• Title/Summary/Keyword: curve expression

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Raw Starch-digesting Amylase is Comprised of two Distinct Domains of Catalytic and Substrate-Adsorbable Domain: Role of the C- Terminal Region in Raw-Starch-Binding

  • Kim, Cheorl-Ho
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2001.06a
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    • pp.40-45
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    • 2001
  • Raw starch-digesting amylase (BF-2A, M.W. 93, 000 Da) from Bacillus circulans F-2 was converted to two components during digestion with subtilisin. Two components were separated and designated as BF-2A' (63, 000 Da) and BF-2B (30, 000 Da), respectively. BF-2A' exhibited the same hydrolysis curve for soluble starch as the original amylase (BF-2A). Moreover, the catalytic activities of original and modified enzymes were indistinguishable in $K_{m}$, Vmax for, and in their specific activity for soluble starch hydrolysis. However, its adsorbability and digestibility on raw starch was greatly decreased. Furthermore, the enzymatic action pattern on soluble starch was greatly different from that of the BF-2A. A smaller peptide (BF-2B) showed adsorb ability onto raw starch. By these results, it is suggested that the larger peptide (BF-2A') has a region responsible for the expression of the enzyme activity to hydrolyze soluble substrate, and the smaller peptide (BF-2B) plays a role on raw starch adsorption. A similar phenomenon is observed during limited proteinase K, thermolysin, and endopeptidase Glu-C proteolysis of the enzyme. Fragments resulting from proteolysis were characterized by immunoblotting with anti-RSDA. The proteolytic patterns resulting from proteinase K and subtilisin were the same, producing 63- and 30-kDa fragments. Similar patterns were obtained with endopeptidase Glu-C or thermolysin. All proteolytic digests contained a common, major 63-kDa fragment. Inactivation of RSDA activity results from splitting off the C-terminal domain. Hence, it seems probable that the protease sensitive locus is in a hinge region susceptible to cleavage. Extracellular enzymes immunoreactive toward anti-RSDA were detected through whole bacterial cultivation. Proteins of sizes 93-, 75-, 63-, 55-, 38-, and 31-kDa were immunologically identical to RSDA. Of these, the 75-kDa and 63-kDa proteins correspond to the major products of proteolysis with Glu-C and thermolysin. These results postulated that enzyme heterogeneity of the raw starch-hydrolysis system might arise from the endogeneous proteolytic activity of the bacterium. Truncated forms of rsda, in which the gene sequence encoding the conserved domain had been deleted, directed the synthesis of a functional amylase that did not bind to raw starch. This indicates that the conserved region of RSDA constitutes a raw starch-binding domain, which is distinct from the active centre. The possible role of this substrate-binding region is discussed.d.

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Functional Role of $^{60}RR^{61}$ in 23S rRNA Methylation, Which is in N-Terminal End Region of ErmSF (ErmSF의 N-Terminal End Region에 존재하는 $^{60}RR^{61}$의 23S rRNA Methylation에서의 역할)

  • Jin, Hyung-Jong
    • Korean Journal of Microbiology
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    • v.44 no.3
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    • pp.193-198
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    • 2008
  • ErmSF is one of the proteins which are produced by Streptomyces fradiae to avoid suicide by its autogenous macrolide antibiotic, tylosin and one of ERM proteins which are responsible for transferring the methyl group to $A_{2058}$ (Escherichia coli coordinate) in 23S rRNA, which reduces the affinity of MLS (macrolide-lincosamide-streptogramin B) antibiotics to 23S rRNA, thereby confers the antibiotic resistance on microorganisms ranging from antibiotic producers to pathogens. ErmSF contains an extra N-terminal end region (NTER), which is unique to ErmSF and 25% of amino acids of which is arginine known well to interact with RNA. Noticeably, arginine is concentrated in $^{58}RARR^{61}$ and functional role of each arginine in this motif was investigated through deletion and site-directed mutagenesis and the activity of mutant proteins in cell R60 and R61 was found to play an important role in enzyme activity through the study with deletion mutant up to R60 and R61. With the site-directed mutagenesis using deletion mutant of 1 to 59 (R60A, R61A, and RR60, 61AA), R60 was found more important than R61 but R61 was necessary for the proper activity of R60 and vice versa. And these amino acids were presumed to assume a secondary structure of $\alpha$-helix.

