• Title/Summary/Keyword: LC/MS analysis

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Quantitative analysis of water-soluble vitamins and polyphenolic compounds in tomato varieties (Solanum lycopersicum L.) (토마토(Solanum lycopersicum L.) 품종 간 수용성 비타민과 폴리페놀계 성분 함량 변이 분석)

  • Kim, Daen;Son, Beunggu;Choi, Youngwhan;Kang, Jumsoon;Lee, Yongjae;Je, Beungil;Park, Younghoon
    • Journal of Plant Biotechnology
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    • v.47 no.1
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    • pp.78-89
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    • 2020
  • Tomato fruit quality is determined by the contents of various functional metabolites in addition to fruit appearance. To develop tomato cultivars with higher amounts of functional compounds, an efficient quantification method is required to identify the natural variations in the compounds in the tomato germplasm. In this study, we investigated tomato varieties, which included 23 inbred lines and 12 commercial F1 cultivars, for their contents of seven watersoluble vitamins (vitamin C, vitamins B1, B2, B3, B5, B6, and B9) and five polyphenolic compounds (quercetin, rutin, kaempferol, myricetin, and naringenin chalcone). The results of high performance liquid chromatography and liquid chromatography-mass spectrometry showed that vitamin C and naringenin chalcone were the major water-soluble vitamins and polyphenolic compounds, respectively, and their abundance was highly variable depending on the cultivar. By contrast, the contents of vitamin B1, quercetin, and kaempferol were lowest among the cultivars. With regard to the relationship between metabolic compounds and fruit characteristics, a significant association was found in fruit size, indicating that cherry tomato varieties contain higher amounts of the compounds compared to large fresh-type varieties. However, no direct association was detected in fruit color, except for naringenin chalcone. The results of this study provide new insights on the quantification of metabolic compounds and the selection of breeding materials, which are prerequisites for the development of functional tomato varieties.

Influence of Microwave Pretreatment on the Formation of Heterocyclic Amines in Fried Beef Patties (Microwave를 이용한 예열 처리가 조리한 쇠고기 패티에서의 Heterocyclic Amines 형성에 미치는 영향)

  • Jung, Kyung-Hee;Shin, Han-Seung
    • Food Science of Animal Resources
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    • v.29 no.6
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    • pp.719-725
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    • 2009
  • Heterocyclic aromatic amines (HCAs) are potent mutagens and possible human carcinogens that are formed during the heating of protein-rich foods. The effects of preheating treatment of beef patties using a microwave prior to frying at $220^{\circ}C$ for 10 min on each side on the reduction of HCAs (amino-carbolines and amino-imidazo-azaarenes) were evaluated. The amount of HCAs was then evaluated by solid-phase extraction and high pressure liquid chromatography (HPLC) analysis. The beef patties were treated by microwaving for various times (0, 1, 1.5, 2, or 3 min) before pan-frying. The results revealed the presence of 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2), 3-amino-1,4-dimethyl-5H-pyrido-[4,3-b] indole (Trp-P-1), 2-amino-6-methyldipyrido[1,2-a:3',2'-d]imidazole (Glu-P-1), 2-aminodipyrido[1,2-a:3',2'-d]imidazole (Glu-P-2), 9H-pyrido[3,4-b]indole (Norharman), 1-methyl-9H-pyrido[3,4-b]indole (Harman), 2-amino-9H-pyrido[2,3-b] indole ($A{\alpha}C$), 2-amino-3,8 dimethylimidazo[4,5-f]-quinoxaline (MeIQx) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]-pyridine (PhIP) in all samples. However, microwave pretreatment for 1 min inhibited the formation of these HCAs by up to 90% when compared to the control.

