Cyanidin-3-glucoside (C3G) and cyanidin-3-rutinoside (C3R) were isolated by high-performance countercurrent chromatography (HPCCC) using a two-phase solvent system composed of tert-butyl methyl ether/n-butanol/acetonitrile/water/trifluoroacetic acid (1 : 3 : 1 : 5 : 0.01, v/v) to give pure C3G (34.1 mg) and C3R (14.3 mg) from 1.5 g crude mulberry fruit extract. Using the pure C3G and C3R, a reliable high-performance liquid chromatography (HPLC) method was developed and validated to determine the C3G and C3R contents in mulberry fruit. C3G and C3R were separated simultaneously using an Eclipse XDB-C18 column ($4.6{\times}250mm$ I.D., $5{\mu}m$) coupled with a photodiode array detector (PDA). The gradient elution of the mobile phase consisting of acetonitrile (0.5% formic acid) and water (0.5% formic acid) was applied (1.0 mL/min), and the detection wavelength was 520 nm. The calibration curves of C3G and C3R showed good linearity (both with $r^2=0.9996$) in the concentration range $15.625-500{\mu}g/mL$, and the relative standard deviations (RSD%) of intra- and inter-day variability were in the ranges 2.1 - 8.2% and 4.1 - 17.1%, respectively. The accuracies were ranged 96.5 - 102.6% for C3G and C3R, respectively. The developed HPLC method was used to determine the contents of C3G and C3R in newly harvested mulberry from eight different provinces of Korea.
Kim, Il-Woung;Sun, Won Suk;Yun, Bong-Sik;Kim, Na-Ri;Min, Dongsun;Kim, Si-Kwan
Journal of Ginseng Research
/
v.37
no.1
/
pp.124-134
/
2013
The authentication of the physico-chemical properties of ginsenosides reference materials as well as qualitative and quantitative batch analytical data based on validated analytical procedures is a prerequisite for certifying good manufacturing practice (GMP). Ginsenoside Rb1 and Rg1, representing protopanaxadiol and protopanaxatriol ginsenosides, respectively, are accepted as marker substances in quality control standards worldwide. However, the current analytical methods for these two compounds recommended by Korean, Chinese, European, and Japanese pharmacopoeia do not apply to red ginseng preparations, particularly the extract, because of the relatively low content of the two agents in red ginseng compared to white ginseng. In manufacturing fresh ginseng into red ginseng products, ginseng roots are exposed to a high temperature for many hours, and the naturally occurring ginsenoside Rb1 and Rg1 are converted to artifact ginsenosides such as Rg3, Rg5, Rh1, and Rh2 during the heating process. The analysis of ginsenosides in commercially available ginseng products in Korea led us to propose the inclusion of the (20S)- and (20R)-ginsenoside Rg3, including ginsenoside Rb1 and Rg1, as additional reference materials for ginseng preparations. (20S)- and (20R)-ginsenoside Rg3 were isolated by Diaion HP-20 adsorption chromatography, silica gel flash chromatography, recrystallization, and preparative HPLC. HPLC fractions corresponding to those two ginsenosides were recrystallized in appropriate solvents for the analysis of physico-chemical properties. Documentation of those isolated ginsenosides was achieved according to the method proposed by Gaedcke and Steinhoff. The ginsenosides were subjected to analyses of their general characteristics, identification, purity, content quantification, and mass balance tests. The isolated ginsenosides showed 100% purity when determined by the three HPLC systems. Also, the water content was found to be 0.534% for (20S)-Rg3 and 0.920% for (20R)-Rg3, meaning that the net mass balances for (20S)-Rg3 and (20R)-Rg3 were 99.466% and 99.080%, respectively. From these results, we could assess and propose a full spectrum of physico-chemical properties of (20S)- and (20R)-ginsenoside Rg3 as standard reference materials for GMP-based quality control.
