• Title/Summary/Keyword: precision validation

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Development and validation of LC-MS/MS for bioanalysis of hydroxychloroquine in human whole blood

  • Park, Jung Youl;Song, Hyun Ho;Kwon, Young Ee;Kim, Seo Jin;Jang, Sukil;Joo, Seong Soo
    • Journal of Biomedical and Translational Research
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    • v.19 no.4
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    • pp.130-139
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    • 2018
  • This study aimed to analyze a high-performance liquid chromatography (HPLC) separation using a pentafluorophenyl column of parent drug hydroxychloroquine (HCQ) and its active metabolite, desethylhydroxchloroquine (DHCQ) applying to determine bioequivalence of two different formulations administered to patients. A rapid, simple, sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for bioanalysis of HCQ and its metabolite DHCQ in human whole blood using deuterium derivative $hydroxychloroquine-D_4$ as an internal standard (IS). A triple-quadrupole mass spectrometer was operated using electrospray ionization in multiple reaction monitoring (MRM) mode. Sample preparation involves a two-step precipitation of protein techniques. The removed protein blood samples were chromatographed on a pentafluorophenyl (PFP) column ($50mm{\times}4.6mm$, $2.6{\mu}m$) with a mobile phase (ammonium formate solution containing dilute formic acid) in an isocratic mode at a flow rate of 0.45 mL/min. The standard curves were found to be linear in the range of 2 - 500 ng/mL for HCQ; 2 - 2,000 ng/mL for DHCQ in spite of lacking a highly sensitive MS spectrometry system. Results of intra- and inter-day precision and accuracy were within acceptable limits. A run time of 2.2 min for HCQ and 2.03 min for DHCQ in blood sample facilitated the analysis of more than 300 human whole blood samples per day. Taken together, we concluded that the assay developed herein represents a highly qualified technology for the quantification of HCQ in human whole blood for a parallel design bioequivalence study in a healthy male.

Nucleotide Analysis in Korean Dairy Products Using High- Performance Liquid Chromatography with Diode Array Detector

  • Won, Jong-Eun;Bang, Han-Yeol;Kwak, Byung-Man;Park, Jong-Su;Kim, Gui-Ran;Kwon, Joong-Ho
    • Food Science of Animal Resources
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    • v.39 no.1
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    • pp.93-101
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    • 2019
  • Nucleotides play important roles in numerous intracellular biochemical processes and are used in infant formulas and other dairy products. However, domestic analytical methods for assessing nucleotide content in products have not yet been established, and therefore, methods for determining nucleotide content are urgently required. A rapid and simple analytical method for determining the content of five types of nucleotides in dairy products was improved using solid phase extraction clean-up and high-performance liquid chromatography with diode array detector. The extraction solvent used in the AOAC method was not well dissolved and was changed to hydrophilic EDTA-Na. In addition, the results obtained using the isocratic elution method and a single wavelength were similar to those obtained using the AOAC method, and the time taken for analysis was shortened from 40 min to 25 min. The process of method validation revealed the following parameters: accuracy (84.69%-102.72%), precision (1.51%-6.82%), linearity (0.999), and limit of detection (cytidine 5'-monophosphate, 0.09 mg/L; uridine 5'-monophosphate, 0.11 mg/L; adenosine 5'-monophosphate, 0.12 mg/L; guanosine 5'-monophosphate, 0.11 mg/L; and inosine 5'-monophosphate, 0.14 mg/L). The method was also used to determine the nucleotide concentration in 25 samples (infant formulas, 1.99-29.39 mg/100 g; and cow milk, 0.28-0.83 mg/100 g). The newly improved method was appropriate for analyzing nucleotides in infant formulas and other dairy products faster when compared to conventional methods.

