Two-dimensional Thin-layer Chromatography Research Articles

Two-Dimensional High-Performance Thin-Layer Chromatography with Bioautography for Distinguishing Angelicae Dahuricae Radix Varieties: Chemical Fingerprinting and Antioxidant Profiling.

Angelicae Dahuricae Radix (ADR) holds a prominent place in traditional medicine for its remarkable antioxidative, anti-allergic, and antiproliferative capabilities. Recognized within the Korean Pharmacopoeia (KP 12th), Angelica dahurica (Hoffm.) Benth. and Hook.f. ex Franch. and Sav. (AD) and Angelica dahurica var. formosana (H. Boissieu) Yen (ADF) serve as the botanical origins for ADR. Differentiating these two varieties is crucial for the formulation and quality control of botanical drugs, as they are categorized under the same medicinal label. This research utilized two-dimensional high-performance thin-layer chromatography (2D-HPTLC) to effectively distinguish AD from ADF. Additionally, a quantitative analysis reveals significant differences in the concentrations of key active constituents such as oxypeucedanin, imperatorin, and isoimperatorin, with AD showing higher total coumarin levels. We further enhanced our investigative depth by incorporating a DPPH bioautography, which confirmed known antioxidant coumarins and unearthed previously undetected antioxidant profiles, including byakangelicin, byakangelicol, falcarindiol in both AD and ADF, and notably, 2-linoleoyl glycerol detected only in AD as an antioxidant spot. This comprehensive approach affords a valuable tool set for botanical drug development, emphasizing the critical need for accurate source plant identification and differentiation in ensuring the efficacy and safety of herbal medicine products.

Characterization of Secondary Metabolites of Leaf Buds from Some Species and Hybrids of Populus by Gas Chromatography Coupled with Mass Detection and Two-Dimensional High-Performance Thin-Layer Chromatography Methods with Assessment of Their Antioxidant Activity.

Poplars provide medicinal raw plant materials used in pharmacy. Leaf buds are one of the herbal medicinal products collected from poplars, having anti-inflammatory and antiseptic properties, but there are no quality standards for their production and there is a need to determine their botanical sources. Therefore, the chemical compositions of the leaf buds from four species and varieties of poplars, Populus balsamifera, P. × berolinensis, P. × canadensis 'Marilandica', and P. wilsonii were investigated and compared using gas chromatography coupled with mass detection (GC-MS) and two-dimensional high-performance thin-layer chromatography (2D-HPTLC) in order to search for taxa characterized by a high content of biologically active compounds and with a diverse chemical composition that determines their therapeutic effects. The presence of 163 compounds belonging to the groups of flavonoids, phenolic acids derivatives, glycerides, and sesquiterpenes was revealed. Moreover, the conditions for the separation and identification of biologically active compounds occurring in analyzed leaf buds using 2D-HPTLC were optimized and used for metabolomic profiling of the studied poplars, enabling their fast and simple botanical identification. The total phenolic (TPC) and flavonoid (TFC) contents of examined extracts were determined and their antioxidant capacities were estimated by spectrophotometric DPPH, ABTS, and FRAP assays. Based on the analysis of phytochemicals and antioxidant activity, P. × berolinensis buds were selected as the raw plant material for medicinal purposes with the highest content of active compounds and the strongest antioxidant activity.

Evaluation of Lipids for the Study of Photosynthetic Membranes.

The biological role of lipids goes far beyond the formation of a structural membrane bilayer platform for membrane proteins and controlling fluxes across the membranes. For example, in photosynthetic thylakoid membranes, lipids occupy well-defined binding niches within protein complexes and determine the structural organization of membrane proteins and their function by controlling generic physicochemical membrane properties. In this chapter, two-dimensional thin-layer chromatography (2D TLC) and gas chromatography (GC) techniques are presented for quantitative analysis of lipid classes and fatty acids in thylakoid membranes. In addition, lipid extraction methods from isolated thylakoid membranes and leaves are described together with a procedure for the derivatization of fatty acids to fatty acid methyl esters (FAME) that is required for GC analysis.

