• Title/Summary/Keyword: High performance Liquid Chromatography

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Stable Isotope Measurement of Ammonium Using HPLC-RTS (high performance liquid chromatography-retention time shift) (HPLC-RTS (high performance liquid chromatography-retention time shift)를 이용한 암모늄 이온의 안정동위원소 측정방법의 개선)

  • An, Soonmo;Lee, Jiyoung;Gardner, Wayne S.
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.18 no.1
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    • pp.47-52
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    • 2013
  • Despite the usefulness of nitrogen isotope tracer experiments in nitrogen cycling studies, there are not such many measurement data mainly due to the difficulties in analytical methods. Although Gardner et al. (1996) developed a relatively simple and accurate method that can measure ammonium isotope using HPLC and used it widely in various N dynamics studies, the technique was not adopted to other laboratories. An HPLC-RTS system using updated HPLC pumps that can perform the same measurements as that of Gardner et al. (1996) was built. The result of standard sample showed linear increase of RTS with the $^{15}N$ proportions. Centroid retention times calculated with Matlab$^{(R)}$ program enhanced the linearity of the response. In a sea water incubation experiment spiked with $^{15}NH_4{^+}$, the uptake and regeneration of ammonium could be separately estimated using the temporal change of $^{15}N/^{14}N$.

Analysis of Cyclic Adenosine Monophosphate (cAMP) Separation via RP-HPLC (reversed-phase high-performance liquid chromatography) by the Moment Method and the van Deemter Equation (역상 크로마토그래피에서 모멘트 방법과 van Deemter 식을 이용한 고리형 아데노신 일인산의 분리특성 연구)

  • Lee, Il Song;Ko, Kwan Young;Kim, In Ho
    • Korean Chemical Engineering Research
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    • v.53 no.6
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    • pp.723-729
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    • 2015
  • The moment analysis of cyclic adenosine monophosphate (cAMP) was performed using chromatograms that were obtained with the pulse input method from an octadecyl silica (ODS) high-performance liquid chromatography (HPLC) column. The general rate (GR) model was employed to calculate the first absolute moment and the second central moment. Three important coefficients for moment analysis, which are molecular diffusivity ($D_m$), external mass transfer coefficient ($k_f$), and intra-particle diffusivity ($D_e$), were estimated by the Wilke-Chang equation, Wilson-Geankoplis equation, and comparing van Deemter equation to theoretical plate number equation, respectively. Experiments were conducted by various conditions of flow rates, methanol volume ratio of the mobile phase, and solute concentration. After the moment analysis, results were organized by van Deemter plots. Also van Deemter coefficients were compared each other to effect $H_{ax}$, $H_f$, and $H_d$ on height equivalent to a theoretical plate (HETP, $H_{total}$). The value of intraparticle diffusion ($H_d$) was the primary factor which makes for HETP whereas external mass transfer ($H_f$) was disregardable factor.

Simultaneous determination and difference evaluation of 14 ginsenosides in Panax ginseng roots cultivated in different areas and ages by high-performance liquid chromatography coupled with triple quadrupole mass spectrometer in the multiple reaction-monitoring mode combined with multivariate statistical analysis

