Differences of lipids, especially total lipid composition, fatty acid and sterol composition of the flesh lipids between three species of cephalopods were investigated, since available researches concerning lipids in flesh tissues of the cephalopod are very limited. Extracted total lipid from the flesh tissues were fractionated by silicic acid column chromatography into three lipid classes of neutral lipids, glycolipids and phospholipids. The lipid compositions of total lipid and neutral lipids were estimated by the method of thin layer chromatography and TLC-scanner. The sterol compositions of unsaponifiable matters from total lipid were determined by using thin layer chromatography and gas-liquid chromatography. The fatty acid composition of each lipid class was also determined by gas-liquid chromatography. Total lipid contents of flesh tissues from three species of the cephalopods were 0.5 in Octopus vulgare, 0.8 in Octopus variabilis and $0.6\%$ in Loligo beka based on wet weight, the contents of total fatty acid in total lipid were 19.3, 47.8 and $38.4\%$, and the contents of unsaponifiable matters were 10.9, 18.8 and $41.1\%$, respectively. Total lipid was mainly composed of sterols and polar lipid-pigments as major components in each sample and the proportion of sterols and polar lipid-pigments to total lipid ranged from 27.0 to $35.5\%$ and 38.3 to $63.4\%$, respectively. The other lipid components of total lipid, e.g. triglycerides, free fatty acids, and carbohydrate-esterified sterols were determined as a minor components. The major component fatty acid in total lipid was palmitic acid and additionaly it chiefly consisted of the other unsaturated acids such as oleic, linoleic, octadecatetraenoic and eicosapentaenoic acid as major components of the acid. The compositions of sterol in three species of cephalopod were found to contain mainly cholesterol for its proportion to total sterols was 82.4 to $89.1\%$. However the other sterols such as 22-dehydrocholesterol and 24-methylenecholesterol were determined in addition to cholesterol as a minor components. The result of fractional composition of lipid class in total lipid was that total lipid had large .amount of polar lipid and small amount of nonpolar lipid i, e, neutral lipid in each sample, and the contents of phospholipid were higher than that of glycolipid in polar lipid. Neutral lipid was mainly composed of free sterol as major components in each sample and its proportion of free sterols to total neutral lipid was 50.0 to $70.5\%$. The other lipid components of neutral lipid showing similar in quantity, esterified sterols, free fatty acids and triglycerides were determined as a minor components. The major components fatty acid in neutral lipid were palmitic, oleic and hexadecadienoic acid. Palmitic acid was the most abundant and additionaly oleic, linoleic, octadecatetraenoic and myristic acid were the major component fatty acid in glycolipid. But, especially, glycolipid of Loligo beka contained a higher amount of arachidonic acid which also consists of major component in addition to those of acids. Palmitic acid was the most abundant and additionaly, oleic, linoleic and octadecatetraenoic acid were the major component fatty acids in phospholipid.