Composition of phospholipids of erythrocyte membranes in newborns from mothers with virus pneumonia caused by SARS-CoV-2 in the third trimester of pregnancy

Aim. To study the composition of phospholipids in erythrocyte membranes of newborns from mothers who had viral pneumonia caused by SARS-CoV-2 in the third trimester of pregnancy.Materials and methods. The study included newborns who were born to mothers diagnosed with COVID-19 and community-acquired pneumonia of viral etiology in the third trimester (main group). Of the total number of newborns (n=67), groups of children were formed from mothers with moderate pneumonia (group 1, n=34), and with severe pneumonia (group 2, n=33). The control group consisted of 35 newborns from practically healthy mothers. The quantitative composition of phospholipids in the membranes of erythrocytes of venous blood of the umbilical cord of newborns was determined by the method of two-dimensional thin-layer chromatography according to Kirchner.Results. The study found a statistically significant decrease in the concentration of phosphatidylethanolamine and phosphatidylcholine in erythrocyte membranes in group 1 by 19 and 20%, respectively (p<0.001), in group 2 – by 31 and 29%, respectively (p<0.001); a significant increase in the concentration of lysophosphatidylcholine, sphingomyelin, phosphatidylserine and phosphatidylinositol in group 1 by 43, 8, 15 and 26%, respectively (p<0.001), in group 2 – by 67, 14, 23 and 35%, respectively (p<0.001), compared with their concentration in newborns of the control group.Conclusion. Infection of the mother with SARS-CoV-2 during pregnancy is accompanied by structural disintegration of the erythrocyte membranes of their newborns, manifested by a change in the phospholipid profile in favor of an increase in the fractions of sphingo-, lyso-, inositoland serine phospholipids, which, ultimately, can disrupt the work of the oxygen transport system of the blood, contributing to the development of hypoxia.

Peculiarities of phospholipiid changes in erythrocyte membranes in parturient women with COVID-19-associated community-acquired pneumonia

Aim. To evaluate the phospholipid composition of erythrocyte membranes in parturient women who had COVID-19-associated community-acquired pneumonia (CAP) in the third trimester.Materials and methods. The material for the study was erythrocytes of peripheral blood of 65 parturient women diagnosed with COVID-19, moderate/severe course, CAP of viral etiology (main group). Patients of the main group, depending on the severity of CAP, were divided into two subgroups: subgroup 1 – moderate course of pneumonia (n=33), subgroup 2 – severe course of pneumonia (n=32). The control group consisted of 35 healthy parturient women. The quantitative composition of phospholipids was studied by two-dimensional thin-layer chromatography according to Kirchner.Results. In subgroup 1, the concentration of phosphatidylethanolamine and phosphatidylcholine in erythrocyte membranes was below the standard values by 38% and 29%, respectively (p<0.001), in subgroup 2, these indicators decreased by 32% and 48%, respectively (p<0.001). At the same time, a significant increase in the concentration of lysophosphatidylcholine was found in patients of subgroup 1 by 92% (p<0.001) and in patients of subgroup 2 by 110% (p<0.001), compared with the group of healthy individuals. In addition, structural changes in the lipid bilayer of erythrocyte membranes under conditions of COVID-19associated CAP were characterized by a pronounced increase in the concentration of minor fractions of phospholipids: phosphatidylserine and phosphatidylinositol in subgroup 1 by 63% and 53%, respectively (p<0.001), in subgroup 2 by 79% and 68%, respectively (p<0.001), compared with similar indicators in the control group.Conclusion. With COVID19-associated CAP in maternity women, structural disorganization of the phospholipid components of erythrocyte membranes is determined, manifested by a decrease in the concentration of phosphatidylethanolamine and phosphatidylcholine with a simultaneous increase in the level of lysophosphatidylcholine, phosphatidylserine and phosphatidylinositol. These disorders increase with increasing severity of pulmonary inflammation. The revealed changes in the lipid spectrum of peripheral blood and the composition of erythrocyte membrane phospholipids in COVID-19-associated CAP indicate the need to develop methods for their correction.

Pseudarthrobacter humi sp. nov., an actinobacterium isolated from soil.