  • Xiu, Yang;Li, Xue;Sun, Xiuli;Xiao, Dan;Miao, Rui;Zhao, Huanxi;Liu, Shuying
    • Journal of Ginseng Research
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    • v.43 no.4
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    • pp.508-516
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    • 2019
  • Background: Ginsenosides are not only the principal bioactive components but also the important indexes to the quality assessment of Panax ginseng Meyer. Their contents in cultivated ginseng vary with the growth environment and age. The present study aimed at evaluating the significant difference between 36 cultivated ginseng of different cultivation areas and ages based on the simultaneously determined contents of 14 ginsenosides. Methods: A high-performance liquid chromatography (HPLC) coupled with triple quadrupole mass spectrometer (MS) method was developed and used in the multiple reaction-monitoring (MRM) mode (HPLC-MRM/MS) for the quantitative analysis of ginsenosides. Multivariate statistical analysis, such as principal component analysis and partial least squares-discriminant analysis, was applied to discriminate ginseng samples of various cultivation areas and ages and to discover the differentially accumulated ginsenoside markers. Results: The developed HPLC-MRM/MS method was validated to be precise, accurate, stable, sensitive, and repeatable for the simultaneous determination of 14 ginsenosides. It was found that the 3- and 5-yr-old ginseng samples were differentiated distinctly by all means of multivariate statistical analysis, whereas the 4-yr-old samples exhibited similarity to either 3- or 5-yr-old samples in the contents of ginsenosides. Among the 14 detected ginsenosides, Rg1, Rb1, Rb2, Rc, 20(S)-Rf, 20(S)-Rh1, and Rb3 were identified as potential markers for the differentiation of cultivation ages. In addition, the 5-yr-old samples were able to be classified in cultivation area based on the contents of ginsenosides, whereas the 3- and 4-yr-old samples showed little differences in cultivation area. Conclusion: This study demonstrated that the HPLC-MRM/MS method combined with multivariate statistical analysis provides deep insight into the accumulation characteristics of ginsenosides and could be used to differentiate ginseng that are cultivated in different areas and ages.

Analysis of Alkaloids in the Seeds of Zizyphus jujuba by High Performance Liquid Chromatography

  • Park, Man-Ki;Park, Jeong-Hill;Shin, Young-Geun;Cho, Kyung-Hee;Han, Byung-Hoon;Park, Myung-Hwan
    • Archives of Pharmacal Research
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    • v.14 no.2
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    • pp.99-102
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    • 1991
  • A hight performance liquid chromatographic methods was developed for the seperation and determination of seven alkaloids in "sanjoin" (the seeds of Zizyphus jujuba Rhamnaceae), a plant with potent sedative activity. A reverse phase system of Lichrosorb RP-Select B column and 0.05 M potassium phosphate buffer (pH = 3.5)-acetonitrile with gradient elution was employed. Two known alkaloids, juzirine and lysicamine, were newly isolated fom "sanjoin"."sanjoin".oin&".ot;.

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Isolation of Herbicidal Compounds from Pulsatilla koreana Roots (백두옹(Pulsatilia koreana Nakai) 뿌리로부터 제초활성물질의 분리)

  • 정형진
    • Korean Journal of Plant Resources
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    • v.9 no.1
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    • pp.47-54
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    • 1996
  • To search herbicidal compounds in Pulsatilla koreana Nakai, methanol extract of P. koreana roots was purified by sequences of XAD-7 column chromatography, silica gel adsorption column chromatography, silica gel flash column chromatography, preparative layer chromatography and preparative high performance liquid chromatography(Prep, HPLC).The final Prep. HPLC gave two herbicidally-active fractions. These fractions treatment at 100ppm inhibited the root length of Echinochloa crus-galli seedlings by 48% and 60% as compared with the control, respectively. Components in the two active fractions were analyzed by GC-MS Spectrometry. These compounds, which were isolated from P. koreana roots, were identified as several fatty esters, hydrocarbons, squalene, evidonol, and a diazepin analogue.

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Production, Purification and Antifungal Activity of Antibiotic Substances Produced by Pseudomonas aeruginosa Strain B5