Differences in lipid composition including fatty acid, lipid class, sterol and especially carotenoid between fleshes of wild and cultured prawn, Penaeus japonicus, were studied. Total lipids were extracted from the flesh during the spawning period and fractionated into two lipid classes of polar and nonpolar lipids by silicic acid column chromatography. The fatty acid composition of each lipid classes, total lipid (TL), nonpolar lipid (NL) and polar lipid (PL) were analyzed by gas liquid chromatography. The sterol and carotenoid composition of total lipids were determined by using thin layer chromatography, gas liquid chromatography and column chromatography using MgO-celite 545 and silicic acid-celite 545 as an absorbent, and by UV spectrophotometry. Total lipid contents of both fleshes from the wild and cultured prawn were about $2.0\%$ on average, but the content of the unsaponifiable matters in the cultured prawn (about $16.2\%$ in total lipid) showed a little higher than that of the wild prawn (about $13.9\%$ in total lipid) and the ratio of NL to PL in total lipid was 1:1.7. In the fatty acid composition of TL, the contents of $Cl_{16:0}\;and\;C_{20:3}$ fatty acids were higher in wild prawn than in cultured prawn, while the contents of $Cl_{18:1}\;and\;C_{20:5}$ fatty acids in cultured prawn were higher than those in wild prawn. The cultured prawn contained higher amounts of monoenoic acids and lower amounts of polyenoic acids than the wild prawn. In the fatty acid composition of NL, the wild prawn showed higher levels in $Cl_{18:0}\;and\;C_{20:1}$ fatty acid contents than the cultured prawn, while the cultured prawn contained much amout of $Cl_{16:0}\;and\;C_{18:1}$ fatty acids. On the other hand, the fatty acid composition of PL showed that $Cl_{16:1}\;and\;C_{17:1}$ fatty acid were higher in the wild prawn than in the cultured prawn, but in $Cl_{16:0}\;and\;C_{18:1}$ fatty acids, the levels were reversed. Consequently, the cultured prawn contained higher amount of monoenoic acids, and similar amounts of saturated acids and polyenoic acids to the wild prawn in NL. And the cultured prawn contained lower amount of monoenoic acids, and similar amounts of saturated acids and polyenoic acids to the wild prawn in PL. In sterol composition of both the wild and cultured prawn, the predominant sterol was cholesterol with the proportion of $78.7{\sim}88.9\%$ to the total sterol. In addition to the cholesterol, the other minor sterols such as 24-methylene cholesterol and sitosterol were detected. Total carotenoid content in flesh of the wild prawn was relatively higher than that of the cultured prawn marking 70 mg/100g of lipid in wild prawn and 40 mg/100 g of lipid in cultured prawn, respectively. The main carotenoids of the both prawns were astaxanthin($54.1{\sim}60.8%$), phoenicoxanthin ($16.5{sim}22.9%$),${\bata}-carotene\;(20.0{\sim}22.0%)$.
The chemical structure of glycolipid of Selenomonas ruminantium cell wall was to be elucidated. The bacterial cells were treated in hot TCA and the glycolipid fractions were extracted by the solvent $CHCl_3\;:\;CH_3OH$ (1 : 3). The extracted glycolipids fraction was further separated by acetone extraction. The acetone soluble fraction was named as the spot A-compound. The acetone insoluble but ether soluble fraction was named as the spot B-compound. These two compounds were examined for elucidation of their chemical structure. The results were as follows: 1. The IR spectral analysis showed that O-acyl and N-acyl fatty acids were linked to glucosamine moiety in the spot A-compound. However in the spot B-compound in addition to O and N-acyl acids phosphorus was shown to be attached to glucosamine. 2. It was recognized by gas liquid chromatography that spot A compound contained beta-OH $C_{13:0}$ fatty acid in predominance in addition to the fatty acid with beta-OH $C_{9:0}$, whereas the spot B compound was composed of the predominant fatty acid of beta-OH $C_{13:0}$ with small amount of beta-OH $C_{9:0}$. 3. According to the paper chromatographic analysis of hydrazinolysis products of the spot A compound, a compound of a similar Rf value as the chitobiose was recognized, which indicated a structure of two molecules glucosamine condensed. The low Rf value of the hydrazinolysis product of the spot B-compound confirmed the presence of phosphorus attached to glucosamine. 4. The appearance of arabinose resulting from. ninhydrin decomposition of the acid hydrolyzate of the spot A compound indicated that the amino group is attached to $C_2$ of glucosamine. 5. The amount of glucosamine in the N-acetylated spot A compound decreased in half of the original content by the treatment. with $NaBH_4$, indicating that there are two molecules of glucosamines in the spot A compound. The presence of 1, 6-linkage between two molecules of glucosamine was suggested by the Morgan-Elson reaction and confirmed by the periodate decomposition test. 6. By the action of ${\beta}-N-acetyl$ glucosaminidase the N-acetylated spot A compound was completely decomposed into N-acetyl glucosamine, whereas the spot B compound was not. This indicated the spot A compound has a beta-linkage. 7. When phosphodiesterase or phosphomonoesterase acted on $^{32}P-labeled$ spot B compound, $^{32}P$ was not released by phosphodiesterase, but completely released by phosphomonoesterase. This indicated that one phosphorus is linked to glucosamine moiety. 8. The spot A compound is assumed to have the following chemical structure: That is glucosaminyl, ${\beta}-1$, 6-glucosamine to which O-acyl and N-acyl fatty acids are linked, of which the predominant fatty acid is beta-OH $C_{13:0}$ fatty acid in addition to beta-OH $C_{9:0}$ fatty acid 9. The spot B compound is likely to have the linkage of $glucosaminyl-{\beta}-1$, 6-glucosamine to which phosphorus is linked in monoester linkage. Furthermore both O-acyl and N-acyl fatty acids contained beta-OH $C_{13:0}$ fatty acid predominantly in addition to beta-OH $C_{9:0}$ fatty acid.
