ABSTRACT
BACKGROUND: Diagnosis of autism spectrum disorder (ASD) is generally made phenotypically and the hunt for ASD-biomarkers continues. The purpose of this study was to compare urine organic acids profiles of ASD versus typically developing (TD) children to identify potential biomarkers for diagnosis and exploration of ASD etiology. METHODS: This case control study was performed in the Department of Pathology and Laboratory Medicine in collaboration with the Department of Pediatrics and Child Health, Aga Khan University, Pakistan. Midstream urine was collected in the first half of the day time before noon from the children with ASD diagnosed by a pediatric neurologist based on DSM-5 criteria and TD healthy controls from August 2019 to June 2021. The urine organic acids were analyzed by Gas Chromatography-Mass Spectrometry. To identify potential biomarkers for ASD canonical linear discriminant analysis was carried out for the organic acids, quantified in comparison to an internal standard. RESULTS: A total of 85 subjects were enrolled in the current study. The mean age of the ASD (n = 65) and TD groups (n = 20) was 4.5 ± 2.3 and 6.4 ± 2.2 years respectively with 72.3% males in the ASD group and 50% males in the TD group. Parental consanguinity was 47.7 and 30% in ASD and TD groups, respectively. The common clinical signs noted in children with ASD were developmental delay (70.8%), delayed language skills (66.2%), and inability to articulate sentences (56.9%). Discriminant analysis showed that 3-hydroxyisovalericc, homovanillic acid, adipic acid, suberic acid, and indole acetic were significantly different between ASD and TD groups. The biochemical classification results reveal that 88.2% of cases were classified correctly into ASD& TD groups based on the urine organic acid profiles. CONCLUSION: 3-hydroxy isovaleric acid, homovanillic acid, adipic acid, suberic acid, and indole acetic were good discriminators between the two groups. The discovered potential biomarkers could be valuable for future research in children with ASD.
Subject(s)
Autism Spectrum Disorder , Autism Spectrum Disorder/diagnosis , Biomarkers , Case-Control Studies , Child , Child, Preschool , Female , Gas Chromatography-Mass Spectrometry , Humans , Male , MetabolomicsABSTRACT
Objective To perform qualitative and quantitative analysis on the volatile components in roots, rhizomes, leaves, and flowers of Asari Radix et Rhizoma derived from Asarum insigne. Methods The volatile components were analyzed by HS-GC-MS, and the relative percentage content of each component was calculated with peak area normalization method. Results There were 58 components separated from four parts of A. insigne, including 27 common components in different parts. The principal constituents was trans-β-farnesene, safrole, and asaricin. Their contents were different in four parts. Especially the contents of safrole in rhizomes, leaves, and flowers were up to 34%, 22%, and 21%; The safrole in rhizomes was over twice higher than that in roots (12%). Because safrole was extremely poisonous, the rhizomes, leaves, and flowers should be used carefully. Conclusion The volatile components in A. insigne can be detected by HS-GC-MS simply and quickly. The research can be helpful for development and quality evaluation of A. insigne.
ABSTRACT
Objective: To establish a rapid analytical method for volatile components in Tibetan medicine Heracleum millefolium and to determine the volatile components from its roots, stems, leaves, and flower parts, respectively. Methods: Headspace sampling incorporation with gas-chromatography-mass spectrum (HS-GC-MS) determination was introduced to analyze the powder directly. Static headspace equilibration was performed at 100℃ for 40 min, and 1 mL of the headspace gas was injected in split mode of 10:1. The split inlet temperature was 260℃. The carrier gas was He at a constant flow rate of 1.8 mL/min. The column oven temperature was initially set at 50℃ for 2 min, then increased to 100℃ at 2℃/min, held for 6 min, then increased to 300℃ at 10℃/min and held for 2 min. The GC/MS interface temperature was maintained at 280℃. The solvent delay time was 3 min (to bypass the solvent peak). The volatile components were confirmed by NIST11.L database, and volatile organic compounds from roots, stems, leaves, and flower parts were compared. Results: The types of compounds in the roots, stems, leaves, and flowers of H. millefolium are mainly aldehyde, benzene, alcohols, and alkene. Octanal, hexanal, and γ-terpinene are the main components in the roots and stems. While o-isopropyltoluene and terpinolene are the main components in the leaves and flowers. Conclusion: HS-GC-MS method is easy, simple, and feasible, and can be widely used in other Chinese materia medica samples for analysis of volatile components.
ABSTRACT
Present study was to characterize the physiochemical properties, free amino acids (FAAs), volatiles and microbial communities of various moromi, respectively sampled from different stages of high-salt dilute-state (HSDS) and low-salt solid-state (LSSS) fermentation, using multiphase analyzing methods. Phospholipid fatty acids (PLFA) analysis indicated that Gram-positive bacteria were dominant bacteria and fungi were principal microbes. For DGGE analysis, dominant microbes in moromi were mainly fell into Weissella, Tetragenococcus, Candida, Pichia, and Zygosaccharomyces. During fermentation, the dominant microbes shifted from nonhalophilic and less acid-tolerant species to halophilic and acid-tolerant species. Total of 15 FAAs and 44 volatiles were identified in moromi, mainly Glu, Asp, Tyr, and acids, alcohols, esters, aldehydes, respectively. Odor activity values analysis suggested that the final moromi of LSSS fermentation had more complicated odors than that of HSDS fermentation. Conclusively, technological parameters, microbial communities, raw materials and fermentation process may result in the discrepancy of HSDS and LSSS moromi.