Rapid separation of Capsicum annuum L. leaf extract using automated HPLC/SPE/HPLC coupling system (Sepbox system) and identification of α-glucosidase inhibitory active substances (자동화 HPLC/SPE/HPLC 시스템(Sepbox system)을 활용한 고추 잎 (leaf of Capsicum annuum L.) 추출물 분리 및 α-glucosidase 억제 활성 물질 탐색)

  • Kim, Min-Seon;Jin, Jong Beom;Lee, Jung Hwan;An, Hye Suck;Pan, Cheol-Ho;Park, Jin-Soo
    • Journal of Applied Biological Chemistry
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    • v.64 no.1
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    • pp.25-32
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    • 2021
  • Phytochemicals include plant-derived natural products that promote and improve the human metabolism and physiological activity, and there is a lot of research to find the value of the molecules is in progress. Likewise, we obtained 288 fractions of Capsicum annuum L. extract in less than 20 h using HPLC/SPE/HPLC coupling experiment through Sepbox system, an effective separation system to search for active substances in natural resources and ensure efficacy and reliability. Therefore, this experiment allowed rapid identification of biologically active molecules from the extract compared to traditional separation processes. Of the above fractions, eight fractions showed the α-glucosidase inhibitory (AGI) activity and subsequent LC-MS analysis revealed one of the active molecules as luteolin 7-O-glucoside. In addition, we proved the increase in AGI activity according to deglycosylation of flavonoid glycoside. Therefore, this study suggests that the Sepbox system can quickly separate and identify active components from plant extract, and is an effective technique for finding new active substances.

Analysis of antioxidant and anti-inflammatory effects of Mongolian wild lingonberry and blueberry, and identification of their bioactive compounds (몽골 야생 링곤베리와 블루베리의 항산화, 항염증 효과 및 생리활성 물질 분석 연구)

  • Lee, Hye Ju;Naranbulag, Batdorj;Jeong, Seung Jin;Seo, Chan;Lee, Sang-Gil
    • Korean Journal of Food Science and Technology
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    • v.54 no.2
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    • pp.147-154
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    • 2022
  • The Mongolian lingonberry and blueberry are two essential food sources found in Mongolia. This study investigated the antioxidant and anti-inflammatory effects of methanol extracts from Mongolian lingonberry (LBE) and blueberry (BBE). Compared to the LBE, the BBE showed higher total phenolic, flavonoid, and anthocyanin contents, as well as antioxidant capacities. The LBE and BBE inhibited the mRNA expression of pro-inflammatory genes, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and cyclooxygenase (COX-2) in lipopolysaccharide-stimulated RAW 264.7 macrophage cells. In addition, the LBE and BBE inhibited NADPH oxidase-2 (Nox2) mRNA expression, indicating that they have cellular antioxidant capacities. Anthocyanin derivatives of the LBE and BBE were analyzed using LC-QTOF/MS. Six anthocyanins were identified in the BBE, while one was detected in the LBE. Our findings demonstrate that the anthocyanin-rich LBE and BBE could be used as functional food sources in Mongolia.

Residual Characteristics and Monitoring of Cyenopyrafen and Cyflumetofen in Strawberries for Export (수출딸기 중 Cyenopyrafen과 Cyflumetofen의 잔류소실 특성평가 및 잔류농약 모니터링)

  • Kim, Yeong-Jin;Kim, Jong-Hwan;Kwon, Young-Sang;Song, Jong-Wook;Seo, Jong-Su
    • Korean Journal of Environmental Agriculture
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    • v.36 no.4
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    • pp.279-287
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    • 2017
  • BACKGROUND: Many farmers who cultivate the strawberries for export have used agricultural chemicals which MRL (Maximum Residue Limits) of main export target countries or simultaneous multi-residue analysis in Korea have not been established. Among them, the cyenopyrafen and cyflumetofen were selected and applied to this study to determine the PHI (pre-harvest interval) which is appropriate to the PLS (Positive List System) criterion (0.01 mg/kg) and to investigate the residual amounts in the samples. In addition, Fifty pesticides were monitored to check up whether it is suitable or not for main export target countries. METHODS AND RESULTS: Cyenopyrafen and cyflumetofen were spayed out to the strawberries. Samples for residual analyses were taken for maximum 60 days. After sampling, they were extracted by the QuEChERS method and analyzed using the LC-MS/MS. Cyenopyrafen and cyflumetofen were detected in a range of 0.0106~2.6517 mg/kg and of 0.0005~1.4480 mg/kg, respectively. From this results, they were found to be suitable for PLS concentration after 30 or 45 days after spray. In addition, they were detected in most samples that were selected at random. Their concentrations were higher than the PLS criterion in the maximum twenty samples. Twelve of pesticides unsuitable for main export target countries have been detected in the monitoring of simultaneous multi-residue analysis. The result indicates they are unsuitable for export since they excesses over PLS criterion. CONCLUSION: The monitoring result showed it is necessary to establish the pesticide standards of safe use suitable for the PLS criterion. In addition, it is considered continues management and inspection are needed to solve problems caused by unsuitable pesticides in export strawberries.