A Gram-stain-positive, facultatively aerobic, rod-shaped and non-motile bacterial strain designated RMG13T was isolated from the soil near Gaetgol Eco Park and collected in Siheung-si, Republic of Korea. It was taxonomically characterized through polyphasic analysis. Phylogenetic analysis based on the 16S rRNA gene revealed that the novel isolate was most closely related to the type strains of species of the genus Pseudarthrobacter. RMG13T shared the highest similarities with Pseudarthrobacter sulfonivorans ALLT (99.2 %) and Pseudarthrobacter psychrotolerans YJ56T (99.0 %). In silico DNA-DNA hybridization values of RMG13T with P. sulfonivorans ALLT and P. psychrotolerans YJ56T were 28.1 and 41.8 %, respectively. The average nucleotide identities of RMG13T with P. sulfonivorans ALLT and P. psychrotolerans YJ56T were 84.2 and 70.3 %, respectively, whilst the average amino acid identities of RMG13T with P. sulfonivorans ALLT and P. psychrotolerans YJ56T were 90.5 and 74.6 %, respectively. Two-dimensional thin-layer chromatography showed that the major polar lipids of RMG13T were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylinositol, and its major cellular fatty acids were anteiso-C15 : 0, iso-C15 : 0 and anteiso-C17 : 0. MK-9 (H2) was the sole menaquinone. Cells grew at 4-37 °C (optimum 30 °C) and pH 5.0-12.0 (optimum pH 8.0) in Reasoner's 2A (MB cell). The cells tolerated 0-5% NaCl (w/v) but not 6 % NaCl. The DNA G+C content of RMG13T was 65.0 %. The results of phenotypic, genotypic, chemotaxonomic and phylogenetic analyses indicated that RMG13T represents a novel member of the genus Pseudarthrobacter, and the proposed name is P. humi sp. nov. The type strain of P. humi is RMG13T (= KACC 22359T = TBRC 15115T).

Identifikasi Komponen Kimia Saponin Greges Otot (Equisetum debile Roxb) Yang Berasal Dari Pallangga Kabupaten Gowa

Has done research on the identification of the chemical components of saponin Greges muscle (Equisetum debile Roxb) derived from Pallangga Gowa. Greges muscle is a plant that is used by people who believed Pallangga empirically nutritious treat fever and diarrhea. This study aims to determine the chemical content of saponin extract from muscle Greges preliminary test to the use of Uv-Vis spectrophotometry. The process of isolation of chemical compounds including reflux extraction with methanol, n-butanol fractionation, isolation by Preparative Thin Layer Chromatography and purity testing.
 Isolation fraction of n-butanol extract of muscle Greges using eluent chloroform-methanol-water (8: 2: 1) yielded isolates 5 isolates were named A, B, C, D, and E. Isolates D and E followed by the separation process using Layer Chromatography thin and Two-dimensional thin Layer Chromatography. The results of a preparative Thin Layer Chromatography isolates D and E give the appearance of a single stain on the purity test that can be said is pure stain. The results of UV-Vis spectrophotometry analysis of Fraction D isolates obtained λ max with a peak wavelength of 536 nm and 550 nm which is characteristic of saponin compounds.

ISOLASI DAN KARAKTERISASI SENYAWA FENOLIK DARI DAUN PUTAT (Planchonia valida Blume) (ISOLATION AND CHARACTERIZATION OF PHENOLIC COMPOUND FROM PUTAT LEAVES (Planchonia valida Blume))

Gallic acid have been isolated from the leaves of the Putat plant (Planchonia valida Blume). This study aimed to isolate and characterize isolates from Putat leaves by phytochemical and FTIR spectrophotometry. The research procedures included extraction by maceration, partitioning by fractionation, vacuum liquid chromatography (KCV), gravity column chromatography (KKG), thin layer chromatography (TLC), and FTIR analysis. A total of 1400 grams of putat leaf powder was macerated to produce 159.243 grams (11.38%) of methanol extract. A total of 101 grams of methanol extract was dechlorophyllated to obtain 21.430 grams (21.21%) of dechlorophyllated extract. The dechlorophyllated extract was partitioned gradually with n-hexane and ethyl acetate to produce n-hexane fractions of 0.341 grams (1.591%), and ethyl acetate of 10.143 grams (47.33%), and methanol of 9.170 grams (42.79%). A phytochemical test showed that in the dechlorophyllated extract, the methanol and ethyl acetate fractions contained alkaloids, phenolics, flavonoids, and terpenoids while the n-hexane fraction contained alkaloids and flavonoids. A total of 7 grams of ethyl acetate fraction was fractionated by vacuum liquid chromatography to produce fractions B1 to B8. The B4 fraction was continued to the separation step by gravity column chromatography so that the fractions S1 to S9 were obtained. The S4 fraction was continued to the separation stage using preparative thin layer chromatography (TLC) to produce isolates S4.1 to S4.5. Isolate S4.1 was tested for purity by two-dimensional TLC showed that the isolate was not pure. The results of the phytochemical test with 5% FeCl3 spray reagent indicated a positive phenolic compound which was strengthened by FTIR characterization of the presence of –OH, -CH aromatic, -CH aliphatic, C=C, C=O and C-O goups. Based on these results, it was concluded that the isolate from the dechlorophyllated extract of Putat leaves belonged to phenolic compounds.

