• Title/Summary/Keyword: HPLC analysis methods

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Inhibitory Effect of Mixed Extracts Obtained from Astragali Radix and Lithospermi Radix on Matrix Metalloproteinases in IL-1β-induced SW1353 Cells and Quantitative Analysis of Active Compounds (황기, 지치 복합물의 연골세포에서의 Matrix Metalloproteinases 저해 효과 및 유효성분의 분석)

  • Choi, Doo Jin;Choi, Bo Ram;Lee, Dae Young;Choi, Soo Im;Lee, Young Seob;Kim, Geum Soog
    • Korean Journal of Medicinal Crop Science
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    • v.27 no.4
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    • pp.247-258
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    • 2019
  • Background: Astragali radix (A) and Lithospermi radix (L) have long been used as traditional medicines due to their known anti-inflammatory effects. This study aimed at evaluating, their optimal mixing ratio and their functional compounds by investigating the inhibitory effects of mixed extracts of A and L and their active compounds on matrix metalloproteinases (MMPs). Methods and Results: A and L extracts were obtained by extraction at $80^{\circ}C$ using 50% and 70% fermented alcohol, respectively, and then mixed at a ratio of 5 : 5, 6 : 4, 7 : 3 and 8 : 2 (w/w). The activities of MMP-1, MMP-3, and MMP-13 were evaluated in interleukin-1beta ($IL-1{\beta}$)-induced SW1353 cells. The extract mixtures showed synergistic inhibitory effects on MMP-3 and MMP-13, higher than the effects of the individual A and L extracts. The 7 : 3 mixture (ALM16) showed the most effective MMPs inhibitory activity, while among the active ingredients, calycosin-7-O-${\beta}$-D-glucoside and lithospermic acid exhibited excellent MMPs inhibitory activity. Additionally, an HPLC method was established for simultaneous quantification of the effective components of the extract mixtures, and validated by measuring the linearity, precision and accuracy of the limit of detection (LOD) and limit of quantification (LOQ). Conclusions: ALM16 showed the most effective MMPs inhibitory activity. Calycosin-O-${\beta}$-D-glucoside, calycosin and lithospermic acid were identified as useful candidates, as they were the major functional compounds in the MMP inhibitory activity. Summarily, ALM16 might be a highly effective in osteoarthritis management, owing to its because it exhibits a protective effect on cartilage via excellent inhibition of MMPs.

Comparison of pharmacokinetic characteristics of sildenafil citrate chewable tablets and film-coated tablets in healthy male subjects

  • Yoo, Hyounggyoon;Cho, Sang Min;Choi, Youn Woong;Lee, Hye Jung;Kwon, Ji-Hye;Kim, Soo-Whan;Kim, Jae Woo;Lee, SeungHwan;Hong, Jang-Hee
    • Translational and Clinical Pharmacology
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    • v.25 no.3
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    • pp.153-156
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    • 2017
  • UI14SDF100CW is a chewable tablet of sildenafil citrate, which was developed to improve compliance through convenience of administration. The purpose of this study was to compare the pharmacokinetic (PK) properties of sildenafil citrate chewable tablets (UI14SDF100CW) and conventional sildenafil citrate film-coated tablets ($Viagra^{(R)}$, Pfizer). A randomized, open-label, single dose, two-treatment, two-period, two-way crossover study was conducted in 60 healthy male volunteers. In each period, the subjects received a single oral dose of UI14SDF100CW or $Viagra^{(R)}$ (both tablets contain 140.45 mg of sildenafil citrate, which is equivalent to 100 mg of sildenafil). Serial blood samples were collected up to 24 h post-dose for PK analysis. The plasma concentration of sildenafil was determined using a validated HPLC-MS/MS assay. PK parameters of sildenafil were calculated using non-compartmental methods. The plasma concentration-time profiles of sildenafil in both formulations were similar. For UI14SDF100CW, the $C_{max}$ and $AUC_{last}$ of sildenafil were $1068.69{\pm}458.25$ (mean${\pm}$standard deviation) mg/L and $3580.59{\pm}1680.29h{\cdot}mg/L$, and the corresponding values for $Viagra^{(R)}$ were $1146.84{\pm}501.70mg/L$ and $3406.35{\pm}1452.31h{\cdot}mg/L$, respectively. The geometric mean ratios (90% confidence intervals) of UI14SDF100CW to $Viagra^{(R)}$ for $C_{max}$ and $AUC_{last}$ were 0.933 (0.853-1.021) and 1.034 (0.969-1.108), respectively, which met the bioequivalence criteria of Korean regulatory agency. In conclusion, UI14SDF100CW and $Viagra^{(R)}$ showed similar PK properties. Therefore, UI14SDF100CW can be an alternative to sildenafil for the treatment of erectile dysfunction, providing better compliance.

