• Title/Summary/Keyword: Liquid Chromatography

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A New Synthetic Medium for Lactic Lactococci: Application to Marine lactic Acid Bacteria

  • KIM Joong K.;BAJPAI Rakesh K.
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.6
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    • pp.812-813
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    • 1995
  • Lactococcal cells are nutritionally fastidious and thus, generally cultured either in milk or M17 medium (Terzaghi and Sandine, 1975). In this study, Lactococcus cremoris wild-type (KH) and its less­proteolytic mutant (KHA1) cells were grown on the M17 medium or with modified M17 medium by replicated parallel experiments. The modified M17 medium had the same composition as M17 medium, except that lactose was replaced by glucose. Analyses of culture-broth samples, in which the M17 and the modified M17 media were used, were conducted by high-performance liquid chromatography (HPLC). But, working with these media created noisy problems in analyses of samples. Therefore, a new semi-synthetic medium was developed on the basis of nutritional requirements (Morishita et al., 1981). The composition of the semi-synthetic medium determined on the basis of the nutritional requirements and the composition of milk, is presented in Table 1. The composition of M17 medium is also presented and compared in the table. L. cremoris KH and KHA1 cells were grown again on the new synthetic medium containing glucose or lactose. The broth samples were then drawn and analyzed by HPLC. Clearer separations of fermented products were achieved from the new medium than those with the M17 and the modified M17 media. In comparison with the M17 or the modified M17 media, growth on the new medium was good (Kim et al, 1993). Additional fermentations were also carried out at a controlled pH of 7.0, where enhanced growth of lactococcal cells was obtained. In the fermentations, samples were also analyzed for the concentrations of sugar and lactic acid. The results showed that the new synthetic medium was as good as or better than the M 17 and the modified M 17 media. This is because casein hydrolysate in the synthetic medium provided a ready supply of amino acids and peptides for L. cremoris KH and KHA1 cells. Lactic acid bacteria (LAB) including Lactococcal cells have been known to be an effective means of preserving foods, at the same time as giving particular tastes in fields of dairy products. LAB also have always occupied an important place in the technology of sea products, and marine LAB have known to be present in traditional fermented products (Ohhira et al, 1988). To apply the new synthetic medium to marine LAB, two different LAB were isolated from pickled anchovy and pollacks caviar and were grown on the new media in which various concentrations of NaCl $(3, 5, 7 and 10\%)$ added. They were also grown on the medium solution in natural seawater $(35\%o\;salinity)$ and on the solution of natural seawater itself, too. As seen in Fig. 1, Marine LAB were grown best on the synthetic medium solution in natural seawater and the higher concentrations of NaCl were added to the medium, the longer lag-phase of growth profile appeared. Marine LAB in natural seawater were not grown well. From these results, the synthetic medium seems good to cultivate cells which are essential to get salted fish aged. In this study, it showed that the new synthetic medium provided adequate nutrition for L. cremoris KH and KHA1 cells, which have been used as cheese starters (Stadhouders et al, 1988). Using this new medium, the acid production capability of starter cultures could be also measured quantitatively. Thus, this new medium was inferior to the M17 or the modified M17 medium in culturing the cheese starters and in measuring fermentation characteristics of the starter cells. Moreover, this new medium found to be good for selected and well-identified marine LAB which are used in rapid fermentations of low-salted fish.

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Effect of Supercritical Carbon Dioxide on Physicochemical Properties and Microbial Reduction of Freeze-Dried Bovine Liver (초임계 이산화탄소 처리가 동결 건조된 소간의 이화학적 특성 및 미생물 저감화에 미치는 영향)

