Harmalan

Integration of LC-MS-Based and GC-MS-Based Metabolic Profiling to Reveal the Effects of Domestication and Boiling on the Composition of Duck Egg Yolks.[Pubmed:36677059]

Metabolites. 2023 Jan 16;13(1):135.

Egg yolks contain abundant lipids, proteins, and minerals that provide not only essential nutrients for embryonic development but also cheap sources of nutrients for consumers worldwide. Previous composition analyses of egg yolks primarily focused on nutrients such as lipids and minerals. However, few studies have reported the effects of domestication and heating on yolk composition and characteristics. The objective of this study was to investigate the impact of domestication and boiling on the metabolite contents of egg yolks via untargeted metabolomics using GC-MS and LC-MS. In this study, eggs were collected from Fenghua teals, captive mallards, and Shaoxing ducks. Twelve duck eggs (half raw and half cooked) were randomly selected from each variety, and the egg yolks were separated for metabolic profiling. The analysis identified 1205 compounds in the egg yolks. Domestication generated more differential metabolites than boiling, which indicated that the changes in the metabolome of duck egg yolk caused by domestication were greater than those caused by boiling. In a comparative analysis of domestic and mallard ducks, 48 overlapping differential metabolites were discovered. Among them, nine metabolites were upregulated in domesticated ducks, including monoolein, emodin, daidzein, genistein, and glycitein, which may be involved in lipid metabolism; some of them may also act as phytoestrogens (flavonoids). Another 39 metabolites, including imethylethanolamine, Harmalan, mannitol, nornicotine, linoleic acid, diphenylamine, proline betaine, alloxanthin, and resolvin d1, were downregulated by domestication and were linked to immunity, anti-inflammatory, antibacterial, and antioxidant properties. Furthermore, four overlapping differential metabolites that included amino acids and dipeptides were discovered in paired comparisons of the raw and boiled samples. Our findings provided new insights into the molecular response of duck domestication and supported the use of metabolomics to examine the impact of boiling on the composition of egg yolks.

Comprehensive Metabolomic Comparison of Five Cereal Vinegars Using Non-Targeted and Chemical Isotope Labeling LC-MS Analysis.[Pubmed:35629931]

Metabolites. 2022 May 10;12(5):427.

Vinegar is used as an acidic condiment and preservative worldwide. In Asia, various black vinegars are made from different combinations of grains, such as Sichuan bran vinegar (SBV), Shanxi aged vinegar (SAV), Zhenjiang aromatic vinegar (ZAV), and Fujian Monascus vinegar (FMV) in China and Ehime black vinegar in Japan (JBV). Understanding the chemical compositions of different vinegars can provide information about nutritional values and the quality of the taste. This study investigated the vinegar metabolome using a combination of GC-MS, conventional LC-MS, and chemical isotope labeling LC-MS. Different types of vinegar contained different metabolites and concentrations. Amino acids and organic acids were found to be the main components. Tetrahydroharman-3-carboxylic acid and Harmalan were identified first in vinegar. Various diketopiperazines and linear dipeptides contributing to different taste effects were also detected first in vinegar. Dipeptides, 3-phenyllactic acid, and tyrosine were found to be potential metabolic markers for differentiating vinegars. The differently expressed pathway between Chinese and Japanese vinegar was tryptophan metabolism, while the main difference within Chinese vinegars was aminoacyl-tRNA biosynthesis metabolism. These results not only give insights into the metabolites in famous types of cereal vinegar but also provide valuable knowledge for making vinegar with desirable health characteristics.

Chromatographic determination of harmalans in the urine of autistic children.[Pubmed:28182283]

Biomed Chromatogr. 2017 Sep;31(9).

