• Title/Summary/Keyword: HPLC quantification

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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 four fluoroquinolones in chicken, pork and beef edible muscle by HPLC (식육 중 항균물질 (플루오르퀴놀론계) 동시 다성분 분석법 개선 연구)

  • Park Dong-Yeob;Hwang Bo-Won;Cho Sung-Suk;Choi Chan-Young;Cho Sang-Lae;Park Ae-Ra;Jung Eun-Hee;Byun You-Sung
    • Korean Journal of Veterinary Service
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    • v.29 no.2
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    • pp.111-122
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    • 2006
  • A direct, accurate and sensitive chromatographic analytical method for quantitative determination of four fluoroquinolones (norfloxacin, cirprofloxacin, danofloxacin and enrofloxacin) in chicken, pork and beef edible muscle is proposed in the present study. The developed method was successfully applied to the determination of enrofloxacin, as the main component of commercially available veterinary drugs. The samples were homogenized and the antimicrobials were added, then they were extracted twice with dichloromethane. Fluoroquinolone antibiotics were separated on an agilent $250x4mm,\;C_{18},\;5{\mu}m$, analytical column, at $25^{\circ}C$. The mobile phase consisted of a mixture of DW : acetonitrile : triethylamine(80:19:1%, v/v, pH 3.0) leading to retention times less than 14 min. at a flow rate 0.5 ml/min. These fluoroquinolones were detected by liquid chromatography with fluorescence at 290 nm excitation and 465 nm emission. The limits of quantification in each edible muscle (chicken, pork, and beef) were 0.32-6.54 ng/g. Using 0.5 g of each sample, average recovery rates at fortification levels of 0.05, 0.1 and 0.2 ${\mu}g/ml$ ranged 70.14-71.71% for NFX, 71.87-73.89% for CFX, 82.16-92.35% for DFX, and 90.13-98.12 for EFX This is a simple and economic method to quantify the presence of NFX, CFX, EFX and DFX in edible muscle of animal origin.

Quality Assessment of Ijung-tang Preparations Using a HPLC Analysis (HPLC 분석법을 이용한 이중탕(理中湯) 제제의 품질평가)

  • Ha, Woo-Ram;Park, Jin-Hyung;Yun, Dong-In;Lee, Jang-Cheon;Kim, Jung-Hoon
    • The Korea Journal of Herbology
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    • v.31 no.3
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    • pp.29-35
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    • 2016
  • Objectives : Ijung-tang (IJT) is a traditional herbal formula and has been used to treat digestive diseases such as abdominal pain, vomiting, and diarrhea. IJT consists of four herbal medicines, Ginseng radix, Atractylodis rhizoma alba, Zingiberis rhizoma, and Glycyrrhizae radix et rhizoma, containing various bioactive compounds. Quality assesment of IJT preparations was performed by analytical method for determining marker compounds.Methods : Determination of seven marker compounds in IJT preparations was quantitatively conducted by high-performance liquid chromatography equipped with a diode-array detector. The marker compounds were separated on a reversed-phase C18 column and the analytical method was successfully validated. Chemometric analysis was performed to compare IJT water extracts and commercial IJT granules.Results : Limit of detection and limit of quantification values were in the ranges of 0.093-2.649 μg/mL and 0.283-8.027 μg/mL, respectively. Precisions were 0.30-3.87% within a day and 0.23-2.35% over three consecutive days. Recoveries of the marker compounds ranged from 87.35-107.05%, with relative standard deviation (RSD) values < 6.15%. Repeatabilities were < 1.20% and < 1.71% of RSD value for retention time and absolute peak area, respectively. The results from quantitative analysis showed that the quantities of seven marker compounds of IJT samples varied, as were found in principal component analysis and hierarchical clustering analysis.Conclusions : The analytical method developed in the present study was precise and reliable to simultaneously determine marker compounds of IJT. Therefore, it can be used for the quality assessment of IJT preparations.