The study of biological active compounds of lipophilic fraction of Schisandra chinensis (Turcz.) Baill seeds

Schisandra chinensis (Turcz.) Baill - Chinese magnolia - a well-known fruit plant intraditional Chinese and Korean medicine. The fruits and seeds of S. chinensis are rich in biologicallyactive substances. According to the literature (Irfan M. etc, 2020; Liu G-Z. etc, 2020; Sun W. etc,2021; Zhao T. etc., 2021; Bodrevich B.B. etc, 2017) the following therapeutic effects of S. chinensis
 fruits and seeds can be called: immunomodulatory, tonic, adaptogenic, anti-inflammatory, hepato-protective, nephroprotective, tonic. The study of the S. chinensis raw materials continues in the future,
 to reveal a wider range of therapeutic actions. The study of S. chinensis seeds is quite relevant toexpand the range of standardized medicinal raw materials, which is not included in the monographsof the State Pharmacopoeia of Ukraine. The main purpose of this study is to obtain the lipophilicfraction of S. chinensis seeds and to study the composition of different groups of lipophilic substancesin the resulting fraction. The main method of obtaining the lipophilic fraction of S. chinensis seeds isa comprehensive extraction in the Soxhlet apparatus. The chemical reaction was used to determinethe presence of tocopherols. The presence of the main groups of biologically active substances in thelipophilic fraction of S. chinensis seeds was established by the method of two-dimensional thin-layerchromatography. The study of fatty acid composition was performed by gas chromatography/massspectrometry (Su L. etc, 2020). The yield of lipophilic fraction of S. chinensis seeds was 35.79±0.7%.The obtained lipophilic fraction of S. chinensis seeds is an oily liquid of deep yellow color with a
 characteristic specific odor. The presence of tocopherols was established. Thin layer chromatogra-phy in the solvent system: I direction - hexane-acetone (8: 2), II direction - hexane-acetone (8: 4),
 development reagent - Stahl reagent, identified at least 14 compounds of terpene nature. As a resultof studies of fatty acids by gas chromatography/mass spectrometry, the quantitative content of 6 fattyacids was identified and established: palmitic, linoleic, oleic, stearic, gondoic and nonadecanoic. The
 quantitative content of unsaturated fatty acids significantly exceeded the quantitative content of satu-rated fatty acids. The results of the study give grounds to draw the following conclusions: the studies
 confirm the rich composition of the lipophilic fraction of S. chinensis seeds: tocopherols, 14 terpenes,6 fatty acids, of which saturated fatty acids (palmitic, stearic, nonadecanoic) and unsaturated fatty
 acids , oleic and gondoin); The results of the study suggest the anti-inflammatory, antioxidant, regen-erating effect of the lipophilic fraction of S. chinensis seeds.

New Mutations in cls Lead to Daptomycin Resistance in a Clinical Vancomycin- and Daptomycin-Resistant Enterococcus faecium Strain

Daptomycin (DAP), a last-resort antibiotic for treating Gram-positive bacterial infection, has been widely used in the treatment of vancomycin-resistant enterococci (VRE). Resistance to both daptomycin and vancomycin leads to difficulties in controlling infections of enterococci. A clinical multidrug-resistant Enterococcus faecium EF332 strain that shows resistance to both daptomycin and vancomycin was identified, for which resistance mechanisms were investigated in this work. Whole-genome sequencing and comparative genomic analysis were performed by third-generation PacBio sequencing, showing that E. faecium EF332 contains four plasmids, including a new multidrug-resistant pEF332-2 plasmid. Two vancomycin resistance-conferring gene clusters vanA and vanM were found on this plasmid, making it the second reported vancomycin-resistant plasmid containing both clusters. New mutations in chromosomal genes cls and gdpD that, respectively, encode cardiolipin synthase and glycerophosphoryl diester phosphodiesterase were identified. Their potential roles in leading to daptomycin resistance were further investigated. Through molecular cloning and phenotypic screening, two-dimensional thin-layer chromatography, fluorescence surface charge test, and analysis of cardiolipin distribution patterns, we found that mutations in cls decrease surface negative charges of the cell membrane (CM) and led to redistribution of lipids of CM. Both events contribute to the DAP resistance of E. faecium EF332. Mutation in gdpD leads to changes in CM phospholipid compositions, but cannot confer DAP resistance. Neither mutation could result in changes in cellular septa. Therefore, we conclude that the daptomycin resistance of E. faecium EF332 is conferred by new cls mutations. This work reports the genetic basis for vancomycin and daptomycin resistance of a multidrug-resistant E. faecium strain, with the finding of new mutations of cls that leads to daptomycin resistance.