  • Kim, Beom-Seok
    • Journal of Microbiology and Biotechnology
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    • v.3 no.1
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    • pp.12-18
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    • 1993
  • Pseudomonas aeruginosa strain B5 with antagonistic activity against Phytophthora capsici and Magnaporthe grisea, was isolated from pepper-growing soil. From the culture of P. aeruginosa strain B5 grown on King's medium B, antibiotic substances were purified using XAD-2 column chromatography. XAD-2 eluates inhibited not only the mycelial growth of P. capsid and M. grisea, but also the development of Phytophthora blight on pepper plants. The crude antibiotic substances were further purified by using silica gel column chromatography, Sephadex LH-20 column chromatography, thin layer chromatography on silica gel plates, and high performance liquid chromatography. Silica gel column chromatogrphy gave good separation of the four antibiotic substances. The pure antibiotics P1, P2, and P3 finally purified by preparative HPLC inhibited the mycelial growth of P. capsici, at concentrations from 7 to 10 $\mu g/ml$. Only P1 and P2 had antifungal activity against M. grisea at 8 $\mu g/ml$. P1 and P3 were highly inhibitory to the mycelial growth of Botryosphaeria dothidea and Botrytis cinerea at relatively low concentrations. However, the three antibiotics had no antifungal activity against Rhizoctonia solani. The chemical structures of these antibiotics are being identified.

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Quantification of Karanjin Using High Performance Liquid Chromatography in Raw and Detoxified Karanj (Pongamia glabra vent) Seed Cake

  • Prabhu, T.M.;Devakumar, C.;Sastry, V.R.B.;Agrawa, D.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.3
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    • pp.416-420
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    • 2002
  • Various products of karanj (Pongamia glabra) are utilized for industrial, health and animal agriculture applications in the Indian subcontinent. Despite a rich source of protein (CP, 28-34%), karanj cake was found to be slightly bitter in taste and toxic owing to the presence of flavonoid (Karanjin), restricting its safe inclusion in the livestock diets. Feeding trials with raw cake revealed its poor palatability and adverse performance among different categories of livestock including poultry. The present study was, therefore, aimed to detoxify karanj cake by various physico-chemical methods like solvent extraction, water washing, pressure cooking and alkali and acid treatments. The level of residual karanjin in raw and variously processed cake was quantified using high performance liquid chromatography (HPLC). The raw expeller karanj cake was found to contain about 0.19% of karanjin. Though a non-polar solvent, soxhlet extraction of expeller pressed cake with petroleum ether drastically reduced karanjin content (0.01%). Soaking of cake for 24 h in 1% NaOH (w/w) solution was found to reduce karanjin to a major extent with little further benefit by increasing alkali level. Milder alkalies like lime and fertilizer grade urea reduced the karanjin levels marginally. Similar was the case with mineral acids such as HCl and glacial acetic acid. It was, therefore, concluded that solvent extraction of karanj seeds would be the best method of detoxification as well as for more recovery of oil and karanjin.

Simultaneous Determination of Amoxicillin and Ampicillin in Fish Meat Using High-Performance Liquid Chromatography (HPLC를 이용한 어류 중의 Amoxicillin과 Ampicillin 항생제 동시 분석법)

  • Jo, Mi-Ra;Kim, Poog-Ho;Lee, Tae-Seek;Oh, Eun-Gyoung;Yu, Hong-Sik;Lee, Hee-Jung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.39 no.6
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    • pp.454-459
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    • 2006
  • A simultaneous high-performance liquid chromatography assay method for amoxicillin and ampicillin in fish products was developed, evaluated, and validated by monitoring these antibiotics in fish samples obtained from aquaculture and distribution. The recovery rate of this method was higher than those of conventional methods and was 95.3-106.6% for amoxicillin and 81.4-92.4% for ampicillin. Our pretreatment procedure sufficiently removed or reduced materials affecting HPLC analysis, such as low-molecular-weight substances. The performance limit of this method was evaluated as 0.01 ppm of amoxicillin and ampicillin in fish muscle. Finally, 171 fish samples, including olive flounder (Paralichthys olivaceus), common sea bass (Lateolabrax japonicus), and black rock fish (Sebastes schlegeli) collected from fish farms in the coastal area between April and September 2005 were analyzed to evaluate the overall efficiency of the method and to monitor the actual of amoxicillin and ampicillin usage in fish farms. The results indicated that the developed method was suitable for analyzing amoxicillin and ampicillin in fish muscle, and determined that those antibiotics were being used for fish farming but were not detected in fish samples during the shipping and distribution stages.