Ahmed, Hanaa H;Abd-Rabou, Ahmed A;Hassan, Amal Z;Kotob, Soheir E
Asian Pacific Journal of Cancer Prevention
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v.16
no.16
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pp.7179-7188
/
2015
Cancer is a major health obstacle around the world, with hepatocellular carcinoma (HCC) and colorectal cancer (CRC) as major causes of morbidity and mortality. Nowadays, there isgrowing interest in the therapeutic use of natural products for HCC and CRC, owing to the anticancer activity of their bioactive constituents. Boswellia serrata oleo gum resin has long been used in Ayurvedic and traditional Chinese medicine to alleviate a variety of health problems such as inflammatory and arthritic diseases. The current study aimed to identify and explore the in vitro anticancer effect of B. Serrata bioactive constituents on HepG2 and HCT 116 cell lines. Phytochemical analysis of volatile oils of B. Serrata oleo gum resin was carried out using gas chromatography-mass spectrometry (GC/MS). Oleo-gum-resin of B. Serrata was then successively extracted with petroleum ether (extract 1) and methanol (extract 2). Gas-liquid chromatography (GLC) analysis of the lipoidal matter was also performed. In addition, a methanol extract of B. Serrata oleo gum resin was phytochemically studied using column chromatography (CC) and thin layer chromatography (TLC) to obtain four fractions (I, II, III and IV). Sephadex columns were used to isolate ${\beta}$-boswellic acid and identification of the pure compound was done using UV, mass spectra, $^1H$ NMR and $^{13}C$ NMR analysis. Total extracts, fractions and volatile oils of B. Serrata oleo-gum resin were subsequently applied to HCC cells (HepG2 cell line) and CRC cells (HCT 116 cell line) to assess their cytotoxic effects. GLC analysis of the lipoidal matter resulted in identification of tricosane (75.32%) as a major compound with the presence of cholesterol, stigmasterol and ${\beta}$-sitosterol. Twenty two fatty acids were identified of which saturated fatty acids represented 25.6% and unsaturated fatty acids 74.4% of the total saponifiable fraction. GC/MS analysis of three chromatographic fractions (I,II and III) of B. Serrata oleo gum resin revealed the presence of pent-2-ene-1,4-dione, 2-methyl- levulinic acid methyl ester, 3,5- dimethyl- 1-hexane, methyl-1-methylpentadecanoate, 1,1- dimethoxy cyclohexane, 1-methoxy-4-(1-propenyl)benzene and 17a-hydroxy-17a-cyano, preg-4-en-3-one. GC/MS analysis of volatile oils of B. Serrata oleo gum resin revealed the presence of sabinene (19.11%), terpinen-4-ol (14.64%) and terpinyl acetate (13.01%) as major constituents. The anti-cancer effect of two extracts (1 and 2) and four fractions (I, II, III and IV) as well as volatile oils of B. Serrata oleo gum resin on HepG2 and HCT 116 cell lines was investigated using SRB assay. Regarding HepG2 cell line, extracts 1 and 2 elicited the most pronounced cytotoxic activity with $IC_{50}$ values equal 1.58 and $5.82{\mu}g/mL$ at 48 h, respectively which were comparable to doxorubicin with an $IC_{50}$ equal $4.68{\mu}g/mL$ at 48 h. With respect to HCT 116 cells, extracts 1 and 2 exhibited the most obvious cytotoxic effect; with $IC_{50}$ values equal 0.12 and $6.59{\mu}g/mL$ at 48 h, respectively which were comparable to 5-fluorouracil with an $IC_{50}$ equal $3.43{\mu}g/mL$ at 48 h. In conclusion, total extracts, fractions and volatile oils of B. Serrata oleo gum resin proved their usefulness as cytotoxic mediators against HepG2 and HCT 116 cell lines with different potentiality (extracts > fractions > volatile oil). In the two studied cell lines the cytotoxic acivity of each of extract 1 and 2 was comparable to doxorubicin and 5-fluorouracil, respectively. Extensive in vivo research is warranted to explore the precise molecular mechanisms of these bioactive natural products in cytotoxicity against HCC and CRC cells.