Bacillus suaedae sp. nov., isolated from the stem of Suaeda aralocaspica in north-west China

A bacterial strain, designated YZJH907-2T, was isolated from the stem of Suaeda aralocaspica, collected from the southern edge of the Gurbantunggut desert, Xinjiang, PR China. Cells of strain YZJH907-2T were Gram-stain-positive, aerobic and rod-shaped. They formed white or colourless circular colonies with smooth convex surfaces. Strain YZJH907-2T grew at 4-50 °C (optimum, 28-30 °C), pH 7.0-10.0 (optimum, pH 8.0-9.0) and with 0-10 % (w/v) NaCl (optimum, 3-7 %). The genomic DNA G+C content of strain YZJH907-2T was 38.1 mol%. Phylogenetic analysis based on 16S rRNA gene sequence similarity showed that the strain was most closely related to Bacillus alcalophilus DSM 485T (97.37 %), Bacillus kiskunsagensis B16-24T (96.87 %) and Bacillus bogoriensis LBB3T (96.71 %). Average nucleotide identity values between YZJH907-2T and B. alcalophilus DSM 485Tand B. bogoriensis LBB3T were 69.2 and 69.0 %, respectively. Digital DNA-DNA hybridization values of YZJH907-2T with B. alcalophilus DSM 485T and B. bogoriensis LBB3T were 19.6 and 20.4 %, respectively. The cell wall of strain YZJH907-2T contained meso-diaminopimelic acid, and the major and secondary isoprenoid quinones were MK-7 and MK-5, respectively. Results of fatty acids showed that anteiso-C15 : 0, iso-C15 : 0 and C16 : 0 were the predominant cellular fatty acids. Two-dimensional thin-layer chromatography analysis indicated that the polar lipids included diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, three unidentified phospholipids and two unidentified glycolipids. Based on the genomic, phylogenetic and phenotypic analyses, strain YZJH907-2T represented a novel species of the genus Bacillus, and thus the name Bacillus suaedae sp. nov. is proposed. The type strain is YZJH907-2T (=CGMCC 1.18763T=KCTC 43335T).

AKTIVITAS ANTIOKSIDAN DAN KARAKTERISASI ISOLAT DARI FRAKSI n-HEKSANA PADA RUMPUT LAUT (Caulerpa sertularioides) ASAL SINGKAWANG KALIMANTAN BARAT (ANTIOXIDANT ACTIVITY AND ISOLATE CHARACTERIZATION OF n-HEXANE FRACTION IN SEAWEED (Caulerpa sertularioides) FROM SINGKAWANG WEST KALIMANTAN)

Seaweed (Caulerpa sertularioides) is one of the marine plants of West Kalimantan, which contains bioactive compounds that can act as antioxidants. The purpose of this study was to determine antioxidant activity and identify components of organic compounds in seaweed C. sertularioides. This research was carried out in several stages, namely extraction, fractionation, antioxidant test (DPPH), vacuum liquid chromatography (KVC), gravity column chromatography (KKG), and two-dimensional thin-layer chromatography (2-Dimensional TLC), and characterization. Antioxidant activity test by DPPH method on methanol extract, n-hexane fraction, and fraction DCM from seaweed C. sertularioides obtained IC50 values of 82.21 g/mL (strong antioxidant), 131.2 g/mL (medium antioxidant), and 192.43 g/mL (weak antioxidant). The results of the FT-IR identification of isolate I4 showed a strong absorption at 3483-3387 cm-1,which was specific for the OH group. The presence of strong absorption of 2926-2854cm-1 C-H stretching group, a wide absorption of 1338 cm-1 of methyl and methylene groups (stretching). There was a strong absorption of 1543-1463 cm-1 C-O ester group (bend), a broad absorption of 1074 cm-1 C-O (stretching), and absorption of 1795-1735 cm-1 specific for the C=O ester group. The results of GC-MS on isolate I4 indicated the main component was compound 1-nonadecene (14.09%), Trifluoroacetoxy hexadecane based on the literature, the compound included the organic component of fatty acids, and esters

Evaluation of in vitro Cholesterol Esterase Inhibitory Activity of Few Selected Vegetables – An Indication of Cardioprotective Property

