• 제목/요약/키워드: AutoML

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Full validation of high-throughput bioanalytical method for the new drug in plasma by LC-MS/MS and its applicability to toxicokinetic analysis

  • Han, Sang-Beom
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 2006년도 추계학술대회
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    • pp.65-74
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    • 2006
  • Modem drug discovery requires rapid pharmacokinetic evaluation of chemically diverse compounds for early candidate selection. This demands the development of analytical methods that offer high-throughput of samples. Naturally, liquid chromatography / tandem mass spectrometry (LC-MS/MS) is choice of the analytical method because of its superior sensitivity and selectivity. As a result of the short analysis time(typically 3-5min) by LC-MS/MS, sample preparation has become the rate- determining step in the whole analytical cycle. Consequently tremendous efforts are being made to speed up and automate this step. In a typical automated 96-well SPE(solid-phase extraction) procedure, plasma samples are transferred to the 96-well SPE plate, internal standard and aqueous buffer solutions are added and then vacuum is applied using the robotic liquid handling system. It takes only 20-90 min to process 96 samples by automated SPE and the analyst is physically occupied for only approximately 10 min. Recently, the ultra-high flow rate liquid chromatography (turbulent-flow chromatography)has sparked a huge interest for rapid and direct quantitation of drugs in plasma. There is no sample preparation except for sample aliquotting, internal standard addition and centrifugation. This type of analysis is achieved by using a small diameter column with a large particle size(30-5O ${\mu}$m) and a high flow rate, typically between 3-5 ml/min. Silica-based monolithic HPLC columns contain a novel chromatographic support in which the traditional particulate packing has been replaced with a single, continuous network (monolith) of pcrous silica. The main advantage of such a network is decreased backpressure due to macropores (2 ${\mu}$m) throughout the network. This allows high flow rates, and hence fast analyses that are unattainable with traditional particulate columns. The reduction of particle diameter in HPLC results in increased column efficiency. use of small particles (<2 urn), however, requires p.essu.es beyond the traditional 6,000 psi of conventional pumping devices. Instrumental development in recent years has resulted in pumping devices capable of handling the requirements of columns packed with small particles. The staggered parallel HPLC system consists of four fully independent binary HPLC pumps, a modified auto sampler, and a series of switching and selector valves all controlled by a single computer program. The system improves sample throughput without sacrificing chromatographic separation or data quality. Sample throughput can be increased nearly four-fold without requiring significant changes in current analytical procedures. The process of Bioanalytical Method Validation is required by the FDA to assess and verify the performance of a chronlatographic method prior to its application in sample analysis. The validation should address the selectivity, linearity, accuracy, precision and stability of the method. This presentation will provide all overview of the work required to accomplish a full validation and show how a chromatographic method is suitable for toxirokinetic sample analysis. A liquid chromatography/tandem mass spectrometry (LC-MS/MS) method developed to quantitate drug levels in dog plasma will be used as an example of tile process.

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그라비올라 잎 추출물의 이화학적 특성 및 항산화 활성 (Antioxidative Effects and Chemical Characteristics of Annona muricata Leaf Extracts)

  • 김영완;김태훈;안희영;조영수
    • 생명과학회지
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    • 제28권5호
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    • pp.540-546
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    • 2018
  • 그라비올라(Annona muricata) 잎으로부터 정제수, 에탄올, 메탄올로 추출하여 수율, 총 페놀성 화합물과 이에 속하는 flavonoid 함량, 미네랄 함량을 측정하고 DPPH free radical scavenging 활성, Cu/Fe 환원력, 간 조직의 microsome 생체막의 지질과산화 억제능 및 linoleic acid를 이용한 지질과산화, ${\beta}$-Carotene 탈색화 정도를 비교검토하였다. 그라비올라 잎의 수율은 수용성 추출물이 가장 높았으며(1.76%), 총 페놀성 화합물 함량은 그라비올라의 메탄올 추출물에서 가장높았으나 페놀성 화합물에 속하는 폴리페놀 함량은 에탄올 추출물에서 가장 높은 결과를 나타내었고, 미네랄 함량은 Ca, K, Mg, Na, Mn의 순으로 높게 측정되었다. DPPH를 이용한 radical 소거능을 측정한 결과, 그라비올라 잎의 에탄올과 메탄올 추출물에서 대조구인 BHT 0.05%와 가장 근접한 활성을 나타내었다. Fe와 Cu의 환원력은 모든 군에서 농도 의존적으로 증가하였고, 그 중 에탄올과 메탄올 추출물에서 Cu/Fe 환원력 활성이 매우 강하였다. 그러나 간 조직을 이용하여 측정하는 지질과산화 억제활성과 TBA법으로 지질과산화를 측정한 결과, 그라비올라 잎의 에탄올 추출물에서 지질과산화 억제능을 보였다. ${\beta}$-Carotene bleaching에서 높은 lipid peroxyl 라디칼 소거 활성이 있음을 확인할 수 있었다. 따라서, 그라비올라 잎은 기능성 식품의 개발을 위한 천연 항산화제 소재로서 활용 될 수 있을 것으로 판단되어진다.

