• Title/Summary/Keyword: HPLC analysis methods

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Development of a Simple Method for Detecting Capsaicinoids Using Gibb's Reagent in Pepper (Gibb's Reagent를 이용한 캡사이시노이드 간이 분석 방법)

  • Jeong, Hee-Jin;Hwang, Do-Yeon;Ahn, Jeong-Tak;Chun, Jin-Young;Han, Ko-Eun;Lee, Woo-Moon;Kwon, Jin-Kyung;Lee, Yong-Jik;Kang, Byoung-Cheorl
    • Horticultural Science & Technology
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    • v.30 no.3
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    • pp.294-300
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    • 2012
  • Capsaicinoids are responsible for the pungency of Capsicum species. Among the several reported methods for quantifying capsaicinoids in pepper, liquid chromatography methods such as TLC and HPLC have been the most widely used due to their precision and reliability. However, they are quite expensive and time consuming to be applied to the field breeding. In this paper, we demonstrated that Gibb's reagent, 2,6-dichloroquinone chlorimide, mediated measurement of capsaicinoids is a simple and reliable method for determining the presence/absence of capsaicinoids, and estimating the amount of capsacinoids in pepper fruits. The capsaicinoids could be also detected via colorimetiric reactions of the Gibb's reagent. This simple method has been verified to be as accurate as the HPLC analysis. We have also modified this method for a high through-put analysis. This method will be useful for measuring capsaicinids in pungency breeding programs in pepper.

Formaldehyde Monitoring of Hygiene Products in Domestic Market (국내 유통 위생용품 중 포름알데히드 잔류량 모니터링)

  • Na, Young-Ran;Kwon, Hyeon-Jeong;Cho, Hyun-Nho;Kim, Hyeon-Jin;Park, Yon-Koung;Park, Sung-Ah;Lee, Seong-Ju;Kang, Jung-Mi
    • Journal of Food Hygiene and Safety
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    • v.35 no.3
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    • pp.225-233
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    • 2020
  • By the standards and specifications for hygiene products, three test methods for formaldehyde are specified for each item type of hygiene product. After derivatization using acetylacetone and 2,4-dinitrophenylhydrazine (2,4-DNPH), formaldehyde is analyzed by spectrophotometer and high-performance liquid chromatography (HPLC). Validation of the three test methods was performed on tissue, diaper lining and waterproof layer, and panty liner products. The results of linearity (R2), limit of detection (LOD), limit of quantification (LOQ), recovery rate (%) and reproducibility (%), showed that all three methods are suitable for analyzing formaldehyde in hygiene products. After derivatization with 2,4-DNPH and cetylacetone, formaldehyde was analyzed at 0, 3, 6, 9, 24 and 48 hours by HPLC. Formaldehyde derivatized with 2,4-DNPH showed no statistically significant change in formaldehyde peak area over time (P>0.05). But, acetylacetone-derivatizated formaldehyde showed a negative correlation coefficient (r) over time (P<0.01). We investigated the residual amounts of formaldehyde in 205 hygiene products distributed in Busan. Among 74 disposable diaper products tested, 73 had low concentrations of formaldehyde (0.13-29.87 mg/kg). Moreover, formaldehyde was not detected in any of 78 tissue, 27 disposable paper towel, 12 disposable dishcloth, 7 paper cup, one brand of paper straw and 6 disposable napkin products.

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.

Mass Spectrometric Analysis for Discrimination of Diastereoisomers

  • Manshoor, Nurhuda;Weber, Jean-Fré
    • Mass Spectrometry Letters
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    • v.6 no.4
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    • pp.99-104
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    • 2015
  • A liquid chromatography mass spectrometry (LC-MS) system was used to identify and distinguish oligostilbene diastereoisomers. A polyphenolic extract from Neobalanocarpus heimii known to be rich in oligostilbenes of various degrees of condensation was used as test material. Fourteen oligostilbenes were isolated from this extract on a fully automated semi-preparative HPLC system. Out of these, two pairs of dimers, one pair of trimers, two pairs of tetramers and a group of four tetramers with similar skeleton were identified as diastereoisomers. Their structures and configurations were established by spectroscopic methods. All isolated compounds were subjected to an LC-MS/MS to study their fragmentation patterns. The experiments were performed on a liquid chromatography-mass spectrometry (LC-MS) with electrospray-ionization (ESI) interface in positive mode. MS/MS spectra of each pure compound were recorded by direct infusion in identical conditions and their product ion spectra were analysed. Some subtle yet significant differences were observed between the spectra of oligostilbenes from the various diastereoisomeric series.

Revers phase HPLC Separation of D-Amygdalin and Neoamygdalin and Optimum Conditions for Inhibition of Racemization of Amygdalin

  • Hwang, Eun-Young;Kim, Dong-Min;Koh, Jung-A;Lee, Sang-Soo;Lee, Je-Hyun;Lee, Yong-Moon;Hong, Seon-Pyo
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.253.3-254
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    • 2002
  • In boiling aqueous solution, D-amygdalin usually begins to convert into neoamygdalin in 3 min and more than 30% of the initial D-amygdalin is found as neoamygdalin after 30 min. In this report, we establish methods for simple HPLC analysis and the inhibition of D-amygdalin conversion. D-Amygdalin and its conversion product, neoamygdalin, were clearly separated on reverse-phase column chromatography by an optimized eluent of 10mM sodium phosphate buffer (pH 3.8) containing 6% acetonitrile. (omitted)

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Isolation and Quantitative Determination of Dioscin from Smilacis Chinae Radix (토복령(土茯令, Smilacis Chinae Radix)으로부터 Dioscin의 분리 및 함량분석)

  • Seo, Jung-Hak;Lee, Joo-Mi;Kwon, Soon-Jin;Chang, Seung-Yeup;Lee, Kyong-Soon;Son, Kun-Ho
    • Korean Journal of Pharmacognosy
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    • v.32 no.2 s.125
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    • pp.153-156
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    • 2001
  • The methods for isolation and quantitative determination of dioscin, a spirostanol triglycoside, from the roots of Smilax china have been developed. Isolation of dioscin was achieved by silica gel and RP-18 chromatography. The HPLC method used for quantitative determination of dioscin enabled the standardization of the crude drug. It suggests that the content of dioscin in Smilax china should be above 0.01%.