Pharmacognosy Research,2022,14,2,188-194.DOI:10.5530/pres.14.2.27Published:April 2022Type:Original ArticleAuthors:Thiyagarajan Sathishkumar, Muruvanda Bhuvan Uthaiah, Ravichandran Vinu Varshith, Kuppamuthu Kumaresan, and Vinohar Stephen Rapheal Author(s) affiliations:Thiyagarajan Sathishkumar, Muruvanda Bhuvan Uthaiah*, Ravichandran Vinu Varshith, Kuppamuthu Kumaresan, Vinohar Stephen Rapheal Department of Biotechnology, Kumaraguru College of Technology, Coimbatore, Tamil Nadu, INDIA. Abstract:Background: Cardiovascular diseases are responsible for severe mortality rate amongst the population. Several drugs available to control and/ or cure the same, cause severe adverse effects amidst human population. Contextually, the present study was devised to screen the in vitro cholesterol esterase (CEase) activity of few selected vegetables as an indicator of cardioprotective property. Objectives: To screen and purify the polyphenolic fractionate from the vegetable(s) extract that record an optimal CEase activity. Materials and Methods: CEase inhibitory activity was investigated using standard spectrophotometric assay. Microwave assisted central composite design based response surface methodology (CCD based RSM) was executed to quantify an improvement in the CEase inhibitory activity of Solanum melongena extract. One dimensional thin layer chromatography (1D TLC) and two-dimensional preparative thin layer chromatography (2D PTLC) analyses were carried out for Brassica oleracea var. botrytis hot water extract to identify and eluate the polyphenolic fractionate, respectively. Results: Significant CEase inhibitory activity was noticed in the hot water extract of Brassica oleracea var. botrytis (84.65 ± 3.29%) than the other vegetable extracts. Even though the optimal process model was validated (at 5% level), no significant improvement was recorded in the CEase inhibition. The presence of polyphenolic content in the hot water extract of Brassica oleracea var. botrytis (Rf = 0.69) was confirmed by comparing with standard quercetin (Rf = 0.60). Strong blue and moderate brownish yellow color spots revealed the presence of phenolic acid and flavonoid eluates respectively, were purified using 2D PTLC technique. The eluate has recorded a near moderate (28.57 ± 2.12%) CEase inhibitory activity. Conclusion: Brassica oleracea var. botrytis possess significant CEase inhibitory activity and proved to be a good diet to protect from various cardiovascular diseases. Key words: Brassica oleracea var. botrytis, Cholesterol esterase, Solanum melongena, Microwave Assisted Response Surface Methodology, Flavonoid. View:PDF (245.05 KB)

EFFECT OF N-STEAROYLETHANOLAMINE ON THE LIPID COMPOSITION OF THE FRONTAL CORTEX AND HIPPOCAMPUS OF THE RAT'S BRAIN AT THE AGING

Aim. To study the possible protective effect of cannabimimetic lipid - N-stearoylethanolamine (NSE) on the lipid composition of the frontal cortex, hippocampus and on the state of episodic memory of old rats. Methods. Extraction of lipids from the tissues of the hippocampus and frontal cortex of rats was performed by the method of Bligh and Dyer. Phospholipids were separated by two-dimensional thin layer chromatography. Methyl esters of fatty acids from lipid extract were obtained by a modified method of Carreau and Dubaco. Quantitative analysis of fatty acid methyl esters was performed by gas-liquid chromatography on an Agilent GC7890 chromatograph with an Agilent 8987 mass detector. The fractions of free and esterified cholesterol were separated by one-dimensional thin layer chromatography. The dry cholesterol residue was analyzed on a Carlo Erba gas-liquid chromatograph. Results. The study of the diacyl (DF) and plasmalogen (PF) forms of phospholipids (PLs) content in the frontal cortex and hippocampus have shown a significant decrease in the plasmalogen form of PE (Phosphatidylethanolamine) (up to 15%) and an increase in its DF, compare to its content in young rats. Administration of NSE to old rats led to a significant increase in PF PE and did not cause significant changes in the content of PF in the composition of other PL of the frontal cortex of the brain and hippocampus. The decrease in the percentage of various phospholipids was found in frontal cortex and hippocampus of old rats: the content of phosphatidylcholine (PC) and phosphatidylinositol (PI) was significantly reduced in the frontal cortex and the decrease of diphosphatidylglycerol (DPG), PI and phosphatidylserine (PS) was found in the hippocampus, compare to the young animals. Administration of NSE to old rats had a different effects on the content of various phospholipids. The increase in the content of PC and PI in the frontal cortex and PS and DPG in the hippocampus is particularly pronounced due to NSE. An increase in the content of saturated fatty acids (FFAs ) and a decrease in the content of unsaturated FFAs in the frontal cortex and hippocampus of old rats also has been found. It has also been found that NSE administration to old rats promoted the growth of the free cholesterol level in the frontal cortex and hippocampus. The results of the New Object Recognition test in old rats have shown that a short-term memory has been improved by NSE. Conclusions. The administration of NSE to old rats causes an increase in PF of PLs in the frontal cortex and hippocampus of the brain, which can be considered as one of the mechanisms of neuroprotective action of NSE in aging. The changes in the phospholipids and fatty acids composition, and free cholesterol level of the frontal cortex and hippocampus of the brain of old rats caused by NSE administration have been shown to be adaptive and restorative. The New Object Recognition Behavioral Test has shown that NSE restores short-term memory in older rats. The obtained results expand the understanding of the mechanisms of biological action of NSE during aging in mammals and create the basis for the development a new drug with geroprotective properties.