잡견에서 조영제 혈관외유출 예방을 위한 스트레인 게이지 기반의 EDA 시스템 성능 평가를 위한 실험적 연구 (An Experimental Study for Performance Evaluation in Dogs of Preventive Contrast Media Extravasation with a Strain Gage Based Prototype Extravasation Detection Accessory System)

  • 권대철;유병규;이종석;조문선;양성환
    • 대한의용생체공학회:의공학회지
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    • 제29권1호
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    • pp.66-72
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    • 2008
  • The major risk associated with the use of automated power injectors is the well known complication of contrast material extravasation at the injection site. Automated injection of computed tomography (CT) contrast media can produce the compartment syndrome. The purpose of this study was to assess the ability of this device during clinically important episodes of extravasation. The extravasation detection accessory (EDA) system was composed of a strain gage, an amplifier and a computer based system. A strain gage pliable adhesive patch was applied to the skin aver the intravenous catheter and the catheter was connected to the power injector with a cable to monitor the resolution data. If the programmed monitoring, which was developed with MS Visual C++, at the extravasation occurred, then the injection was interrupted the auto injector. CT was used to demonstrate the clinically important extravasation. This study was a prospective, observational study in which the EDA system was used to monitor the automated mechanical injection of contrast material in 7 dogs. There were two true-positive cases (range of extravasation volumes: $18{\sim}22ml$), twenty three true-negative cases, three false-positive cases and no false-negative cases. The EDA system had a sensitivity of 100% and a specificity of 88% for the detection of clinically important extravasation. The EDA system had good sensitivity for the detection of clinically important extravasation and the EDA system has the clinical potential for the early detection of extravasation of the contrast medium that is administered with power injectors. The EDA system is easy to use safe and accurate for the monitoring extravasation of the intravenous injections, and this system may prove especially useful in CT applications.

박하의 in vitro 항산화활성 (In vitro Antioxidant Activity of Mentha viridis L. and Mentha piperita L.)

  • 이승은;한희선;장인복;김금숙;신유수;손영득;박충범;성낙술
    • 한국약용작물학회지
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    • 제13권6호
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    • pp.255-260
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    • 2005
  • 새로운 기능성 소재를 발굴하기 위한 목적으로 2종의 박하(M. viridis (spicata), M. piperita)를 대상으로 수행된 몇 가지 in vitro 항산화계에서의 활성, 총페놀 함량 및 수율을 분석한 결과는 다음과 같다. 1. Superoxide 라디칼에 대한 박하의 소거능은 M. viridis가 M. piperita 보다 우수하였으며 특히 물 추출물은 대조물질인 ascorbic acid보다 우수하였다. 2. DPPH 라디칼에 대한 소거능은 메탄올 추출물 및 물 추출물 모두에서 M. viridis가 M. piperita보다 우수하였으며 특히 M. viridis의 메탄올 추출물은 롤${\alpha}-tocopherol$과 비슷한 수준이었다. 3. 사람의 저밀도지단백질의 산화에 대한 박하의 저해활성을 분석한 결과는 M. viridis가 M.piperita 보다 우수하였으며 M. viridis의 모든 추출물 ${(65.88%{\sim}77.59%,\;37.69%{\sim}87.21%)}$${{\alpha}-tocopherol\;(28.37%{\sim}66.54%)}$보다 월등하게 우수하였다. 4. Linoleic acid의 산화에 대한 저해효과는 M. viridis가 M. piperita보다 높았으며 ${\alpha}-tocopherol$의 활성에 근접하는 수준이었다. 5. 총페놀 함량은 M. viridis (17.95% & 10.18%)가 M. piperita (15.44% & 9.19%)보다 높은 값을 나타내었으며 메탄올 추출물이 물 추출물보다 그 함량이 높았다. 6. 추출물의 수율은 메탄을 추출물 및 물 추출물 모두에서 M. piperita (14.1% & 14.6%)가 M. viridis (13.3% & 13.5%)보다 높게 나타나 항산화활성 및 총페놀 함량과는 상반되는 결과를 나타내었다. 이상의 실험결과를 종합할 때, 실험된 박하 중에서 M. viridis가 M. piperita보다 우수한 산업화 후보 자원이라고 사료된다.