This paper presents a new approach to autism - a complex and still enigmatic condition. We present the results of our preliminary research which was based on the detection of the hallucinogenic substance 6- (or 10-)methoxyHarmalan in the urine samples of autistic children with the use of chromatographic methods. Additionally, we aim to describe the relationship between the level of tryptophan and Harmalan, and the influence of supplementation on the level of this compound. We applied HPLC-UV/vis, HPLC-DAD and LC-MS in order to determine McIsaac's compound in the urine samples obtained from autistic children (n = 132) and healthy individuals (n = 10). The level of tryptophan was quantified with the use of GC-MS. Our research shows the presence of the McIsaac's compound in 110 samples of ASD children contrary to healthy children, where it was not found. No relationship between the level of tryptophan and 6-methoxyHarmalan was noticed. The study shows a strong influence of melatonin supplementation on the presence of the McIsaac's compound. We believe that the results of our research can contribute to a better understanding of autism spectrum disorders. Moreover, our findings can form the basis for other studies focused on autism, eventually making it possible to understand its etiology.

Synthesis and antiviral and fungicidal activity evaluation of beta-carboline, dihydro-beta-carboline, tetrahydro-beta-carboline alkaloids, and their derivatives.[Pubmed:24460429]

J Agric Food Chem. 2014 Feb 5;62(5):1010-8.

Six known beta-carboline, dihydro-beta-carboline, and tetrahydro-beta-carboline alkaloids and a series of their derivatives were designed, synthesized, and evaluated for their anti-tobacco mosaic virus (TMV) and fungicidal activities for the first time. All of the alkaloids and some of their derivatives (compounds 3, 4, 14, and 19) exhibited higher anti-TMV activity than the commercial antiviral agent Ribavirin both in vitro and in vivo. Especially, the inactivation, curative, and protection activities of alkaloids Harmalan (62.3, 55.1, and 60.3% at 500 mug/mL) and tetrahydroharmane (64.2, 57.2, and 59.5% at 500 mug/mL) in vivo were much higher than those of Ribavirin (37.4, 36.2, and 38.5% at 500 mug/mL). A new derivative, 14, with optimized physicochemical properties, obviously exhibited higher activities in vivo (50.4, 43.9, and 47.9% at 500 mug/mL) than Ribavirin and other derivatives; therefore, 14 can be used as a new lead structure for the development of anti-TMV drugs. Moreover, most of these compounds exhibited good fungicidal activity against 14 kinds of fungi, especially compounds 4, 7, and 11.

Molecular analysis and structure-activity relationship modeling of the substrate/inhibitor interaction site of plasma membrane monoamine transporter.[Pubmed:21816955]

J Pharmacol Exp Ther. 2011 Nov;339(2):376-85.

Plasma membrane monoamine transporter (PMAT) is a new polyspecific transporter that interacts with a wide range of structurally diverse organic cations. To map the physicochemical descriptors of cationic compounds that allow interaction with PMAT, we systematically analyzed the interactions between PMAT and three series of structural analogs of known organic cation substrates including phenylalkylamines, n-tetraalkylammonium (n-TAA) compounds, and beta-carbolines. Our results showed that phenylalkylamines with a distance between the aromatic ring and the positively charged amine nitrogen atom of approximately 6.4 A confer optimal interactions with PMAT, whereas studies with n-TAA compounds revealed an excellent correlation between IC(50) values and hydrophobicity. The five beta-carbolines that we tested, which possess a pyridinium-like structure and are structurally related to the neurotoxin 1-methyl-4-phenylpyridinium, inhibited PMAT with high affinity (IC(50) values of 39.1-65.5 muM). Cytotoxicity analysis further showed that cells expressing PMAT are 14- to 15-fold more sensitive to Harmalan and norharmanium, suggesting that these two beta-carbolines are also transportable substrates of PMAT. We then used computer-aided modeling to generate qualitative and quantitative three-dimensional pharmacophore models on the basis of 23 previously reported and currently identified PMAT inhibitors and noninhibitors. These models are characterized by a hydrogen bond donor and two to three hydrophobic features with distances between the hydrogen bond donor and hydrophobic features ranging between 5.20 and 7.02 A. The consistency between the mapping results and observed PMAT affinity of a set of test compounds indicates that the models performed well in inhibitor prediction and could be useful for future virtual screening of new PMAT inhibitors.

