• Title/Summary/Keyword: high performance liquid chromatography

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Development and Validation of an HPLC-PDA Method for Quantitation of Ten Marker Compounds from Eclipta prostrata (L.) and Evaluation of Their Protein Tyrosine Phosphatase 1B, α-Glucosidase, and Acetylcholinesterase Inhibitory Activities

  • Nguyen, Duc Hung;Le, Duc Dat;Ma, Eun Sook;Min, Byung Sun;Woo, Mi Hee
    • Natural Product Sciences
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    • v.26 no.4
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    • pp.326-333
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    • 2020
  • The aerial parts of Eclipta prostrata is used as a traditional medicine and vegetable. In traditional folk medicine, it is used for treatment of hemorrhages, hepatic, disease, renal injuries, hair loss, tooth mobility, and viper bites. In this study, ten compounds (1 - 10) were isolated from the aerial parts of E. prostrata. A reliable high performance liquid chromatography equipped with photometric diode array detector (HPLC-PDA) method was developed to simultaneously quantitate 10 marker compounds [chlorogenic acid (1), paratensein 7-O-��-ᴅ-glucoside (2), quercetin 7-O-��-ᴅ-glucoside (3), luteolin 7-O-��-ᴅ-glucoside (4), apigenin 7-O-��-ᴅ-glucoside (5), apigenin 4'-O-��-ᴅ-glucoside (6), apigenin (7), luteolin (8), wedelolactone (9), and paratensein (10)]. In addition, compounds 5 and 6 showed considerable inhibitory effects against protein-tyrosine phosphatase 1B (PTP1B) enzyme. Moreover, compounds 6 - 8, and 10 exhibited potent α-glucosidase inhibitory effects with IC50 values of 24.5 ± 1.9, 33.0 ± 0.5, 45.5 ± 0.1, and 23.8 ± 1.0 µM, respectively. All compounds (1 - 10) showed considerable acetylcholinesterase (AChE) inhibitory effects with IC50 ranging from 30.1 to 75.2 µM.

Determination of cyromazine in commercial insecticides using HPLC-DAD

  • Kim, Young-Wook;Han, Bok Hee;Kang, Young Eun;Rhee, Chae Hong;Seo, Sang-Ji;Kim, Soohee;Jeong, Wooseog;Her, Moon
    • Korean Journal of Veterinary Service
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    • v.43 no.4
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    • pp.261-265
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    • 2020
  • Each commercial cyromazine insecticide has different HPLC conditions. The aim of this study was to establish a standardized chromatographic method for the determination of cyromazine in commercial insecticides. The separation was achieved on two C18 columns - Waters® Bondapak C (4×300 nm i.d., 10 ㎛) and X bridge (4.6×250 nm i.d., 5 ㎛) using a mobile phase composed of water/methanol/ethanolamine (76:24:0.1, v/v), with UV detection at wavelengths 230 nm and 254 nm. A total of six commercial cyromazine insecticides were analyzed. In this study, the optimal high-performance liquid chromatography conditions for the analysis of cyromazine were as follows: a mobile phase of water/methanol/ethanolamine (76:24:0.1, v/v) at a flow rate of 1.0 mL/min and a detection wavelength of 230 nm using a X bridge C18 column (4.6×250 nm i.d., 5 ㎛) at a column temperature of 25℃. The calibration curve was linear in the concentration range of 5~50 ㎍/mL, with a correlation coefficient of 0.99995. The cyromazine detection limit was 0.2 ㎍/mL, and the limit of quantification was 0.59 ㎍/mL. The percentage recovery ranged from 99.8% to 101.0% for cyromazine, and the relative standard deviation was not over 2.0%. The cyromazine concentration ranged from 92.7% to 109.4% and was within the acceptable range (90~120%) for the percent of the labeled amount. This method was found to be suitable for determining cyromazine in commercial insecticides.