Park, Eun-Jeong;Lim, Ji-Huen;Lee, Sung-Mo;Im, Jeong-Soo;Oh, Dae-Kyu
Korean Journal of Veterinary Service
/
v.31
no.3
/
pp.385-395
/
2008
This study was carried out to investigate the residue level of fluoroquinolones in hen's general eggs and specific eggs by microbiological assay method and high performance liquid chromatography (HPLC) method. HPLC separation was carried out by reversed phase chromatography on a Symmetry $C_{18}$ (250${\times}$4.6 mm, $5{\mu}m$ particle size) with a phase composed of distilled water (containing 0.4% triethylamine and phosphoric acid) : Methanol (780 : 220, v/v), pumped isocratically at a flow rate of 1.0ml/min. A fluorescence detector was utilized with an excitation wavelength of 278nm and an emission wavelength of 456nm. The calibration curves were linear $({\gamma}^2{\geq}0.999)$ over a concentration range of $0.025{\sim}0.4{\mu}g/ml$. Average recoveries of the five fluoroquinolones in whole eggs at fortified levels of $0.05{\sim}0.2{\mu}g/g$ were ranged mean $78.1{\sim}91.7%$ and low coefficient of variation was less than 10% for all analysed samples. The limits of detection and limits of quantification for whole eggs were $1.2{\sim}6.0ng/g$ and $2.3{\sim}9.1ng/g$, respectively. Only one hen's general eggfrom chicken farm in Incheon was detected with the residual fluoroquinolones (Microbiological assay method; 1 of 47 general eggs) ; the range of residual concentration enrofloxacin was 0.12ppm. Those in food stores were detected with the residual fluoroquinolones (Microbiological assay method; 4 of 88 general eggs) ; the ranges of residual concentration enrofloxacin were $0.15{\sim}2.2 ppm$, ciprofloxacin $0.01{\sim}0.06ppm$, and hen's specific eggs (40) in food stores were not detected. For the microbiological assay method of fluoroquinolones in hen's eggs, as the results of comparative analysis, the disc diffusion method with E coli may be a little highly detected for the residual fluoroquinolones.
Kim, Young-Ju;Ko, Jae-Ho;Kim, Eun-Hye;Nam, Hyuk-Jin;Jo, So-Hee;Kim, Heon-Woong;Kim, Jung-Bong;Han, Byoung-Su
The Korean Journal of Food And Nutrition
/
v.25
no.3
/
pp.499-504
/
2012
We analyzed the contents of ${\gamma}$-oryzanol, which is contained in brown rice of the nation rice varieties Chucheong, Heukjinju and Sindongjin, by HPLC. Furthermore we also performed experiments on its biological activity, to prove the effectiveness of rice bran. The contents of ${\gamma}$-oryzanol contained in hulled rice showed 1,587 ppm for Heukjinju, followed by Chucheong(1,038 ppm), and by Sindongjin(472 ppm). In anti-oxidative activity, we performed an experiment, by measuring the radical scavenging activity of DPPH. Heukjinju showed the best effect, and Chucheong showed the worst effect. In cholesterol lowering activity, Heukjinju showed the best activity and Sindongjin showed the worst effect. In anti-bacterial activity as well, Heukjinju showed the best activity, and Sindongjin showed the worst effect. Through these experiments, we compared the contents of ${\gamma}$-oryzanol, which is contained in hulled rice(Chucheong, Heukjinju, Sindongjin). Also, we found the anti-oxidation effect, cholesterol lowering effect, and anti-bacterial activity of the ${\gamma}$-oryzanol extracts. Based on our research, we expect that ${\gamma}$-oryzanol can work as a new drug, or nutritional supplement.
Park, Jung-Sun;Kim, Hye-Kyung;Lee, Hye-Won;Lee, Mi-Hyun;Kim, Hyun-Gi;Chae, Soo-Wan;Chae, Han-Jung
Korean Journal of Clinical Pharmacy
/
v.16
no.1
/
pp.23-27
/
2006
Doxorubicin (DXR) is a type of anti-cancer drug called an 'anthracycline glycoside', It works by impairing DNA synthesis, a crucial feature of cell division, and thus is able to target rapidly dividing cells. Doxorubicin is a very serious anti-cancer medication with definite potential to do great harm as well as great good. A liquid chromatography-tandem mass spectroscopy (LC-MS/MS) method was developed to identify and quantify DXR in small-volume biological samples. After the addition of internal standard (IS, $5{\mu}L\;of\;1{\mu}M/ml$ daunorubicin methanol solution) into the serum sample, the drug and IS were extracted by methanol. Following vortex for a 1min and centrifugation at 15,000g for 10 min the organic phase was transferred and evaporated under a vacuum. The residue was reconstituted with $350{\mu}L$ of mobile phase and $10{\mu}L$ was injected into C18 column with mobile phase composed of 0.05M ammonium acetate (0.1 M acetic acid adjusted to pH 3.5) and acetonitrile (40:60, v/v). The flow rate was kept constant at $350{\mu}L/min$. The ions were quantified in the multiple reaction mode (MRM), using positive ions, on a triple quadrupole mass spectrometer. The lower limits of quantification for Doxorubicin in plasma and small tissues were approximately 0.5 ng/mL and 0.5 ng/mL respectively. Intra- and inter-assay accuracy (% of nominal concentration) and precision (% CV) for all analytes were within 15%, respectively.