Two-Dimensional Thin Layer Chromatography of Melatonin and Related Compounds.

Two-dimensional thin layer chromatography has been used by workers in the field to separate radiolabeled serotonin derivatives from complex mixtures of culture media and homogenates of glands. The compounds resolved include N-acetylserotonin, melatonin, hydroxytryptophol, methoxytryptophol, hydroxyindole acetic acid, and methoxyindole acetic acid. The method requires either radiolabeled tryptophan or serotonin, if an investigator wants to study conversion. It is also useful in the chemical synthesis of serotonin metabolites because it is relatively fast. It pointed to the enzyme that converts serotonin to N-acetylserotonin as being key in controlling the nocturnal increase in vertebrate melatonin production. This enzyme, arylalkylamine N-acetyltransferase (E.C. 2.3.1.87), has been the focus of hundreds of papers which probed its biology, biochemistry, molecular biology, structural biology, neural regulation, development, evolution, and genetics.

Isolation and identification of flavonoids from avocado leaves (Persea americana Mill)

Leaves of avocado (Persea americana Mill) contains bioactive compounds that can act as natural antioxidants. This study was conducted to isolate and identify the flavonoid compounds of avocado leaves. Research stages includes maceration, fractionation, thin-layer chromatography analysis, isolation and purification of compounds also identification of isolates in two-dimensional thin-layer chromatography and by using three different types of solvents. Isolates were identified using ultraviolet-visible spectrophotometry and infrared spectrophotometry. From the results obtained one isolate FA6 showed a single compound. Based on the results by testing the purity of isolates using two-dimensional thin-layer chromatography and by using three different types of solvents, isolates FA6 is a pure compound. Identification by using ray-visible spectrophotometry FA6 isolates showed maximum wavelength at 280.6 nm and 342.0 nm on bands II and on band I. Infrared spectrum ofisolate FA6 showed aromatic OH groups, aliphatic CH, C = O stretching, C = C aromatic, and C-O-C stretch. From the results obtained isolates FA6 is a class of flavonoids group flavones or flavonol (3-OH substitional).

Polyisoprenoid composition in chengham (Scyphiphora hydrophyllacea, Gaertn. f., Rubiaceae)

Chengham (Scphyphora hydrophyllacea Gaertn. f., Rubiaceae) is a mangrove shrub or tree commonly in landward mangrove zones having some biological activities. All parts of S. hydrophyllacea have shown some components benefits to biological and pharmaceutical properties. This present study aimed to analyze the distribution and composition of polyisorenoid in senesnece leaves, fruits and branches of S. hydrophyllacea. Polyisoprenid patter was evaluated by two-dimensional thin layer chromatography (2D-TLC) approach. The total lipid (TL) content of five samples ranged from 5.7 mg/g in branches to 10.0 mg/g in the old leaves. Additionally, the percentage of polyprenol of senescence leaves slightly abundance compare to dolichol in the polyisoprenoid presence. Overall, polyprenol was found in higher content than dolichol in the yellow leaves. Results of this study found the type-II of S. hydrophyllacea fruits was in line with earlier reports on the plant fruits. The present study provided the occurrence of polyprenols and dolichols in S. hydrophyllacea tested organs.

