• 제목/요약/키워드: phosphate buffer

검색결과 684건 처리시간 0.02초

용출규격 설정을 위한 생물약제학적분류체계 개념 활용 (Implementation of Biopharmaceutics Classification System Concepts in Developing Dissolution Tests)

  • 사홍기;이경신;백민선
    • Journal of Pharmaceutical Investigation
    • /
    • 제36권3호
    • /
    • pp.161-167
    • /
    • 2006
  • The objective of this study was to investigate the dissolution patterns of variety of orally administered drug products available on the market. It aimed to understand their dissolution behaviors on the basis of the biopharmaceutics classification system (BCS) concept. On the tenets of BCS, several active pharmaceutical ingredients were selected: fluoxetine hydrochloride (class I), naproxen sodium (class ll), pyridostigmine bromide (class III), furosemide (class IV) and simvastatin (class IV). Typical dissolution media used in this study were pH 1.2, pH 4 & 6.8 phosphate buffers, and water. In cases, particular dissolution media specified in the KP and/or USP were used. Dissolution patterns of fluoxetine hydrochloride and pyridostigmine bromide products were characterized by their rapid release In addition, their dissolution characteristics were relatively unaffected by the type of a dissolution medium. Similar dissolution patterns were observed with pH 1.2, pH 4 & 6.8 phosphate buffers and water. By sharp contrast, poor dissolution patterns were noticed with naproxen sodium products, when pH 1.2 and pH 4 phosphate buffer were used. Improvements in its dissolution were achieved by switching the dissolution media to pH 6.8 phosphate buffer or water. Unsatisfactory dissolution data also were observed with a simvastatin product, when it was subject to dissolution tests by use of a surfactant-free pH 1.2, pH 4 & 6.8 phosphate buffers and water. All the release patterns reported in this study were best understood when BCS concepts were implemented. Our results demonstrated that a BCS-based drug classification should be considered first to choose a dissolution test/method and set up dissolution specification.

RP-HPLC에서 Buffer와 메탄올의 조성에 의한 Mononucleotides 체류인자의 조절 (Modification of Retention Factor of Mononucleotides by Compositions of Buffers and Methanol in RP-HPLC)

  • 강덕희;이주원;노경호
    • KSBB Journal
    • /
    • 제15권5호
    • /
    • pp.452-457
    • /
    • 2000
  • Buffer의 특성에 맞는 체류인자에 관한 모텔식을 얻기 위 해서는 buffer 9} 양이온의 농도에 관한 관련식을 실험으로 결 정해야 한다 또한 model 식에서 비이온과 음이온의 체류인 자를 나타내는 ko와 k의 값이 일정한 경향을 나타내고 있기 때문에 buffer의 농도와 modifier의 농도를 동시에 고려한 model 식이 가능하게 된다. 본 연구에서 제안한 모델식을 이용하여 임의의 buffer의 농도에 따른 시료의 체류시간을 예측할 수 있다. 이러한 모텔식을 이용하여 buffer를 이 용한 RP-HPLC의 분석 및 분리조건을 예측할 수 있고 시행 오차적인 방법보다 빠른 시간 내에 최적의 분석 및 분리조건 을 얻을 수 있다

  • PDF

In Vitro Anticholinesterase 스크리닝을 위한 집파리 및 3종 곤충으로부터의 Acetylcholinesterase의 추출 (Extraction of Acetylcholinestrase from the Housefly and Three Other Insect Species for In Vitro Anticholinesterase Screening)