  • Kim, Hye-Min;Woo, Sung-Woon;Kim, Ah-Na;Heo, Ho-Jin;Chun, Ji-Yeon;Choi, Sung-Gil
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.12
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    • pp.1847-1855
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    • 2015
  • Supercritical carbon dioxide ($SC-CO_2$) treatment has been becoming an important method for substituting the use of organic solvents for samples extraction prior to analysis due to its low toxicity, ease of handling, low cost of disposal etc. Freeze-dried bovine liver was treated with $SC-CO_2$ under different pressures (200, 300, and 450 bar) in order to investigate effects on physicochemical properties and reduction of microbial load. The yield of lipid extraction from bovine liver by $SC-CO_2$ treatment increased with increasing pressure, with values of 84, 86, and 90% in response to 200, 300, and 450 bar, respectively. Results of high performance liquid chromatography analysis showed that vitamin A and coenzyme $Q_{10}$ ($CoQ_{10}$), which is soluble in lipid, were almost removed from bovine liver by $SC-CO_2$ treatment. Saturated fatty acids ratio of bovine liver decreased with increasing pressure, whereas polyunsaturated fatty acids increased with increasing pressure. Total content of amino acids in bovine liver treated by $SC-CO_2$ was less than that of the control sample without treatment. The number of aerobic bacteria in bovine liver, which was stored at $5^{\circ}C$ for 5 days and freeze-dried, decreased from 6.2 to 4.2 log CFU/g by $SC-CO_2$ treatment at 100 bar for 3 h. Interestingly, coliform bacteria were not found in the bovine liver sample by $SC-CO_2$ at 100 bar for 3 h under all storage conditions. This indicates that $SC-CO_2$ treatment can effectively reduce coliform bacteria in the food matrix even at low moisture. In conclusion, freeze-dried bovine liver by proper $SC-CO_2$ treatment may be used as a potential high protein source, with increasing microbial safety and stability of lipid oxidation.

Variation in Taxol Content of Bark, Shoots and Needles of Seven Populations of Three Native Taxus Species (한국자생(韓國自生) 주목(朱木), 설악(雪岳)눈주목(朱木), 회율(檜率)나무의 집단(集團) 및 채취부위(採取部位)에 따른 탁솔함량(含量)의 변이(變異))

  • Lee, Yong Wook;Lee, Kyung Joon
    • Journal of Korean Society of Forest Science
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    • v.83 no.3
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    • pp.365-371
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    • 1994
  • This study was conducted to investigate variations in taxol contents of bark, shoots, needles of three native Taxus species in Korea. Sample materials were collected in early October of 1992 from five individual, trees each of the five populations of Taxus cuspidata located at Mt. Halla, Mt. Jiri, Mt. Deokyu, Mt. Sobaek, and Mt. Taebaek and two individual trees of Taxus cuspitosa in Mt. Seolak and in March of 1993 from the five individual trees of Taxus cuspidata var. latifolia in Ulleung Island. The collected materials were analyzed by HPLC to determine the amount of taxol content. Regardless of species and population, the taxol content was highest in bark and followed by shoots and needles. In Taxars cuspidata the taxol content in the bark was highest for the population of Mt. Halla (0.017%) and decreased in the order of Mt. Jiri, Mt. Sobaek, Mt. Deokyu, and Mt. Taebaek. The taxol content in the shoots was highest for the population of Mt. Halla(0.0053%) and decreased in the order of Mt. Sobaek, Mt. Jiri, Mt. Deokyu, and Mt. Taebaek. The taxol content in the needles was higher for the population of Mt. Halla(0.0013%) than any other populations and decreased in the order of Mt. Taebaek, Mt. Sobaek, Mt. Deokyu and Mt. Jiri. The taxol contents of Taxus caespitosa(0.0035%) were lower than that of Taxus cuspidate in all of the three plant parts. The taxol content of Taxus cuspidate var. latifolia(0.0064%) was higher than that of Taxus cuspidate for shoots and needles, but lower than that of Taxus cuspidate. There was a tendency of increasing taxol contents of bark and shoots of Taxus cuspidate, as latitude decreased. This study indicated that taxol content was relatively high in the populations of Taxus cuspidate. Particularly taxol content in the bark of the Taxus cuspidate in Mt. Halla population was about the same as that of pacific yew, indicating a high potential for commercial use.

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Studies on the Organic Acids Composition in Shellfishes 2. Nonvolatile Organic Acids Composition of Oyster, Sea-mussel, Baby Clam, Hen Clam and Their Boiled-dried Products (패류의 유기산 조성에 관한 연구 2. 굴, 홍합, 바지락, 개량조개 및 그 자건품의 비휘발성유기산 조성)