Development of Highly Sensitive Analytical Method for Evaluation of Evening Primrose Oil's Enhancing Effect in Prostaglandin E1(OP 1206) Biosynthesis

  • Lee, Sung-Hoon
    • Journal of People, Plants, and Environment
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    • v.21 no.6
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    • pp.485-492
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    • 2018
  • This study aimed to develop and validate highly sensitive determination method of a prostaglandin ($PGE_1$, OP 1206) in human plasma by LC-MS/MS using column switching. Plasma stored at $-30^{\circ}C$ and treated with methanol effectively inhibited interferences synthesized post-sampling. Samples were added with internal standard and were separated by reversed-phase HPLC with a cycle time of 30min. The method was selective for OP 1206 and the regression models, based on internal standard, were linear across the concentration range 0.5-50 pg/mL with the limit of quantification of 0.5 pg/mL (limit of quantitation, LOQ) for OP 1206. The calibration curve of OP 1206 standards spiked in five individual plasma samples was linear ($r^2=0.9999$). Accuracy and precision at the concentrations of 0.5, 1.5, 5.0 and 40 pg/mL, and at the lower LOQ of 0.5 pg/mL were excellent at 20%. OP120 < 6 was stable in plasma samples for at least 24 hours at room temperature, 24 hours frozen at $-70^{\circ}C$, 24 hours in an auto sampler at $6^{\circ}C$, and for two freeze/unfreezing cycles. The validated determination method successfully quantified the concentrations of OP 1206 in plasma samples from simulated administrating a single $5{\mu}g$ OP 1206 formulation. Thus, this novel LC-MS/MS technique for drug separation, detection and quantitation is expected to become the standard highly-sensitive detection method in bioanalysis and to be applied to many low dose pharmaceutical products.

Comparison of Marker Components and Biological Activities of Socheongryong-tang by Different Extract Methods (소청룡탕 제조방법에 따른 성분 및 생리활성 비교)

  • Baek, Ka Yeon;Kim, Se Jin;Kim, Jong Beom;Lee, Jeong Hyeon;Moon, Sung Ok;Lee, Hwa Dong
    • Korean Journal of Pharmacognosy
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    • v.49 no.4
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    • pp.349-361
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    • 2018
  • Socheongryong-tang (SCRT) was one of the major traditional herbal medicines wildly used in the treatment of respiratory disease. SCRT is being commercially produced in the form of mix extracts powder and soft dry extract by different extract methods in the Korean Herbal Pharmacopeia (KHP). In this study, the contents of marker components and biological activities of the SCRT mix extract powder were compared with those of the SCRT decoction. To analyze the marker components of SCRT, nine marker from eight herbal preparations were chosen. And the method using high performance liquid chromatography (HPLC) with diode-array detector method was established for the simultaneous analysis. Method validation was accomplished by linearity, precision test, and recovery test. The contents of nine marker components in this extract was ascertained by ratio. The biological activities were examined the effect of SCRT on anti-oxidation and pro-inflammation mediated by LPS-stimulation. We confirmed that both of SCRT mix extrct powder and decoction have the similar contents on total polyphenol and flavonoid and inhibited the secretion of nitric oxide (NO), $IL-1{\beta}$, IL-6, tumor necrosis factor $(TNF)-{\alpha}$ and the expression of iNOS, COX-2, $IL-1{\beta}$, IL-6, $TNF-{\alpha}$. These results suggest that SCRT mix extract powder and decoction have a significant correlation.

Evaluation and application of pretreatment methods for pharmaceuticals and personal care products in the solid phase of sewage samples (하수처리시설 고상시료 중 잔류의약물질 분석을 위한 전처리법 평가 및 적용)

  • Park, Junwon;Kim, Changsoo;Ju, Byoungkyu;Lee, Wonseok;Chung, Hyenmi;Jeong, Dong-Hwan
    • Journal of Korean Society of Water and Wastewater
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    • v.32 no.6
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    • pp.559-572
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    • 2018
  • The aim of this study was to evaluate pretreatment methods for 27 pharmaceuticals and personal care products (PPCPs) in various sewage samples using a modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) and online solid-phase extraction with LC-MS/MS. Extraction efficiencies of PPCPs in the solid phase under different experimental conditions were evaluated, showing that the highest recoveries were obtained with the addition of sodium sulfate and ethylenediaminetetraacetic acid disodium salt dehydrate in acidified conditions. The recoveries of target compounds ranged from 91 to 117.2% for liquid samples and from 61.3 to 137.2% for solid samples, with a good precision. The methods under development were applied to sewage samples collected in two sewage treatment plants (STPs) to determine PPCPs in liquid and solid phases. Out of 27 PPCPs, more than 19 compounds were detected in liquid samples (i.e., influent and effluent) of two STPs, with concentration ranges of LOQ-33,152 ng/L in influents and LOQ-4,523 ng/L in effluents, respectively. In addition, some PPCPs such as acetylsalicylic acid, ibuprofen, and ofloxacin were detected at high concentrations in activated sludge as well as in excess sludge. This methodology was successfully applied to sewage samples for the determination of the target compounds in STPs.