Park, Shin-Min;Do, Jung-Ah;Lim, Seung-Hee;Yoon, Ji-Hye;Pak, Won-Min;Shin, Hye-Sun;Kuk, Ju-Hee;Chung, Hyung-Wook
Journal of Food Hygiene and Safety
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v.33
no.4
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pp.296-305
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2018
Spiroxamine, one of fungicides, is used to control powdery mildew in various crops and black yellow sigatoka in bananas. The major strength of spiroxamine is to control powdery mildew in various crops and bananas yellow sigatoka in bananas. The compound has shown a high level of activity, good persistence and crop tolerance. Besides powdery mildew, good control of rust, net blotch and Rhynchosporium diseases been indicated in cereals, together with a complementary activity against Septoria diseases. In 2017, the maximum residue limit (MRL) of spiroxamine established in Korea. According to Ministry of ood and rug afety) regulations, spiroxamine residues defined only parent compound. Thus, a analytical method is needed to estimate the residue level of the parent compound. The objective of this study was to develop and validate analytical method for spiroxamine in representative agricultural commodities. Samples were extracted with acetonitrile and partitioned with dichloromethane to remove the interfering substances. The analyte were quantified and confirmed liquid chromatograph-tandem mass spectrometer (LC-MS/MS) in positive-ion mode using multiple reaction monitoring (MRM). Matrix matched calibration curves were linear over the calibration ranges ($0.0005{\sim}0.1{\mu}g/mL$) for the analyte in blank extract with coefficient of determination ($r^2$) > 0.99. For validation purposes, recovery studies will be carried out at three different concentration levels (LOQ, 10LOQ, and 50LOQ) performing five replicates at each level. The recoveries 70.6~104.6% with relative standard deviations (RSDs) less than 10%. All values were consistent with the criteria ranges in the Codex guidelines (CAC/GL40, 2003) and MFDS guidelines. proposed analytical method be used as an official analytical method in the Republic of Korea.
