2,2-Diphenyl-1-picrylhydrazyl-ultra-high performance liquid chromatography-Q-time-of-flight mass spectrometry (DPPH-UPLC-Q-TOF/MS), as a rapid and efficient means, now was used for the first time to screen antioxidants fromSelaginella doederleinii. The nine biflavone compounds were screened as potential antioxidants. The biflavones were structurally identified and divided into the three types, that is, amentoflavone-type, robustaflavone-type, and hinokiflavone-type biflavonoids. Among the compounds bilobetin (3) and putraflavone (8) were found fromSelaginella doederleiniifor the first time and others including amentoflavone (1), robustaflavone (2), 4′-methoxy robustaflavone (4), podocarpusflavone A (5), hinokiflavone (6), ginkgetin (7), and heveaflavone (9) were identified previously in the plant. Moreover, nine biflavones possessed a good antioxidant activity via their DPPH free radical scavenging. It demonstrates that DPPH-UPLC-Q-TOF/MS exhibits strong capacity in separation and identification for small molecule. The method is suitable for rapid screening of antioxidants without the need for complicated systems and additional instruments.

As new green solvents, ionic liquids (ILs) have been generally applied in the extraction and separation of natural product. In this study, microwave assisted extraction based on IL (IL-MAE) was firstly employed to extract total biflavonoids from Selaginella doederleinii. Based on single-factor experiment, microwave power (300–700 W), extract time (30–50 min) and extract temperature (40–60 °C) on total bioflavonoids and antioxidant activities of the extracts were further investigated by a Box-Behnken design of response surface methodology (RSM) selecting total bioflavonoids yields and IC₅₀ of radical scavenging as index. Besides antioxidant activity of the extract was evaluated by a 2,2-diphenyl-1-picrylhydarzyl (DPPH) and 2,2′-azinobis-(3-ethylbenzthiazoline-6-sulphonate (ABTS) radical scavenging assay, ferric reducing power assay and chelation of ferrous ions assay, and then anticaner activity was also researched against A549 cell line and 7721 cell line. The results illustrated that three factors and their interactions could be well suited for second-order polynomial models (p < 0.05). Through process parameters, optimization of the extract (460 W, 40 min, and 45 °C) and detection of bioactivity, the yield of total bioflavonoids was 16.83 mg/g and IC₅₀ value was 56.24 μg/mL, respectively, indicating the extract has better anti-oxidation effect and antitumor activity. Furthermore, IL-MAE was the most efficient extracting method compared with MAE and Soxhlet extraction, which could improve extraction efficiency in a shorter time and at a lower temperature. In general, ILs-MAE was first adopted to establish a novel and green extraction process on the yields of total biflavonoids from S. doederleinii. In addition, the extract of containing biflavones showed potent antioxidant and anticancer capacity as a utilized valuable bioactive source for natural medicine.

In this study, the deep eutectic solvent based ultrasound-assisted extraction (DES-UAE) was investigated for the efficient and environmentally friendly extraction of Selaginella chaetoloma total biflavonoids (SCTB). As an extractant for optimization, tetrapropylaminium bromide-1,4-butanediol (Tpr-But) was employed for the first time. 36 DESs were created, with Tpr-But producing the most effective results. Based on response surface methodology (RSM), the greatest extraction rate of SCTB was determined to be 21.68 ± 0.78 mg/g, the molar ratio of HBD to HBA was 3.70:1, the extraction temperature was 57 °C, and the water content of DES was 22 %. In accordance with Fick's second rule, a kinetic model for the extraction of SCTB by DES-UAE has been derived. With correlation coefficients 0.91, the kinetic model of the extraction process was significantly correlated with the general and exponential equations of kinetics, and some important kinetic parameters such as rate constants, energy of activation and raffinate rate were determined. In addition, molecular dynamics simulations were used to study the extraction mechanisms generated by different solvents. Comparing the effect of several extraction methods on S.chaetoloma using ultrasound-assisted extraction and conventional methods, together with SEM examination, revealed that DES-UAE not only saved time but also enhanced SCTB extraction rate by 1.5-3 folds. SCTB demonstrated superior antioxidant activity in three studies in vitro. Furthermore, the extract could suppress the growth of A549, HCT-116, HepG2, and HT-29 cancer cells. Alpha-Glucosidase (AG) inhibition experiment and molecular docking studies suggested that SCTB exhibited strong inhibitory activity against AG and potential hypoglycemic effects. The results of this study indicated that a Tpr-But-based UAE method was suitable for the efficient and environmentally friendly extraction of SCTB, and also shed light on the mechanisms responsible for the increased extraction efficiency, which could aid in the application of S.chaetoloma and provide insight into the extraction mechanism of DES.

A novel mitochondria-targeted ratiometric fluorescent probe was developed via condensation of carbazole with two indolium units, which could realize the detection of bisulfite in a PBS buffer solution. The probe exhibited good mitochondrial location ability and could selectively respond to bisulfite among other sulfur-containing species in living cells.

Flavonoids are the main active ingredient in Citri Reticulatae Pericarpium Viride (CRPV).