Study on Effect of Micro Tooth Shape Modification on Power Transmission Characteristics based on the Driving Gear of Rotating Machining Unit (마이크로 치형수정이 선회가공 유닛 구동기어의 동력전달 특성에 미치는 영향에 관한 연구)

  • Jang, Jeong-Hwan;Qin, Zhen;Kim, Dong-Seon;Wu, Yu-Ting;Lyu, Sung Ki
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.18 no.6
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    • pp.91-97
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    • 2019
  • Rotating machining unit is a revolutionary product that can process worm shaft or spiral shaft with fast and precise, a rotary type cutting tool, which is attached to automatic lathe and processes spiral groove on outer circumference of round bar. In this work, a study on micro tooth shape modification method of driving gear train in the rotating machining unit was presented. To observe the effect on power transmission characteristics of the driving gear pair, visualize the gear meshing condition and the load distribution on the gear teeth by using the professional gear train analysis program RomaxDesigner. By comparing the repeated analysis results, the effect of micro tooth shape modification on power transmission characteristics on driving gear can be summarized. The optimized gears were fabricated and measured by precision tester as a validation in this research.

Power Transmission Optimization Based on the Driving Gear of a Cross Drilling/Milling Unit using a Micro Geometry Method (마이크로 지오메트리 방법을 이용한 크로스 드릴링/밀링 유닛 구동기어의 동력전달 최적화에 관한 연구)

  • Kim, Dong-Seon;Zhen, Qin;Beak, Gwon-In;Wu, Yu-Ting;Jeon, Nam-Sul;Lyu, Sung Ki
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.18 no.4
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    • pp.93-99
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    • 2019
  • A cross drilling/milling Unit is an important mechanical part which is widely used in many kinds of machining tool, and various gear trains with good accuracy and reliability features are widely used in power transmission systems. A study on a novel power transmission optimization method for driving gear trains in cross drilling/milling units is presented in this paper. A commercial program for gear system simulation, Romax Designer, was used in this research to intuitively observe the gear meshing and the load distribution conditions on the gear teeth. We obtained the optimal modification value through comparing the results of repeated experiments. For validation, optimized gears were fabricated and then measured with a precision tester.

Rapid determination and quantification of hair-growth compounds in adulterated products by ultra HPLC coupled to quadrupole-orbitrap MS

  • Lee, Ji Hyun;Park, Han Na;Kang, Gihaeng;Kim, Nam Sook;Park, Seongsoo;Lee, Jongkook;Kang, Hoil
    • Analytical Science and Technology
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    • v.32 no.2
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    • pp.56-64
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    • 2019
  • Recently, a number of adulterated products, which are advertised as hair-growth enhancer have been emerged among those who suffer hair loss disease. For continuous control of illegal products, in this study, a rapid and sensitive method for simultaneous screening of 12 compounds that enhance hair-growth was established to protect public health by ultrahigh-performance liquid chromatography coupled to quadrupole-orbitrap mass spectrometry (UHPLC-Q-Orbitrap-MS). Fragmentation pathways of them were proposed based on $MS^2$ spectral data obtained using the established method. In this analysis, the LODs and LOQs ranged from 0.05 to 50 ng/mL and from 0.17 to 167 ng/mL, respectively. The square of the linear correlation coefficient ($R^2$) was determined as more than 0.995. The intra- and inter-assay accuracies were respective 88-112 % and 88-115 %. Their precision values were measured within 5 % (intra-day) and 10 % (inter-day). Mean recoveries of target compounds in adulterated products ranged from 84 to 115%. The relative standard deviation of stability was less than 12 % at $4^{\circ}C$ for 48 h. The method was employed to screen 14 dietary supplements advertised to be effective for the treatment of hair loss. Some of the products (~21 %) were proven to contain synthetic drugs that promote hair growth such as triaminodil, minoxidil, and finasteride.

