• 제목/요약/키워드: L20 method

검색결과 2,327건 처리시간 0.027초

Large-scale Culture of Plant Cell and Tissue by Bioreactor System

  • Son, Sung-Ho;Park, Sung-Mee;Park, Seung -Yun;Kwon, Oh-Woung;Lee, Yun-Hee;Paek, Kee-Yoeup
    • Journal of Plant Biotechnology
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    • 제1권1호
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    • pp.1-7
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    • 1999
  • Large-scale cultures of plant cell, tissue, and organ have been achieved by using BTBB. When different sized BTBBs (5 L, 20 L, 100 L, 300 L, and 500 L) were tested for the culture of yew cells (Taxus cuspidata Sieb. et Zucc.), cell growth increment reached to 94.5% in SCV after 24 days of culture with 30% of inoculation cell density. However, there were some variations in the production of taxol and its derivatives among the BTBBs of different size. Approximate 4 ㎎/l of taxol and 84 ㎎/l of total taxanes were obtained by using a 500L BTBB after 6 weeks of culture. With a 20L BTBB, about 20,000 cuttings of virus-free potatoes (cv. Dejima) could be obtained by inoculating 128 explants and maintaining 8 weeks under 16 hr light illumination. The frequency of ex vitro rooting of the cuttings revealed as more than 99% under 30% shade. By incorporating two-stage culture process consisting of multiple bulblet formation in solid medium and bulblet development in liquid medium, mass propagation of lily through bioreactor seemed to be possible. In the case of 'Marcopolo', the growth of mini-bulblets in BTBB was nearly 10 folds faster than that of the solid medium. Time course study revealed that maximum MAR yield of ginseng (Panax ginseng C. A. Meyer) in a 5 L and 20 L BTBB after 8 weeks of culture was 500 g and 2.2 ㎏, respectively. By cutting the MAR once and/or twice during the culture, the yield of root biomass could be increased more than 50% in fresh weight at the time of harvest. With initial inoculum of 500 g of sliced MAR in a 500 L BTBB, 74.8 ㎏ of adventitious root mass was obtained after 8 weeks of culture. The average content of total ginseng saponin obtained from small-scale and/or pilotscale BTBBs was approximately 1% per gram dry weight. Based on our results, we suggest that large-scale cultures of plant cell, tissue, and organ using BTBB system should be quite a feasible approach when compared with conventional method of tissue culture.

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안질환세균에 대한 도꼬마리 추출물의 항균 활성 (Antibacterial Activity of Xanthium strumarium L. Extract Against Bacteria Causing Eye Disease)

  • 한선희;김봉환
    • 한국안광학회지
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    • 제13권2호
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    • pp.69-73
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    • 2008
  • 목적: 이 논문은 안질환세균에 대한 도꼬마리 추출물의 항균활성을 알아보아 콘택트렌즈 관리용액에 첨가되어 있는 합성보존제를 대체할 천연 보존제로서의 가능성을 제시하기 위함이다. 방법: 안질환세균에 대한 도꼬마리 추출물의 항균활성은 한천 확산법(agar diffusion test)으로 측정했으며, 항균활성은 종이원반(paper disc) 주위의 선명지대(clear zone)의 생성유무와 선명지대(clear zone)의 크기(mm)로서 알아보았다. 실험에 사용한 세균은 포도상구균 (Staphylococcus)과 녹농균(Pseudomonas)이었다. 결과: Staphylococcus aureus KCCM 40050에 대한 도꼬마리 추출 물 $1000{\mu}g/20{\mu}l$농도의 선명지대는 11.5 mm로 나타났다. 또한 Pseudomonas aeruginosa KCCM 11803 균주에 대한 도꼬마리 추출물 $1000{\mu}g/20{\mu}l$농도의 선명지대는 9.0 mm, Pseudomonas fluorescens KCCM 40223 균주에 대해서는 7.0 mm로 나타났다. 결론: 이 실험을 통해 안질환세균에 대한 도꼬마리 추출물의 항균활성이 입증되었다. 따라서 도꼬마리 추출물이 콘택트렌즈 관리용액에 첨가될 천연 보존제로서의 가치가 충분히 있다고 판단되어진다.

