• Title/Summary/Keyword: quantitation

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A New Method for Analysis of Capsaicinoids Content in Microcapsule. (미세캡슐내의 캡사이시노이드의 새로운 분석법)

  • Jung, Jong-Min;Kang, Sung-Tae
    • Korean Journal of Food Science and Technology
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    • v.32 no.1
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    • pp.42-49
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    • 2000
  • A new method for the quantitative determination of capsaicinoids in microcapsule has been developed. Among seventeen solvents tested for solubilizing wall material (gum arabic and modified starch) of microcapsule, dimethyl sulfoxide (DMSO) was selected as an optimal solvent. The most appropriate mixing ratio of microcapsule to DMSO for solubilizing wall material was 1 to 10(w/v). Appropriate carriersolubilizing temperature and time were $55^{\circ}C$ and 30 min, respectively. Also conditions for extracting oleoresin from the solubilized microcapsule were studied. The mixing ratio of ethanol to DMSO was optimal at 8 to 1(v/v). Optimized vortexing time was 5 min at 40㎐. Pecipitant was obtained by centrifugation at 21000 rpm for 15 min. The precipitant was reextracted with ethanol. The extracted supernatants were combined and adjusted to final volume of 25 ml. Extracted solutions were analyzed for quantitation of total capsaicinoids by employing HPLC and for quantitation of total carotenoids by spectrophotometric method. This method can be used to monitor changes of capsacinoid during manufacturing or storage of red pepper oleoresin microcapsule powder.

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Biochemical Quantitation of PM2 Phage DNA as a Substrate for Endonuclease Assay

  • Joo, Yoo-Jin;Kim, Hee-Ju;Lee, Jae-Yung;Kim, Joon
    • Journal of Microbiology
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    • v.42 no.2
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    • pp.99-102
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    • 2004
  • Bacteriophage PM2 has a closed circular form of double stranded DNA as a genome. This DNA from the phage is a useful source for nick-circle endonuclease assay in the fmol range. Due to difficulties in the maintenance of viral infectivity, storage conditions of the phage should be considered for the puri-fication of PM2 DNA. The proper condition for a short-term storage of less than 2 months is to keep the PM2 phage at 4$^{\circ}C$; whereas the proper condition for a long-term storage of the PM2 phage for over 2 months is to keep it under liquid nitrogen in 7.5 % glycerol. The optimal conditions for a high yield of phage progeny were also considered with the goal to achieve a successful PM2 DNA preparation. A MOI(Multiplicity Of Infection) of 0.03, in which the OD$\sub$600/ of the host bacteria was between 0.3 and 0.5, turned out to be optimal for the mass production of PM2 phage with a burst size of about 214. Considerations of PM2 genome size, and the concentrations and radiospecific activities of purified PM2 DNA, are required to measure the endonuclease activity in the fmol range. This study reports the proper quantitation of radioactivity and the yield of purified DNA based on these conditions.

Identification of Pitfalls Related to the Analysis of Liquid Chromatography-Tandem Mass Spectrometry and Liquid Chromatography-Time of Flight Mass Spectrometry (액체크로마토그래프-질량분석기를 이용한 정성 및 정량 오류의 확인)

  • Kwon, Jin-Wook;Cho, Yoon-Jae;Rhee, Gyu-Seek
    • Korean Journal of Environmental Agriculture
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    • v.34 no.3
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    • pp.230-237
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    • 2015
  • BACKGROUND: To identify the sources of inaccuracy in LC/MS/MS methods used in the routine quantitation of small molecules are described and discussed. METHODS AND RESULTS: Various UPLC coupled to triple quadrupole mass spectrometer and time of flight (TOF) were used to identify the potential sources of inaccuracy and inducing the pitfalls of qualification and quntitation during the veterinary drug residue analysis. Some of stable isotope labelled veterinary drugs, which were used as internal standards, presented "cross-talk", regardless of manufactures of mass spectrometer and types of spectrometer. Group of sulfonamides also presented inaccuracy qualification and quantitation due to the multi-residue analytical method with the same fragment ions at the close retention times. CONCLUSION: The phenomena of "cross-talk" occurring between subsequently monitored transition from stable isotope labelled and isotope non-labelled authentic chemical were identified. To prevent errors and achieve more accurate data during the analysis of small molecules by LC/MS/MS SRM method, Followings should be taken care of and kept checking; purity and concentration of stable isotope as an internal standard, prevention of carry-over during the separation in column, minimizing the ion suppression by matrix effect, identification of retention time, precursor ion and product ion, and full knowledge of data processing including smoothing and peak integration.