  • 이시혁;이준호;조광연
    • 한국응용곤충학회지
    • /
    • 제30권1호
    • /
    • pp.18-28
    • /
    • 1991
  • 집파리(Musca domestica L.)외 3종 곤충으로부터 0.01 M sodium phosphate buffer를 이용하여 AChE를 분리, 추출할 때 AChE의 안정성 및 buffer capacity를 고려한 적정 pH는 7.5였으며, 0.1M sidium phosphate buffer의 기질 수용액으로 AChE의 활성을 측정할 경우 효소의 활성화와 buffer capacity를 고려했을 때 pH는 7.5가 적합한 조건이었다. AChE 분리.추출을 위한 조직의 마쇄시에는 Potter Elvehjem homogenizer를 사용하였으며, 마쇄액을 원심분리하여 AChE현탁액을 조제할 때 집파리의 경우에는 성충 두부700g 상등액을, 벼멸구(Nilaparvata lugens Stal)는 5령 약충 전충체의 700g 상등액을, 배추좀나방 (Plutella xylostella L.)의 경우에는 5령 유충 전충체의 지질을 제거한 10,000g 상등액을, 그리고 담배거세미나방(Spodoptera litura F.)의 경우에는 4령 유충.두부의 700g 상등액을 AChE 0.2~0.5U 정도로 조정하여 효소원으로 사용하는 것이 대량조제, 추출수율 및 활성 측정상의 안정성등을 고려할 때 합리적인 것으로 조사되었다. 4종 곤충의 AChE현탁액을 $-18^{\circ}C$ 조건에서 냉동보관할 경우 3주까지는 90% 이상의 활성이 유지되었다. 집파리, 벼멸구 및 배추좀나방 AChE의 Km 값은 각각 0.042, 0.037 및 0.043 mM이었으며, 고농도에서 AChE 특이적 기질 저해 현상이 관찰되었으나 담배거세미나방 ChE의 경우에는 1.15 mM의 Km값을 가지며 BuChE적 특성을 보이는 듯 하나 추가실험이 요구된다. 각 곤충의 AChE를 대상으로 한 저해실험시의 적정 기질농도는 집파리, 벼멸구, 배추좀나방의 경우에는 공히 0.5 mM 그리고 담배거세미나방의 경우는 12 mM 이었다.

  • PDF

About Chromium (VI) Extraction from Fertilizers and Soils

  • Sager Manfred
    • 자원환경지질
    • /
    • 제38권6호
    • /
    • pp.657-662
    • /
    • 2005
  • Extractions fro fertilizer and soil samples were performed to yield the operationally defined fractions 'soluble' chromate (extractable with $NH_4NO_3$), 'exchangeable' chromate (extractable with phosphate buffer pH 7.2), and these results were compared with the data obtained by extractions with ammonium sulfate, borate buffer pH 7.2, saturated borax pH 9.6, and polyphosphate (Graham's salt). In order to maintain the pH of extractant solution about constant, the concentration of extractant buffer had to be raised to at least 0.5 M. The results strongly depended on the kind of extractant, and the solid: liquid ratio. For most of the samples investigated, the extraction efficiency increased in the order borate-sulfate-nitrate-phosphate. Whereas the recovery of $K_2CrO_4\;and\;CaCrO_4$ added to the samples of basic slags prior to the extraction was about complete, the recovery of added $PbCrO_4$ was highly variable. In soil extracts, the color reaction was interfered from co-extracted humics, which react with the chromate in weak acid solution during the time period necessary for color reaction (1 hour). However, this problem can be overcome by standard addition and subtraction of the color of the extractant solution. In soil extract of about pH < 7, organic material reduced chromate during the extraction period also, and standard addition of soluble chromate is recommended to prove recovery and the stability of chromate in the samples. In admixtures of soils and basic slags, results for hexavalent chromium were lower than from the mere basic slags. This effect was more pronounced in phosphate than in nitrate extracts. As a proficiency test, samples low in organic carbon from contaminated sites in Hungary were tested. The results from $NH_4NO_3$ extracts satisfactorily matched the results of the Hungarian labs obtained from $CalCl_2$ extractants.

