• Title/Summary/Keyword: UHPLC

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Specific urinary metabolites in canine mammary gland tumors

  • Valko-Rokytovska, Marcela;Ocenas, Peter;Salayova, Aneta;Titkova, Radka;Kostecka, Zuzana
    • Journal of Veterinary Science
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    • v.21 no.2
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    • pp.23.1-23.10
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    • 2020
  • The identification of biomarkers that distinguish diseased from healthy individuals is of great interest in human and veterinary fields. In this research area, a metabolomic approach and its related statistical analyses can be useful for biomarker determination and allow non-invasive discrimination of healthy volunteers from breast cancer patients. In this study, we focused on the most common canine neoplasm, mammary gland tumor, and herein, we describe a simple method using ultra-high-performance liquid chromatography to determine the levels of tyrosine and its metabolites (epinephrine, 3,4-dihydroxy-L-phenylalanine, 3,4-dihydroxyphenylacetic acid, and vanillylmandelic acid), tryptophan and its metabolites (5-hydroxyindolacetic acid, indoxyl sulfate, serotonin, and kynurenic acid) in canine mammary cancer urine samples. Our results indicated significantly increased concentrations of three tryptophan metabolites, 5-hydroxyindolacetic acid (p < 0.001), serotonin, indoxyl sulfate (p < 0.01), and kynurenic acid (p < 0.05), and 2 tyrosine metabolites, 3,4-dihydroxy-L-phenylalanine (p < 0.001), and epinephrine (p < 0.05) in urine samples from the mammary gland tumor group compared to concentrations in urine samples from the healthy group. The results indicate that select urinary tyrosine and tryptophan metabolites may be useful as non-invasive diagnostic markers as well as in developing a therapeutic strategy for canine mammary gland tumors.

Multiclass Method for the Determination of Anthelmintic and Antiprotozoal Drugs in Livestock Products by Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry

  • Hyunjin Park;Eunjung Kim;Tae Ho Lee;Sihyun Park;Jang-Duck Choi;Guiim Moon
    • Food Science of Animal Resources
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    • v.43 no.5
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    • pp.914-937
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    • 2023
  • The objective of this study was to establish a multi-residue quantitative method for the analysis of anthelmintic and antiprotozoal drugs in various livestock products (beef, pork, and chicken) using ultra-high-performance liquid chromatography-tandem mass spectrometry. Each compound performed validation at three different levels i.e., 0.5, 1, and 2× the maximum residue limit according to the CODEX guidelines (CAC/GL 71-2009). This study was conducted according to the modified quick, easy, cheap, effective, rugged, and safe procedure. The matrix-matched calibrations gave correlation coefficients >0.98, and the obtained recoveries were in the range of 60.2%-119.9%, with coefficients of variation ≤32.0%. Furthermore, the detection and quantification limits of the method were in the ranges of 0.03-3.2 and 0.1-9.7 ㎍/kg, respectively. Moreover, a survey of residual anthelmintic and antiprotozoal drugs was also carried out in 30 samples of beef, pork, and chicken collected in Korea. Toltrazuril sulfone was detected in all three samples. Thus, our results indicated that the developed method is suitable for determining the anthelmintic and antiprotozoal drug contents in livestock products.

Comprehensive Lipid Profiling Recapitulates Enhanced Lipolysis and Fatty Acid Metabolism in Intimal Foamy Macrophages From Murine Atherosclerotic Aorta

  • Jae Won Seo;Kyu Seong Park;Gwang Bin Lee;Sang-eun Park;Jae-Hoon Choi;Myeong Hee Moon
    • IMMUNE NETWORK
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    • v.23 no.4
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    • pp.28.1-28.20
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    • 2023
  • Lipid accumulation in macrophages is a prominent phenomenon observed in atherosclerosis. Previously, intimal foamy macrophages (FM) showed decreased inflammatory gene expression compared to intimal non-foamy macrophages (NFM). Since reprogramming of lipid metabolism in macrophages affects immunological functions, lipid profiling of intimal macrophages appears to be important for understanding the phenotypic changes of macrophages in atherosclerotic lesions. While lipidomic analysis has been performed in atherosclerotic aortic tissues and cultured macrophages, direct lipid profiling has not been performed in primary aortic macrophages from atherosclerotic aortas. We utilized nanoflow ultrahigh-performance liquid chromatography-tandem mass spectrometry to provide comprehensive lipid profiles of intimal non-foamy and foamy macrophages and adventitial macrophages from Ldlr-/- mouse aortas. We also analyzed the gene expression of each macrophage type related to lipid metabolism. FM showed increased levels of fatty acids, cholesterol esters, phosphatidylcholine, lysophosphatidylcholine, phosphatidylinositol, and sphingomyelin. However, phosphatidylethanolamine, phosphatidic acid, and ceramide levels were decreased in FM compared to those in NFM. Interestingly, FM showed decreased triacylglycerol (TG) levels. Expressions of lipolysis-related genes including Pnpla2 and Lpl were markedly increased but expressions of Lpin2 and Dgat1 related to TG synthesis were decreased in FM. Analysis of transcriptome and lipidome data revealed differences in the regulation of each lipid metabolic pathway in aortic macrophages. These comprehensive lipidomic data could clarify the phenotypes of macrophages in the atherosclerotic aorta.