A Study on the Saponin Contents and Antioxidant Activity of the Ginseng and Extruded Ginseng by Using Different Solvents for Extraction (추출 용매에 따른 인삼과 압출 성형 인삼의 사포닌 함량 및 항산화 활성 연구)

  • Kim, Sung-Hwan
    • The Korean Journal of Food And Nutrition
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    • v.24 no.4
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    • pp.528-534
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    • 2011
  • This study was conducted to investigate the changes in saponin content and antioxidant activity of crude ginseng and extruded ginseng by using different solvent extraction methods. Each of the fractions was first extracted by 80% ethanol followed by ether treatment to remove the lipid components. Water soluble components were separated by ethylacetate and water saturated butanol. Four fraction, including 80% ethanol, ethylacetate, butanol and water were obtained from crude and extruded ginsengs to analyze saponin content and antioxidant activity. Saponin content and antioxidant capacity of each of the four fractions were measured by LC/MS analysis and ORAC(Oxygen Radical Absorbance Capacity) assay, respectively. It was found that a major portion of saponin was present in ethyl acetate and water saturated butanol fractions. When extracted by 80% ethanol, ginsenoside Rb1 and Rg1 were mostly found in crude ginseng, while ginsenoside Re and Rb1 were detected in extruded ginseng. Even though Rh1 and Rg3 were found in a very small quantity in crude ginseng, there was a significant quantity of both in extruded ginseng when extracted by 80% ethanol. Similar tendency was also observed in extruded ginseng fraction when extracted with ethyl acetate and butanol. In crude ginseng, the level of Rg1 was the highest among other ginsenosides upon extraction by ethyl acetate, while Rh1 and Rg3 were predominantly found by employing similar solvent extraction in the extruded ginseng. Also, Rg1, Re and Rb1 were also found in the extruded ginseng with small quantity. Rg1, Re and Rb1 were found in crude ginseng by butanol extraction, while Rb1 and Re were extracted from the extruded ginseng. Overall, there was no difference in the saponin content between crude ginseng and extruded ginseng when extracted by butanol and water, but twice as much of saponin was obtained by 80% ethanol extraction and 6 times more saponin were obtained in ethyl acetate fraction in the extruded ginseng. Antioxidant capacity of crude ginseng as determined by ORAC assay was higher in 80% ethanol(high in many different kinds of biological compounds) and water saturated butanol(high in polar saponin) fractions than the ethyl acetate and water fractions. No difference in antioxidant capacity was observed between crude and extruded ginseng. However, antioxidant capacity of ethyl acetate and water fractions in extruded ginseng was significantly higher than crude ginseng($P$ >0.05). All the fractions in both, crude and extruded ginseng possessed antioxidant capacity and even water fractions that contained almost no saponin had some antioxidant capacity. While determining correlation coefficient between fractions in extruded ginseng by Pearson correlation, it was observed that 80% ethanol fraction was in correlation with ethyl acetate($P$ >0.01) and ethanol($P$ >0.001) and in the case of ethylacetate, correlation was observed only with butanol fraction($P$ >0.05).

Antioxidative, and Inhibitory Activities on Melanogenesis of Vitex negundo L. Leaf Extract (목형 잎 추출물의 항산화 활성과 멜라닌 생합성에 대한 저해활성)