Competitive Enzyme-Linked Immunosorbent Assay for Glucose-6-Phosphate Dehydrogenase

  • Kim, Moon-Hee
    • BMB Reports
    • /
    • 제30권5호
    • /
    • pp.326-331
    • /
    • 1997
  • To construct a competitive ELISA standard curve for the detection of glucose-6-phosphate debydrogenase (G6PD), we used highly purified native G6PD (nG6PD) as both immobilized and soluble antigens and anti-G6PD serum raised against nG6PD as antibody. The polystyrene cuvettes coated with nG6PD were challenged with a mixture of a limiting amount of anti-G6PD serum and various doses of nG6PD as competitors followed by incubation with alkaline phosphatase-anti-IgG conjugate. The competitive ELISA did not exhibit the typical sigmoidal dose-response curve characteristic of competition immunoassays under the optimal concentrations of antigen and antibody. The soluble nG6PD used as competitor failed to effectively inhibit the binding of antibodies to the immobilized nG6PD. The addition of NADP, a cofactor of G6PD enzyme, to coating buffer used for immobilizing nG6PD to the cuvettes and PBS-Tween-BSA buffer for diluting competitors did not improve the inhibition of antibody binding to immobilized nG6PD by soluble n/G6PD. The addition of BSA to coating buffer did not increase inhibition, either. Surprisingly, when partially active G6PD (paG6PD), obtained by repeated freeze-thawing, was used as competitor, the antibody binding to either immobilized nG6PD or immobilized paG6PD was inhibited 49-58%. We conclude that an effective competitive ELISA system with nG6PD enzyme and anti-G6PD serum for the detection of G6PD may not be established due to the poor inhibition of antibody binding to immobilized nG6PD by soluble nG6PD under the present assay conditions and that the inhibition may be improved by using an inactivated enzyme as competitor regardless of the type of immobilized antigen used. These results imply that the immobilized nG6PD may undergo denaturation upon binding to the polystyrene cuvettes and that our anti-G6PD serum may recognize denatured enzyme better than active enzyme.

  • PDF

고온성 사상균의 효소에 관한 연구-(제3보) 고온성 사상균의 Amylase- (Studies on Enzyme of the Thermophilic Mold-Part. 3. Thermophilic mold amylase-)

  • 정동효;이계호
    • Applied Biological Chemistry
    • /
    • 제13권3호
    • /
    • pp.231-235
    • /
    • 1970
  • 1. 내열성 사상균 Humicola속의 Amylase를 정제하였다. 2. DEAE-Cellulose Column Chromatography로 2개의 활성구분으로 나누어 졌다. 3. 즉 pH 6.0, 0.05M와 0.5M의 인산염완충용액으로 용출시킬때 전자에서 당화형, 후자에서 호정화형 Amylase가 각각 분리되었다. 4. 이 당화형 Amylase의 최적 pH는 $4.5{\sim}5.5$이고, 안정 pH 범위는 $4.0{\sim}9.0$범위였다. 최적온도는 $60{\sim}65^{\circ}C$로 다른 사상균의 그것보다 훨씬높고 $80^{\circ}C$ 이상에서는 불활성화 되었다.

  • PDF

Cytosine 정량을 위한 Proteus mirabilis와 Citrobacter freundii 박테리아전극의 개발과 그 비교 (Preparation and Comparison of Proteus mirabilis and Citrobacter freundii Bacterial Electrodes for the Determination of Cytosine)

  • 인권식;김봉원
    • 대한화학회지
    • /
    • 제32권4호
    • /
    • pp.333-341
    • /
    • 1988
  • Cytosine에 대한 bio-electrode는 $NH_3$ 기체감응기에 Proteus mirabilis 와 Citrobacter freundii 박테리아를 고정하여 조립하였다. cytosine deaminase를 포함하는 박테리아는 cytosine 1분자를 $NH_3$ 1분자로 전환시킨다. Proteus mirabilis 박테리아 전극의 감응은 0.2M phosphate 완충용액, pH 8.4에서 $1.0{\times}10^{-3}\;-\;7.0{\times}10^{-3}$M직선범위와 45-48 mV/decade의 감응기울기를 가진다. Citrobacter freundii박테리아 전극의 감응은 0.05M phosphate완충용액, pH 7.6에서 $7.0{\times}10^{-5}\;-\;7.0{\times}10^{-3}$M 직선범위와 48 mV/decade의 감응기울기를 가진다. 이 전극을 pH, 온도, 완충용액, 박테리아의 양, 방해물질, 무기염류의 영향과 전극의 수명을 조사하였다.