Antioxidant and Antiaging Assays of Hibiscus sabdariffa Extract and Its Compounds

  • Widowati, Wahyu;Rani, Andani Puspita;Hamzah, R. Amir;Arumwardana, Seila;Afifah, Ervi;Kusuma, Hanna Sari W.;Rihibiha, Dwi Davidson;Nufus, Hayatun;Amalia, Annisa
    • Natural Product Sciences
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    • v.23 no.3
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    • pp.192-200
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    • 2017
  • Skin aging is a complex biological process due to intrinsic and extrinsic factors. Free radical oxidative is one of extrinsic factors that induce activation of collagenase, elastase and hyaluronidase. Natural product from plants has been used as antioxidant and antiaging. This study aimed to evaluate antioxidant and antiaging properties of Hibiscus sabdariffa extract (HSE) and its compounds including myricetin, ascorbic acid, and ${\beta}$ carotene. The phytochemical of H. sabdariffa was determined using modified Farnsworth method and presence of phenols, flavonoids and tannins were in moderate content, whereas triterpenoids and alkaloids were in low content. Total phenolic content performed using Folin-Ciocalteu method, was $23.85{\mu}gGAE/mg$. Quantitative analysis of myricetin, ${\beta}-carotene$, and ascorbic acid of HSE was performed with Ultra-High Performance Liquid Chromatography (UHPLC) that shows $78.23{\mu}g/mg$ myricetin, $0.034{\mu}g/mg$ ${\beta}-carotene$, whilst ascorbic acid was not detected. HSE has lower activity on DPPH ($IC_{50}=195.73{\mu}g/mL$) compared to ${\beta}-carotene$, the lowest in ABTS assay ($IC_{50}=74.58{\mu}g/mL$) and low activity in FRAP assay ($46.24{\mu}MFe(II)/{\mu}g\;$) compared to myricetin, ${\beta}-carotene$. Antiaging was measured through inhibitory activity of collagenase, elastase, and hyaluronidase. HSE had weakest collagenase inhibitory activity ($IC_{50}=750.33{\mu}g/mL$), elastase inhibitory activity ($103.83{\mu}g/mL$), hyaluronidase inhibitory activity ($IC_{50}=619.43{\mu}g/mL$) compared to myricetin, ${\beta}-carotene$, and ascorbic acid. HSE contain higher myricetin compared to ${\beta}-carotene$. HSE has moderate antioxidants and lowest antiaging activities. Myricetin is the most active both antioxidant and antiaging activities.

Chemical transformation and target preparation of saponins in stems and leaves of Panax notoginseng

  • Wang, Ru-Feng;Li, Juan;Hu, Hai-Jun;Li, Jia;Yang, Ying-Bo;Yang, Li;Wang, Zheng-Tao
    • Journal of Ginseng Research
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    • v.42 no.3
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    • pp.270-276
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    • 2018
  • Background: Notoginsenoside Ft1 is a promising potential candidate for cardiovascular and cancer disease therapy owing to its positive pharmacological activities. However, the yield of Ft1 is ultralow utilizing reported methods. Herein, an acid hydrolyzing strategy was implemented in the acquirement of rare notoginsenoside Ft1. Methods: Chemical profiles were identified by ultraperformance liquid chromatography coupled with quadruple-time-of-flight and electrospray ionization mass spectrometry (UPLC-Q/TOF-ESI-MS). The acid hydrolyzing dynamic changes of chemical compositions and the possible transformation pathways of saponins were monitored by ultrahigh-performance LC coupled with tandem MS (UHPLC-MS/ MS). Results and conclusion: Notoginsenoside Ft1 was epimerized from notoginsenoside ST4, which was generated through cleaving the carbohydrate side chains at C-20 of notoginsenosides Fa and Fc, and vinaginsenoside R7, and further converted to other compounds via hydroxylation at C-25 or hydrolysis of the carbohydrate side chains at C-3 under the acid conditions. High temperature contributed to the hydroxylation reaction at C-25 and 25% acetic acid concentration was conducive to the preparation of notoginsenoside Ft1. C-20 epimers of notoginsenoside Ft1 and ST4 were successfully separated utilizing solvent method of acetic acid solution. The theoretical preparation yield rate of notoginsenoside Ft1 was about 1.8%, which would be beneficial to further study on its bioactivities and clinical application.