The bioequivalence of two triflusal products was evaluated with 20 healthy volunteers following single oral dose according to the guidelines of Korea Food and Drug Administration (KFDA). Trisa $l^{R}$ capsule (Whanin Pharm. Corp., Korea) and Disgre $n^{R}$ capsule (Myung-In Pharm. Corp., Korea) were used as test product and reference product, respectively. Both products contain 300 mg of trifusal. One capsule of test product or reference product was orally administered to the volunteers, respectively, by randomized two period crossover study (2$\times$2 Latin square method). Blood samples were taken at predetermined time intervals for 4 hours and the determination of trifusal was accomplished using semi-microbore HPLC equipped with automated column switching system. The analytical method with HPLC was validated according to the Bioanalytic Method Validation guideline by F7A prior to determining the plasma samples. The pharmacokinetic parameters (AU $C_{0-4h}$$C_{max}$ and $T_{max}$) were calculated and ANOVA test was utilized for statistical analysis of parameters. As a result of the assay validation, the limit of quantification of trifusal in human plasma by current assay procedure was 50 ng/ml using 500 $\mu$l of plasma. The accuracy of the assay was from 97.76% to 116.51% while the intra-day and inter-day coefficient of variation of the same concentration range was less than 15%. Average drug concentration at the designated time intervals and pharmacokinetic parameters calculated were not significantly different between two products (p>0.05). The difference of mean AU $C_{olongrightarrow4hr}$, $C_{max}$, and $T_{max}$ between the two products (2.92, 4.39, and -2.44%, respectively) were less than 20%. The power (1-$\beta$) and treatment difference ($\Delta$) for AU $C_{olongrightarrow4hr}$ and $C_{max}$ were more than 0.8 and less than 0.2, respectively. Although the power for $T_{max}$ was under 0.8, $T_{max}$ of the two products was not significantly different from each other (p>0.05). These results satisfied the criteria of KFDA guideline for bioequivalence, indicating the two products of triflusal were bioequivalent.quivalent.ent.ent.
Kim, Dan-Bi;Kim, Sunyoung;Lee, Sanghee;Yoo, Miyoung
Analytical Science and Technology
/
v.30
no.5
/
pp.295-302
/
2017
This study presents a method validation for extraction and quantitative analysis of levulinic acid in soy sacues using high performance liquid chromatograph-photodiode array detector (HPLC-PDA). The levulinic acid in samples were extracted with distilled water, and then purified with C18 Sep-Pak cartridge. The calibration curves showed good linearity (R > 0.999) in a relatively wide concentration range ($2.5-400{\mu}g/mL$). Mean recoveries and relative standard deviation (RSD) of levulinic acid spiked in soy sauce samples at different spiking levels ($2.5-400{\mu}g/mL$; 6 point). Recoveries were 87.58-97.26 % with RSD less than 15 %, and limit of detection (LOD) and limit of quantification (LOQ) were 0.64 and $1.64{\mu}g/mL$, respectively. According to monitoring result with the established method, levulinic acid was found in 43 of 59 domestic commercial soy sauces, soy sauce based sauces and seasoned meats. The contamination levels were 0.44-1.23 mg/mL for soy sauces, 0.03-0.83 mg/mL for soy sauce based sauces and 8.43-38.94 mg/mL for seasoned meats. The results indicated to be rapidly and accurately qualifying levulinic acid and can be used as a suitable quality control method for soy sauce and soy sauce related commodities.