Variations of glucosinolates in kale leaves (Brassica oleracea var. acephala) treated with drought-stress in autumn and spring seasons (수분스트레스에 의한 케일 내 글루코시놀레이트 변화)

  • Jeong, Na-Rae;Chun, Jin-Hyuk;Park, Eun-Jae;Lim, Ye-Hoon;Kim, Sun-Ju
    • Korean Journal of Agricultural Science
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    • v.42 no.3
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    • pp.167-175
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    • 2015
  • The present study aimed to investigate the effects of drought stress on the accumulation of glucosinolates (GSLs) in the leaves of Kale cultivated in autumn and spring. HPLC analysis guided to identify seven GSLs including progoitrin, glucoraphanin, sinigrin, gluconapin, glucobrassicin, 4-methoxyglucobrassicin and neoglucobrasscin. Quantification of GSLs revealed that the contents of sigirin was the highest (45%) followed by the level of progoitrin (24%) in terms of total GSLs. The ranges of total GSL contents was 1.16 (84)-15.88 (89 DAS, ${\mu}mol/g$ dry wt. (DW)) in treatment plot and 1.23 (84)-7.05 (74 DAS, ${\mu}mol/g$ dry wt.) in control plot showed the enhancement in the contents of GSLs in treatment than in the control plot. The present results evidenced that the variation of total GSL contents were depending on the harvest period. In 105 DAS, comparatively no differences in the GSL contents on each sample in autumn season, whereas in spring season, although there was decrease in the GSLs tendency from 74 DAS to 84 DAS in both control and treatment plot, the GSL contents of treatment plot was dramatically increased in 89 DAS. In treatment plot, the GSL contents on 89 DAS (1.16) was 15 fold higher to 84 DAS ($15.88{\mu}mol/g$ DW). The variation in the contents of GSL in spring and autumn did not documented significant differences because of their differences in the growth time and cultivation conditions. In conclusion, the GSL contents in kale was likely to be affected by drought stress treatment. Scrutiny and further research for exact relation between drought stress and GSL contents in kale should be needed.

Quantitative Changes in Phenolic Compounds of Safflower (Carthamus tinctorius L.) Seeds during Growth and Processing

  • Kim, Eun-Ok;Lee, Jun-Young;Choi, Sang-Won
    • Preventive Nutrition and Food Science
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    • v.11 no.4
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    • pp.311-317
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    • 2006
  • Phenolic compounds in safflower seeds were recently found to stimulate bone formation and increase plasma HDL cholesterol levels in estrogen deficient rats, and to inhibit melanin synthesis. Nine phenolic compounds: $N-feruloylserotonin-5-O-{\beta}-D-glucoside,\;8'-hydroxyarctigenin-4'-O-{\beta}-D-glucoside,\;luteolin-7-O-{\beta}-D-glucoside$, N-(p-coumaroyl)serotonin, N-feruloylserotonin, 8'-hydroxy arctigenin (HAG), luteolin (LT), $acacetin-7-O-{\beta}-D-glucuronide$ (ATG) and acacetin (AT), were quantified by HPLC in safflower (Carthamus tinctorius L.) seeds during growth and processing. During growth, levels of the nine phenolic compounds in the seeds increased progressively with increasing growth stages, reached a maximum on July 30 (42nd day after flowering), and then remained relatively constant. During the roasting process, levels of phenolic compounds, except HAG, LT and AT, generally decreased with increased roasting temperature and time, whereas those of HAG, LT and AT increased progressively with increased roasting temperature and time. During the steaming process, levels of other phenolic compounds except HAG and AT generally tended to increase with increased steaming time, whereas those of HAG and AT were scarcely changed. During the microwave treatment, quantitative changes of phenolic compounds were similar to the roasting process, although there were some differences in levels of phenolic compounds between two heat treatments. These results suggest that the steamed safflower seeds after harvesting on late July may be useful as potential dietary supplement source of phenolic compounds for prevention of several pathological disorders, such as atherosclerosis and osteoporosis and aging.

Simultaneous Determination of Creatine, Dicyandiamide and Dihydrotriazine in Dietary Supplements by High Performance Liquid Chromatography (고성능 액체크로마토그래피를 이용한 식이보충제에서 크레아틴, 디시안디아마이드, 디하이드로트리아진의 동시분석)