HPLC/UV Quantification of (+)-Catechin in Filipendula glaberrima from Different Regions and Flowering Stages (터리풀의 채집장소 및 채집시기에 따른 카테킨 함량 HPLC/UV 분석)

  • Lee, Hak-Dong;Lee, Yunji;Kim, Hoon;Kim, Hangeun;Park, Chun-Gun;Lee, Sanghyun
    • Korean Journal of Pharmacognosy
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    • v.51 no.4
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    • pp.291-296
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    • 2020
  • Filipendula glaberrima (FG) is a plant endemic to South Korea. It is economically important as a food source and used as a medicine in treating ailments. Filipendula flowers are characterized by the presence of several polyphenolic constituents. The aim of this study is to determine the content of (+)-catechin in Filipendula glaberrima collected from different regions at different flowering stages. High-performance liquid chromatography with a gradient elution system (0.5% acetic acid in water : acetonitrile = 95 : 5 to 0 : 100 for 35 min) was used. A reverse-phase INNO column with UV detection at 278 nm was employed. The results revealed that F. glaberrima from Mt. Odae has the highest (+)-catechin content (10.600 mg/g). Furthermore, its content was the lowest in samples collected during the pre-flowering period and the highest at the early-flowering stage. This study provides a basis in establishing the optimal period and the best region for collecting F. glaberrima with maximized (+)-catechin yield.

Antibacterial compounds against fish pathogenic bacteria from a combined extract of Angelica gigas and Artemisia iwayomogi and their quantitative analyses

  • Lim, Jae-Woong;Kim, Na Young;Seo, Jung-Soo;Jung, Sung-Hee;Kang, So Young
    • Fisheries and Aquatic Sciences
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    • v.24 no.10
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    • pp.319-329
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    • 2021
  • In the search for antibiotic alternatives from safe and effective medicinal plants against fish pathogenic bacteria, we found that a combined extract (CE) of 1:1 (w/w) ratio of Angelica gigas Nakai roots and aerial parts of Artemisia iwayomogi Kitamura showed antibacterial activity against the fish pathogenic bacteria. By antibacterial activity-guided fractionations and isolations, five compounds were isolated and identified as decursinol angelate (1), decursin (2), xanthotoxin (3), demethylsuberosin (4), and 2,4-dihydroxy-6-methoxyacetophenone (5) through spectroscopic analyses, such as nuclear magnetic resonance (NMR) and mass spectrometry (MS). Among the compounds, 1 and 2 showed the highest antibacterial activities against Streptococcus iniae and Vibrio anguillarum, showing minimum inhibitory concentrations (MICs) of 62.5-250 ㎍/mL. Compounds 3, 4, and 5 were also found to be active, with MICs of 31.25-1,000 ㎍/mL for those strains. Furthermore, active compounds, 1 and 2 in CE were simultaneously quantified using high-performance liquid chromatography-tandem MS (HPLC-MS/MS). The average contents of 1 and 2 in CE was 3.68% and 6.14%, respectively. The established method showed reliable linearity (r2 > 0.99), good precision, accuracy, and specificity with intra- and inter-day variations of < 2 % and recoveries of 90.13%-108.57%. These results may be helpful for establishing the chemical profile of CE for its commercialization as an antibiotic alternative in aquaculture.

Underlying mechanisms of phosphodiesterase 10A and glutamate-ammonia ligase genes that regulate inosine monophosphate deposition and thereby affect muscle tenderness in Jingyuan chickens

  • Wang, Weizhen;Zhang, Juan;Hu, Honghong;Yu, Baojun;He, Jintong;Yao, Tingting;Gu, Yaling;Cai, Zhengyun;Xin, Guosheng
    • Animal Bioscience
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    • v.35 no.11
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    • pp.1771-1786
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    • 2022
  • Objective: Inosine monophosphate (IMP) is a key factor that imparts of meat flavor. Differences in the IMP content in the muscles were evaluated to improve chicken meat quality. Methods: For this study, the IMP content was detected by high performance liquid chromatography. The gene expression profiles of Jingyuan chickens with different feeding patterns and different sexes were analyzed by RNA-sequencing (RNA-seq). Results: Breast muscle IMP content in free-range chickens was extremely significantly higher than that of caged chickens (p<0.01). Breast muscle IMP content in hens was also higher than that of cocks, but the difference was not significant. Correlation analysis showed that the breast muscle IMP content in caged hens and cocks was negatively correlated with the shear force, and the breast muscle IMP content in free-range hens was significantly negatively correlated with the shear force (p<0.05). The two key genes associated with IMP synthesis in chickens with different feeding patterns were glutamate-ammonia ligase (GLUL) and phosphodiesterase 10A (PDE10A). Bioinformatics analysis revealed that the GLUL and PDE10A genes are involved in glutamine biosynthesis and purine salvage pathways respectively. In addition, GLUL expression was positively correlated with the IMP content in caged and free-range chickens, and PDE10A expression was significantly positively correlated with the IMP content in caged and free-range chickens (p<0.05). Conclusion: These findings will facilitate the comprehension of the deposition of IMP in the muscles and thereby aid the process of selection and breeding of good quality local chickens.