  • Kim, A Reum;Park, Su Ah;Ha, Ji Hoon;Park, Soo Nam
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.135-144
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    • 2013
  • The aim of this study was to evaluate various aspects of Vitex negundo L. leaf extract, such as the antioxidative activity, tyrosinase inhibitory effects, and inhibitory activities on ${\alpha}$-MSH induced melanogenesis, and active component analysis. The DPPH (1, 1-diphenyl-2-picrylhydrazyl) scavenging activities ($FSC_{50}$) of the ethyl acetate fraction and aglycone fraction of V. negundo L. leaf extract were $14.51{\mu}g/ml$ and $13.96{\mu}g/ml$, respectively. A luminol-dependent chemiluminescence assay revealed that the reactive oxygen species (ROS) scavenging activity ($OSC_{50}$) of the aglycone fraction of V. negundo L. leaf extract on ROS generated in an $Fe^{3+}$-$EDTA/H_2O_2$ system was the most prominent at $0.22{\mu}g/ml$. The protective effects of the extracts fractions of V. negundo L. leaf against the rose-bengal sensitized photohemolysis of human erythrocytes were increased in a concentration dependent manner ($1{\sim}50{\mu}g/ml$). In particular, there were greater protective effects of the aglycone fraction on the cellular membrane than that of the fat-soluble antioxidant (+)-${\alpha}$-tocopherol. The inhibitory effects ($IC_{50}$) on mushroom tyrosinase were the highest for the ethyl acetate fraction ($IC_{50}$ = $48.58{\mu}g/ml$). The inhibitory effect on ${\alpha}$-MSH induced melanogenesis in B16 melanoma cells was 41.80% at $50{\mu}g/ml$ of ethyl acetate fraction. Active component analyses by TLC, HPLC and LC/ESI-MS revealed luteolin and isoorientin. These results indicate that V. negundo L. leaf extract can be used as an antioxidant for ROS scavenging. Particularly, the luteolin and isoorientin of the ethyl acetate fraction may be applicable to new whitening cosmetics because of its inhibitory effect on mushroom tyrosinase and ${\alpha}$-MSH induced melanogenesis in B16 melanoma cells.

Use of Dried Stoned Olive Pomace in the Feeding of Lactating Buffaloes: Effect on the Quantity and Quality of the Milk Produced

  • Terramoccia, S.;Bartocci, S.;Taticchi, A.;Di Giovanni, S.;Pauselli, M.;Mourvaki, E.;Urbani, S.;Servili, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.7
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    • pp.971-980
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    • 2013
  • Dried stoned olive pomace (DSOP) was administered to dairy water buffaloes, and their productive performance and milk composition were analysed. Sixteen pluriparous lactating buffaloes were divided into two uniform groups (control and experimental), taking into consideration the following parameters: milk production (2,192 and 2,102 kg) and duration of lactation (254 and 252 d) of the previous year, distance from calving (51 and 43 d), milk production (9.71 and 10.18 kg/d), body condition score (BCS) (6.44 and 6.31) and weight (617 and 653 kg) at the beginning of the trial. Both diets had the same formulation: second cut alfalfa hay 20%, corn silage 42%, concentrate 38% but the two concentrates differed in their formulation, the experimental one contained 15.50% of DSOP as fed. The employed DSOP showed high amounts of secoiridoids, such as 3,4-dihydroxyphenylethanol (3,4-DHPEA) (1.2 g/kg DM), 3,4-dihydroxyphenylethanol-elenolic acid di-aldehyde (3,4-DHPEA-EDA) (12.6 g/kg DM), p-hydroxyphenylethanol-elenolic acid di-aldehyde (p-HPEA-EDA) (5.6 g/kg DM) and lignans, which are known to be powerful bioactive compounds. The control diet had an energy-protein content of 0.86 Milk FU/kg DM and 143.3 g/kg DM of crude protein, whereas the experimental diet of 0.87 Milk FU/kg DM and 146.6 g/kg DM of crude protein. Each animal of the two groups received 17 kg DM/d and each buffalo of the experimental group, by way of the concentrate, ingested 1.05 kg DM/d of DSOP. The trial lasted 40 days. No significant difference was found between the BCS (6.41 and 6.53), live weight (625.93 and 662.50 kg) and milk production (9.69 and 10.08 kg/d) of the two groups, as was the case for fat, protein, lactose, pH and coagulating parameters of the two milks. The milk fat of the experimental group had a significantly higher content of total tocopherols (10.45 vs $8.60{\mu}g/g$, p<0.01) and retinol (3.17 vs $2.54{\mu}g/g$, p<0.01). The content of the reactive substances with tiobarbituric acid (TBARs) was significantly lower in the milk fat of the experimental group (12.09 vs $15.05{\mu}g$ MDA/g, p<0.01). The acid content of the milk fat of the experimental group had a significantly higher content (p<0.05) of C18:0 and of $C18:3{\omega}6$. LC-MS/MS analysis showed the presence of 3,4-DHPEA ($36.0{\mu}g/L$) in the milk of the DSOP-fed buffaloes, while other phenols were not found. DSOP, in the quantity utilized, can be used in the feeding of the lactating buffalo; the dietetic-nutritional characteristics of the milk are improved due to a greater contribution of tocopherols, retinol and the presence of hydroxytyrosol.