  • PDF

한국산(韓國産) 대두(大豆)의 단백질(蛋白質)에 관(關)한 연구(硏究) (제2보(第2報)); 여지전기영동법(濾紙電氣泳動法)에 의(依)한 대두단백질(大豆蛋白質)의 분석(分析) (Studies on the Protein of Korean Soybeans Part 2; Paper Electrophoresis of the Proteins of Korean Soybeans)

  • 변시명;김재욱;이춘영
    • Applied Biological Chemistry
    • /
    • 제7권
    • /
    • pp.85-91
    • /
    • 1966
  • 한국(韓國) 재배(栽培) 대두(大豆) 25품종(品種)을 선택하여 각(各) 품종(品種)의 단백질(蛋白質) 패턴을 여지전기영동법(濾紙電氣泳動法)으로 시행하여 품종간(品種間)의 차이(差異)를 비교(比較)해 보았다. 이 방법(方法)으로 5종(種)의 단백질(蛋白質)을 분리(分離)할 수 있었으며 각(各) 품종간(品種間)의 정량적(定量的)인 함율차(含率差)를 찾아 볼 수 있었다. 각(各) 단백질(蛋白質) 성분간(成分間)의 평균(平均) 함량비(含量比)는 다음과 같다. I 14.47% II 7.22% III 4.43% IV 71.70% V 2.18% 상기(上記) 각획분(各劃分)을 동정(同定)하여 본 결과(結果)는 다음과 같다.

  • PDF

Optimization of 1D 1H Quantitative NMR (Nuclear Magnetic Resonance) Conditions for Polar Metabolites in Meat

  • Kim, Hyun Cheol;Ko, Yoon-Joo;Kim, Minsu;Choe, Juhui;Yong, Hae In;Jo, Cheorun
    • 한국축산식품학회지
    • /
    • 제39권1호
    • /
    • pp.1-12
    • /
    • 2019
  • The objective of this study was to establish an optimized 1D $^1H$ quantitative nuclear magnetic resonance (qNMR) analytical method for analyzing polar metabolites in meat. Three extraction solutions [0.6 M perchloric acid, 10 mM phosphate buffer, water/methanol (1:1)], three reconstitution buffers [20 mM 3-morpholinopropane-1-sulfonic acid, 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid, phosphate buffer], and two pulse programs (zg30, noesypr1d) were evaluated. Extraction with 0.6 M perchloric acid and 20 mM phosphate resulted in a stable baseline and no additional overlap for quantifying polar metabolites in chicken breast. In qNMR analysis, zg30 pulse program (without water-suppression) showed smaller relative standard deviation (RSD) and faster running time than noesypr1d (water-suppression). High-performance liquid chromatography was compared with qNMR analyses to validate accuracy. The zg30 pulse program showed good accuracy and lower RSD. The optimized qNMR method was able to apply for beef and pork samples. Thus, an optimized 1D $^1H$ qNMR method for meat metabolomics was established.

L-Ascorbic Acid-2-Phosphate Mg염의 합성 및 응용

  • 양창모
    • 대한화장품학회지
    • /
    • 제13권1호
    • /
    • pp.29-35
    • /
    • 1987
  • Purely synthesized L-ascorbic acid 2 phosphate Mg salt (1 AsA PMg) improved the weak point of ascorbic acid which is easily decomposed in water solution. This compound is hydrolyzed with phosphatase of skin to corresponding ascorbic acid giving Vitamine C activities. The buffer solution of potassium acetate 0.5% and citric acid 0.005% and the sodium sulfite respectively showed good stabilizing effect of the AsA PMg solution. Compared to the other ascorbic acid derivatives the good solubility of AsA PMg gives broad application to cosmetic field.

  • PDF