TLC and GC-MS analysis of petroleum ether fraction of fermented wood "Nikhra

10.31195/ejejfs.472038 Eurasian Journal of Forest Science 2019 7(1): 23-29 http://dergipark.gov.tr/ejejfs TLC and GC-MS analysis of petroleum ether fraction of fermented wood "Nikhra" of Acacia seyal Noha Fadle1, Abdalbasit Adam Mariod2, , Hiba Abdel Rahman Ali3 , Alfatih Ahmed Hasan4 1 The National Centre for Research, Environment and Natural Resources Research Institute, Khartoum, Sudan. 2 Indigenous Knowledge Center, Ghibaish College of Science and Technology, Ghibaish, Sudan. 3 Commission for Biotechnology and Genetic Engineering (CGEB), National Centre for Research 4 Department of Organic Chemistry, College of Science, Sudan University of Science and Technology, P.O. Box , Khartoum, Sudan. Corresponding author: Abstract This study aims to detect organoleptically Acacia seyal fractions (petroleum ether, chloroform, methanol and aqueous) fermented wood "Nikhra" ofthe fractions accumulating the strongest sweet fragrance these fragrances were mainly accumulated in the petroleum ether fermented wood "Nikhra" fraction and analysis fraction by using chromatographic and spectroscopic analysis. Petroleum ether fermented wood "Nikhra" fraction analysis with TLC and spray TLC with vanillin H2SO4 (pink) (C1, C2,C3,C4,C5,C6,C7), Rf values (0.92, 0.86, 0.71, 0.64, 0.57, 0.50, 0.36) ,were expected to be phenolic,with vanillin HCL (red) compounds spots (C5,C6,C7) with Rf values (0.57, 0.50, 0.37) was expected to be catechin and with vanillin H3PO4, blue-violet zones compounds spots (C6, C7), with Rf values (0.50, 0.36), were expected to be lignans. Petroleum ether fermented wood"Nikhra"fraction was divided into two types of compounds classes aromatic and non-aromatic by http://research.easybib.com and hence compounds were classified to phenolics and terpenoids compounds by GC/MS. Fragrant aromatics or terpenoids were targeted in this part of study. GC-MS analysis gave a spectrum of fragrance aromatic compounds (phenolics) in the petroleum ether fermented wood "Nikhra"fractions of A.seyal, was Petadecanoic acid (5.64%) and Tetracosamethyl-cyclododecasiloxane (4.17%) total fragrance aromatic compounds (44.57%), and main terponoids compounds was Octadecanoic acid (2.52) % total fragrance aromatic compounds (11.87%). Keywords: Acacia seyal, Gas Chromatography (GC), Thin-layer chromatography (TLC) Introduction The resinous heartwood of A. seyaltrees are usually used in Sudanese fragrances. The wood of A. seyal is pale yellow to medium brown, with localized pinkish-brown patches and some dark mahogany-red heartwood in larger or older individuals. A. seyal wood has potential in rural areas as timber. A. seyal, also produces a gum which, in spite of being of an inferior quality than that of A. senegal, is still marketed in Sudan 36.000-40.000 tons. The gum is edible when fresh, with a slightly acidic taste. Talh gum is attractive because of its clarity and solubility, gum is mixed with soot and powdered Nubian sandstone for black and red ink (Kimaroet al., 2011). Phytochemically A. seyal was characterized with high contents of proteins, phenolics, flavonoids and anthocyanins. The bark contains 18-30 % tannins and is a source of red dye (Orwa et al., 2012). The bark of A. seyal is the most valuable part of A. seyal. It is, extensively, used for feeding cattle, goats and sheep during the dry season. In human medicine A. 23 Eurasian Journal of Forest Science – TLC and GC-MS analysis of petroleum ether fraction by Fadle et al. 2019 seyal leaves, gum and bark are used in phytotherapy for haemorrhage, colds, diarrhoea, gastro-intestinal disorders, jaundice, biliary diseases, syphilis, and headaches and as emollient, astringent, for burns and ophtalmia (Orwa et al., 2012). Materials and Methods Collection of plant materials Fermented hardwood "Nikhra"of A. seyal were collected in March 2011 from Kordofan state, Sudan. They were, carefully, washed, oven-dried for 1 h at 50°C and put in the shade in an aerated place till complete drying, then were ground into a fine powder. Plant materials preparation and extraction A hundred g of ground powder of each plant was extracted using methanol and a soxhlet apparatus. The methanolic extract was fractionated, sequentially, using solvents of increasing polarity namely petroleum ether, chloroform, and aqueous. Fractions were dried using an evaporator and stored at 4 oC for further analysis (Fyhrquist et al., 2002). Fermented (Nikhra) wood A B Figure 1. (A).Tree of Acacia seyal (Talh) (B). Fermented wood (Nikhra) of Acacia seya Thin layer Chromatographic analysis TLC was performed on a pre-coated silica gel TLC plates grade F254 (E-Merck, Darmstadt, German) to determine the number of compounds present in petroleum ether fermented wood "Nikhra" fraction.sample was spotted at 1 cm from the bottom ofsilica gel plates using capillary tubes. Development of the chromatogram was done in closed tanks, in which the atmosphere has been saturated with eluent vapour by wetting a filter paper lining. The chromatogram was visualized under UV light (366 nm and 254 nm), Natural Product Reagent (NPR), H2SO4, HCL and H3PO4 acid reagent spray. The Rf values of the compounds were calculated using the following formula. Rf = distance travelled by the compound/Distance travelled by the solvent front GC-MS Analysis Petroleum ether fermented wood "Nikhra" fraction was subjected to GC MS analysis to identify the various bioactive compounds present. The sample was analyzed in GCMS-QP2010 Plus from Delhi 24 Eurasian Journal of Forest Science – TLC and GC-MS analysis of petroleum ether fraction by Fadle et al. 2019 University (DU) India. ACQ Mode Scan: 40m/z to 600m/z, Column flow is 1.21mL/min and total flow is 16.3ml/min. Flow control with linear velocity which 40.9cm/sec. The identification of compounds was done using computer matching of mass spectra with those of standards (WILEY8. LIB. and NIST11.library). RESULTS AND DISCUSSION Thin layer Chromatographic analysis The presence of flavonoids was confirmed by their color change from quenching fluorescence (254nm) to yellow or orange color for flavonoid and prominent blue color in case of flavonoidal acids or other phenolic acids (366 nm) after spraying with Natural Product Reagent (NPR). Polyphenols were mainly accumulated in the petroleum ether fraction has been detected using NPR. Fluorescence behavior of flavonoids in response to NPR is structure dependent. Flavonoids e.g. quercetin and myrecitin develops orange color and those of kaempferol and isorhamntin yellow to green colors. Flavones glycosides of luteolin develops orange colors and those apigenin yellow to green (Wagner and Bladt, 1996). Vanillin H2SO4 is a universal reagent that detects components of the petroleum ether fraction, terpenoids, phenols etc., typical pink to purple colors were developed upon spraying with vanillin H2SO4 (heat 110ºC). All phenolic at UV 254 nm show prominent quenching, and they give blue fluorescence at UV 366 nm (Wagner an (...truncated)