Pharmacokinetics of thalidomide in dogs: can feeding affect it? A preliminary study

  • Pierini, Alessio;Sartini, Irene;Giorgi, Mario;Lebkowska-Wieruszewska, Beata;Lisowski, Andrzej;Poapolathep, Amnart;Marchetti, Veronica
    • Journal of Veterinary Science
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    • v.21 no.5
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    • pp.60.1-60.11
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    • 2020
  • Background: Tumor-associated neoangiogenesis is a crucial target for antitumor therapies. Thalidomide (TAL) is a promising anti-neoangiogenetic drug that has recently been used in the treatment of several malignancies in dogs. Objectives: The aim of the study was to assess the pharmacokinetics of TAL after single oral administration in dogs. Additionally, the influence of feeding on the pharmacokinetic profile of TAL in dogs has been preliminarily investigated. Methods: Six healthy adult female Labradors were enrolled according to a randomized single-dose, 2-treatment, 2-phase, paired 2 × 2 cross-over study design. The dogs were administered a single 400 mg capsule of TAL in fasted and fed conditions. Blood was collected from 15 min to 48 h after dosing, and TAL quantified in plasma by a validated high-performance liquid chromatography method. The pharmacokinetics of TAL were analyzed using a non-compartmental approach. Results: TAL concentration was quantifiable up to 10 h and 24 h after fasted and fed conditions, respectively. Cmax (fasted, 1.34 ± 0.12 ㎍/mL; fed, 2.47 ± 0.19 ㎍/mL) and Tmax (fasted, 3 h; fed, 10 h) differed substantially between the 2 groups. AUC and t1/2λz were significantly higher in fed (42.46 ± 6.64 mg × h/L; 17.14 ± 4.68 h) compared to fasted (12.38 ± 1.13 mg × h/L; 6.55 ± 1.25 h) dogs. The relative oral bioavailability of TAL for the fasted group was low (36.92% ± 3.28%). Conclusions: Feeding affects the pharmacokinetics of oral TAL in dogs, showing a delayed, but higher absorption with different rate of elimination. These findings are of importance in clinical veterinary settings, and represent a starting point for further related studies.

Allometric analysis of tylosin tartrate pharmacokinetics in growing male turkeys

  • Pozniak, Blazej;Tikhomirov, Marta;Motykiewicz-Pers, Karolina;Bobrek, Kamila;Switala, Marcin
    • Journal of Veterinary Science
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    • v.21 no.3
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    • pp.35.1-35.11
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    • 2020
  • Background: Despite common use of tylosin in turkeys, the pharmacokinetic (PK) data for this drug in turkeys is limited. Within a few months of growth, PK of drugs in turkeys undergoes changes that may decrease their efficacy due to variable internal exposure. Objectives: The objective of this study was to investigate the influence of age on the PK of a single intravenous (i.v.) and oral administration of tylosin to turkeys at a dose of 10 and 50 mg/kg, respectively. Methods: Plasma drug concentrations were measured using high-performance liquid chromatography with UV detection. The PK parameters were assessed by means of non-compartmental approach and were subjected to allometric analysis. Results: During a 2.5-month-long period of growth from 1.4 to 14.7 kg, the median value for area under the concentration-time curve after i.v. administration increased from 2.61 to 7.15 mg × h/L and the body clearance decreased from a median of 3.81 to 1.42 L/h/kg. Over the same time, the median elimination half-life increased from 1.03 to 2.96 h. For the oral administration a similar trend was noted but the differences were less pronounced. Bioavailability was variable (5.76%-21.59%) and age-independent. For both routes, the plasma concentration of the major tylosin metabolite, tylosin D, was minimal. Protein binding was age-independent and did not exceed 50%. Allometric analysis indicated a relatively poor predictivity of clearance, volume of distribution and elimination half-life for tylosin in turkeys. Conclusions: Age has a significant impact on tylosin PK in turkeys and dosage adjustment may be needed, particularly in young individuals.