Polyisoprenoid Profiling of Mangrove Litters–based Zonations and Salinity Groups in North Sumatra, Indonesia

The polyprenols and dolichols in mangrove litter–based salinity groups and zonations in Lubuk Kertang, North Sumatra, Indonesia, was performed using two-dimensional thin-layer chromatography. Eight sites with twenty-four samples consisting of 0, 2, and 3% salt concentrations and five zonations (<em>Avicennia spp, Bruguiera spp, Nypa fruticans</em> community, <em>Rhizophora </em>spp<em>,</em> and <em>Sonneratia </em>spp) were analyzed. In the zonations, two types concerning the distribution of polyprenols and dolichols were detected. Type-I, showing predominance of dolichols over polyprenols, was observed in <em>Avicennia </em>spp<em>, Bruguiera spp, Nypa fruticans,</em> and <em>Rhizophora </em>spp<em>.</em> Type-II, having both polyprenols and dolichols, was observed in <em>Sonneratia </em>spp<em>.</em> In contrast, no type-I distribution was found in the salinity group. A type-II distribution was also observed in 0, 2, and 3% salt concentrations. The diversity of polyisoprenoid composition in the mangrove litters of salinity groups was noted, whereas dolichols predominated in the zonations (80%). In <em>Avicennia spp</em> litter, dolichols were found to be longer than other types of community litter (<em>Bruguiera </em>spp<em>, Nypa, and Rhizophora </em>spp). These conditions can be caused by leaf litter factors that have different ages and environments. A dendrogram was constructed using the Unweighted-Pair Group Method with Arithmetic mean (UPGMA) method to confirm these findings. The dendrogram demonstrated that the zonations and salinity groups were generally clustered according to appropriate species and families. The study suggested that dominated dolichols function as chemotaxonomic markers, useful in identifying and classifying mangroves, and in phylogenetic studies.

Polyisoprenoid Distribution in Stems and Leaves of Pinus Merkusii Strains

Living things produce secondary metabolites, one of which is abundant in plants, namely polyisoprenoid alcohol compounds. Polyisoprenoids are also found in plants with different chain lengths. The distribution of polyisoprenoid compounds (dolichol and polyprenol) was found in the leaf and stem tissues of Pinus merkusii strains derived from Aceh, Tapanuli and Kerinci. The extracted samples were analyzed by the two-dimensional thin-layer chromatography (2D-TLC) method. The distribution of dolichol and polyprenol compounds on the stems and leaves of Aceh, Tapanuli and Kerinci pines provided a type II categorization, both dolichol and polyprenol were traced in pine tissues. Total lipids in stem tissue ranged from 564.4-58.9 mg/g dw, polyisoprenoid values ranged from 1.34-2.44 mg/g dw. In the leaves, total lipids ranged from 590 to 669.43 mg/g dw while polyisoprenoid values ranged from 1.29 to 5.70 mg/g dw. The dolichol carbon chain length in the Pine stem of strain composed C70-C90, C65-C90, and C65-C95. Meanwhile, carbon chain-length of C85-C105, C70-C95, and C50-C65 were found in leaf tissues. The length of the polyprenol carbon chain in the stem was C60-C90, C50-C90, and C70-C90 respectively, while the chain lengths on the leaves were C60-C115, C50-C95, and C30-C90. The present study suggested the presence of both dolichol and polyprenols in Pinus merkusii without predomination either dolichol or polyprenol.

Polyisoprenoid Compounds from Tropical Fruit Trees in Universitas Sumatera Utara Campus

Tropical trees are a source of secondary metabolite compounds that have various biological activities that can help human life both for industrial and pharmaceutical needs. One of the secondary metabolites is polyisoprenoid. This study aims to identify and analyze polyisoprenoid compounds quantitatively from tropical fruit trees at the Universitas Sumatera Utara. Nine samples of tropical fruit were obtained, namely: Psidium guajava, Tamarindus indica, Manilkara kauki, Morinda citrifolia, Mangifera indica, Artocarpus communis, Artocarpus heterophyllus, Gmelina arborea, and Syzygium aqueum. Each sample was extracted and isolated the polyisoprenoid alcohol, then analyzed using two-dimensional thin-layer chromatography. The polyisoprenoid in fruit samples found at the Universitas Sumatera Utara was a type II and III polyisoprenoid compound. The total lipid values ranged from 48.7 mg/g dw to 262.9 mg/g dw, polyisoprenoids ranged from 1.2 mg/g dw to 9.5 mg/g dw, and polyprenols ranged from 0.5 mg/g dw to 5.7 mg/g dw. Carbon chain length and dolichol polyprenol lengths of each fruit were collected (Psidium guajava L.) (C70 C80), (Tamarindus indica L.) (C80 C95), (Manilkara kauki L.) (C80 C95 and C70 C90), (Morinda citrifolia L.) (C75 C100), (Mangifera indica L.) (C80 C90 and C75 C90), (Artocarpus communis) (C75 C95), (Artocarpus heterophyllus) (C70 C75 and C70 C100), (Gmelina arborea Roxb.) (C90 C95), and (Syzygium aqueum) (C65 C90 and C75 C90).