Validation of Gamma Knife Perfexion Dose Profile Distribution by a Modified Variable Ellipsoid Modeling Technique

  • Hur, Beong Ik;Jin, Seong Jin;Kim, Gyeong Rip;Kwak, Jong Hyeok;Kim, Young Ha;Lee, Sang Weon;Sung, Soon Ki
    • Journal of Korean Neurosurgical Society
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    • v.64 no.1
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    • pp.13-22
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    • 2021
  • Objective : High precision and accuracy are expected in gamma knife radiosurgery treatment. Because of the requirement of clinically applying complex radiation and dose gradients together with a rapid radiation decline, a dedicated quality assurance program is required to maintain the radiation dosimetry and geometric accuracy and to reduce all associated risk factors. This study investigates the validity of Leksell Gamma plan (LGP)10.1.1 system of 5th generation Gamma Knife Perfexion as modified variable ellipsoid modeling technique (VEMT) method. Methods : To verify LGP10.1.1 system, we compare the treatment plan program system of the Gamma Knife Perfexion, that is, the LGP, with the calculated value of the proposed modified VEMT program. To verify a modified VEMT method, we compare the distributions of the dose of Gamma Knife Perfexion measured by Gafchromic EBT3 and EBT-XD films. For verification, the center of an 80 mm radius solid water phantom is placed in the center of all sectors positioned at 16 mm, 4 mm and 8 mm; that is, the dose distribution is similar to the method used in the x, y, and z directions by the VEMT. The dose distribution in the axial direction is compared and analyzed based on Full-Width-of-Half-Maximum (FWHM) evaluation. Results : The dose profile distribution was evaluated by FWHM, and it showed an average difference of 0.104 mm for the LGP value and 0.130 mm for the EBT-XD film. Conclusion : The modified VEMT yielded consistent results in the two processes. The use of the modified VEMT as a verification tool can enable the system to stably test and operate the Gamma Knife Perfexion treatment planning system.

The Effect of Type of Input Image on Accuracy in Classification Using Convolutional Neural Network Model (컨볼루션 신경망 모델을 이용한 분류에서 입력 영상의 종류가 정확도에 미치는 영향)

  • Kim, Min Jeong;Kim, Jung Hun;Park, Ji Eun;Jeong, Woo Yeon;Lee, Jong Min
    • Journal of Biomedical Engineering Research
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    • v.42 no.4
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    • pp.167-174
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    • 2021
  • The purpose of this study is to classify TIFF images, PNG images, and JPEG images using deep learning, and to compare the accuracy by verifying the classification performance. The TIFF, PNG, and JPEG images converted from chest X-ray DICOM images were applied to five deep neural network models performed in image recognition and classification to compare classification performance. The data consisted of a total of 4,000 X-ray images, which were converted from DICOM images into 16-bit TIFF images and 8-bit PNG and JPEG images. The learning models are CNN models - VGG16, ResNet50, InceptionV3, DenseNet121, and EfficientNetB0. The accuracy of the five convolutional neural network models of TIFF images is 99.86%, 99.86%, 99.99%, 100%, and 99.89%. The accuracy of PNG images is 99.88%, 100%, 99.97%, 99.87%, and 100%. The accuracy of JPEG images is 100%, 100%, 99.96%, 99.89%, and 100%. Validation of classification performance using test data showed 100% in accuracy, precision, recall and F1 score. Our classification results show that when DICOM images are converted to TIFF, PNG, and JPEG images and learned through preprocessing, the learning works well in all formats. In medical imaging research using deep learning, the classification performance is not affected by converting DICOM images into any format.