioxidant components from the different parts of Citrus reticulata , including its peels, leaves, vascular bundies and seeds. After method validation (system adaptability, extraction yield and repeatability), the four samples of different botanical parts were analyzed. As a result, among 49 found chromatographic peaks, 33 peaks displayed varying degrees of antioxidant activity and 39 peaks were tentatively or unambiguously identified based on their retention time, UV spectrum and mass spectrometry information and the comparison with reference substance and data in literature, including 10 phenolic acids, 9 polymethoxy flavonoids, 7 flavonoid- O -glycosides and 13 other ingredients. It is concluded that the established method is simple, fast, and reliable, which not only saves the materials and solvents as well as time, but also reduces the risk of degradation of chemical components during the extraction process, suitable for rapid screening and identification of antioxidant components of complex mixture.

CAS / MDL: 7727-21-1
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Journal of Environmental Chemical Engineering · 2023 · IF 7.7

Rapid spectrophotometric determination of peroxydisulfate in water via oxidative coloration of 2,2′azino-bis(3-ethylbenzothiazoline)-6-sulfonate (ABTS) using ferrous ion as catalyst

Yixin Huang,Jing Zou,Jiawen Li,Hongfang Ma,Siying Chen,Lingying Chen,Xiaobin Liao,Jun Ma

In this study, a rapid, sensitive and reliable spectrophotometric method for peroxydisulfate (PDS) determination in water was established. The proposed spectrophotometric method was based on the oxidation of 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonate) (ABTS) to form stable and green-colored ABTS radical (ABTS • + ) by ferrous ion (Fe(II))-catalyzed PDS. The formed ABTS • + performed well linear relationship with PDS concentration (R 2 > 0.999) in the range of 0–60 μM at four wavelengths of 415, 650, 732, and 820 nm under optimized conditions. The proposed Fe(II)-ABTS spectrophotometric method was highly sensitive with the absorptivity of 3.595 × 10 4 M −1 cm −1 , 1.530 × 10 4 M −1 cm −1 , 1.945 × 10 4 M −1 cm −1 and 1.768 × 10 4 M −1 cm −1 at four detection peaks of 415, 650, 732 and 820 nm, respectively. Additionally, the proposed Fe(II)-ABTS spectrophotometric method possessed well anti-interference capacity to common natural water constituents and was reliable to determine PDS concentration in water. What’s more, the proposed Fe(II)-ABTS spectrophotometric method was successfully applied in monitoring the change of PDS concentration during the degradation of diclofenac in heat-activated PDS process. In a word, the proposed Fe(II)-ABTS spectrophotometric method could be employed as a promising approach for the rapid and accurate measurement of PDS in water, and this study provided a new perspective for the measurement of PDS by the visible spectrophotometry .

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CHEMOSPHERE · 2021 · IF 7.086

Multi-wavelength spectrophotometric determination of peracetic acid and the coexistent hydrogen peroxide via oxidative coloration of ABTS with the assistance of Fe2+ and KI

Jinbin Lin,Junyang Xiao,Hengyu Cai,Yixin Huang,Jiawen Li,Haoyu Yang,Tao Li,Jing Zou

In this study, a multi-wavelength spectrophotometric method for simultaneous determination of peracetic acid (PAA) and coexistent hydrogen peroxide (H 2 O 2 ) was presented. This method was based on the oxidation of 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) with the assistance of Fe 2+ /KI to produce a stable green radical (ABTS ●+ ), which could be determined at four characteristic peaks (i.e., 415 nm, 650 nm, 732 nm, and 820 nm). The absorbances of ABTS ●+ at four peaks were well linear (R 2  > 0.999) with concentrations of both total peroxides (PAA + H 2 O 2 ) and PAA in the range of 0–40 μM under optimized conditions. The sensitivities for determining total peroxides at 415 nm, 650 nm, 732 nm and 820 nm were determined to be 4.248 × 10 4  M −1  cm −1 , 1.682 × 10 4  M −1  cm −1 , 2.132 × 10 4  M −1  cm −1 , and 1.928 × 10 4  M −1  cm −1 , respectively. For determining PAA, the corresponding sensitivities were 4.622 × 10 4  M −1  cm −1 , 1.895 × 10 4  M −1  cm −1 , 2.394 × 10 4  M −1  cm −1 and 2.153 × 10 4  M −1  cm −1 , respectively. The concentration of coexistent H 2 O 2 was gained by deducting PAA concentration from total peroxides concentration. The ABTS method was accurate enough to determine PAA concentration in natural water samples. Moreover, the ABTS method was successfully used to determine the changes of PAA and coexistent H 2 O 2 and to distinguish their role on naproxen degradation in heat-activated PAA process. Overall, the ABTS method could be used as an alternative method for the convenient, rapid and sensitive determination of PAA and the coexistent H 2 O 2 in water samples.

