• Title/Summary/Keyword: HPLC Chromatography

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A Lateral Flow Immunoassay Kit for Detecting Residues of Four Groups of Antibiotics in Farmed Fish (어류 중 4계열 잔류 항생물질 검출을 위한 Lateral Flow Immunoassay Kit 개발)

  • Jo, Mi Ra;Son, Kwang Tae;Kwon, Ji Young;Mok, Jong Soo;Park, Hong Jae;Kim, Hyun Yong;Kim, Gyung Dong;Kim, Ji Hoe;Lee, Tae Seek
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.2
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    • pp.158-167
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    • 2015
  • A lateral flow immunoassay kit based on antigen-antibody interactions was developed to detect residues of beta-lactams, quinolones, tetracyclines, and sulfonamides in farmed fish. Group-specific antibodies showing cross-reactivity with other antibiotics in the same group were produced in rabbits. The rabbits were immunized eight times to obtain the maximum titers. Antibodies were extracted from the antisera collected from the immunized rabbits and produced group-specific reactions with antibiotics from the four groups. A kit was prepared that optimize conditions for the antigen-antibody reaction, using colloidal gold conjugated antibodies, and was designed to detect the four groups of antibiotics simultaneously. The kit enabled the detection of antibiotics in the four groups at below maximum residue limits (MRLs), which were $200{\mu}g/kg$ for tetracyclines, $100{\mu}g/kg$ for sulfonamides, $50{\mu}g/kg$ for beta-lactams, and $100{\mu}g/kg$ for quinolones. The cross-reactivity of the antibodies ranged from 10-80% for the sulfonamides, 20-100% for tetracyclines, 38-100% for quinolones, and 20-100% for the beta-lactams, confirming that the antibodies were group specific. The test kit was used 30 times to examine spiked antibiotics at the limits of detection (LODs) and all produced positive results, indicating high sensitivity. The LODs for the assay ranged from 4-20 ng/mL for beta-lactams, 25-50 ng/mL for sulfonamides, 20-100 ng/mL for tetracyclines, and 30-80 ng/mL for quinolones, and there were no false negative reactions at above these LODs. In addition, all of the LODs of the developed kit were correlated with high-performance liquid chromatography (HPLC) data. Our lateral flow immunoassay kit can simultaneously detect antibiotic residues from a large number of fish samples rapidly, strengthening the safety of domestic farmed and imported fish.

Effects of Aspergillus oryzae Strains on Quality of KOCHUJANG (국균(麴菌)이 고추장 품질(品質)에 미치는 영향(影響))

  • Park, Chang Hee;Park, Yoon Joong
    • Korean Journal of Agricultural Science
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    • v.11 no.1
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    • pp.8-24
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    • 1984
  • In this study, investigated the effects of four strains belonging to Aspergillus oryzae on the quality of Kochujang. In the Koji and Kochujang making, investigated the difference of enzyme production of each strain, the change of each component and color during the aging, and tested the sensory taste. The results obtained were as follows; 1. Protease activity (acid, neutral) in wheat flour Koji was high in the case of Aspergillus $oryzae-S_1$ and Aspergillus oryzae - M of short stalked type. The strain Aspergillus $oryzae-S_1$ showed maximum activity after two days of Koji making, while the strain Aspergillus oryzae - M showed low activity till two days, but showed maximum activity after three days-four days of Koji making. 2. In $\alpha$-amylase activity, strain Aspergillus $oryzae-S_1$, Aspergillus $oryzae-S_2$ and Aspergillus oryzae - M showed high activity after two days of Koji making. Aspergillus oryzae-NB strain showed slower ${\alpha}$ - amylase activity than that strains. 3. In glucoamylase activity, all strain tested showed high activity after three days of Koji making, but st rain Aspergillus oryzae - NB showed slower activity than ot - hers. 4. In protease activity (acid, neutral) during the aging of Kochujang, strain Aspergillus $oryzae-S_1$ and Aspergillus oryzae - M of short stalked type showed higher activity than that of long stalked type. 5. Amino type nitrogen contents during the aging of Kochujang was very higher in the case of strains Aspergillus $oryzae-S_1$ and Aspergillus oryzae - M of short stalked type than other strains, and each contents was 315mg% and 337mg% after aged for ninty days. 6. The results that analysed free sugar of Kochujang aged for ninty days with HPLC were; glucose 5.84~7.13%, fructose 4.13~5.00%, rhamnose 0.91~1.04%, maltose 0.72~0.92% and presence of xylose was recognized. 7. The results that analysed alcohols of Kochujang aged for ninty days with gas chromatography were; ethanol 1.51~1.78%, n-propyl alcohol 1.13~1.20mg%, iso-amyl alcohol 3.5~4.4mg%. 8. In the sensory test of Kochujang aged for sixty days and for ninty days, the case of strains Aspergillus oryzae-M and Aspergillus $oryzae-S_1$ of short stalked type showed good taste, while the case of strains Aspergillus $oryzae-S_2$ and Aspergillus oryzae-NB of long stalked type showed good flavor and color.