Contents of vitamin B9 (folate) and B12 (cobalamins) in commonly consumed seafood menus in Korea (한국인 상용 수산물 식단의 비타민 B9과 B12 함량)

  • Park, Eun-Young;Jeong, Bomi;Chun, Jiyeon
    • Journal of Nutrition and Health
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    • v.54 no.2
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    • pp.211-223
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    • 2021
  • Purpose: A total of 39 seafood menus were prepared according to the Korean standard recipe, and analyzed for vitamin B9 (folate) and B12 (cobalamins) contents, using validated applied analytical methods. The menus included Guk/Tang/Jjigae (boiled or stewed dishes, n = 10), Bokkeum (stir-fried dishes, n = 10), Jjim/Jorim (braised or steamed dishes, n = 7), Gui (baked or grilled dishes, n = 7), Twigim (deep-fried dishes, n = 2) and Muchim (dried or blanched-seasoned dishes, n = 3). Methods: The contents of vitamin B9 and B12 in all food samples were determined by the trienzyme extraction-Lactobacillus casei and immunoaffinity-high-performance liquid chromatography/photodiode array detection methods. Analytical quality control was performed in order to assure reliability of the analysis. Results: Accuracy (97.4-100.6% recoveries) and precision (< 6% relative standard deviations for repeatability and reproducibility) of vitamin B9 and B12 analyses were determined to be excellent. The vitamin B9 and B12 contents of the 39 seafood menus evaluated, varied in the range of 1.83-523.08 ㎍/100 g and 0.11-38.30 ㎍/100 g, respectively, depending on the ingredients and cooking methods. The vitamin B9 content was highest in Jomi-gim (523.08 ㎍/100 g), followed by Geonsaeu-bokkeum (128.34 ㎍/100 g) and Janmyeolchi-bokkeum (121.53 ㎍/100 g). Vitamin B12 was detected in all seafood menus, with highest level obtained in Kkomack-jjim (41.58 ㎍/100 g). The seaweed dish was found to have high levels of both vitamin B9 and B12. All assays were performed under strict quality control. Conclusion: Guk and Tang menus, which contain a large amount of water, were relatively lower in the vitamin B9 and B12 contents than the other menus. Bokkeum menus containing various vegetables were high in the vitamin B9 content, but the vitamin B12 content was dependent on the type of seafood used in the menu.

Antioxidant activity of partially purified extracts isolated from Acanthopanax sessiliflorum Seeman (오가피 분획 추출물의 항산화효과)