  • Park, Sang-Wook;Yoo, Myung-Sang;Lee, Wonjae
    • KSBB Journal
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    • v.29 no.4
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    • pp.232-238
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    • 2014
  • The simultaneous determination of creatine monohydrate (CrM), dicyandiamide and dihydrotriazine in dietary supplements using reversed-phase high performance liquid chromatography (HPLC) was developed. Chromatography was performed on a Nuclosil 100-5 SA ($4.6{\times}250mm$) column with a mobile phase of 2.3% ammonium phosphate (pH 5.5), and UV detection at 224 nm, 212 nm, and 237 nm, respectively. The performance characteristics of HPLC were determined in terms of selectivity, linearity, precision, recovery, limit of detection (LOD), and limit of quantification (LOQ). The calibration curves were linear within the concentration range of $40.0{\sim}500.0{\mu}g/mL$ for creatine, $0.1{\sim}12.8{\mu}g/mL$ for dicyandiamide, and $0.05{\sim}6.4{\mu}g/mL$ for dihydrotriazine. The detection limits of the method were 1.09, 0.01, and $0.08{\mu}g/mL$ for creatine, dicyandiamide, and dihydrotriazine, respectively. The recoveries of creatine, dicyandiamide, and dihydrotriazine were 97.2~100.9, 92.3~106.5, and 97.2~105.5%, respectively. It is expected that the chromatographic analytical method developed in this study will be usefully applicable to simultaneous determination of creatine, dicyandiamide, and dihydrotriazine contained in dietary supplements.

Simultaneous Quantitative Analysis of Mixtures of Ixeridium dentatum (Thunb.) Tzvelev, Plantago asiatica L. and Rumex crispus L. Ethanol Extracts (씀바귀, 질경이, 소리쟁이 에탄올 추출 혼합물에 대한 주요 성분 동시 분석)

  • Kim, Ga-Ram;Kim, Eun-Nam;Birasuren, Bayarmaa;Min, Yeonhong;Jeong, Gil-Saeng
    • Korean Journal of Pharmacognosy
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    • v.51 no.3
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    • pp.222-229
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    • 2020
  • Recently, consume for functional cosmetics containing natural products has been greatly increased. In order to develop it as a natural cosmetic material, we selected Ixeridium dentatum (Thunb.) Tzvelev, Plantago asiatica L. and Rumex crispus L. that have antioxidant and anti-inflammatory effects. In this study, simultaneous quantitative analysis of the isolated compounds and natural product complexes (Mix.) were validated using high performance liquid chromatography (HPLC). The isolated six compounds were shown in a large linearity with a correlation coefficient (R2) of 0.99. The limit of detection (LOD) of chlorogenic acid, plantamajoside, acteoside, emodin chrysophanol and physcion were 0.36 ㎍/mL, 0.36 ㎍/mL, 0.37 ㎍/mL, 0.30 ㎍/mL, 0.22 ㎍/mL and 0.12 ㎍/mL, respectively. The limit of quantification (LOQ) of chlorogenic acid, plantamajoside, acteoside, emodin chrysophanol and physcion were 1.10 ㎍/mL, 1.08 ㎍/mL, 1.12 ㎍/mL, 0.99 ㎍/mL, 0.66 ㎍/mL and 0.35 ㎍/mL, respectively. Content analysis showed chlorogenic acid (0.19 ± 0.02%), plantamajoside (0.48 ± 0.01%), acteoside (0.65 ± 0.01%), emodin (1.15 ± 0.11%), chrysophanol (0.73 ± 0.01%) and physcion (0.69 ± 0.09%). Therefore, the results of this study may provide for basic data of standardization research natural cosmetic material development on the I. dentatum, P. asiatica and R. cripus.

Molecular Identification and Chemical Analysis of Aconiti Kusnezoffii Tuber on the Domestic Markets (국내 시장에서 유통되는 초오의 DNA 감별과 화학적 분석)