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Content Analysis and Classification for Polygonati Odorati Rhizoma and Polygonati Rhizoma by Steroidal Saponin (Steroidal Saponin을 이용한 위유, 황정의 분류 및 함량 분석법 개발)

  • Kim, Sun-Gun;Shin, So-Young;Moon, Ye-Ji;Seo, Ji-Yoon;Kim, Ho-Kyoung;Whang, Wan-Kyunn
    • YAKHAK HOEJI
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    • v.54 no.6
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    • pp.441-448
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    • 2010
  • In present study, classification and quality control of Genus Polygonatum were developed using the isolated from Polygonati Odorati Rhizoma and Polygonati Rhizoma. 3 components were isolated from Butanol fractions of Polygonati Rhizoma, and 2 components were isolated from Hexane and Butanol fractions of Polygonati Odorati Rhizoma. All the components were obtained using silica gel and ODS column chromatography. The compounds were identified as adenosine, 14-hydroxylfurost-5-ene-3-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}2$)-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}4$)-O-${\beta}$-D-galactopyranosyl-26-O-${\beta}$-D-glucopyranoside, 22-O-methyl-14-hydrocxyfurost-5-ene-3-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}2$)-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}4$)-O-${\beta}$-Dgalactopyranosyl-26-O-${\beta}$-D-glucopyranoside, ${\beta}$-Sitosteryl-3-O-${\beta}$-D-D-glucopyranoside, 14-hydoxylfurost-5-ene-3-O-${\beta}$-Dglucopyranosyl-($1{\rightarrow}2$)-O-[${\beta}$-D-xylopyranosyl-($1{\rightarrow}3$)]-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}4$)-O-${\beta}$-D-galactopyranoside through physicochemical data, spectroscopic methods ($^1H$-NMR, $^{13}C$-NMR, Mass) according references. The quality control of genus Polygonatum were conducted using HPLC quantitative analysis of 14-hydroxylfurost-5-ene-3-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}2$)-O-${\beta}$-D-glucopyranosyl-($1{\beta}4$)-O-${\beta}$-D-galactopyranosyl-26-O-${\beta}$-D-glucopyranoside, 14-hydoxylfurost-5-ene-3-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}2$)-O-[${\beta}$-D-xylopyranosyl-($1{\rightarrow}3$)]-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}4$)-O-${\beta}$-D-galactopyranoside in 30 samples collected throughout Korea and China. This method provided a tool for standardization of mix or misusing the commercial Odorati Rhizoma and Polygonati Rhizoma. As a result, contained quantity of 14-hydroxylfurost-5-ene-3-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}2$)-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}4$)-O-${\beta}$-D-galactopyranosyl-26-O-${\beta}$-D-glucopyranoside was measured $0.008{\pm}0.006%$ and 14-hydoxylfurost-5-ene-3-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}2$)-O-[${\beta}$-D-xylopyranosyl-(13)]-O-${\beta}$-D-glucopyranosyl-($1{\rightarrow}4$)-O-${\beta}$-Dgalactopyranoside was measured $0.026{\pm}0.012%$.