Comparative Analysis of Antioxidant, Anti Aging and Phenolic Compounds of Different Solvent Extracts from Saccharina japonica and Costaria costata (추출용매에 따른 쇠미역과 다시마의 항산화, 항노화 활성과 페놀화합물 비교분석)

  • Hyun-Hwa Lee;Jin-Sol Kim;Jun-Han Jeong;Chun Sung Kim;Sook Young Lee
    • Korean Journal of Plant Resources
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    • v.36 no.2
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    • pp.107-121
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    • 2023
  • This study analyzed the polyphenol, flavonoid contents, antioxidant activity, anti-aging activity and phenol component contents of Saccharina japonica (SJ), Costaria costata (CC) extracts with hot water, 95% methanol, 95% prethanol for investigating possible utilization of SJ and CC extracts. The result revealed that the SJ and CC methanol extracts showed the highest polyphenol and flavonoid contents, 4.63 mg TAN/g, and 4.19 mg QUE/g respectively. Also, the SJ and CC methanol extracts showed higher antioxidant activity than prethanol and hot water extracts, whereas the ABTS radical scavenging activities were the highest in prethanol extracts (IC50 = 15.4, 10.3 ㎍/µL). In anti-aging activity for evaluating the anti-wrinkle activity and skin whitening activity, the CC methanol extracts had high collagenase inhibitory activity (88.3%), and the SJ prethanol extracts showed higher elastase inhibitory activity (19.0%) compared to other extracts. Then the tyrosinase inhibitory activity was significantly higher in the SJ and CC methanol extracts (41.8, 30.3%, respectively), whereas prethanol extracts were the lowest. To identify the phenol component contents of SJ and CC extracts, 4-hydroxybenzoic acid, naringenin, naringin and nicotinic acid were measured using LC-MS/MS. As a result, the phenol contents were the highest in SJ methanol extract (4-hydroxybenzoic acid), SJ and CC prethanol extract (naringin and naringenin) and CC prethanol extract (nicotinic acid). Lastly, the antioxidant activity of SJ and CC showed high correlations with polyphenol and flavonoid contents (R = -0.946~0.883). These results suggest that prethanol or methanol extracts of SJ and CC have higher antioxidant activities, anti-aging activity and the potential to be used as material for health functional food and cosmetics.

Establishment of an Analytical Method for Determination of Fungicide Oxathiapiprolin in Agricultural Commodities using HPLC-UV Detector (HPLC-UVD를 이용한 농산물 중 살균제 Oxathiapiprolin의 잔류분석법 확립)

  • Jang, Jin;Kim, Heejung;Do, Jung Ah;Ko, Ah-Young;Lee, Eun Hyang;Ju, Yunji;Kim, Eunju;Chang, Moon-Ik;Rhee, Gyu-Seek
    • Journal of Food Hygiene and Safety
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    • v.31 no.3
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    • pp.186-193
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    • 2016
  • An analytical method was developed for the determination of oxathiapiprolin in agricultural commodities. Oxathiapiprolin is a new oomycide (fungicide of piperidinyl thiazole isoxazoline class) which controls downy mildew in cucurbits caused by Pseudoperonospora cubensis (oomycete plant pathogen). Agricultural commodities were extracted with acetonitrile and partitioned with dichloromethane to remove the interference, adjusting pH between 9 and 10 by 1 N sodium hydroxide. After purification by silica SPE cartridge to clean up the interference of organic compounds, they were finally quantified by HPLC-UVD (high performance liquid chromatograph ultraviolet detector) using a wavelength at 260 nm and confirmed by LC-MS (liquid chromatograph mass spectrometer) in electro-spray ionization positive ion mode. The standard calibration curve was linear with coefficients of determination ($r^2$) 1.00 over the calibration ranges (0.025-2.5 mg/L). Recoveries were ranged between 86.7 to 112.7%, with relative standard deviations less than 10% at three concentration levels (LOQ, 10LOQ, and 50LOQ) performing five replicates. The overall results were determined and estimated according to the CODEX guidelines (CAC/GL40). The proposed method for determination of oxathiapiprolin residues in agricultural commodities can be used as an official method.