Application of effect-directed analysis using TLC-bioautography for rapid isolation and identification of antidiabetic compounds from the leaves of Annona cherimola Mill.

INTRODUCTION: Type 2 diabetes mellitus is a globally prevalent chronic disease characterised by hyperglycaemia and oxidative stress. The search for new natural bioactive compounds that contribute to controlling this condition and the application of analytical methodologies that facilitate rapid detection and identification are important challenges for science. Annona cherimola Mill. is an important source of aporphine alkaloids with many bioactivities. OBJECTIVE: The aim of this study is to isolate and identify antidiabetic compounds from alkaloid extracts with α-glucosidase and α-amylase inhibitory activity from A. cherimola Mill. leaves using an effect-directed analysis by thin-layer chromatography (TLC)-bioautography. METHODOLOGY: Guided fractionation for α-glucosidase and α-amylase inhibitors in leaf extracts was done using TLC-bioassays. The micro-preparative TLC was used to isolate the active compounds, and the identification was performed by mass spectrometry associated with web-based molecular networks. Additionally, in vitro estimation of the inhibitory activity and antioxidant capacity was performed in the isolated compounds. RESULTS: Five alkaloids (liriodenine, dicentrinone, N-methylnuciferine, anonaine, and moupinamide) and two non-alkaloid compounds (3-methoxybenzenepropanoic acid and methylferulate) with inhibitory activity were isolated and identified using a combination of simple methodologies. Anonaine, moupinamide, and methylferulate showed promising results with an outstanding inhibitory activity against both enzymes and antioxidant capacity that could contribute to controlling redox imbalance. CONCLUSIONS: These high-throughput methodologies enabled a rapid isolation and identification of seven compounds with potential antidiabetic activity. To our knowledge, the estimated inhibitory activity of dicentrinone, N-methylnuciferine, and anonaine against α-glucosidase and α-amylase is reported here for the first time.

Advances in analytical techniques coupled to in vitro bioassays in the search for new peptides with functional activity in effect-directed analysis.

Bioactive peptides derived from food proteins have important biological effects. The search for new bioactive peptides from natural sources needs to base their decisions on evidence from methods providing structural information, where classic biological assays do not provide it. In this context, high-performance thin-layer chromatography (HPTLC) coupled with bioassays in effect-directed analysis (EDA) has been used recently to detect the bioactivities of peptides. In EDA, peptides are separated and the biological effects of individual fractions can be determined. HPTLC-EDA coupled with MS represents an emerging technology, complementary to the more common HPLC and electrophoresis analytical methods used for peptide analysis. In addition, thin-layer chromatography (TLC) can be used to identify the origins of substances causing biological effects. This review provides an overview of the status of analytical methods in these systems as well as new strategies for the detection and characterization of bioactive peptides.

Antioxidant Compound Adsorption in Polyvinylpolypyrrolidone from Chilean Carménère, Cabernet Sauvignon, and Merlot Grape Pomaces as Potential By-Products.

Grape pomace (GP) is a by-product resulting from the winemaking process and its potential use as a source of bioactive compounds is well known. The GP bioactive compounds can be retained in the well-known polyvinylpolypyrrolidone (PVPP), industrially used in the clarification and stabilization of wine and other drinks. Thus, the polyphenolic compounds (PC) from the Chilean Carménère, Cabernet Sauvignon, and Merlot GP were extracted, and their compositions and antioxidant capacities (ORAC-FL) were determined. In addition, the retention capacity of the PC on PVPP (PC-PVPP) was evaluated. The bioactivities of GP extracts and PC-PVPP were estimated by the agar plate inhibition assay against pathogenic microorganisms. Results showed a high amount of TPC and antioxidant capacity in the three ethanolic GPs extracts. Anthocyanins, flavan-3-ol, and flavonols were the most abundant compounds in the GP extract, with retentions between 70 and 99% on PVPP. The GP extracts showed inhibition activity against B. cereus and P. syringae pv. actinidiae but the GP-PVPP had no antimicrobial activity. The high affinity of the identified PCs from GPs on PVPP polymer could allow the design of new processes and by-products for the food or cosmeceutical industry, promoting a circular economy by reducing and reusing wastes (GPs and PVPP) and organic solvents.

Phycobiliproteins: Structural aspects, functional characteristics, and biotechnological perspectives.

Phycobiliproteins (PBPs) are fluorescent proteins of various colors, including fuchsia, purple-blue and cyan, that allow the capture of light energy in auxiliary photosynthetic complexes called phycobilisomes (PBS). PBPs have several highly preserved structural and physicochemical characteristics. In the PBS context, PBPs function is capture luminous energy in the 450-650 nm range and delivers it to photosystems allowing photosynthesis take place. Besides the energy harvesting function, PBPs also have shown to have multiple biological activities, including antioxidant, antibacterial and antitumours, making them an interesting focus for different biotechnological applications in areas like biomedicine, bioenergy and scientific research. Nowadays, the main sources of PBPs are cyanobacteria and micro and macro algae from the phylum Rhodophyta. Due to the diverse biological activities of PBPs, they have attracted the attention of different industries, such as food, biomedical and cosmetics. This is why a large number of patents related to the production, extraction, purification of PBPs and their application as cosmetics, biopharmaceuticals or diagnostic applications have been generated, looking less ecological impact in the natural prairies of macroalgae and less culture time or higher productivity in cyanobacteria to satisfy the markets and applications that require high amounts of these molecules. In this review, we summarize the main structural characteristics of PBPs, their biosynthesys and biotechnological applications. We also address current trends and future perspectives of the PBPs market.