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Structural Analysis of Black Common Bean (Phaseolus vulgaris L.) Anthocyanins

  • Choung, Myoung-Gun
    • Food Science and Biotechnology
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    • v.14 no.5
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    • pp.672-675
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    • 2005
  • Two anthocyanins were isolated from 1% HCl-20% methanol extracts of KG 97287 black common bean (Phaseolus vulgaris L.) using semipreparative, high-performance liquid chromatography (HPLC). The anthocyanins were identified using a combination of LC/ES-mass spectrometry (MS) and spectroscopic methods of UV-Vis, $^1H-$ and $^{13}C-$ nuclear magnetic resonance (NMR). The chemical structures of these two anthocyanins were elucidated as delphinidin 3-glucoside and petunidin 3-glucoside and their contents in KG 97287 black common bean seed coats were determined to be $2.614{\pm}0.11$ and $0.167{\pm}0.01\;mg/g$, respectively. These contents were lower than reported internationally and we recommend the introduction into Korea of high anthocyanin varieties of black common bean.

Separation of Tetrodotoxin, DHA and EPA from Pufferfish Liver Waste (복어간 폐기물로부터 Tetrodotoxin, DHA 및 EPA의 분리)

  • 차병윤;최진석;임정규;이동익;이원갑;이은열;김희숙;김동수
    • Journal of Life Science
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    • v.10 no.2
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    • pp.115-124
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    • 2000
  • The present study was undertaken to separate the available components effectively, such as tetrodotoxin(TTX), docosahexaenoic acid(DHA, C22:6,ω-3) and eicosapentaenoic acid (EPA, C20:5,ω -3) from pufferfish liver waste, which are known to have high values as bioactive materials. By using ultrafiltration, it was possible to separate high contents of 68mg TTX from pufferfish liver waste. In contrast, by activated charcoal column, it was to obtain about 54mg TTX. The recovering ratios were 65.3% and 45.0% in the two different methods of ultrafiltration and activated charcoal column, respectively. From the results of HPLC and gas chromatography-mass spectrometry(GC-MS), the obtained toxins were identified to be TTX and its derivatives. In addition, it was also possible to obtain 72.3g DHA and 11.4g EPA from 1kg of pufferfish liver by high performance liquid chromatography (HPLC). These amounts of DHA and EPA were also 17.70% and 1.04% in the total lipid of pufferfish liver oil from analysis of gas chromatography(GC), respectively.

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Production of Ginseng Saponins with Cell Culture(II) (세포배양에 의한 고려인삼 성분의 생산 연구(II))

  • Chi, Hyung-Joon;Shin, Kuk-Hyun;Kim, Hyun-Soo;Cho, Hee-Jae
    • Korean Journal of Pharmacognosy
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    • v.20 no.3
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    • pp.162-169
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    • 1989
  • Tissue culture of the roots of Panax ginseng was carried out to enhance the production of ginseng callus as well as to increase its contents of ginsenosides. A long cylinder type callus mass was formed when cultured IK callus by rotary shaking culture method, the growth ratio of the callus being 7.71 which was approximately 4 fold higher than those obtained by other culture methods. Ginsenosides $Rg_1$, Re and $Rb_1$ could be detected from the callus mass by TLC, however, their total contents were revealed to be approximately 9% compared to that of the fresh ginseng root equivalent by HPLC analysis,.

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Quantitative analysis of Cynanchum wilfordii Hemsley (Cynanchum wilfordii의 성분함량 분석)

  • Lee, Hye-Won;Park, So-Young;Lee, A-Young;Chae, Sung-Wook;Choi, Go-Ya;Choo, Byung-Kil;Kim, Ho-Kyoung
    • Korean Journal of Oriental Medicine
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    • v.14 no.1
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    • pp.107-111
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    • 2008
  • Objectives : Cynanchum wilfordii(Asclepiadaceae) has been traditionally used as a tonic, the prevention and treatment of various geriatric diseases in Korea. Acetophenone derivatives from C. wilfordii showed neuroprotective activity. In this study, two acetophenones were isolated and quantitative determination of acetophenones from C. wilfordii has been developed for quality stand. Methods : Three acetophenone derivatives were isolated from methanol extract of C. willfordii by the chromatographic separation. Their structures were identified as cynandione A, 2,5-dihydroxy acetophenone and cynanchone A on the basis of spectral data(MS, $^1H-NMR$, $^{13}C-NMR$) and chemical analysis. HPLC analysis was performed to determine the contents of cynandione A and 2,5-dihydroxyacetophenone in C. wilfordii. Results : According to the results, the contents of cynandione A and 2,5- dihydroxyacetophenone were 0.274%, 0.035% by HPLC, respectively. Conclusions : In these results, we have determined the contents of cynandione A and 2,5-dihydroxyacetophenone in Cynanchum wilfordii, respectively. We hope that this study will contribute to the standardization and quality control of herbal medicine.

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