Evaluation of Serum and Urine Protein Electrophoresis in Patients with Renal Disease (신장질환 환자에서 혈청과 요단백 전기영동 분획 평가)

  • Lim, Hyun-Jin;Back, Seong-Ok;Lee, Bum-Hee;You, Seon-Woo
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.2
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    • pp.111-116
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    • 2006
  • One of the cardinal findings of the renal diseases is proteinuria, which appears in the early phase of kidney diseases and is very important in diagnosis, prognosis and decision making in the treatment process and results of the treatment. The study subjects were 126 patients who visited the nephrology department of Kangbuk Samsung Hospital. Serum was requested for urine protein electrophoresis. Total protein was measured with Bayer Advia 1650 (Biuret). Quantitation of each fraction was done by multiplying the percentage of each fraction by the total protein. Serum creatinine and BUN were also measured with Bayer Advia 1650 (Jaffe and Urease). Serum protein EP was done with REP(rapid electrophoresis) using Helena Kit reagents (REP Ultra SPE Kit, Ponceau S stain, Acetic acid, Methanol, EP Control). Concentrated urine was used for urine protein EP. The SPSS package was used for statistics analysis. Percentage and quantitation of the level of albumin in renal diseases were significantly lower than those in healthy controls. Total protein was correlated with albumin. In terms of proportion, ${\alpha}1$-globulin, ${\alpha}2$-globulin, ${\beta}$-globulin, and ${\gamma}$-globulin fractions were increased in the disease group. But, in the quantified level, ${\alpha}2$-globulin was increased and ${\beta}$-globulin and ${\gamma}$-globulin were decreased. ESRD patients showed an increased secretion of high molecular proteins in urine protein EP. A decreased level in serum total protein correlated with the decreased level of serum albumin and the total amount of urine total protein. This study revealed the variety in the level of serum and urine proteins and their subgroups by EP.

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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.

A Simple and Efficient Method to Determine Rivaroxaban in Rat Plasma Using Liquid-Liquid Extraction and LC-MRM

  • Lee, Hyo Chun;Kim, Dong Yoon;Choi, Min-Jong;Jin, Sung Giu;Choi, Yong Seok
    • Mass Spectrometry Letters
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    • v.10 no.2
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    • pp.66-70
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    • 2019
  • Rivaroxaban (RRN) is the first available active direct factor Xa inhibitor (anticoagulant) with oral administration. Due to its success in market, there have been efforts to develop various RRN formulations, and the development of good analytical methods for its in vivo evaluation is an essential prerequisite. Thus, here, a simple and efficient method to determine RRN in rat plasma using liquid-liquid extraction (LLE) and liquid chromatography and multiple reaction monitoring (LC-MRM) was presented. The use of ethyl acetate as the LLE solvent results appropriate extraction and purification of RRN and it also helps the significant reduction of rat plasma volume required for RRN quantitation. The developed method showed good analytical performance including specificity, linearity ($r^2{\geq}0.999$ within 0.5 - 500 ng/mL), sensitivity (the lower limit of quantitation at 0.5 ng/mL), accuracy (89.3 - 107.0%), precision (${\geq}12.7%$), and recovery (89.2 - 105.7%). Additionally, RRN in sample extracts showed good stability. Finally, the applicability of the validated method to the PK evaluation of RRN was confirmed after its oral administration to normal rats. The present method is the first analytical method employing LLE for the simple and efficient extraction and purification of RRN in rat plasma. Therefore, the present method can contribute to the development of new RRN formulations as well as to the monitoring of RRN in special clinical situations through its efficient determination in various samples with or without minor modification.

Objective Quantitation of EGFR Protein Levels using Quantitative Dot Blot Method for the Prognosis of Gastric Cancer Patients

  • Xin, Lei;Tang, Fangrong;Song, Bo;Yang, Maozhou;Zhang, Jiandi
    • Journal of Gastric Cancer
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    • v.21 no.4
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    • pp.335-351
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    • 2021
  • Purpose: An underlying factor for the failure of several clinical trials of anti-epidermal growth factor receptor (EGFR) therapies is the lack of an effective method to identify patients who overexpress EGFR protein. The quantitative dot blot method (QDB) was used to measure EGFR protein levels objectively, absolutely, and quantitatively. Its feasibility was evaluated for the prognosis of overall survival (OS) of patients with gastric cancer. Materials and Methods: Slices of 2×5 ㎛ from formalin-fixed paraffin-embedded gastric cancer specimens were used to extract total tissue lysates for QDB measurement. Absolutely quantitated EGFR protein levels were used for the Kaplan-Meier OS analysis. Results: EGFR protein levels ranged from 0 to 772.6 pmol/g (n=246) for all gastric cancer patients. A poor correlation was observed between quantitated EGFR levels and immunohistochemistry scores with ρ=0.024 and P=0.717 in Spearman's correlation analysis. EGFR was identified as an independent negative prognostic biomarker for gastric cancer patients only through absolute quantitation, with a hazard ratio of 1.92 (95% confidence interval, 1.05-3.53; P=0.034) in multivariate Cox regression OS analysis. A cutoff of 208 pmol/g was proposed to stratify patients with a 3-year survival probability of 44% for patients with EGFR levels above the cutoff versus 68% for those below the cutoff based on Kaplan-Meier OS analysis (log rank test, P=0.002). Conclusions: A QDB-based assay was developed for gastric cancer specimens to measure EGFR protein levels absolutely, quantitatively, and objectively. This assay should facilitate clinical trials aimed at evaluation of anti-EGFR therapies retrospectively and prospectively for gastric cancer.