Free radical scavenging activity and characterization of the extrcts from Alpinia katsumadai and Areca catechu (초두구와 빈량 추출물의 자유라디칼 소거 활성 효과와 특성에 관한 연구)

  • 강순옥;이건국;최정도
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.27 no.1
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    • pp.133-150
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    • 2001
  • 자유라디칼은 반응성이 커서 세포의 구성 물질인 단백질, 지질, 당, DNA등과 반응하여 세포 및 조직을 손상시킴으로서 노화를 가져온다. 이러한 자유라디칼들은 자외선, 일반 대사, 스트레스, 질병, 흡연 등에 의해 생성되고 특히, 활성 산소 종인 hydroxyl radical, superoxide radical, lipid peroxide radical등은 피부 노화를 발생시키는 원인이 된다. 따라서 본 연구에서는 한방 식물로부터 유해한 활성 산소종을 소거하는 활성을 가진 물질을 기 위해 1차 스크리닝 하였고, 이중 육두구, 관중, 초두구, 빈랑, 금은화, 포공영, 우롱차, 황금, 녹차 추출물이 효과가 높게 나타났으며, 그중 초두구와 빈랑 90% 메탄올 추출물에서 가장 우수한 소거 활성을 나타냄으로써 이 추출물에 대하여 자유라디칼 소거 활성에 대하여 여러 가지 생물학적 활성을 조사하였다. 초두구와 빈랑추출물에 대한 hydroxyl radical, superoxide radical, lipid peroxide radical을 소거하는 활성을 핵산, 클로로포름, 에틸아세테이트, 물층으로 분획하여 조사한 결과 에틸아세테이트층에서 가장 우수한 활성을 보였다. 에틸아세테이트 분획에 대한 lipid peroxide radical 소거에 있어서 빈랑 추출물이 $IC_{50}$/ 값 50 $\mu\textrm{g}$/ml, 초두구 추출물은 $IC_{50}$/ 값 59 $\mu\textrm{g}$/ml로서 기준물질로 사용되는 vitamic C (120 $\mu\textrm{g}$/ml)나 butylated hydroxyl toluene (80 $\mu\textrm{g}$/ml) 보다 더 우수한 소거 효과를 보여주었고, hydroxyl radical을 소거하는 능력은 25 $\mu\textrm{g}$/ml와 150 $\mu\textrm{g}$/ml로 hydroxyl radical의 소거 능이 좋은 vitamin C (180 $\mu\textrm{g}$/ml) 보다 뛰어난 소거 활성을 나타내었다. Superoxide radical을 소거하는 효과는 초두구 추출물의 $IC_{50}$/ 값 10 $\mu\textrm{g}$/ml, 빈랑 추출물이 15 $\mu\textrm{g}$/ml을 나타냈고, 이는 기준 물질인 vitamin C (35 $\mu\textrm{g}$/ml)보다 좋은 소거 활성을 보여주었으며, gallic acid 9 $\mu\textrm{g}$/ml과 유사한 효과를 나타내었다. 사람 섬유아세포를 배양하여 hydroxyl radical과 superoxide radical를 발생시킨 후 초두구와 빈랑 추출물의 세포 보호 효과를 실험한 결과 25 $\mu\textrm{g}$/ml의 농도로 처리하였을 때 각각 85% 이상의 우수한 세포 보호 효과를 나타내었다. 초두구로부터는 자유라디칼 소거 활성이 있는 물질을 분리하기 위하여 분획한 후 가장 높은 소거 활성을 보인 에틸아세테이트 층에 대하여 silica column chromatography, preparative TLC를 수행하였다. 초두구로부터 분리된 물질은 HPLC를 이용한 분리에서 phenol성 물질인 gallic acid와 동일한 retention time을 보여줌으로써 초두구로부터 분리된 물질은 gallic acid와 유사한 phenol성 물질이거나 그의 유도체일 것으로 추측된다. 따라서, 초두구와 빈랑 추출물은 피부 노화의 주요인이 되고 있는 lipid radical, hydroxyl radical, superoxide radical을 소거하는 활성이 뛰어나 자유라디칼에 의하여 발생되는 피부노화를 방지할수 있는 물질로서의 효과가 기대된다.가 기대된다.