Phytochemical and pharmacological study of roots and leaves of Guiera senegalensis J.F. Gmel (Combretaceae).[Pubmed:16443341]

J Ethnopharmacol. 2006 Jun 30;106(2):173-8.

The chemical composition of total alkaloids from leaves and roots of Guiera senegalensis was investigated. Three beta-carboline alkaloids were purified: in addition to harman and tetrahydroharman, known in roots and leaves, Harmalan (dihydroharman) was isolated for the first time from roots of Guiera senegalensis. Guieranone A, a naphthyl butenone, was also purified from leaves and roots. The in vitro antiplasmodial activity and the cytotoxicity of extracts and pure compounds were evaluated. Each total alkaloid extract and beta-carboline alkaloids presented an interesting antiplasmodial activity associated with a low cytotoxicity. Harmalan was less active than harman and tetrahydroharman. Guieranone A showed a strong antiplasmodial activity associated with a high cytotoxicity toward human monocytes. Its cytotoxicity was performed against two cancer cell lines and normal skin fibroblasts in order to study its anticancer potential: guieranone A presented a strong cytotoxicity against each cell strains. Finally, we evaluated the potent synergistic antimalarial interaction between Guiera senegalensis and two plants commonly associated in traditional remedies: Mitragyna inermis and Pavetta crassipes. Three associations evaluated were additive. A synergistic effect was shown between total alkaloids extracted from leaves of Guiera senegalensis and those of Mitragyna inermis. This result justified the traditional use of the plants in combination to treat malaria.

Harmane and harmalan are bioactive components of classical clonidine-displacing substance.[Pubmed:15610033]

Biochemistry. 2004 Dec 28;43(51):16385-92.

Elucidation of the structure of the endogenous ligand(s) for imidazoline binding sites, clonidine-displacing substance (CDS), has been a major goal for many years. Crude CDS from bovine lung was purified by reverse-phase high-pressure liquid chromatography. Electrospray mass spectrometry (ESMS) and nuclear magnetic resonance ((1)H NMR) analysis revealed the presence of L-tryptophan and 1-carboxy-1-methyltetrahydro-beta-carboline in the active CDS extract. Competition radioligand binding studies, however, failed to show displacement of specific [(3)H]clonidine binding to rat brain membranes for either compound. Further purification of the bovine lung extract allowed the isolation of the beta-carbolines harmane and Harmalan as confirmed by ESMS, (1)H NMR, and comparison with synthetic standards. Both compounds exhibited a high (nanomolar) affinity for both type 1 and type 2 imidazoline binding sites, and the synthetic standards were shown to coelute with the active classical CDS extracts. We therefore propose that the beta-carbolines harmane and Harmalan represent active components of classical CDS. The identification of these compounds will allow us to establish clear physiological roles for CDS.

Interaction of beta-carbolines with central dopaminergic transmission in mice: structure-activity relationships.[Pubmed:7609916]

Neurosci Lett. 1995 Apr 14;189(2):121-4.

Although several beta-carboline alkaloids display hallucinogenic properties in humans, their mechanism of action remains unclear. To ascertain their influence on central dopaminergic transmission, in this study we investigated the facilitating effect of low doses of various beta-carbolines on L-DOPA (250 mg/kg)-induced stereotypy in mice. Harmaline (0.075, 0.15 and 0.35 mg/kg) and harmine (0.15, 0.35 and 0.75 mg/kg) powerfully enhanced the degree of stereotypy, whereas 6-methoxy-Harmalan, 6-methoxy-harman and harman were far less potent, augmenting stereotypy only at much higher doses (3 mg/kg). 6-Methoxy-tetrahydro-beta-carbolin (6-MeO-THBC) had only a weak effect at the dose of 3 mg/kg and tetrahydro-beta-carbolin (THBC) had no effect up to the dose of 3 mg/kg. The Ca(2+)-channel blocker nimodipine (2.5 mg/kg) only slightly antagonized harmaline (0.15 mg/kg) facilitation of L-DOPA-induced stereotypy.