Evaluation of the Physiological Activity and Identification of the Active Ingredients of Crab Apple (Malus prunifolia Borkh.) Extracts (꽃사과(Malus prunifolia Borkh.) 추출물의 생리활성 평가 및 활성 성분의 규명)

  • Shin, Hyun Young;Kim, Hoon;Jeong, Eun-Jin;Kim, Hyun-Gyeong;Lee, Kyung-Haeng;Bae, Yun-Jung;Kim, Woo Jung;Lee, Sanghyun;Yu, Kwang-Won
    • The Korean Journal of Food And Nutrition
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    • v.34 no.5
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    • pp.477-486
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    • 2021
  • To utilize Malus pruniforia Borkh. as a functional material, cold-water (CW), hot-water (HW), and 70% ethanol (EtOH) extracts were prepared, and their antioxidant and anti-inflammatory activities were compared. The antioxidant activity of the HW extract evaluated by ABTS and DPPH radical scavenging and FRAP activity was significantly effective. The total polyphenol content of the HW extract was also higher by 15.5±0.7 mg GAE/g extract compared to other extracts. The EtOH extract showed significantly decreased TNF-α (39.8%), IL-6 (65.5%), and NO (34.9%) levels in RAW 264.7 cells compared to the LPS-induced control group. The levels of IL-6 (21.1%) and IL-8 (19.3%) were significantly decreased by treatment of EtOH extract in HaCaT keratinocytes induced with TNF-α and IFN-γ. The UHPLC-MS results indicated that the EtOH extract might have chlorogenic acid and phlorizin as the major compounds. This was validated using HPLC-DAD, which showed that the EtOH extract had higher levels of chlorogenic acid and phlorizin (1,185±58 and 470±10 ㎍/g extract, respectively). In conclusion, the present study suggested that the anti-inflammatory activity of the EtOH extract was more effective than the CW and HW extracts, and chlorogenic acid and phlorizin could be used as indicator compounds and functional substances.

A Survey on the Residual Pesticides on Agricultural Products on the Markets in Incheon from 2016 to 2018 (인천광역시 유통 농산물의 최근 3년간 잔류농약 실태조사)

  • Park, Jeong-Eun;Lee, Mi-Yeon;Kim, Sun-Hoi;Song, Sung-Min;Park, Byung-Kyu;Seo, Soon-Jae;Song, Jae-Yong;Hur, Myong-Je
    • Korean Journal of Environmental Agriculture
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    • v.38 no.3
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    • pp.205-212
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    • 2019
  • BACKGROUND: This survey conducted on agricultural products in Incheon for the past three years from 2016 to 2018 to detect residual pesticides and to monitor the use of pesticides. METHODS AND RESULTS: Residual pesticides were analyzed for a total of 5,937 agricultural products in Samsan wholesale market, traditional market and large retailers in Incheon. Samples were analyzed by multi class pesticide method using GC-MS/MS, LC-MS/MS, GC-ECD/NPD, and UHPLC. In 59 cases (1.0%) residual pesticides were detected improperly and these cases exceeded maximum residue limits (MRLs). CONCLUSION: The ratios of violative agricultural products were similar each year at approximately 1.0% per year, but the residual pesticide detection rates with the limit included in the samples were shown to be gradually decreasing to 17.9%, 12.6%, and 11.2% annually. The frequency of violative residual pesticides was high in order of Diazinon, Chlorpyrifos, Iprodione, etc. Residual pesticide monitoring might be needed in the future continuously, as violations in agricultural products distributed in Incheon are shown at a similar level every year.