Jung, Sam Ju;Hwang, Young Sook;Choi, Chae Man;Park, Ae Suk;Kim, Su Un;Kim, Hyun Jung;Kim, Jung Hun;Jung, Kweon
Journal of the Society of Cosmetic Scientists of Korea
/
v.41
no.3
/
pp.219-227
/
2015
This study was conducted to determine 10 preservatives (benzyl alcohol (BAl), phenoxyethanol (PE), benzoic acid (BA), sorbic acid (SA), methyl paraben (MP), ethyl paraben (EP), propyl paraben (PP), isopropyl paraben (IPP), butyl paraben (BP), isobutyl paraben (IBP)) levels in 125 cosmetics (n = 125) for children by the simultaneous analysis of HPLC. The detection ranges were as follows; 0.01 ~ 0.91% (n = 35) for PE, 0.01 ~ 0.48% (n = 28) for BA, 0.01 ~ 0.78% (n = 9) for BAl, 0.01 ~ 0.11% (n = 3) for SA, 0.04 ~ 0.21% (n = 8) for MP, 0.02 ~ 0.09% (n = 8) for PP, and 0.04% (n = 1) for EP. The order of detection rates was cleanser (63%) > cream (48%) > sunscreen (46%) > lotion (38%) > oil (13%). At least one of target preservatives was contained in 50% (63/125) of samples and the content of the detected preservatives was within maximum allowed amount established by KFDA. Phenoxyethanol and benzoic acid were used more frequently than paraoxybenzoate esters (parabens) in cosmetics for children and the detected parabens was mainly the mixture of methyl paraben and propyl paraben.
Jeong, Min Kyu;Park, Chan Uk;Park, Min Hee;Yeo, JuDong;Park, SeungKwan;Kim, SoHee;Shin, Tae-Sun;Baek, Hyung Hee;Lee, JaeHwan
Food Engineering Progress
/
v.15
no.1
/
pp.75-79
/
2011
Analytical methods for food antioxidants including ascorbic acid, erythorbic acid, ascorbyl palmitate (AP), and ascorbyl stearate (AS), were validated using high performance liquid chromatography. Validation parameters such as linearity, limit of detection (LOD), limit of quantification (LOQ), and recovery were tested using lard and cider as food model systems. Linearity of ascorbic acid and erythorbic acid were both higher than ($R^2$> 0.99), LOD of these compounds were 0.46 and 0.48 ${\mu}g/mL$, respectively and LOQ were 1.39 and 1.45 ${\mu}g/mL$, respectively. The recovery rates of these compounds were 86.35-94.78% and 84.76-95.02%, respectively. However, the concentration of AP and AS decreased in methanol stock solution. Four other solvents including ethanol, acetonitrile, mixture of methanol and acetonitrile, and mixture of ethanol and acetonitrile were tested to increase the stability of AP and AS under room temperature and refrigerated temperature. Ethanol provided better stability of AP and AS under both room and refrigerated temperature. This study can help to accurately analyze the content of ascorbic acid and its derivatives in processed foods.
Yeon Suk Kim;Hyun Young Shin;Eun Ji Ha;Ja Pyeong Koo;Se Bin Jeong;Gaeuleh Kim;Mi Yeun Joung;Kwang-Won Yu
The Korean Journal of Food And Nutrition
/
v.36
no.2
/
pp.93-102
/
2023
Centella asiatica (C. asiatica) has been widely used in food, cosmetics, and pharmaceutical industry as a functional material. In a previous study, we have investigated not only pharmacological effects such as antioxidative and anti-inflammatory effects, but also analyzed various functional ingredients. In this study, triterpenoids were analyzed using HPLC-DAD to determine marker compounds among functional ingredients. When triterpenoids were analyzed, asiaticoside from C. asiatica was determined as an optimal marker compound. Next, specificity, linearity, limited of detection (LOD), limited of quantification (LOQ), precision, accuracy, and range were evaluated using HPLC-DAD to determine asiaticoside contents in C. asiatica juice and extracts. The specificity was elucidated by chromatogram and retention time using an established analytical method. The coefficient of correlation obtained was 0.9996. LOD was 4.99 ㎍/mL and LOQ was 15.12 ㎍/mL. Intra- and inter-day precision of asiaticoside were determined to be 0.48~1.68% and 0.08~1.09%, respectively. Furthermore, the recovery rate of asiaticoside was 98.88% and the analytical range of Field-70E was determined to be 0.625~10 mg/mL. As a results of evaluating ABTS, DPPH, and FRAP antioxidative effect, Field-70E showed potent antioxidant activities. Results of this study could be used as basic data for quality standardization of C. astiatica juice and extracts.
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