  • Jang, Hyeri;Joe, Kyeong-Hwa;Song, Kwangho;Lee, Kyoung Jin;Park, Sait Byul;Lee, Chaemin;Ha, In Jin;Lee, Kyungjin;Suh, Youngbae;Kim, Yeong Shik
    • Korean Journal of Pharmacognosy
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    • v.49 no.2
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    • pp.145-154
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    • 2018
  • Aconiti Kusnezoffii Tuber has been traditionally used to treat the symptoms of rheumatoid arthritis and joint pain. The main constituents are diterpenoid alkaloids such as benzoylmesaconine, benzoylaconine, mesaconitine, aconitine, and hypaconitine. In Korea, Aconiti Kusnezoffii Tuber is officially defined as the tubers of Aconitum kusnezoffii Reichb., A. ciliare Decasisne, and A. triphyllum Nakai. On the other hand, only the tuber of A. kusnezoffii is to be used in China. In order to identify the botanical origin of Aconiti Kusnezoffii Tuber circulated in Korea, we analyzed 24 samples of Aconiti Kusnezoffii Tuber obtained from local markets for comparative DNA analysis. The sequence analysis of nrRNA ITS 1 was useful to distinguish Aconitum species and revealed that the roots of A. karakolicum were circulated in Korean markets without discretion. HPLC quantitative analysis showed that aconitine was detected at the highest amount in A. karakolicum. Authentic diterpenoid alkaloids were coinjected for quantification of aconitine-type ingredients. All data were statistically grouped by Principal Component Analysis (PCA). This study suggests that both molecular and chemical analyses should be utilized for the standardization and the quality control for Aconiti Kusnezoffii Tuber.

Qualitative Analysis of the Major Constituents in Traditional Oriental Prescription Bang-poong-tong-sung-san by Liquid Chromatography/Ultraviolet Detector/Ion-Trap Time-of-Flight Mass Spectrometry

  • Eom, Han Young;Kim, Hyung-Seung;Han, Sang Beom
    • Mass Spectrometry Letters
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    • v.5 no.1
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    • pp.24-29
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    • 2014
  • An advanced and reliable high performance liquid chromatography (HPLC)/ultraviolet detector (UV)/ion-trap time-of-flight (IT-TOF) mass spectrometry was developed for the simultaneous quantification of 19 marker compounds in Bang-poong-tong-sung-san (BPTS), a traditional oriental prescription. Various parameters affecting HPLC separation and IT-TOF detection were investigated, and optimized conditions were identified. The separation was achieved on a Capcell PAK C18 column ($1.5mm{\times}250mm$, $5{\mu}m$ particle size) using a gradient elution of acetonitrile and water containing 0.1% formic acid at a flow rate of 0.1 mL/min. The column temperature was maintained at $40^{\circ}C$ and the injection volume was $2{\mu}L$. IT-TOF system was equipped with an electrospray ion source (ESI) operating in positive or negative ion mode. The optimized electrospray ionization parameters were as follows: ion spray voltage, +4.5 kV (positive ion mode), or -3.5 kV (negative ion mode); drying gas ($N_2$), 1.5 L/min; heat block temperature, $200^{\circ}C$. Automatic $MS^n$ (n = 1~3) analyses were carried out to obtain structural information of analytes. Elemental compositions and their mass errors were calculated based on their accurate masses obtained from a formula predictor software. The marker compounds in BPTS were identified by comparisons between $MS^n$ spectra from standards and those from extracts. Moreover, the libraries of $MS^2$ and $MS^3$ spectra and accurate masses of parent and fragment ions for marker compounds were constructed. The developed method was successfully applied to the BPTS extracts and identified 17 out of 19 marker compounds in the BPTS extracts.

Simultaneous analysis and occurrences of six pharmaceuticals in surface water by LC/ESI-MS/MS (LC/ESI-MS/MS를 이용한 하천수 중 잔류 6종 의약물질의 동시분석 및 모니터링)

  • Kim, Byung-Ju;Myung, Seung-Woon
    • Analytical Science and Technology
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    • v.23 no.6
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    • pp.572-578
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    • 2010
  • The extraction/clean-up and concentrating of pharmaceuticals from surface water were performed by HLB (Hydrophilic-Lipophilic Balanced) cartridge. The method allows for the simultaneous determination of six pharmaceuticals by HPLC/ESI(+)-MS/MS. Recoveries of the pharmaceutical were between 71.1 to 92.6% (except fenbendazole) and the overall variability of the method was below 11.2% (RSD). The calibration curves for the pharmaceuticals from blank surface water showed good linearities (above $r^2$ = 0.99) in the concentration range of 0.007~1.2 ng/mL. The limit of detection (LOD) and the limit of quantification (LOQ) were 7.2~128.7 pg/mL and 23.8~429.1 pg/mL, respectively. The present analytical method can be useful for monitoring residual pharmaceuticals in surface water and other aquatic samples. High concentrations of iopromide and fenbendazole were detected in a few samples of surface water.