  • Im, Kyung-Ran;Kim, Mi-Jin;Jung, Teak-Kyu;Yoon, Kyung-Sup
    • KSBB Journal
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    • v.23 no.4
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    • pp.329-334
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    • 2008
  • The antioxidant activity and the qualitative analysis of Acanthopanax sessiliflorum Seeman were studied by partially purified extract using various methods: extraction by using ethanol solutions and temperatures, and absorption to Diaion HP20 column chromatography using 10%, 20%, 40%, 60% ethanol solutions. Major constituents, chlorogenic acid, caffeic acid, eleutheroside E, was determinated by HPLC method in Acanthopanax sessiliflorum S. 10% and 20% ethanol solutions contain chlorogenic acid (3.020$\pm$0.080%, 20.500$\pm$1.150%, respectively). 40% ethanol solution contains caffeic acid and eleutheroside E (12.270$\pm$0.360%, 1.670$\pm$0.140%, respectively). Diaion HP20 fractions (10%, 20%, 40%, 60% ethanol solutions) showed the scavenging activity of radicals and reactive oxygen species with the $IC_{50}$ values of $81.534{\pm}0.992{\mu}g/ml$, $1.748{\pm}0.098{\mu}g/ml$, $11.487{\pm}1.768{\mu}g/ml$, $21.960{\pm}0.547{\mu}g/ml$ against 1,1-diphenyl-2-picrylhydrazly radical and $1713.548{\pm}34.565{\mu}g/ml$, $131.419{\pm}2.235{\mu}g/ml$, $200.681{\pm}2.444{\mu}g/ml$, $757.897{\pm}6.868{\mu}g/ml$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. Especially, 20% and 40% ethanol fractions showed more antioxidant activity than dl-$\alpha$-tocopherol. These results suggest that Acanthopanax sessiliflorum S. extract and Diaion HP20 fractions may be useful as a potential source of nutraceutical and cosmetic products.

Analysis of PAHs (polycyclic aromatic hydrocarbons) in Ground Coffee Using GC-tandem Mass Spectrometry and Estimation of Daily Dose (GC-tandem mass spectrometry를 이용한 분쇄원두커피 중 PAHs(polycyclic aromatic hydrocarbons) 분석법 연구 및 인체노출량 평가)

  • Jung, So-Young;Park, Ju-Sung;Son, Yeo-Joon;Choi, Su-Jeong;Lee, Yun-Jeong;Kim, Mi-Sun;Park, So-Hyun;Lee, Sang-Me;Chae, Young-Zoo
    • Korean Journal of Food Science and Technology
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    • v.43 no.5
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    • pp.544-552
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    • 2011
  • The purpose of this study was to develop an analytical method for determining 15 polycyclic aromatic hydrocarbons (PAHs) of EU priority using gas chromatography (GC)-tandem mass spectrometry (MS). The PAHs in ground coffee were analyzed after being extracted using methods such as saponification-liquid-liquid extraction, Soxhlet extraction, and solid-liquid extraction. The solid-liquid extraction method showed the greatest repeatability and most efficient reduction of the matrix effect. GC-tandem MS for the quantification of the 15 PAHs showed better resolution and lower limit of detections (LODs) than GC-MS-selected ion monitoring (SIM) and high performance liquid chromatography with fluorescence detector. LODs of this method for the ground coffee types were 0.002-0.1 ${\mu}g/kg$ and limit of quantifications (LOQs) were 0.006-0.2 ${\mu}g/kg$ The recoveries ranged from 52.6 to 93.3%. Forty-six commercial types of ground coffee were analyzed to determine their PAHs contamination levels. PAHs concentration ranged from ND to 5.988 ${\mu}g/kg$. This study was conducted with toxicity equivalence factors, the U.S. EPA recommendation to identify dietary risks for PAHs in different types of coffee. The estimated average daily dose of PAHs was $5.24{\times}10^{-8}$ mg/kg body weight/day.

Extraction Characteristics of Flavonoids from Lonicera flos by Supercritical Fluid Carbon Dioxide ($SF-CO_2$) with Co-solvent (초임계유체 $CO_2$ 및 Co-solvent 첨가에 따른 금은화(Lonicera fles)의 Flavonoid류 추출특성)

  • Suh, Sang-Chul;Cho, Sung-Gill;Hong, Joo-Heon;Choi, Yong-Hee
    • Korean Journal of Food Science and Technology
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    • v.37 no.2
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    • pp.183-188
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    • 2005
  • Effects of co-solvent polarity, citric acid, pressure, temperature, run time, and co-solvent ratio on extraction of major flavonoids from Lonicera Flos were investigated using supercritical fluid $CO_{2}(SF-CO_{2})$. HPLC analysis revealed addition of pure methanol resulted in low extraction yield of major flavonoids, luteoloin (Lu), Quercetin (Qu), Apigenin (Ap). Under same condition, as co-solvent polarity increased, yields of major flavonoids increased gradually, At optimum co-solvent extraction condirion of 60% aqueous methanol (10%, v/v), yields of Lu, Qu, and Ap were 42.09, 28.18, and 3.49 mg/100 g, respectively. Addition of citric acid to 60% aqueous methanol gave higher, with addition of 1% citrie acid resulting in highest yields of 63.2 (Lu), 39.35 (Qu), and 5.79 (Ap) mg/100 g. Optimum extraction conditions of major flavonoids were 200 bar, $50^{\circ}C$, 60 min, and $CO_{2}$-methanol-water(20: 1.8: 1.2).