CAS / MDL: 7681-11-0 28752-68-3
✓ 已验证 产品关联 DOI PubMed 高影响因子
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BIOMEDICAL CHROMATOGRAPHY · 2024 · IF 1.8

Comparison of the Antioxidant Capacity of Cell Wall-Broken Decoction Pieces and Traditional Decoction Pieces of Astragli Radix Based on HPLC-ABTS Analytical Method

Yonglin Ma,Yue Zhang,Yu Zhao,Jiwen Wang,Qianqian Hu,Lianlin Yang,Shuzhen Chen,Yong Diao,Hongliang Ma

In this study, an online antioxidant assay based on HPLC-ABTS was mainly developed for screening the antioxidants of flavonoids from Astragali Radix (AR), and comparing the antioxidant capacity between traditional decoction pieces (TDP) of AR and cell wall-broken decoction pieces (CDP) of AR. The experimental results showed that the overall antioxidant capacity of CDP of AR was about twice as much as that of TDP of AR, which was specifically expressed as the antioxidant capacity of the screened antioxidants extracted from CDP was equivalent to 1.9–5.1 times that of those extracted from TDP, and three antioxidants were successfully screened, which were calycosin-7-O-β-D-glucoside, calycosin, and formononetin. The method established in this study is characterized by high efficiency and accuracy, which can simultaneously accomplish the screening of antioxidant components and the comparison of antioxidant capacity between samples, and provides a new method for the quality evaluation of AR from the perspective of antioxidant activity.

✓ 已验证 产品关联 DOI PubMed 高影响因子
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BMC Complementary Medicine and Therapies · 2020 ·Vol. 20 ·期 1 ·1-10 · IF 0

“Quantity-effect” research strategy for comparison of antioxidant activity and quality of Rehmanniae Radix and Rehmannia Radix Praeparata by on-line HPLC-UV-ABTS assay

Li Hong-Ying,Fang Jiang-Ji,Shen Hua-Dan,Zhang Xue-Qiong,Ding Xiao-Ping,Liu Jun-Feng

Background Quantitation analysis and chromatographic fingerprint of multi-components are frequently used to evaluate quality of herbal medicines but fail to reveal activity of the components. It is necessary to develop a rational approach of chromatography coupled with activity detection for quality assessment of herbal medicines. Methods An on-line HPLC-ultraviolet detection-2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) free radical scavenging (HPLC-UV-ABTS) method was developed to obtain the chromatographic fingerprints and ABTS +• inhibition profiles (active fingerprints) of Rehmanniae Radix (Dihuang) and Rehmannia Radix Praeparata (Shu Dihuang). Eighteen compounds showing ABTS +• inhibition activity were identified by HPLC-fourier-transform mass spectrometry (HPLC-FTMS). Verbascoside was used as a positive control to evaluate the total activities of the samples and the contribution rate of each compound. The similarities of the chromatographic and active fingerprints were estimated by the vectorial angle cosine method. Results The results showed that the HPLC-UV-ABTS method could efficiently detect antioxidant activity of the herbal medicine samples. The antioxidants were different between the two herbs and several new antioxidants were identified in Shu Dihuang. A function equation was generated in terms of the negative peak area (x) and the concentrations of verbascoside (y, μg/mL), y = 2E-07 ×  4 - 8E-05 ×  3  + 0.0079 ×  2  + 0.5755x + 1.4754, R 2  = 1. Iridoid glycosides were identified as main antioxidants and showed their higher contributions to the total activity of the samples. The total contributions of the three main active components in the Dihuang and Shu Dihuang samples to the total activity, such as echinacoside, verbascoside and an unknown compound, were 39.2–58.1% and 55.9–69.4%, respectively. The potencies of the main active components in the Shu Dihuang samples were two to ten times those in the Dihuang samples. Similarity values for S12 in the chromatographic fingerprints and S03, S12 and P03 in the active fingerprints were less than 0.9. The three batches of samples might show their different quality with the other samples. Conclusions The results suggested that the combination of “quantity-effect” research strategy and the HPLC-UV-ABTS analysis method could comprehensively evaluate the active components and quality of Dihuang and Shu Dhuang.

CAS / MDL: 64-18-6
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