  • JO Kil-Suk;PARK Yeung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.18 no.5
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    • pp.417-423
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    • 1985
  • In succession to the previous paper, the present study was directed to investigate the nonvolatile organic acids composition in raw and belied-dried products of oyster, sea-mussel, baby clam and hen clam by gas liquid chromatography. The results obtained are summarized as follows : In four kinds of the samples examined, eight kinds of organic acids were identified and determined in oyster, sea-mussel and baby clam, and nine kinds in hen clam. The major organic acids in oyster were pyroglutamic, succinic and malic acid which was $94.2\%$ of total quantity of organic acid, and those in sea-mussel, baby clam and hen clam were succinic and malic acid which were $90.8\%,\;89.7\%\;and\;86.4\%$ of total acids, respectively. The most abundant organic acid in sea-mussel, baby clam and hen clam was succinic acid that was $80.6\%,\;84.9\%\;and\;73.2\%$ in total acids, repectively. And that of oyster was pyroglutamic acid which marked $38.8\%$ in total acids, and the next one was succinc acid marked $34.4\%$. In the total quantity of organic acid, the highest was 913.0 mg/100g in oyster which showed 4.5 times as much as in hen clam, followed by 478.4 mg/100g in sea-mussel, 246.3mg/100g in baby clam, and the least was 201.2 mg/100g in hen clam. The decreasing rate of total quently of organic acids by boiled-dried procersing was the highest in oyster, $54.7\%$, followed by $46.5\%$ in sea-mussel, $37.1\%$ in hen clam and $29.4\%$ in baby clam. The decreasing rate of each organic acid shelved much difference according to the samples examined, in general, great in malic, fumaric and proglutamic acid ana less in succinic, lactic and oxalic acid.

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Effect of Relative Humidity on the Changes of Lipids in Freeze-Dried Fish during Storage (동결건조 어육지질의 변화에 미치는 상대습도의 영향)

  • LEE Hyeung-Il;PARK Yeung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.18 no.6
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    • pp.519-528
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    • 1985
  • Fillets of mackerel (Scomber japonicus) and flounder (Xystrias grigorjewi) which, are representatives in red fleshed fish and white fleshed fish, respectively, were freeze-dried and stored in tightly sealed containers which were controlled to different relative humidity at $25^{\circ}C$. The changes of lipids were examined periodically by measuring the peroxide value (POV), the thiobarbituric acid (TBA) and the acid value (AV). And the fatty acid composition of lipids was investigated by gas-liquid chromatography (GLC). The results obtained are summarized as foollows: From the changes of POV and TBA value during storage, the oxidation of lipids was distinct at the lower relative humidities, $0\%\;and\;23\%$, while inhibited at the higher relative humidities, $52\%\;and\;81\%$. The changes in acid value during storage were more prominent at the hifger relative himidites than at the lower relative humidities. The content of $C_{16:0},\;C_{18:0}\;and\;C_{22:6}$ acids in the fatty acid composition of total lipids was abundant in both fleshed fishes. The content of $C_{18:1}$ acid in the nonpolar lipid and that of $C_{16:0}$ acid in the polar lipid were higher than other fatty acids. In the fatty acid composition of total lipids during storage, polyenoic acids decreased with storage period at $0\%\;and\;23\%$ relative humidities, while the fatty acid composition didn't show a great change at $52\%\;and\;81\%$ relative humidities. In the non-polar lipid, polyenoic acids coherently decreased under all the conditions of relative humidities but the saturated acids and the monoenoic acids increased. In the polar lipid, polyenoic acids decreased at $0\%\;and\;23\%$ relative humidities, while the saturated acids and monoenoic acids decreased at $52\%\;and\;81\%$ relative humidities. On the other hand, the oxidation of lipids was more significant in mackerel than in the flounder, and the changes of fatty acid composition were shown a similar pattern.

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Characterization of Grain Amino Acid Composition and Proteome Profile of a High-lysine Barley Mutant Line M98 (고-Lysine 보리 돌연변이 계통 M98 종실의 아미노산 조성 및 Proteome Profile 특성)