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체외에서 ESI-MS/MS 탐지에 연결된 HPLC에 의한 Entacapon의 수량화: 생물학적 동등성 연구에 적용 (Quantification of Entacapone in Human Plasma by HPLC Coupled to ESI-MS/MS Detection: Application to Bioequavalence Study)

  • Balasekhara Reddy., Ch.;Baburao., Ch.;Chandrasekhar., K.B.;Kanchanamala., K.;RihanaParveen., S.K.;Ravikumar., Konda
    • 대한화학회지
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    • 제54권5호
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    • pp.523-532
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    • 2010
  • 제안된 방법은 국제표준(IS)인 Entacapone-d10(EAD10)을 사용하여 체외에서 Entacapon(EA)의 정량화를 위한 간단하고, 감도가 좋고, 명확한 액체 크로마토그래피-직렬 질량 분석법(LC-ESI-MS/MS)이다. 크로마토그래피 분리는 Zorbax SB-C18에서 수행되었고, $2.1{\times}50\;mm$, $5\;{\mu}m$ 컬럼과 10 mM Ammonium formate (pH 3.0)로 구성된 이동상에서 수행되었다: 0.7 mL/min 유속의 아세토나이트릴(60:40 v/v)은 액체-액체 추출을 따른다. EA와 EAD10은 다중 반응 탐색법(MRM)에서 수소부가물을 가지고 상대적으로 포지티브 모드인 m/z $306.1{\rightarrow}233.1$$316.3{\rightarrow}233.0$에서 수소부과물을 가지고 측정되었다. 그 방법은 상관계수($r^2$) 0.993 이상을 갖는 1.00 - 2000.00 ng/mL의 선형 농도 범위 이상으로 입증되었다. 하루 중과 하루 이내에 3.60에서 7.30과 4.20에서 5.50% 이내의 정밀성과 97.30에서 104.20과 98.30에서 105.80% 이내의 정확도는 EA를 위해 입증되었다. 이러한 방법은 건강한 인도인 자원자들의 생물학적 동등성 연구에서 성공적으로 적용되었다.

HPLC/DAD/ESI-MS를 이용한 혈장시료 중 갑상선 호르몬 분석 (Determination of thyroid hormones in plasma samples by high performance liquid chromatograph/diode array detector/electrospray ionization mass spectrometer)

  • 곽선영;문명희;표희수
    • 분석과학
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    • 제20권5호
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    • pp.424-433
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    • 2007
  • 혈장시료 중의 갑상선 호르몬을 고체상 추출법을 이용하여 추출한 후 HPLC/DAD/ESI-MS (high-performance liquid chromatograph/diode array detector/electrospray ionization-mass spectrometer)를 사용하여 분석하였다. 7종의 thyroid hormones의 HPLC 분리조건은 Hypersil ODS 컬럼(4.6 mm I.D., 250 mm length, particle size $5{\mu}m$)을 사용하고 ammonium formate buffer와 아세토나이트릴을 이동상으로 하여 기울기 용리한 결과 완전 분리가 가능하였으며, UV spectra 및 질량스펙트럼을 확인할 수 있었다. 고체상추출법에 의한 전처리 최적 조건을 조사한 결과 시료를 pH 3으로 한 후 C18 고체상을 사용하여 4 mL의 메탄올로 용리할 경우 회수율이 74.5-115.7%로 나타났다. HPLC/ESI-MS를 이용하여 20-500 ng/mL범위에서 검량선을 작성한 결과 $r^2$값은 0.9939-0.9978으로 나타났으며 검출한계는 20-50 ng/mL (38.1-162.8 pmol/mL)로 계산되었다. 정상인의 혈장에서 thyroxine의 정량이 가능하였으며 그 결과 50.98-112.97 ng/mL로 검출되었다.

기체크로마토그래프/질량분석계에 의한 물시료 중 1,4-dioxane의 분석 및 위해성 평가 (The Analysis of 1,4-Dioxane in Water Sample by Gas Chromatograph/Mass Spectrometer and Risk Assessment)

  • 홍지은;표희수;박송자
    • Environmental Analysis Health and Toxicology
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    • 제18권3호
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    • pp.219-224
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    • 2003
  • 1,4-Dioxane is used as a solvent for lacquers, paints, varnish removers, dye baths and printing compositions. And it is also used for detergent preparations, cosmetics, deodorants and fumigants. A method is described for the determination of 1,4-dioxane in water samples by GC/MS. The extraction recoveries were studied for some solvents and solvent volume ratio were investigated using r-butyl methyl ether (MTBE). Optimum condition was obtained by the liquid-liquid extraction using the 10 mL of MTBE for 10 mL of water. Method detection limit of 1,4-dioxane in the 20 mL of water samples was 0.05 ng/mL. It could be determined in the range of 0.24∼240 ng/mL in treated water, and in the range of 0.69∼81.9 ng/mL in raw water, respectively. Risk assessments with 1,4-dioxane exposure by drinking water ingestion were carried out. Based on the results of analysis, chronic daily intake of 1,4-dioxane was 2.22${\times}$10$\^$-4/ mg/kg/day and excess cancer risk was calcu-lated to be 2.44${\times}$10$\^$-6/.