The purpose of this in vitro study was to compare the effects of root canal cleanness following two Ni-Ti rotary instruments with different rake angle. Thirty-six sound, extracted human premolars with single root were randomly divided into three groups. The used rotary instruments were HEROShaper (Group 1, Micro-Mega, Besancon, France, n=12) and ProFile (Group 2, Maillefer, Ballaigues, Switzerland, n=12). Control group (n=12) was only extirpated with barbed broach (Mani, Matsutani Seisakusho Co., Japan) Group 1 & 2 teeth were prepared to a #40/.04 taper at the apex followed by 1 mm using crown-down technique. After canal preparation and frequent irrigation with 5.25% sodium hypochlorite, the roots split longitudinally into a bucco-lingual direction. Root halves were cross-sectioned in apical third portion again. All root specimens were processed for SEM investigation and photographed. Separate evaluations by one endodontist were undertaken for smear layer on prepared walls with a five score-index for each using reference photograph in root halves. The penetration depth of smear layer into dentinal tubules was also estimated in the other halves. Following results were obtained: 1. Smear layer was observed on all the prepared walls with two experimental groups except control group. 2. Smear layer characteristics in two experimental groups; 1) HEROShaper group showed snowy, dusty appearance and were shown open dentinal tubuli on the prepared walls of almost specimens, and the thickness of smear layer covering onto dentinal surfaces was within 1-2 ${\mu}m$ in a few specimens. 2) ProFile group showed shiny, burnished appearance and complete root canal wall covered by a homogenous smear layer with no open dentinal tubuli in all specimens. The penetration of smear layer into dentinal tubules was found in all specimens and the thickness was at 2-4 ${\mu}m$ in all specimens. These results demonstrated that a completely clean root canal could not be achieved regardless of positive or negative rake angle, which is in accordance with the majority of previous studies on root canal cleanliness In conclusion, through irrigation with antibacterial solutions or chelating agents is recommended to remove the smear layer on prepared canal wall in spite of Ni-Ti instrumentation.
The water-soluble materials extracted from fruit bodies and mycelium of H. erinaceum were prepared. In-vitro anticancer activities on cancer cells and In-vivo proliferation effect on mouse peritoneal exudate cell and spleen cell of samples were investigated. Also, nitric oxide (NO) generation of peritoneal exudate cell, IL-2 production capacity of spleen cells and phagocytic activity of peritoneal macrophages were examined. The water extracts of H. erinaceum suppressed the proliferation of cancer cell (HeLa, Raw264.7, Jurkat, KATO3, EL4, LyD9) with concentration-dependent. The water extract from fruit body showed better suppression effect than that from mycelium in most of cancer cells used. The anticancer effect of water extract of fruits body in the range of 0.01 and 10 mg/ml for Raw 264.7 and EL4 cell lines were the same as the Taxol with one thousandth equivalent of fruit body concentration. Water extracts of fruit body and liquid-cultured products of H. erinaceum induced nitric oxide (NO) generation of peritoneal exudate cell and increased NO generation by stimulus of lipopolysaccharide. Water extracts alone did not induce the proliferation and IL-2 production capacity of spleen cells. However, spleen's proliferation and IL-2 production were induced significantly by the addition of lipopolysaccharide and Con A (concanavalin A) or Con A alone, and the effectiveness of mycelium extract with water were more active than those from fruit body.
Shin, Jung-Ah;Chun, Ji Yeon;Lee, Junsoo;Shin, Ki Yong;Lee, Soon Kyu;Lee, Ki-Teak
Journal of the Korean Society of Food Science and Nutrition
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v.42
no.12
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pp.1949-1957
/
2013
The contents of ${\beta}$-carotene and retinol in processed and restaurant foods, such as Korean noodles, mandus, rice cakes and bread products, were quantified by high-performance liquid chromatography (HPLC) with UV/visible and fluorescence detector, respectively. Samples were collected from different local areas (i.e. Gangwon-do, Gyeonggi-do, Gyeongsang-do, Seoul, Jeolla-do, and Chungcheong-do). After homogenization, samples were hydrolyzed by direct alkali saponification; thereafter, fat-soluble components were extracted by a mixture of n-hexane/ethylacetate (85:15, v/v), containing 0.01% butylated hydroxytoluene (BHT). ${\beta}$-carotene and retinol contents in infant formula used as an in-house material for the analytical quality control. Among 14 Korean noodles, high contents of ${\beta}$-carotene were found in Bibim-Guksu (average 442.43 ${\mu}g/100g$) and Jjolmyeon (average 301.39 ${\mu}g/100g$). In 4 Korean mandus, the highest contents of ${\beta}$-carotene was determined in Kimchi-mandu (average 197.64 ${\mu}g/100g$), resulting in 33.3 RE of the converted vitamin A. Among 12 Korean rice cakes, Maeun-Tteokbokki and Modm-Chaltteok contained relatively high content of ${\beta}$-carotene with 205.11 and 41.33 ${\mu}g/100g$, respectively, while retinol was detected only in Maeun- Tteokbokki (1.65~10.45 ${\mu}g/100g$). In addition, among 8 bread products, 77.3 RE of pastry, 51.2 RE of buttercream- bread, and 41.4 RE of morning roll were found as the contents of the converted vitamin A.