  • Kim, Dea-Wook;Kim, Hong-Sik;Park, Hyoung-Ho;Hwang, Jong-Jin;Kim, Sun-Lim;Lee, Jae-Eun;Jung, Gun-Ho;Hwang, Tae-Young;Kim, Jung-Tae;Kim, Si-Ju;Rakwal, Randeep;Kwon, Young-Up
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.57 no.2
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    • pp.171-181
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    • 2012
  • Lysine is the first limiting essential amino acid in cereals for humans and monogastric animals, although its content is generally low. A chemically induced high-lysine barley mutant, M98, has an agronomically undesirable shrunken endosperm trait. In order to obtain detailed insight into the atypical traits of M98 grains, we characterized amino acid composition and protein profiles of M98 and its parent cultivar Chalssalbori. Among a total of 16 amino acids, the percentage of each of the 7 amino acids, including lysine, was 1.2~1.8 times higher in M98, comparing to Chalssalbori. The percentage of proline and its precursor, glutamic acid, in M98 was about the half of that of the amino acids in Chalssalbori, but arginine synthesized from glutamic acid was 1.8 times higher in M98, compared that in the parent cultivar. Theses results indicated that the mutation in M98 grains might alter the proportion of amino acids linked to each other in a biosynthetic pathway. A comparison of grain proteome profiles between Chalssalbori and M98 revealed 70 differentially expressed protein spots, where 45 protein spots were up-regulated and 25 protein spots down-regulated in M98 compared to those in Chalssalbori. Of these changed protein spots, 53 were identified using nano-electrospray ionization liquid chromatography mass spectrometry. Most of these identified proteins were involved in various biological processes. In particular, 28 protein spots such as ${\beta}$-amylase, serpins and B3-hordein were identified as proteins associated with the atypical traits of M98. It was thought that a genetic study on the unique protein profile of M98 would be needed to develop an agronomically feasible barley cultivar with high-lysine trait.

Determination of cyromazine residues in agricultural commodities using HPLC-UVD/MS (HPLC-UVD/MS를 이용한 농산물 중 Cyromazine의 잔류분석법)

  • Song, Lee-Seul;Kim, Young-Hak;Lee, Su-Jin;Hwang, Young-Sun;Kwon, Chan-Hyeok;Do, Jung-Ah;Oh, Jae-Ho;Im, Moo-Hyeog;Chang, Woo-Suk;Lee, Young-Deuk;Choung, Myoung-Gun
    • The Korean Journal of Pesticide Science
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    • v.16 no.3
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    • pp.202-208
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    • 2012
  • A high-performance liquid chromatographic (HPLC) method was developed to determine residues of cyromazine, a triazine insecticide, in agricultural commodities. Cyromazine was extracted with 90% aqueous methanol from representative crops which comprised brown rice, oyster mushroom, oriental melon, watermelon, and Chinese cabbage. Following to evaporation of methanol in the extract, the aqueous concentrate was acidified to form the protonated cyromazine. Dichloromethane partition was then applied to remove nonpolar co-extractives in the aqueous phase. Strong cation-exchange chromatography using Dowex 50W-X4 resin was employed for final purification of the extract. Cyromazine was successfully separated on a Zorbax SB-Aq $C_{18}$ column showing high retention for polar compounds. Cyromazine was sensitively quantitated by ultraviolet absorption at 214 nm. Limit of quantitation (LOQ) of the method was 0.04 mg/kg irrespective of sample types. Each crops were fortified at 3 different concentrations of cyromazine for recovery test. Mean recoveries from samples fortified at LOQ~2.0 mg/kg in triplicate ranged 80.2~103.3% in five agricultural commodities. Relative standard deviations in recoveries were all less than 6%. A selected-ion monitoring LC/MS method with electrospray ionization in positive-ion mode was also provided to confirm the suspected residue. The proposed method was reproducible and sensitive enough to routinely determine and inspect the residue of cyromazine in agricultural commodities.

Pharmacokinetics of Florfenicol and its Metabolite, Florfenicol Amine, in Broiler Chickens (육계에서의 플로르페니콜 및 그의 대사체인 플로르페니콜 아민의 약물동태학적 연구)