Studies on the Anti-aging Action of Korean Ginseng (I) Comparative Study of Red and White Ginsengs on Anti-aging Action (고려인삼(高麗人蔘)의 노화억제작용(老化抑制作用)에 관한 연구(硏究) 제1보(第1報) 홍삼(紅蔘)과 백삼(白蔘)의 노화억제작용(老化抑制作用) 비교연구(比較硏究))

  • Choi, Jin Ho;Oh, Sung Ki
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.12 no.4
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    • pp.323-335
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    • 1983
  • The inhibitory effects of the extract and crude saponin of red and white ginsengs on lipoperoxide formation in vitro and in vivo were studied and correlated with anti-aging. To this end, antioxidant activity, induction period and lipoperoxide were measured by the methods of EDA, POV and TBA value. And also superoxide dismutase and peroxidase activity were measured by pyrogallol autoxidation method (${\Delta}A$ 420/min) and initial velocity(${\Delta}A$ 436/min), respectively. From HPLC analysis, the PT/PD ratio of red and white ginsengs was found to be 0.561% and 0.401%, respectively, and red ginseng increased the PT/PD ratio in comparison with white ginseng. The EDA activity of red ginseng was higher than that of white ginseng; red ginseng showed stronger antioxidative effect than white ginseng. The inhibitory effect of red ginseng was lower than that of white ginseng during the induction period. It was proved that high molecular coloring substance was deeply related to the initial stage of lipoperoxidation. There was no significant difference between red and white ginsengs in both in vitro and intraperitoneal administration experiments, and red ginseng was more effective than white ginseng in longterm administration. And also inhibitory effect on lipoperoxide formation was mainly occurred in liver, suggesting that the function of liver played an important role in anti-aging actions. From the measurement of superoxide dismutase(SOD) activity for both ginseng groups intraperitoneally and orally administered, it was found that red ginseng group administered extract and crude saponin showed remarkable inhibitory effects in comparison with white ginseng. In particular, orally administered group showed more stronger inhibitory effect on lipid peroxidation in comparison with intraperitoneally administered group. It was also found that the continuous oral administration was more effective than temporary administration. Red ginseng was more notable anti-aging effect in comparison with white ginseng in vivo, and this may be due to the increase of SOD activity in rat-liver. Peroxidase activity also showed similar trend to SOD activity in vitro and in vivo experiments. Red ginseng was not only superior to white ginseng for preservation but also for biochemical and pharmaceutical efficacy.

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Changes in the constituents and UV-photoprotective activity of Astragalus membranaceus caused by roasting (황기의 볶음 조건에 따른 성분 및 자외선 광보호 활성 변화)