The alterations of $Ca^{2+}$-activated $K^+$ channels in coronary artery during cardiac hypertrophy

  • Kim, Nari;Lee, Sang-Kyeong;Chung, Joon-Yong;Seog, Dae-Hyun;Kim, Euiyong;Jin Han
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.23-23
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    • 2003
  • It has been suggested that the impairment of smooth muscle cell (SMC) function by alterations in the $Ca^{2+}$-activated $K^{+}$ ( $K_{Ca}$ ) channels accounts for the reduction in coronary reserve during left ventricular hypertrophy (LVH). However, this hypothesis has not been fully investigated. The main goal of this study was to assess whether the properties of $K_{Ca}$ channels in coronary SMCs were altered during LVH. New Zealand white rabbits (0.8-1.0 kg) and Sprague-Dawley rats (300-400 g) were randomly selected to receive either an injection of isoproterenol (300 $\mu\textrm{g}$/kg body weight) or an equal volume of 0.9% saline (1 mL/kg body weight). The animals developed LVH 10 days after injection. In patch-clamp experiments, the unitary current amplitude and open probability for the $K_{Ca}$ channels were significantly reduced in LVH patches compared with control patches. The concentration-response curve of the $K_{Ca}$ channel to [C $a^{2+}$]$_{i}$ was shifted to the right. Inhibition of the $K_{Ca}$ channels with TEA was more pronounced in LVH cells than in the control cells. The whole-cell currents of $K_{Ca}$ channels were reduced during LVH. Western blot analysis indicated no differences in $K_{Ca}$ channel expression between the control and LVH coronary SM membranes. In contraction experiments, the effect of a high $K^{+}$concentration on the resting tension of the LVH coronary artery was greater than on that of the control. The effect of TEA on the resting tension of the LVH coronary artery was reduced as compared with the effect on the control. Our findings imply a novel mechanism for reduced coronary reserve during LVH.ing LVH.

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Beneficial effect of Combination with Korean Red Ginseng, Gastrodia Rhizoma and Polygoni Multiflori on Cholesterol and Erectile Dysfunction in Hyperlipidemia rats (홍삼, 천마, 적하수오 병용투여에 의한 고지혈증 랫드에서의 콜레스테롤 및 발기부전 개선효과)

  • Lee, Yun Jung;Kho, Min Chul;Tan, Rui;Lee, Jae Yun;Hwang, Jin Seok;Cha, Jeong Dan;Choi, Kyung Min;Kang, Dae Gill
    • The Korea Journal of Herbology
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    • v.30 no.6
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    • pp.69-75
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    • 2015
  • Objectives : This study was designed to investigate effects of the combination with Korean Red Ginseng (Panax ginseng C.A. Meyer), Gastrodia Rhizoma (Gastrodia elata Blume) and Polygoni Multiflori Radix (Polygonum multiflorum Thunberg) on metabolic disorders including cholesterol and erectile dysfunction in hyperlipidemia rats.Methods : Animals were divided into six groups; Control with normal diet, high fat/cholesterol-diet (HFCD), fluvastatin, Korean Red Ginseng treated (KRG), and the combination treated (Korean Red Ginseng, Gastrodia Rhizoma and Polygoni Multiflori Radix; 1:1:1 for KGP1 and 2:1:1 for KGP2). The experimental groups initially received HFCD for 10 weeks and then treated orally with fluvastatin, KRG, KGP1 and KGP2 during the final 6 weeks. Erectile function was determined by the measurements of intracavernosal pressure (ICP) and maximal arterial pressure (MAP) after electrical stimulation of the cavernosal nerve.Results : KGP2 decreased the level of total cholesterol and LDL cholesterol in the sera of HFCD rats without no changes of body weights. KRG, KGP1 and KGP2 decreased the level of C-reactive protein (CRP) levels except of fluvastatin, synthetic HMG-CoA reductase inhibitor. KRG, KGP1 and KGP2 significantly increased the ICP, ICP/MAP ratio, area under the curve (AUC) compared with those of normal rat. Morphometric analyses showed that KRG, KGP1 and KGP2 increased the volume of smooth muscle and the regular arrangement of collagen fibers in corpus cavernosum of HFCD rats. The penile expression of eNOS was increased by KRG, KGP1 and KGP2.Conclusions : Based on these results, we suggest that the combination with Korean Red Ginseng, Gastrodia Rhizoma and Polygoni Multiflori may improve hyperlipidemia through regulating the lipid profiles and erectile dysfunction in rats.

ᴅ-Xylose as a sugar complement regulates blood glucose levels by suppressing phosphoenolpyruvate carboxylase (PEPCK) in streptozotocin-nicotinamide-induced diabetic rats and by enhancing glucose uptake in vitro