불산 중 극미량 음이온 분석을 위한 고상 추출법 및 이온크로마토그래프를 이용한 동시분석법 확립 (Optimization of solid phase extraction and simultaneous determination of trace anions in concentrated hydrofluoric acid by ion chromatography)

  • 윤석환;조동호;김현지;신호상
    • 분석과학
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    • 제29권5호
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    • pp.219-224
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    • 2016
  • A sensitive method to detect trace anions in hydrofluoric acid (HF) by solid-phase extraction (SPE) clean-up and ion chromatography (IC) was described. Fluoride in HF solution was eliminated with solid-phase extraction, and residual fluoride, acetate, chloride, bromide, nitrate, phosphate and sulfate were consecutively separated with IC. The SPE parameters (selection of adsorbent, sample volume and pH, elution solvent and its volume) were optimized and selected. The removal effect of fluoride in HF solution was the best on Oasis WAX column, and the optimum conditions (1.0 mL of 25 % HF solution and 50 mM ammonium acetate 5 mL as elution solvent) were established by the variation of parameters. Under the established condition, the method detection limits of chloride, bromide, nitrate, phosphate, and sulfate were 0.04~0.30 µg/L in 25 % HF solutions (w/w) and the relative standard deviation was less than 5 % at concentrations of 20.0 and 40.0 µg/L. The concentrations of anions in a 25 % HF had detectable levels of 4.2 to 47.5 µg/L. The method was sensitive, reproducible and simple enough to permit the reliable routine analysis of anions in HF solution used in the process of producing semiconductors.

고속액체크로마토그라피법을 이용한 사람 혈장 중 알리벤돌(Alibendol)의 정량 및 검증 (Determiniation and Validation of Alibendol using High Pressure Liquid Chromatography in Human plasma)

  • 송현호;유지영;김보겸;박현주;최광식;권영이
    • 약학회지
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    • 제54권4호
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    • pp.295-299
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    • 2010
  • The aim of this study was to develop and validate for determination of alibendol in human plasma by HPLC method. After precipitation of 500 ${\mu}l$ plasma samples by 50% methanol 50 ${\mu}l$ and 60% perchloric acid 30 ${\mu}l$ and the supernatant 50 ${\mu}l$ was injected into HPLC. The assay was performed isocratically using 10 mM potassium phosphate (pH 3.0) and acetonitrile (80 : 20, v/v) as mobile phase. The $C_{18}$ column (particle size $3.5{\mu}m$, $4.6{\times}50$ mm, Zorbax Eclipse) was used as a solid phase. The mobile phase was delivered at a flow-rate of 1.7 ml/min, detection was by ultraviolet absorption at 232 nm and concentrations were calculated on the basis of peak areas. In these conditions, alibendol can be separated from ethylparaben, the internal standard, and endogenous substances. The retention times of alibendol and ethylparaben were just about 2.6 and 3.5 minutes, respectively. This rapid HPLC method was validated by examining the precision and accuracy for inter- and intra-day analysis. The standard curve was linear ($R^2$=1.0000) over the concentration range of 0.05~20 ${\mu}g$/ml. The inter-day relative standard deviation (R.S.D.) and accuracy were 0.2~12.2% and 94.4~101.2% (82.7% at the lower limit of quatitation). The intra-day R.S.D. and accuracy were 0.1~11.8% and 98.8~102.5%, respectively. The method was successfully applied to the determination of alibendol in plasma for a pharmacokinetic study.