  • Park, Jeong-Yong;Lee, Ji Yeon;Kim, Hyung Don;Jang, Gwi Yeong;Seo, Kyung Hye
    • Journal of Nutrition and Health
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    • v.52 no.5
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    • pp.413-421
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    • 2019
  • Purpose: Astragalus membranaceus (AM) is an important traditional medicinal herb. Pharmacological research has indicated that AM has various physiological activities such as antioxidant, anti-inflammatory, immunoregulatory, anticancer, hypolipidemic, antihyperglycemic, and hepatoprotective activities. The bioactive substances responsible for the physiological activities in AM, including many antioxidant substances, change during the roasting process. This study investigated and compared the changes in the antioxidant constituents of AM caused by roasting. Methods: DPPH (1,1-diphenyl-2-picryl hydrazyl) and $ABTS^+$ (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt) radical scavenging activities and their total phenolic content (TPC) were measured. High-performance liquid chromatography (HPLC) analysis was performed to confirm any changes in the isoflavonoids of roasted AM (R-AM),. The cell viability of UVB-induced HDF (Human dermal fibroblast) cells treated with AM and R-AM extracts was investigated. The comet assay was used to examine the inhibitory effects of R-AM extracts on DNA damage caused by oxidative stress. Results: The DPPH and $ABTS^+$ radical scavenging activities were $564.6{\pm}20.9$ and $108.2{\pm}3.1$ ($IC_{50}$ value) respectively, from the 2R-AM. The total phenol content was $47.80{\pm}1.40mg$ GAE/g from the 1R-AM. The values of calycosin and formononetin, which are the known isoflavonoid constituents of AM, were $778.58{\pm}2.72$ and $726.80{\pm}3.45{\mu}g/g$ respectively, from the 2R-AM. Treatment of the HDF cells with R-AM ($50{\sim}200{\mu}g/mL$) did not affect the cell viability. Furthermore, the R-AM extracts effectively protected against UVB-induced DNA damage. Conclusion: The findings of this study indicate that R-AM increases its isoflavonoid constituents and protects against UVB-induced DNA damage in HDF cells.

Analysis of Components in the Different Parts of Lythrum salicaria L. (털부처꽃의 부위별 성분 분석)

  • Kim, Hee-Young;Park, Yea-Jin;Lee, Ju-Yeon;Kim, Ki-young;Shin, Su;Choi, Min-Woo;Hong, Eun-Jin;Kim, Min-jeong;Yeo, Sujung;Park, In-hwa;Jerng, Ui Min;An, Hyo-Jin;Cha, Yun-Yeop
    • The Korea Journal of Herbology
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    • v.37 no.5
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    • pp.89-96
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    • 2022
  • Objectives : This research was performed to analyze the components in the different parts of Lythrum salicaria L. and to compare which parts of L. salicaria L. are appropriate for food development. Methods : L. salicaria L. was extracted in 20% EtOH at 100 ℃ for 4 hours. Cytotoxicity was investigated in 3T3-L1 cells after treatment of 10-500 ㎍/ml L. salicaria L. for 24 hours. Total polyphenol content (TPC) was estimated using 1 N Folin-ciocateu reagent. 2,2-Diphenyl-1-picryhydrazyl (DPPH) radical scavenging activity was estimated using DPPH reagent and gallic acid. The chemical composition was analyzed by high-performance liquid chromatography (HPLC). 1) Results : The half maximal inhibitory concentration (IC50) in the extracts of the whole plant, aerial parts, and root parts was 350 ㎍/ml, over 500 ㎍/ml, and 150 ㎍/ml, respectively. The TPC in the extracts of the whole plant, aerial parts, and root parts was 527.1 mg/g, 422.6 mg/g, and 781.1 mg/g, respectively. The averages of vitexin contents in the aerial parts, and root parts were 256.7 ± 154.9 ㎍/g and 266.1 ± 63.2 ㎍/g, respectively. The averages of TPC in the leaves, roots, flower stalks and stems were 224.0 ± 53.7 tannin acid (TA) mg/g, 221.8 ± 70.2 TA mg/g, 249.8 ± 34.4 TA mg/g, and 67.7±8.9 TA mg/g, respectively. The averages of DPPH radical scavenging activity in the leaves, roots, flower stalks, and stems were 282.01 ± 43.3 gallic acid equivalent (GAE) 𝜇mole/g, 260.16 ± 44.1 GAE 𝜇mole/g, 288.0 ± 9.3 GAE 𝜇mole/g, and 97.6 ± 10.7 GAE 𝜇mole/g, respectively. Conclusions : There were no significant differences in the content of components or antioxidant activity in the aerial parts compared to those in the whole plant of L. salicaria L. Furthermore, the root parts had low extract yield, cytotoxicity, and quality control problems, therefore our results suggest that the use of the aerial part of L. salicaria L. would be the most appropriate for food development.