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Residues of Thiamphenicol in Blood of Cultured Olive Flounder and Black Rockfish Treated by Single Oral Administration (Thiamphenicol의 단일 경구투여에 따른 넙치와 조피볼락의 혈액내 잔류 분석)

  • JUNG, Sung-Hee;SEO, Jung-Soo
    • Journal of Fisheries and Marine Sciences Education
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    • v.28 no.3
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    • pp.692-700
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    • 2016
  • This study was aimed at investigating the residual patterns of thiamphenicol (TP) in the cultured olive flounder (Paralichthys olivaceus) and black rockfish (Sebastes schlegeli) following oral administration. TP concentration were detected by high performance liquid chromatography with UV detector. The recovery rates of TP in serums ranged 77.05~97.23% (olive flounder) and 89.96~97.11% (black rockfish) for the concentration of 0.1, 1.0, $10{\mu}g/mL$, respectively. After single administration of TP (100 mg/kg body weight) by oral route in olive flounder ($700{\pm}50g$, $23{\pm}1.5^{\circ}C$) and black rockfish ($500{\pm}30g$, $23{\pm}1.5^{\circ}C$), the concentration in the serum was determined at 1, 5, 10, 15, 24, 30, 50, 168, 264 and 432 h post-dose. Two-compartment model was applied to analyze in the pharmacokinetics of TP administered to the fishes. In the serum of olive flounder, TP was detected on 10 and 15 hours after treatment were $10.08{\mu}g/mL$ and $10.06{\mu}g/mL$ as maximum level, respectively. In the serum of black rockfish, TP was detected on 15 hours after treatment were $8.88{\mu}g/mL$ as maximum level. Concentrations of TP to the fishes were not measurable at 432 hours (18 days) after treatment in all samples. Similar residual patterns of TP were revealed between the fishes. These results are helpful for estimating withdrawal time of TP which has been already in use for farmed fish treatment.

Analysis and Risk Assessment of Aflatoxin M1 in Infant Formula (분유 중 아플라톡신 M1 분석 및 위해평가)

  • Kang, YoungWoon;Song, Jeong-Eon;Suh, Junghyuck;Park, Sung Kug;Kim, Meehye
    • Korean Journal of Food Science and Technology
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    • v.45 no.2
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    • pp.235-240
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    • 2013
  • To analyze aflatoxin $M_1$ ($AFM_1$), we dissolved infant formula in warm water and cleaned it by using an immunoaffinity column (IAC). The amount of $AFM_1$ was determined by high-performance liquid chromatography coupled with fluorescence detection. $AFM_1$ was detected in 281 of 439 samples. Thus, the detection rate of $AFM_1$ was 64.0%. The average concentration of $AFM_1$ in positive samples was 2.6 ng/kg (of prepared formula). The estimated daily intake (EDI) of $AFM_1$ through infant formula was 0.087-0.646 ng/kg body weight/day and the additional number of cases of liver cancer associated with exposure to $AFM_1$ would be 0.003-0.020 cancer cases/1,000,000. Because there is less than 1 cancer case/1,000,000 per year, the exposure to $AFM_1$ through infant formula in Korea is considered to be an unlikely human health concern.