Methyl-beta-carbolinium analogs of MPP+ cause nigrostriatal toxicity after substantia nigra injections in rats.[Pubmed:7796140]

Brain Res. 1995 Mar 27;675(1-2):279-88.

Eleven beta-carbolinium compounds (beta C+s) and MPP+ were stereotaxically injected (40-200 nmol in 5 microliter of vehicle) unilaterally into the substantia nigra of anesthetized adult male Sprague-Dawley rats. The rats were sacrificed after three weeks. The ipsilateral striatum was analyzed for dopamine and DOPAC levels with HPLC. The brainstem injection site was fixed and cut coronally. The largest lesion area in each animal was measured using NIH IMAGE. Three beta C+s produced lesions whose mean areas were nearly as large as that produced by MPP+ (defined as 100%): 2,9-Me2-harman (94%), 2-Me-harmol (74%), and 2,9-Me2-norharman (57%). Three other compounds produced somewhat smaller lesions: 2-Me-harmaline (34%), 6-MeO-2-Me-harman (29%), and 2-Me-harmine (25%). The remaining compounds were ineffective (< or = 12%): norharman, 2-Me-norharman, 2-Me-harman, harmine, and 2-Me-6-MeO-Harmalan. A 40 nmol dose of MPP+ reduced ipsilateral striatal dopamine to 0.6% of control. None of the beta C+s approached this, although several did significantly reduce striatal dopamine at doses of either 40 nmol (2,9-Me2-harman (37%), 2,9-Me2-norharman (42%), and 2-Me-harman (63%)) or 200 nmol (2-Me-harmaline (23%), norharman (63%), and 2-Me-norharman (64%)). There was a moderate negative correlation between lesion size and dopamine level (r = -0.65). There were also moderately strong correlation between lesion size and dopamine level (r = -0.65). There were also moderately strong correlations (r = 0.39-0.78) between the beta C+ nigral lesion area or striatal dopamine level potencies and their previously described IC50 values for inhibiting mitochondrial respiration or their toxicity to PC12 cells in culture. Interestingly, our correlation analysis revealed a remarkably strong correlation between beta C+ Ki MAO-A values and their toxicity to PC12 LDH release (r = -0.84) or PC12 protein loss (r = 0.79). Although beta C+s appear to be less specific toxins than MPP+, their levels in human substantia nigra are 8-20-fold higher than in cortex, making their role as relatively selective nigral toxins in Parkinson's disease plausible.

Formation of 1-methyl-beta-carbolines in rats from their possible carboxylic acid precursor.[Pubmed:3412494]

Naunyn Schmiedebergs Arch Pharmacol. 1988 May;337(5):566-71.

In vivo metabolism of 1-methyl-1,2,3,4-tetrahydro-beta-carboline-1-carboxylic acid (1-CTHH), a possible precursor of the endogenous beta-carbolines tetrahydro-harman (THH) and harman was investigated in rats. Following intraperitoneal injection of [4-14C]1-CTHH, a rapid distribution of the radioactivity in the tissues was observed. The highest radioactivity was measured in the kidney and the lowest in the brain as well as in the fat tissue. Approximately 55% of the administered dose was excreted in the urine within 90 min. The radioactivity in the urine consisted of unchanged 1-CTHH (greater than 90%) besides Harmalan and trace amounts of harman. Harmalan represents the major degradation product of 1-CTHH; it could be identified in all tissues examined and in the urine. The concentration in the blood, however, was low at all time points investigated. The peak concentration of Harmalan in most tissues was measured between 15-30 min after injection. A time-dependent formation of THH was found in the lung and spleen indicating an important role of these organs in the biosynthesis of THH. Furthermore, the metabolism of [4-14C]1-CTHH in the brain was studied following intracerebroventricular injection. The formation of Harmalan in the brain was not affected by pretreatment with the aromatic amino acid decarboxylase inhibitor NSD 1015. Determination of the Harmalan concentration in several brain regions revealed a high level in the hippocampus and hypothalamus and a small concentration in pons, corpus striatum, cerebellum and cerebral cortex 20 min after injection. The analyses of the

Biotransformation of 1-methyl-1,2,3,4-tetrahydro-beta-carboline-1-carboxylic acid to harmalan, tetrahydroharman and harman in rats.[Pubmed:3431747]

Neurosci Lett. 1987 Oct 29;81(3):325-30.

