Optimization of Extraction Solvent and Fast Blue BB Assay for Comparative Analysis of Antioxidant Phenolics from Cucumis melo L.

Melon (Cucumis melo L.) fruits contain multiple health-promoting compounds, including phenolic compounds, which are antioxidants. Accurate measurement of antioxidant activities and total phenolic contents (TPCs) require an efficient solvent extraction. In this study, we evaluated free radical scavenging activity and TPC of melon extracts extracted with 22 different solvent combinations. The DPPH scavenging activities were high in 100% methanolic (39.48 ± 0.36 µg g-1) and 80% methanolic extracts (38.99 ± 0.44 µg g-1). Similarly, the ABTS scavenging activities were high in 100% methanolic (315.11 ± 10.38 µg g-1) and 80% methanol extracts (297.39 ± 14.98 µg g-1). The Folin-Ciocalteu (F-C) assay is typically used to measure TPC but may be affected by interference from sugars and other compounds. Therefore, we optimized an assay for TPC using Fast Blue (FB) salt and developed a standard operating procedure for microplate analysis using FB. Our analysis of standard samples and comparisons with the F-C assay suggested that the optimized FB assay could be used to measure TPC in fruit and juice samples. Moreover, we successfully detected six phenolic compounds in methanol extracts of melon by LC-HR-QTOF/MS.

Manganese Oxide Nanoparticles as Safer Seed Priming Agent to Improve Chlorophyll and Antioxidant Profiles in Watermelon Seedlings.

The use of nanoscale nutrients in agriculture to improve crop productivity has grown in recent years. However, the bioefficacy, safety, and environmental toxicity of nanoparticles are not fully understood. Herein, we used onion bulb extract to synthesize manganese oxide nanoparticles (MnO-NPs). X-ray diffraction, X-ray photoelectron spectroscopy, and high-resolution transmission electron microscopy were used for the structural and morphological characterization of synthesized MnO-NPs. The MnO-NPs were oval shape crystalline nanoparticles of Mn2O3 with sizes 22-39 nm. In further studies, we assessed the comparative toxicity of seed priming with MnO-NPs and its bulk counterparts (KMnO4 and Mn2O3), which showed seed priming with MnO-NPs had comparatively less phytotoxicity. Investigating the effect of seed priming with different concentrations of MnO-NPs on the hormonal, phenolic acid, chlorophyll, and antioxidant profiles of watermelon seedlings showed that treatment with 20 mg·L-1 MnO-NPs altered the chlorophyll and antioxidant profiles of seedlings. At ≤40 mg·L-1, MnO-NPs had a remarkable effect on the phenolic acid and phytohormone profiles of the watermelon seedlings. The physiological outcomes of the MnO-NP seed priming in watermelon were genotype-specific and concentration-dependent. In conclusion, the MnO-NPs were safer than their bulk counterparts and could increase crop productivity.

In vitro digestion with bile acids enhances the bioaccessibility of kale polyphenols.

Kale (Brassica oleracea) is a leafy green vegetable belonging to the Brassicaceae family, and kale leaves have large amounts of dietary fiber and polyphenolics. Dietary fiber can bind bile acids, thus potentially decreasing cholesterol levels; however, whether the polyphenols from kale contribute to in vitro bile acid binding capacity remains unclear. In the present study, kale was extracted with hexane, acetone, and MeOH : water and the dried extracts, as well as the fiber-rich residue, were tested for their bile acid binding capacity. The fiber-rich residue bound total bile acids in amounts equivalent to that bound by raw kale. The lyophilized acetone extract bound significantly more glycochenodeoxycholate and glycodeoxycholate and less of other bile acids. To test whether bile acid binding enhanced the bioaccessibility of polyphenolic compounds from kale, we used ultra-performance liquid chromatography coupled with electrospray ionization/quadrupole-time-of-flight mass spectrometry to identify chemical constituents and measure their bioaccessibility in an in vitro digestion reaction. This identified 36 phenolic compounds in kale, including 18 kaempferol derivatives, 13 quercetin derivatives, 4 sinapoyl derivatives, and one caffeoylquinic acid. The bioaccessibility of these phenolics was significantly higher (69.4%) in digestions with bile acids. Moreover, bile acids enhanced the bioaccessibility of quercetin by 25 times: only 2.7% of quercetin derivatives were bioaccessible in the digestion without bile acids, but with bile acids, their accessibility increased to 69.5%. Bile acids increased the bioaccessibility of kaempferol from 37.7% to 69.2%. The extractability and biostability of total phenolics in the digested residue increased 1.8 fold in the digestions with bile acids. These results demonstrated the potential use of kale to improve human health.

In vivo induction of phase II detoxifying enzymes, glutathione transferase and quinone reductase by citrus triterpenoids.

BACKGROUND: Several cell culture and animal studies demonstrated that citrus bioactive compounds have protective effects against certain types of cancer. Among several classes of citrus bioactive compounds, limonoids were reported to prevent different types of cancer. Furthermore, the structures of citrus limonoids were reported to influence the activity of phase II detoxifying enzymes. The purpose of the study was to evaluate how variations in the structures of citrus limonoids (namely nomilin, deacetyl nomilin, and isoobacunoic acid) and a mixture of limonoids would influence phase II enzyme activity in excised tissues from a mouse model. METHODS: In the current study, defatted sour orange seed powder was extracted with ethyl acetate and subjected to silica gel chromatography. The HPLC, NMR and mass spectra were used to elucidate the purity and structure of compounds. Female A/J mice were treated with three limonoids and a mixture in order to evaluate their effect on phase II enzymes in four different tissues. Assays for glutathione S-transferase and NAD(P)H: quinone reductase (QR) were used to evaluate induction of phase II enzymatic activity. RESULTS: The highest induction of GST against 1-chloro-2,4-dinitrobenzene (CDNB) was observed in stomach (whole), 58% by nomilin, followed by 25% isoobacunoic acid and 19% deacetyl nomilin. Deacetyl nomilin in intestine (small) as well as liver significantly reduced GST activity against CDNB. Additionally isoobacunoic acid and the limonoid mixture in liver demonstrated a significant reduction of GST activity against CDNB. Nomilin significantly induced GST activity against 4-nitroquinoline 1-oxide (4NQO), intestine (280%) and stomach (75%) while deacetyl nomilin showed significant induction only in intestine (73%). Induction of GST activity was also observed in intestine (93%) and stomach (45%) treated with the limonoid mixture. Finally, a significant induction of NAD(P)H: quinone reductase (QR) activity was observed by the limonoid mixture in stomach (200%). In addition, the deacetyl nomilin treatment group displayed an increase in QR activity in liver (183%) and intestine (22%). CONCLUSION: The results of the present study suggests that, dietary intake of citrus limonoids may provide a protective effect against the onset of various cancers by inducing the activity of certain phase II detoxifying enzymes in specific organs.