  • Park Byung-Kwon;Lim Jong-Hwan;Kim Myoung-Seok;Hwang Youn-Hwan;Yun Hyo-In
    • Journal of Veterinary Clinics
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    • v.23 no.2
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    • pp.114-118
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    • 2006
  • A study on bioavailability and pharmacokinetics of florfenicol was conducted in broilers following a single intravenous (i.v.) and oral (p.o.) doses of 20 mg/kg body weight (b.w.). Florfenicol concentrations in plasma were determined by a high-performance liquid chromatography/mass spectrometry. Plasma concentration-time data after i.v. administration were analyzed by a non-compartmental analysis. Following i.v. injection, the total body clearance was $0.74{\pm}0.25L/kg/h$ and the volume of distribution at steady-state was $1.16{\pm}0.19L/kg$. Florfenicol was rapidly distributed and eliminated following i.v. injection with $1.15{\pm}1.06h$ of elimination half-life. After oral administration, the calculated $C_{max}$ values ($8.18{\pm}0.97{\mu}g/mL$) were reached at $1.33{\pm}0.29h$ in broilers. The elimination half-life of florfenicol was $1.31{\pm}0.27h$ and the absolute bioavailability (F) was 75.46% after oral administration of florfenicol. Florfenicol amine, a major metabolite of florfenicol, was detected in all broilers after i.v. and p.o. administration of florfenicol. The observed $C_{max}$ values of florfenicol amine ($3.96{\pm}2.60\;and\;2.22{\pm}1.71{\mu}g/mL$) were reached at $0.16{\pm}0.19\;and\;1.61{\pm}1.02h$ after i.v. and p.o. administration of florfenicol, respectively. Florfenicol amine was eliminated with $1.88{\pm}0.39\;and\;2.64{\pm}1.39h$ of the elimination half-life after i.v. and p.o. administration of florfenicol, respectively.

Development of Rapid Analytical Method of Forbidden Medicines in Dietary Supplements Using LC-ESI-Tandem MS (LC-ESI-tandem MS를 이용한 기능성표방식품 중 부정유해물질 신속검사체계 개발)

  • Kim, Hee-Yun;Jang, Young-Mi;Joo, Hyun-Jin;Jung, Young-Hyun;Lee, Myoung-Sook;Park, Jong-Seok;Lee, Kwang-Ho;Lee, Hwa-Mi
    • Korean Journal of Food Science and Technology
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    • v.39 no.4
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    • pp.372-379
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    • 2007
  • A high-performance liquid chromatography-electrospray ionization (HPLC-ESI) tandem MS was developed for the rapid and simultaneous determination of forbidden medicines in dietary supplements. Thirteen medicinal components such as PDE-5 inhibitors and their analogues, and the newly identified dimethylsildenafil and xanthoanthrafil, were included in this study. After tentative standardization of molecular ions in both polarities using thirteen references on the mass spectrometer, with ESI-continuous infusion via the syringe pump method, the relative intensity of the ions present in the resulting spectra was quantitatively compared. From the results, the ion mode was selected depending on each reference's characteristics. A HPLC method coupled with the ESI mode was developed considering the matrix effect and interference depending on the type of sample. The validation test of the developed method was followed by carrying out precision, accuracy, recovery, sensitivity and linearity, etc. The method showed sufficiently high sensitivity, reproducibility, and specificity, and produced 4 times faster results when compared with the existing HPLC/UV method for the determination of forbidden compounds in dietary supplements.

Antioxidant capacity and hepatoprotective effect of ethyl acetate fraction from shoot of Aralia elata on alcohol-induced cytotoxicity (두릅 아세트산 에틸 분획물의 산화방지 효과 및 알코올에 대한 간세포 보호효과)

  • Kwon, Bong Seok;Park, Seon Kyeong;Kim, Jong Min;Kang, Jin Yong;Park, Sang Hyun;Kang, Jeong Eun;Lee, Chang Jun;Park, Su Bin;Yoo, Seul Ki;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.50 no.2
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    • pp.216-224
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    • 2018
  • To evaluate physiological effect of Aralia elata, in vitro antioxidant activity and hepatic protective effects were investigated. Ethyl acetate fraction from Aralia elata (EFAE) had higher total phenolic content than other fractions (n-hexane, chloroform, and distilled water layers). EFAE also showed significantly greater radical scavenging activity against 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH), than other fractions. Moreover, EFAE showed dose-dependent inhibitory effect of malondialdehyde (MDA). Hepatoprotective effects of EFAE against ethanol- and $H_2O_2$-induced oxidative stress and cytotoxicity in H4IIE and HepG2 hepatic cells were examined using 2',7'-dichlorofluorescein diacetate (DCF-DA) and 3-[4,5-dimethythiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. The results showed that EFAE reduced cellular oxidative stress, and increased hepatic cell viability. In addition, EFAE inhibited ethanol-induced lipid accumulation in HepG2 cells. Finally, physiological substances of EFAE were analyzed using high performance liquid chromatography (HPLC), and the major bioactive compounds identified were 3,5-dicaffeoylquinic acid and chlorogenic acid.