Kim, Hee-Yun;Park, Se-Jong;Chung, So-Young;Choi, Seon-Hee;Oh, Sun-Woo;Lee, Jin-Sook;Choi, In-Sun;Shin, Min-Su;Song, Jae-Sang
Korean Journal of Food Science and Technology
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v.43
no.1
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pp.58-64
/
2011
The purpose of this study was to analyze the content and consumption of total sugar from school lunches at Incheon and Chuncheon. The samples were collected from eight elementary schools and eight middle schools in Incheon and Chuncheon for 15 days. The analysis of total sugar content was performed for 1334 main dishes, side dishes, and desserts, which were supplied by elementary and middle school foodservices. Total sugar content was extracted from various types of food with 50% ethanol after defatting. We simultaneously analyzed sugars such as fructose, glucose, sucrose, maltose, and lactose by a high performance liquid chromatography with refractive index detector. The average lunch intake for elementary school and middle school students was $372.6{\pm}72.2g$ and $449.2{\pm}81.1g$, respectively. The total sugar content in desserts was $7.21{\pm}6.32g/100g$, $1.69{\pm}2.44g/100g$ in side dishes, and $0.32{\pm}0.77g/100g$ in main dishes. Among side dishes, fried foods, stir-fried foods, and sauces contained the highest amounts of total sugar. The total sugar intake was $4.63{\pm}5.11g$ in desserts, $0.64{\pm}1.01g$ in side dishes, and $0.55{\pm}1.48g$ in main dishes. Main and side dishes with a high total sugar intake included cooked rice with seasoning, fried foods, stir-fried foods, and preserved foods. The average total sugar intake per meal for elementary school, middle school, and all students was $4.03{\pm}3.67g$, $6.97{\pm}6.59g$, and $5.50{\pm}5.53g$, respectively. We have provided useful information to decrease the intake of total sugar in school lunches. It is recommended that total sugar intake be continuously monitored.
Kim, Ji Yoon;Woo, Min Ji;Hur, Kyung Jin;Manoharan, Saravanan;Kwon, Chan-Hyeok;Hur, Jang Hyun
The Korean Journal of Pesticide Science
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v.19
no.2
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pp.81-87
/
2015
The present study was aimed to predict the pre-harvest residue limits (PHRLs) of pyrimethanil (fungicide) and methoxyfenozide (insecticide) in grape, and to estimate their biological half-lives and residual characteristics. The pesticides were sprayed once on grape in two different fields 10 days before harvest. At the end of 0, 1, 2, 3, 5, 7 and 10 days after application, samples were harvested for further analysis. The residual pesticides were extracted with acetonitrile and partitioned with dichloromethane, and the high-performance liquid chromatography with diode array detector (HPLC/DAD) was employed for the residue analysis. The results obtained in the present study show that the limit of detection of both pesticides were found to be $0.01mg\;kg^{-1}$. The recoveries of these pesticides were ranged between 80.6% and 102.5% with coefficient of variation lower than 10%. The biological half-lives of both pesticides were observed in field 1 and field 2 which shows 7.7 and 7.4 days for pyrimethanil and 5.1 and 6.1 days for methoxyfenozide, respectively. Further, the PHRL of pyrimethanil and methoxyfenozide was found to be $8.90mg\;kg^{-1}$ and $5.51mg\;kg^{-1}$, respectively at 10 days before harvest. Consequently, the present study suggests that the residual amounts of both pesticides will be lower than the maximum residue limits (MRLs) when grape is harvested.
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