Metabolites profiling and hypolipidemic/hypocholesterolemic effects of persimmon (Diosyros kaki Thumb.) by different processing procedures: in vitro and in vivo studies (제조방법에 따른 떫은감 (Diosyros kaki Thumb.)의 대사체 프로파일링과 중성지질/콜레스테롤 대사 관련 유전자발현 연구 : in vitro 및 in vivo 연구)

  • Park, Soo-Yeon;Oh, Eun-Kyung;Lim, Yeni;Shin, Ji-Yoon;Jung, Hee-Ah;Park, Song-Yi;Lee, Jin Hee;Choe, Jeong-Sook;Kwon, Oran
    • Journal of Nutrition and Health
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    • v.51 no.4
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    • pp.275-286
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    • 2018
  • Purpose: Our previous study demonstrated that persimmon (Diospyros kaki Thumb.) at different stages of ripening provided different protective effects against high-fat/cholesterol diet (HFD)-induced dyslipidemia in rats. In this study, we compared the metabolites profile and gene expressions related to triglyceride (TG)/cholesterol metabolism in vitro and in vivo after treating with persimmon water extracts (PWE) or tannin-enriched persimmon concentrate (TEP). Methods: Primary and secondary metabolites in test materials were determined by GC-TOF/MS, UHPLC-LTQ-ESI-IT-MS/MS, and UPLC-Q-TOF-MS. The expression of genes related to TG and cholesterol metabolism were determined by RT-PCR both in HepG2 cells stimulated by oleic acid/palmitic acid and in liver tissues obtained from Wistar rats fed with HFD and PWE at 0, 150, 300, and 600 mg/d (experiment I) or TEP at 0, 7, 14, and 28 mg/d (experiment II) by oral gavage for 9 weeks. Results: PLS-DA analysis and heatmap analysis demonstrated significantly differential profiling of metabolites of PWE and TEP according to processing of persimmon powder. In vitro, TEP showed similar hypolipidemic effects as PWE, but significantly enhanced hypocholesterolemic effects compared to PWE in sterol regulatory element-binding protein 2 (SREBP2), HMG-CoA reductase (HMGCR), proprotein convertase subtilisin/kexin type 9 (PCSK9), cholesterol $7{\alpha}-hydroxylase$ (CYP7A1), and low density lipoprotein receptor (LDLR) gene expression. Consistently, TEP and PWE showed similar hypolipidemic capacity in vivo, but significantly enhanced hypocholesterolemic capacity in terms of SREBP2, HMGCR, and bile salt export pump (BSEP) gene expression. Conclusion: These results suggest that column extraction after hot water extraction may be a good strategy to enhance tannins and long-chain fatty acid amides, which might cause stimulation of hypocholesterolemic actions through downregulation of cholesterol biosynthesis gene expression and upregulation of LDL receptor gene expression.

Analysis of Antioxidant Activity and Serotonin Derivatives in Safflower (Carthamus tinctorius L.) Germplasm Collected from Five Countries (국외 수집 홍화 유전자원의 항산화 활성 및 세로토닌 유도체 함량 분석)

  • Jung, Yi Jin;Assefa, Awraris Derbie;Lee, Jae Eun;Lee, Ho Sun;Rhee, Ju Hee;Sung, Jung Sook
    • Korean Journal of Plant Resources
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    • v.32 no.5
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    • pp.423-432
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    • 2019
  • In order to select potential plant resources as functional materials and natural antioxidants, we evaluated antioxidant activity and serotonin derivatives of safflower germplasm collected from five countries. N-(p-Coumaroyl) serotonin (CS) and N-feruloylserotonin (FS) were analyzed by using Ultra Performance Liquid Chromatography (UPLC). Total polyphenol content (TPC) was determined by Folin-Ciocalteu method and antioxidant activities were estimated by 2,2-diphenyl-1-picryl-hydrazil (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS), Ferric reducing antioxidant power (FRAP) and Reducing power (RP) assays. The TPC showed a range of 28.25 to $90.53{\mu}g$ gallic acid equivalent (GAE)/mg dried extract (DE). DPPH, ABTS, FRAP and RP assay were in the range of 18.76 to 93.98, 48.91 to 163.73, 3.80 to 132.29 and 26.32 to $80.08{\mu}g$ ascorbic acid equivalent (ASCE)/mg DE, respectively. Among the five countries, safflower seed collected from Iran had the highest levels of serotonin derivatives and antioxidant activities than other countries (p<0.05). CS showed high correlation with TPC, ABTS and DPPH (r=0.673,0.727,0.820), and FS showed high correlation with DPPH (r=0.740). Accessions IT321214 and IT321215 could be useful for development of new functional materials and could be used as a source of valuable natural antioxidant materials.