1-Methyl-1,2,3,4-tetrahydro-beta-carboline-1-carboxylic acid (1-carboxytetrahydroharman, 1-CTHH) has been detected in the brain of rats following intracerebroventricular injection of tryptamine and pyruvic acid. We now report the metabolism of this compound. Following intraperitoneal injection of 1-CTHH into rats, Harmalan was found to be the major metabolite besides tetrahydroharman (THH) and harman. A high concentration of THH was measured in the lung while most of harman was found in the urine. Harmalan and THH could be detected in the brain in low concentrations. The products were separated following extraction from tissues by high-performance liquid chromatography (HPLC) on a reversed phase C18-DB column. The identity of the metabolites was confirmed by mass spectrometry (MS) analysis. The results demonstrate the role of 1-CTHH as a precursor of the biologically active compounds Harmalan, THH and harman.

Harman induces preference for ethanol in rats: is the effect specific for ethanol?[Pubmed:2885856]

Pharmacol Biochem Behav. 1987 Apr;26(4):749-55.

Increasing concentrations of either ethanol, etonitazene, clomethiazole or midazolam were offered to male Wistar rats for 21 days. Between day 8 and day 21, the animals were treated with several doses of harman, Harmalan, and tetrahydronorharman (tetrahydro-beta-carboline) by means of continuous intraventricular infusion. Harman and THN induced a significant preference for ethanol in a dose-dependent manner. Harman was approximately three times more potent than THN. The amount of ethanol consumed during the second and third weeks of the experimental period correlated with the harman concentration in the brain after the cessation of the treatment (p less than 0.01). Harman infusion attenuated the clomethiazole intake, whereas that of etonitazene and midazolam was not affected as compared with CSF-treated rats. By counting licking movements, it was found that the rats drank ethanol and water at distinct time periods with the pattern dependent on the concentration of the ethanol solution offered. The intervals between the maxima were 6 to 8 hours at low ethanol concentrations. Relatively high concentrations caused a disruption of the regular rhythms in favour of shorter ones with increasing intervals between the maxima (3 hr, 4 hr, 5 hr intervals). Harman treatment (27 nmol/hr) disturbed the regular rhythms at lower ethanol concentrations but mimicked the ultradian rhythm which was observed at high ethanol concentrations in CSF-treated animals. The observed coincidence of water and ethanol intake was uncoupled if the highest ethanol concentration in both treatments was offered. Thus, treatment with harman changed the rhythm of fluid intake in a direction which was detected in CSF-treated rats only at relatively high ethanol concentrations.

Pharmacology of harmalan (1-methyl-3,4-dihydro-beta-carboline).[Pubmed:2580722]

Eur J Pharmacol. 1985 Mar 12;109(3):363-71.

Harmalan is presumably formed in vivo as an intermediate product of the biosynthesis of harman as well as tetrahydroharman. The pharmacological effects of Harmalan as well as its affinity towards benzodiazepine, 5-HT2 and tryptamine binding sites were investigated in the present study. Harmalan induced clonic convulsions which were antagonized by diazepam. Receptor binding experiments as well as combined treatments with antagonists point to an interaction which involves neither benzodiazepine nor 5HT2 receptor sites but rather tryptamine binding sites. Good agreement was found between the potency of Harmalan to increase spontaneous motor activity and the affinity to the tryptamine binding sites when compared with the effects of tryptamine in both tests. In the light-dark-chamber test for predicting anxiolytic effects of drugs, Harmalan elicited opposite effects to diazepam. The results of combined treatment also suggested an interaction of both compounds not involving benzodiazepine receptors. Tryptamine binding sites seemed to play no role since the amine was inactive under these conditions. Thus, Harmalan induces several tryptamine agonistic effects and others not involving tryptamine binding sites.