Lipids and Free Sugar Composition in Ginseng Classified by Years (한국(韓國) 인삼(人蔘)의 연근별(年根別) 지질(脂質) 및 유리당조성(遊離糖組成))

  • Sohn, K.M.;Sung, T.S.;Cho, Y.J.;Lee, K.S.;Choi, C.
    • Applied Biological Chemistry
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    • v.31 no.2
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    • pp.169-176
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    • 1988
  • Lipid contents, fatty acid compositions and free sugar contents of dried ginsengs grown for different years were studied. The lipid and free sugar were separated and quantified by silicic acid column chromatography, thin layer chromatography, gas liquid chromatography and high performance liquid chromatography. Ginseng contained $1.07{\sim}67%$ crude lipid and the hightest lipid content was shown in 3 year old root. The lipid fractions obtained by silicic acid column chromatography were mainly composed of neutral lipid$(51.35{\sim}72.30%)$. The contents of phospholipid and glycolipid in total lipids were $15.03{\sim}34.59%$ and $11.83{\sim}20.72%$, respectively. As the ginseng grew, neutral lipid content increased gradually but glycolipid decreased to the half of the one year old root. Seven components of neutral lipid separated by thin layer chromatography were identified but the other two components were not able to be identified. Triglyceride content was $14.42{\sim}23.91%$, sterol ester and unidentified material(II) were $13.58{\sim}21.26%$ and $7.13{\sim}15.83%$ respectively. The major fatty acids were linoleic acid, palmitic acid, oleic acid and linolenic acid. Linoleic acid content of total and neutral lipid was $61{\sim}65%$ palmitic acid content in phospholipid was $26{\sim}39%$ and one year old root showed the hightest contents. The fatty acid composition of neutral lipid was similar to the pattern of total lipid. Linolenic acid content of glycolipid decreased gradually. Free sugars were composed of rhamnose, fructose, glucose, sucrose, maltose and an unidentified material. The content of sucrose was $92{\sim}94%$ of total free sugars. Two year old root showed the highest sugar content and it decreased since then.

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Determination of mandipropamid residues in agricultural commodities using high-performance liquid chromatography with mass spectrometry (고성능액체크로마토그래피를 이용한 농산물 중 Mandipropamid의 잔류분석법 확립)

  • Kwon, Chan Hyeok;Chang, Moon Ik;Im, Moo Hyeog;Choi, Hoon;Jung, Da I;Lee, Su Chan;Yu, Jin Young;Lee, Young Deuk;Lee, Jong Ok;Hong, Moo Ki
    • Analytical Science and Technology
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    • v.21 no.6
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    • pp.518-525
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    • 2008
  • Mandipropamid is a new mandelamide-type fungicide to control foliar Oomycete pathogens in some vegetables. An analytical method was developed to determine mandipropamid residues in agricultural commodities using high-performance liquid chromatography (HPLC) and liquid chromatography/mass spectrometry (LC/MS). Mandipropamid was extracted with methanol from grape, tomato, green pepper, Chinese cabbage and potato samples. The extract was diluted with saturated sodium chloride solution and distilled water, and dichloromethane partition was followed to recover the mandipropamid from the aqueous phase. Florisil column chromatography was employed to further remove interfering co-extractives prior to HPLC analysis. Reverse-phased HPLC was successfully applied to determine mandipropamid in sample extracts with the detection at its ${\lambda}_{max}$ (223 nm). Overall recoveries of mandipropamid from fortified samples averaged $99.8{\pm}1.7$ (n=6), $89.3{\pm}5.3$ (n=6), $98.7{\pm}2.2$ (n=6), $99.7{\pm}6.8$ (n=6) and $91.1{\pm}3.1$ (n=6) for grape, tomato, green pepper, Chinese cabbage and potato, respectively. Limit of quantification of the method was 0.02~0.04 mg/kg for all samples. A LC/mass spectrometry with selected-ion monitoring was also provided to confirm the suspected residue. The proposed method was reproducible and sensitive enough to determine the terminal residue of mandipropamid in agricultural commodities.