Studies on the theory of Oriental Medicine Diagnosis and applicatin of Moire topography (한의학적(韓醫學的) 진단원리(診斷原理)와 모아레 토포그래피의 응용(應用))

  • Lee, Jae-Won
    • Korean Journal of Oriental Medicine
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    • v.1 no.1
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    • pp.273-287
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    • 1995
  • Moire topography, a simple technique for three-dimensional quantitation, was used to provide interference fringe photographs of the human back with sufficient accuracy to be used for detecting patient with asymmetry due to scoliosis, the disease of cervix and lumbar, muscle dysfunction. Contour lines are a suitable and widely accepted method of describing a three-dimensional surface. In the moire technique, contour lines of an object are produced as interference fringes while the object is illuminated by a spotlight through a special grating. The fringe pattern is produced by the interference of the grating and its shadow on the object. A photograph of a moire pattern on the human back will permit an assessment of the overall body shape and the symmetry of the back. This study uses shadow moire topography. Moire topography provides a non-invasive technique for quantifying the shape of the human body. In the use of moire topography for the Oriental Medicine Diagnosis, the strength of moire lies in the ablility to detect change due to deformity of human body.

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Effects of Korean Ginseng Saponin Fraction on the Biosynthesis of Spermidine and Spermine from Bat Prostate and Testis

  • 조윤식;조영대
    • Journal of Ginseng Research
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    • v.22 no.4
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    • pp.316-323
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    • 1998
  • To study e(feces of Korean Binseng (Parfax ginseff C. A. hfeyer) total saponin fraction on spermidine and spermine metabolism in rat reproductive systems, we administrated the saponin fractation to rats for 2 years. Then, we determined the activities of S-adenosylmethionine decarboxylase (SAMDC), the quantitation of the enzyme protein and the amounts of spermidine and spermine contents In prostate and testis. In young sexually immature stage, administration of Korean ginseng saponin fraction showed no effect on SAMDC activities. The stimulatory effect on the activities of SAMDC gradually increased and reached maximal activities in test groups of prostate and testis at sexually mature stage. The amounts of SAMDC protein in test groups were paralleled by the changes of SAMDC activities in test groups, indicating that all of the increased activity occurring in administration of ginseng saponin fraction was not due to the activation of SAMDC activity but to the Increase in enzyme protein. However, the spermidine and spermine contents of test groups showed small increase in compared to that of control groups. From these results, we suggest that administration of ginseng saponin fraction alter the spermidine and spermine metabolism in sexually mature and aged reproductive systems in rats.

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Quantitation of Pulmonary Nodule's Border Structure by Means of Fourier Transform by using Chest X-ray CT Images (흉부 X선 CT 화상을 이용한 푸리에 변환에 의한 폐종류의 경계 형상의 정량화)

  • 김응규;권영도
    • Proceedings of the IEEK Conference
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    • 2000.06d
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    • pp.135-138
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    • 2000
  • 흉부 X선 CT 화상을 이용한 폐종류의 경계 형상을 정량적으로 평가하기 위하여 푸리에 변환된 폐종류 음영의 윤곽선 내 power spectrum 고주파 성분의 총합이 폐종류 음영의 경계 형상에 관한 유효한 평가 값이 되는지의 여부를 검토하였다. 이 평가 값은 폐종류 음영의 CT 화상 위의 특징을 명확히 반영한다고 판단된다. 다시 말해서, 윤곽선은 양성 혹은 악성 종류에 있어서 각각 명확하거나 불투명하다. 양성 IS명과 악성 16명인 환자 31명에 대해서 이 평가 값을 계산하여 통계적 처리를 행한 결과 양성과 악성 간에 뚜렷한 차이를 인식할 수 있었다. 이러한 제안된 평가 방법에 의해, power spectrum 고주파 성분의 총합이 폐종류 경계 형상의 평가치가 되어, 정량적인 폐종류의 양성과 악성 감별을 행할 때 유용한 값이 될 가능성을 시사한다고 볼 수 있다.

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