Comparison of Vitamin B5 Content and True Retention in Commonly Consumed Vegetables by Different Cooking Methods (국내 다소비 채소류의 조리에 따른 비타민 B5 함량 및 잔존율 비교)

  • Jin Ju, Park;Arin, Park;Eunji, Park;Youngmin, Choi
    • Journal of the Korean Society of Food Culture
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    • v.37 no.6
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    • pp.540-546
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    • 2022
  • This study aimed to determine the changes in the vitamin B5 content of raw and cooked vegetables. The nineteen vegetables were subjected to different cooking methods, viz. blanching, boiling, pan-broiling, and steaming. Vitamin B5 was quantified by reversed-phase high-performance liquid chromatography (HPLC) using photodiode-array (PDA) detection (200 nm). The standard reference materials (SRM) were used to validate the accuracy of vitamin B5 measurement method used in this study. The cooking yields ranged from 82.63 to 107.62% and decreased in most of the vegetables except bitter melon, curled mallow, and eggplant. The raw kabocha squash, Danhobak, had the highest vitamin B5 content (0.671 mg/100 g) among the samples. All cooked vegetables showed lower vitamin B5 content compared to the raw samples. The true retention ranged from 0% (crown daisy, blanching) to 84.49% (kabocha squash, steaming). These results indicate that vitamin B5 is degraded after cooking. Pan-broiling and steaming are better cooking methods than the others for retaining vitamin B5. The true retention of vitamin B5 in the samples markedly depends on the cooking method and food matrix. These results can be used as important basic data for nutritional evaluation of meals.

Comparison of Bioactivities from Centella asiatica Cultivated in Smart Farm and Field (스마트팜과 노지에서 재배한 병풀의 생약학적 비교)

  • Jin Hong, Park;Da Hee, Lee;Seong Min, Jo;Jeong Hwan, Choi;Nam Jun, Kim;Min Su, Kim;Youngmin, Park;Kiman, Lee
    • Korean Journal of Pharmacognosy
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    • v.53 no.4
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    • pp.192-201
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    • 2022
  • This study aimed to compare bioactivities of Centella asiatica (CA) cultivated in smart farms and fields. Component analysis, cell viability, anti-inflammatory activity, neuroprotection activity, and antioxidant activity were examined with 70% ethanol extracts of CA cultivated in smart farm (SEE) and field (FEE), respectively. Asiaticoside was analyzed by high performance liquid chromatography (HPLC) and as a result, SEE had more asiaticoside content than FEE. After treatment of RAW 264.7 cells with SEE and FEE, there was no cytotoxicity within the treated concentrations. SEE and FEE showed nitric oxide (NO), tumor necrosis factor (TNF)-α, and interleukin (IL)-6 inhibitory activities in a dose-dependent manner in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Moreover, SEE inhibited more NO, TNF-α, and IL-6 production levels than FEE. SEE and FEE reversed the H2O2-induced SH-SY5Y cell death. Especially, SEE was more effective in changing the H2O2-induced SH-SY5Y cell death than FEE. The antioxidant activity was confirmed by various methods such as total phenol content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and superoxide dismutase (SOD). As a result, SEE showed the most potent antioxidant activities about TPC, DPPH, and SOD methods. This study suggested that SEE has higher bioactivities such as effect of anti-inflammation, neuroprotection, and antioxidation than FEE.

Compositional differences of Bojungikgi-tang decoctions using pressurized or non-pressurized extraction methods with variable extraction times

  • Kim, Jung-Hoon;Seo, Chang-Seob;Kim, Seong-Sil;Shin, Hyeun-Kyoo
    • The Korea Journal of Herbology
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    • v.28 no.4
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    • pp.1-6
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    • 2013
  • Objectives : In other to determine the optimal extraction conditions, the various Bojungikgi-tang (BJIGT) decoctions prepared by different pressure levels and different extraction times were compared and evaluated in terms of the extract yield and the total soluble solid content. Methods : Decoctions were prepared by the pressure levels of 0 (non-pressurized) and 1 $kgf/cm^2$ (pressurized) for 60, 120 and 180 min. The extract yield and the total soluble solids content of decoctions were measured, and the amounts of the reference compounds in decoctions were investigated by the analysis using high performance liquid chromatography. Results : The extract yield and the total soluble solid content were higher in decoctions extracted by the pressurized method than those from decoction with non-pressurized method. The patterns of yield and contents showed a proportional increase to the extraction time. In analysis of the linear regression for four reference compounds such as liquiritin, nodakenin, hesperidin, and glycyrrhizin, the good linearity with the correlation coefficient more than 0.9999 was observed. The highest contents for four reference compounds were observed at 180 min of both the pressurized method and the non-pressurized method. Conclusions : This study suggests that the pressure in extraction method and the extraction time affect the compositional constituents in BJIGT decoctions. The extraction time of 180 min could be chosen in both pressurized and non-pressurized method as optimal extraction condition.