Preparation and biological evaluation of 99mTc tricarbonyl cysteine (테크네슘-99엠 트리카보닐 시스테인의 제조 및 생물학적 특성 평가)

  • Jang, Beom-su;Park, Kyung-bae;Yun, Hyo-in
    • Korean Journal of Veterinary Research
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    • v.44 no.1
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    • pp.15-21
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    • 2004
  • This paper describes the development of $^{99m}Tc$ tricarbonyl cysteine as potential renal function diagnostic radiopharmaceutical and evaluation of its biological characteristics using experimental animals. l-Cysteine was labeled efficiently with $^{99m}Tc$ tricarbonyl precursor $([^{99m}Tc(CO)_3(H_2O)_3)]^{+})$ under 30 min heating at ${75^{\circ}C}$. Labeling yield and stability were analyzed by high performance liquid chromatography (HPLC). The biodistribution property of $^{99m}Tc$ tricarbonyl cysteine in mice and its dynamic imaging profiles in rabbits were carried out. To investigate the excretion mechanism of $^{99m}Tc$ tricarbonyl cysteine, tubular transport inhibition test with probenecid was adopted. $^{99m}Tc$ tricarbonyl cysteine was obtained with a high labeling yield under the moderate condition. The results of biodistribution experiments of $^{99m}Tc$ tricarbonyl cysteine in ICR mice at 3 and 90 min provided that $^{99m}Tc$ tricarbonyl cysteine was very highly accumulated in the kidney and bladder, thereby almost 99% of $^{99m}Tc$ tricarbonyl cysteine was excreted within 90 min post injection. The same results were confirmed by the whole body dynamic images for 30 minutes and static images in rabbits at given time intervals after injection. Renogram of $^{99m}Tc$ tricarbonyl cysteine in rabbits showed that its $T_{max}$ and $T_{1/2}$ of $^{99m}Tc$ tricarbonyl cysteine were $2.33{\pm}0.56$ and $4.30{\pm}0.79$ min, respectively. The $T_{max}$ of $^{99m}Tc$ tricarbonyl cysteine with probenecid pretreatment was $2.30{\pm}0.17$ min, whereas $T_{1/2}$ of that with probenecid pretreatment was $17.0{\pm}32.47$ min. $T_{1/2}$ of $^{99m}Tc$ tricarbonyl cysteine with probenecid pretreatment was significantly different, as compared to the result without probenecid (p<0.0001). The results showed that the excretion of $^{99m}Tc$ tricarbonyl cysteine was extremely affected by probenecid. Therefore, $^{99m}Tc$ tricarbonyl cysteine was rapidly excreted from the kidney principally by the tubular secretion.

Determination of Preservatives in Pharmaceuticals and Personal Care Products (의약품 및 개인위생·생활용품 중 보존제 함유량 분석)

  • Kim, Boyoung;Lee, Joohee;Kim, Seonyeob;Lee, Eunji;Choi, Chorong;Kho, Younglim
    • Journal of Environmental Health Sciences
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    • v.42 no.1
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    • pp.53-60
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    • 2016
  • Objectives: Preservatives are commonly used in pharmaceuticals, cosmetics and other products to extend the expiration date and prevent the growth of microorganisms. Preservatives are generally effective in controlling mold and inhibiting yeast growth, and against a wide range of bacterial attacks as well. They also adversely affect the quality of sperm and cause precocious puberty in children. This study was performed to analyze seven preservatives used in pharmaceuticals and personal care products. Methods: Five kinds of pharmaceuticals and personal care products (PPCPs) were examined for analysis with a high performance liquid chromatography-diode array detector. Each sample was homogenized and the targeted compounds were extracted with methanol. The suspended particulate was removed by syringe filter. Next, the sample was injected into an HPLC system. The separation of the seven preservatives was achieved with a C18 column and gradient mode. The accuracies were between 73% and 120% and precision was lower than 11.58% (RSD). Results: All of the calibration curves showed good linearity with a coefficient of determination ($r^2$) over 0.999. Among the PPCP samples, the detection rate of preservatives was 32.5% for pharmaceuticals, 44.8% for toothpaste, 76.9% for mouthwash, 40.0% for body lotion and 56.0% for wet tissues. The average concentrations of the preservatives in PPCPs were BA 1141.0 mg/kg, MP 709.8 mg/kg, EP 624.9 mg/kg, PP 216.9 mg/kg, BP 167.8 mg/kg, and TCS 538.2 mg/kg. The most frequently detected preservatives in pharmaceuticals and personal care products were BA, MP and PP. The concentrations of preservatives exceeded Korean regulatory standards in 11 samples of medicines, three of mouthwash and two of body lotion. Conclusion: We found that most of the PPCP samples contained various preservatives. It is necessary to identify which preservatives were used and to determine the level of preservatives in PPCPs and to assess the health risk to susceptible populations such as children.

