Ion-pairing chromatography and amine derivatization provide complementary approaches for the targeted UPLC-MS analysis of the polar metabolome

Liquid chromatography coupled to mass spectrometry is a key metabolomics technology. Reversed-phase liquid chromatog-raphy (RPLC) is very widely used as a separation step, possessing excellent characteristics with respect to reproducibility and reliability, but typically has poor retention of highly polar metabolites. Here, we evaluated the combination of two alter-native methods for improving retention of polar metabolites based on 6-aminoquinoloyl-N-hydroxysuccinidimyl carbamate derivatization for amine groups, and ion-pairing chromatography (IPC) using tributylamine as an ion-pairing agent to retain acids. We compared both of these methods to RPLC and also to each other, for targeted analysis using a triple-quadrupole mass spectrometer, applied to a library of ca. 500 polar metabolites. IPC and derivatization were complementary in terms of their coverage: combined, they improved the proportion of metabolites with good retention to 91%, compared to just 39% for RPLC alone. We detected 132 metabolites for real biological samples (liver extracts) with good reproducibility (based on coefficients of variation in pooled biological quality control samples). Finally, we tested the combination of methods with real-world samples by analyzing a set of liver extracts from aged male and female mice that had been treated with the poly-phenol compound ampelopsin. Furthermore, we also compared the results of these LC-MS methods to 1H NMR spectrosco-py as an orthogonal method (also termed statistical heterospectroscopy (SHY)), and found a strong correlation between the results of these different analytical approaches. By these means, not only were a number of significantly changed metabolites detected, but also it could be shown that there was a clear interaction between ampelopsin treatment and sex, in that the di-rection of metabolite change was opposite for males and females.

Antioxidant Activity Test Of Endophyte Fungi Extract From Kayu Jawa (Lannea coromandelica (Houtt.) Merr.) Roots Using Dpph Method (2,2-Diphenyl-1-1-Picrylhydrazyl)

Indonesia has abundant and diverse natural resources from land and sea. These biological natural resources can be explored as medicinal materials well-sourced from plants, animals, or microorganisms. Endophytic fungus is a microbial resource that grows in plant tissues that can produce compounds that have the same efficacy as the host plant. Zulfa (2016) has successfully isolated the endophytic fungi from kayu Jawa (Lannea coromandelica) roots and extracted the extracts with given code A11KA, A11KB, A12KC, A12KD, A21KK, A22KJ, AP12A, AP13L, AP21C, and AP32I. Other activities in these extracts besides antibacterial activity need to be tested. So this study was conducted to test the antioxidant activity of the extract with the DPPH method and used vitamin C positive control. Fifteen samples were obtained in this study, they are MeOH A11KA; MeOH A11KB; MeOH A12KC; MeOH A12KD; MeOH A21KK; MeOH AP12A; MeOH AP13L; MeOH AP12C; MeOH AP321; EA A11KA; EA A12KC; EA AP321; EA AP12A; EA AP13L; EA A22KJ from methanol fractions (MeOH) and ethyl acetate fraction (EA). The antioxidant activity test results showed that only six samples were potential as antioxidants. Value of AAI (Antioxidant Activity Index) EA A12KC; EA A11KA; MeOH A11KA; EA AP321; MeOH AP21C; EA A22KJ and vitamin C are 2.27 (very strong), 2.07 (very strong), 1.78 (strong), 1.19 (strong), 0.81 (medium), 0.53 (medium) and 11.67 (very strong) respectively. The result of antioxidant analysis using GC-MS showed only EA A11KA showing the presence of phenol compound, the largest phenol compound in this sample was phenol, 2,4-bis (1,1-dimethyl ethyl).

Jumlah Trombosit pada Mencit Diabetes setelah Pemberian Ekstrak Rumput Laut Merah (Rhodophyceae)

Oxidative stress in diabetes mellitus triggers activation and increases in platelet counts that play a role in the formation of platelet plugs in the blood clotting process. If excessive blood clots occur, there is a risk of cardiovascular disease. Red seaweed extract contains carotenoids, phenols and their derivatives, fikobilin pigments, polysaccharide sulfates, and vitamins. The biggest phenol compound is flavonoids which are compounds that can produce platelet antia-gregation effects. The purpose of this study was to examine the effect of giving red seaweed extract to platelet counts in diabetic mice. The study group consisted of the treatment group of red seaweed extract, normal control, positive control and negative controls. The dosage of seaweed extract as much as 10 mg / 20 grBB is given in condition. The calculation of platelet count is carried out directly with the Brecher Cronkite method using a calculated room Improved Neubauer. The One Way ANOVA test results showed a significance value (p <0.05), that is, the group of diabetic mice given red seaweed extract had a lower platelet count of 280000 μl compared to diabetic mice given metformin as much as 379000 μl. The decrease is due to flavonoids, saponins, alkaloids, β carotene, vitamins and minerals able to stabilize the production of megakaryocytes and platelets, prevent the destruction of pancreatic β cells and prevent free radicals. It was concluded that red seaweed extract can reduce platelet counts of diabetic mice near normal values.