단백질 glycation 저해효과가 있는 식품소재 (Anti-glycation Activities from Various Agricultural Products)

  • 최희돈;최인욱;김윤숙
    • 한국식품과학회지
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    • 제39권4호
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    • pp.458-463
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    • 2007
  • 식품소재의 단백질 glycation 저해효과를 조사하기 위하여 곡류, 채소류 및 생약재 등 83종 에탄올 추출물에 대한 단백질 가교결합 저해효과를 $[^{14}C]$-N-formyl-lysine incorporation method를 이용하여 측정하였다. 대부분의 추출물에서 저해효과가 나타났으나 일부 추출물은 단백질의 가교를 촉진하는 것으로 나타났다. 상대적으로 높은 활성을 나타낸 시료 중 20개를 선별하여 형광광도법을 이용한 AGE형성 억제능을 조사한 결과, 1일 발아한 메밀의 추출물 (GB-l)이 가장 높은 저해활성을 가지는 것으로 나타났다. 차후 GB-0l은 콜로로포름, 에틸아세테이트, 부탄올 및 물로 분획되었으며 그 중 에틸아세테이트 분획이 가장 높은 단백질 glycation 저해활성(95.2% inhibitory activity at 100 ug/mL addi- tion level)을 보였다. HPLC를 이용한 폴리페놀 분석결과 발아 및 추출, 분획과정을 거친 메밀의 아세테이트 분획은 rutin 및 quercetin 함량이 상당히 증가한 것으로 나타나서 부분정제물의 rutin 및 quercetin 함량과 저해활성사이에 높은 상관성을 나타내었다.

HPLC Determination and Steady-State Bioavailability Study of Levodropropizine Sustained-release Tablets in Dogs

  • Yan, Lin;Li, Tongling;Zhang, Rongqin;Xu, Xiaohong;Zheng, Pengcheng
    • Archives of Pharmacal Research
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    • 제29권6호
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    • pp.514-519
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    • 2006
  • A simple HPLC method using UV detection was developed and validated for the determination of levodropropizine (LDP) In dog plasma. The sample was prepared for injection using a liquid-liquid extraction method with 1-phenypiperazine as the internal standard. The mobile phase was methanol - diethylamine solution (0.05 M) (20:80, v/v, pH adjusted to 3.0 with $H_3PO_4$) with a detection wavelength of 240 nm. The limit of quantitation (LOQ) of LDP in a biological matrix was determined to be 25.25 ng/mL. The calibration curve was linear across the concentration range of 25.25 to 2020 ng/mL. The intra-day and inter-day precision values (CV%) were within 7% and accuracy (R.E. %) was within 6% of the nominal values for medium (252.5 ng/mL) and high (2020 ng/mL) LDP concentrations. For the LDP concentration at the LOQ, the intra-day and inter-day precision and accuracy were within 20% and 10%, respectively. The average absolute recovery for LDP was 70.28%. This method was successfully used to analyze plasma samples in a steady-state bioavailability study of a newly developed sustained-release LDP tablets (SR) using immediate-release tablets (IR) as the reference. The relative bioavailability of the SR was determined to be $106.3\;{\pm}\;12.8%$ (n=6). The $C_{max}$ of the SR was significantly lower (p<0.05), and the $t_{max}$ was significantly longer than that of the IR (p<0.05). The results of ANOVA and two one-sided tests indicated that the SR exhibited acceptable sustained release properties and was bioequivalent to the IR.

제품폐기물 중 폴리브롬화에테르류 (PBDEs) 분석방법 비교 연구 (Evaluation of analytical method for polybrominated diphenyl ethers (PBDEs) in manufactured products waste)

  • 신선경;김형섭;전태완;김태승;김종하;이정아
    • 분석과학
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    • 제20권2호
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    • pp.109-114
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    • 2007
  • 본 연구에서는 폴리머 재질로부터 시료를 분리하기 위하여 속실렛 추출방법과 용해-침전법이 적용되어졌다. 속실렛 추출의 경우 DCM, Toluene, THF, Acetone/Hexane (1:4, v/v)의 용매가 높은 효율을 나타내는 반면, ethyl ether, acetone/ethyl ether (1:4, v/v), acetone/hexane (1:1, v/v), DCM/hexane (1:1, v/v) 용매는 낮은 효율을 보였다. 용해-침전 방법의 경우 용해를 위해 사용된 용매에 따른 차이는 없었다. 다층실리카겔 컬럼과 플로리실 컬럼의 분획실험 결과 각각 hexane 250 mL와 70 mL로 최적 용래량이 결정되었다. 실제 폐 플라스틱 분석 결과 N.D.~1028 ppm인 것으로 조사되었다.