Synthesis and Characterization of Structured Lipids from Evening Primrose Seeds Oil and Rice Bran Oil (달맞이꽃 종자유와 미강유로부터 효소적 합성한 재구성 지질의 이화학적 특성 분석)

  • Kim, Hyo-Jin;Lee, Kyung-Su;Lee, Ki-Teak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.8
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    • pp.1156-1164
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    • 2010
  • Structured lipids (SLs) were synthesized by enzymatic interesterification with evening primrose oil (EPO) and rice bran oil (RBO) in a batch-type reactor. The interesterification was performed using a water shaker for 24 hr at $55^{\circ}C$. Mixing speed was set at 200 rpm and Lipozyme RM IM (immobilized lipase from Rhizomucor miehei, 10% by weight of total substrates) was used as a biocatalyst. Rice bran oil and evening primrose oil were interesterified with various molar ratios (RBO : EPO, 1:3, 1:4, and 1:5 mol/mol). Reversed-phase high performance liquid chromatography connected with evaporative light-scattering detector was performed to separate the triacylglycerol (TAG) species of SLs. In the fatty acid analysis, $\gamma$-linolenic acid (7.9 mol%), linoleic acid (67.3 mol%) and oleic acid (13.2 mol%) were the most abundant fatty acids in the SLs. During 24 hr reaction, most of the reaction occurred within 3 hr. TAG compositions, tocopherols and phytosterols were also analyzed. In the TAG species analysis, LLL (ECN=42, L=linoleic acid) dramatically decreased when the reaction time increased.

Ochratoxin A Contamination of Red Chili Peppers from Chile, Bolivia and Peru, Countries with a High Incidence of Gallbladder Cancer

  • Ikoma, Toshikazu;Tsuchiya, Yasuo;Asai, Takao;Okano, Kiyoshi;Ito, Naoko;Endoh, Kazuo;Yamamoto, Masaharu;Nakamura, Kazutoshi
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.14
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    • pp.5987-5991
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    • 2015
  • Our previous study detected aflatoxins in red chili peppers from Chile, Bolivia, and Peru, each of which have a high incidence of gallbladder cancer (GBC). Since the aflatoxin B1 concentration was not so high in these peppers, it is important to clarify the presence of other mycotoxins. Here we attempted to determine any associations between the concentrations of aflatoxins and ochratoxin A (OTA) in red chili peppers, and the corresponding GBC incidences. We collected red chili peppers from three areas in Peru: Trujillo (a high GBC incidence area), Cusco (an intermediate GBC incidence area), and Lima (a low GBC incidence rate), and from Chile and Bolivia. Aflatoxins and OTA were extracted with organic solvents. The concentrations of aflatoxins B1, B2, G1, and G2, and OTA were measured by high-performance liquid chromatography. The values obtained were compared with the incidence of GBC in each area or country. All of the red chili peppers from the three areas showed contamination with aflatoxins below the Commission of the European Communities (EC) recommended limits ($5{\mu}g/kg$), but the OTA contamination of two samples was above the EC recommended limit ($15{\mu}g/kg$). The mean concentrations of OTA in the peppers from Chile (mean $355{\mu}g/kg$, range < $5-1,059{\mu}g/kg$) and Bolivia (mean $207{\mu}g/kg$, range $0.8-628{\mu}g/kg$), which has a high incidence of GBC, were higher than that in Peru ($14{\mu}g/kg$, range < $5-47{\mu}g/kg$), which has an intermediate GBC incidence. The OTA contamination in the red chili peppers from Chile, Bolivia, and Peru was stronger than that of aflatoxins. Our data suggest that OTA in red chili peppers may be associated with the development of GBC.