Associated Laboratory for Green Chemistry - Clean Technologies and Processes

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Azadirachta indica A. Juss (neem) phenolic extract inhibits human B-lymphoblastoid cells growth via cell cycle arrest, apoptosis induction, and DNA damage

Publication . Santos, Klebson Silva; Costa, Carla; Bessa, Maria João; Teixeira, João Paulo; Muniz, Ana Veruska Cruz da Silva; Padilha, Francine Ferreira; Dariva, Cláudio; Oliveira, Maria Beatriz Pinto Prior

Aim: As far as is known, the pharmaceutical effects of neem on human B-lymphoblastoid (TK6) cells have not been studied until now. Hence, the present study aimed to obtain neem phenolic extracts for inhibits the proliferation of TK6 cells and explore some possible underlying mechanisms involved in these effects. Methods: Hexane extract (HE) was obtained in the first step. After that, the residual hexane was removed from the neem. The dried neem sample was used in a new extraction for obtaining the ethyl acetate extract (EAE). Total phenolic compounds (TPC) and total flavonoid contents (TFC) were determined by spectrophotometric methods. Lactate dehydrogenase (LDH) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) tests were used to evaluate the cytotoxicity in TK6 cells. The stop at G0/G1 cell cycle and inducing apoptosis in the TK6 cells were analyzed by flow cytometry. For deoxyribonucleic acid (DNA) damage evaluation, the alkaline comet test was used. Results: The higher TFC (65.50 mg/g of extract ± 1.17 mg/g of extract) and TPC (52.08 mg of extract ± 0.88 mg of extract) were obtained in EAE compared to HE that was obtained TFC of 14.61 mg/g of extract ± 0.60 mg/g of extract and TPC of 3.20 mg/g of extract ± 1.20 mg/g of extract. EAE was more significantly cytotoxic to TK6 cells than HE. The apoptosis induction was higher after exposure to 15.0 µg/mL of EAE (11.29%) in comparison to 15.0 µg/mL of HE (2.52%). The G0/G1 phase increased from 72% negative control (NC) to 83% after treatment with neem extracts (15 µg/mL). Neem extracts were also able to cause DNA strand breaks in TK6 cells. Conclusions: The extraction residue from neem leaf after hexane extraction is a source important of cytotoxic and genotoxic molecules against TK6 cells, the results also can suggest that the toxic effects in TK6 cells can be provided most likely due to the presence of high content of TPC from neem extracts.

2023-08-29Journal article

Open access

Are chloropropanols and glycidyl fatty acid esters a matter of concern in palm oil?

Publication . Albuquerque, T.G.; Costa, H.S.; Silva, M.A.; Oliveira, M.B.P.P.

Background: Palm oil is the most consumed worldwide, being evident its importance due to several applications by the food industry. The presence of toxic and mutagenic compounds, such as chloropropanols and glycidyl fatty acid esters has increased the attention on this oil. Very recently, there have been several alerts about the safety of certain foods containing palm oil in their composition. Consequently, the European Commission has demanded the evaluation of these hazardous compounds in several food matrices, including palm oil, turning this issue into a public health concern. Scope and approach: This review focuses on chloropropanols and glycidyl fatty acid esters in palm oil, their precursors, occurrence and mitigation strategies, as well as scientific opinions, current legislation and analytical approaches. Key findings and conclusions: Liquid chromatography coupled to mass spectrometry detection is preferred in the analysis of chloropropanols and glycidyl esters in palm oil. Concerning the precursors, the presence of chlorides and mono- and diglycerides is related to high amounts of these compounds, especially glycidyl esters. Several differences in the occurrence of these compounds in palm oil were reported, namely in what concerns to applied processing (refining/deodorization conditions; temperature/time of frying in the presence or absence of food). There is still much work to be done to implement efficient mitigation strategies without compromising the safety of palm oil. Furthermore, regarding the maximum levels in foodstuffs, European Commission is finalising the new EU legislation which will include values for the maximum levels of glycidyl fatty acid esters in vegetable oils.

2020-11Journal article

Restricted access