  • Kim, Eunju;Kim, Yoo-Sun;Kim, Kyung-Mi;Jung, Sangwon;Yoo, Sang-Ho;Kim, Yuri
    • Nutrition Research and Practice
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    • v.10 no.1
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    • pp.11-18
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    • 2016
  • BACKGROUND/OBJECTIVES: Type 2 diabetes (T2D) is more frequently diagnosed and is characterized by hyperglycemia and insulin resistance. $\small{D}$-xylose, a sucrase inhibitor, may be useful as a functional sugar complement to inhibit increases in blood glucose levels. The objective of this study was to investigate the anti-diabetic effects of $\small{D}$-xylose both in vitro and stretpozotocin (STZ)-nicotinamide (NA)-induced models in vivo. MATERIALS/METHODS: Wistar rats were divided into the following groups: (i) normal control; (ii) diabetic control; (iii) diabetic rats supplemented with a diet where 5% of the total sucrose content in the diet was replaced with $\small{D}$-xylose; and (iv) diabetic rats supplemented with a diet where 10% of the total sucrose content in the diet was replaced with $\small{D}$-xylose. These groups were maintained for two weeks. The effects of $\small{D}$-xylose on blood glucose levels were examined using oral glucose tolerance test, insulin secretion assays, histology of liver and pancreas tissues, and analysis of phosphoenolpyruvate carboxylase (PEPCK) expression in liver tissues of a STZ-NA-induced experimental rat model. Levels of glucose uptake and insulin secretion by differentiated C2C12 muscle cells and INS-1 pancreatic ${\beta}$-cells were analyzed. RESULTS: In vivo, $\small{D}$-xylose supplementation significantly reduced fasting serum glucose levels (P < 0.05), it slightly reduced the area under the glucose curve, and increased insulin levels compared to the diabetic controls. $\small{D}$-xylose supplementation enhanced the regeneration of pancreas tissue and improved the arrangement of hepatocytes compared to the diabetic controls. Lower levels of PEPCK were detected in the liver tissues of $\small{D}$-xylose-supplemented rats (P < 0.05). In vitro, both 2-NBDG uptake by C2C12 cells and insulin secretion by INS-1 cells were increased with $\small{D}$-xylose supplementation in a dose-dependent manner compared to treatment with glucose alone. CONCLUSIONS: In this study, $\small{D}$-xylose exerted anti-diabetic effects in vivo by regulating blood glucose levels via regeneration of damaged pancreas and liver tissues and regulation of PEPCK, a key rate-limiting enzyme in the process of gluconeogenesis. In vitro, $\small{D}$-xylose induced the uptake of glucose by muscle cells and the secretion of insulin cells by ${\beta}$-cells. These mechanistic insights will facilitate the development of highly effective strategy for T2D.

Identification and Validation of Circulating MicroRNA Signatures for Breast Cancer Early Detection Based on Large Scale Tissue-Derived Data

  • Yu, Xiaokang;Liang, Jinsheng;Xu, Jiarui;Li, Xingsong;Xing, Shan;Li, Huilan;Liu, Wanli;Liu, Dongdong;Xu, Jianhua;Huang, Lizhen;Du, Hongli
    • Journal of Breast Cancer
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    • v.21 no.4
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    • pp.363-370
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    • 2018
  • Purpose: Breast cancer is the most commonly occurring cancer among women worldwide, and therefore, improved approaches for its early detection are urgently needed. As microRNAs (miRNAs) are increasingly recognized as critical regulators in tumorigenesis and possess excellent stability in plasma, this study focused on using miRNAs to develop a method for identifying noninvasive biomarkers. Methods: To discover critical candidates, differential expression analysis was performed on tissue-originated miRNA profiles of 409 early breast cancer patients and 87 healthy controls from The Cancer Genome Atlas database. We selected candidates from the differentially expressed miRNAs and then evaluated every possible molecular signature formed by the candidates. The best signature was validated in independent serum samples from 113 early breast cancer patients and 47 healthy controls using reverse transcription quantitative real-time polymerase chain reaction. Results: The miRNA candidates in our method were revealed to be associated with breast cancer according to previous studies and showed potential as useful biomarkers. When validated in independent serum samples, the area under curve of the final miRNA signature (miR-21-3p, miR-21-5p, and miR-99a-5p) was 0.895. Diagnostic sensitivity and specificity were 97.9% and 73.5%, respectively. Conclusion: The present study established a novel and effective method to identify biomarkers for early breast cancer. And the method, is also suitable for other cancer types. Furthermore, a combination of three miRNAs was identified as a prospective biomarker for breast cancer early detection.

A study on the origin and transformation of the image of earth goddess wring her hair (머리카락에서 물을 짜내는 지모신 형상의 기원과 변모에 관한 연구)

  • Noh, Jang Suh
    • The Southeast Asian review
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    • v.20 no.3
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    • pp.223-262
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    • 2010
  • This paper has been written to find facts about the image of earth goddess broadly found in the Southeast Asia. The research findings are as follows: Firstly, the image of earth goddess wringing her hair is phenomenally discovered in both Buddhist temple murals and independent shrines in Thailand. This phenomenon is common in other Indochinese Buddhist countries such as Myanmar, Laos and Cambodia. Secondly, the life of Buddha including the story of the victory over Mara is found in such Buddhist canons as Mahavastu, Buddhacarita, Lalitavistara, Nidanakatha and Patamasambodhi. Among the canons, the story of the victory over Mara is described in differently ways. Earth becomes personified as the goddess in later version. The main cause to expel Mara's army also changes from sound to water. Patamasambodhi is most closely associated with the iconography of the earth goddess of Southeast Asia. Thirdly, Vessantara Jataka and Indian ancient customs tell us that a merit maker performs a rite of pouring water on the earth as an evidence for merit-making. This rite is a key to understanding the meaning of the scene where the earth goddess expels Mara's army into the flood by wringing her hair. The earth goddess is personified from the earth upon which the merit water is poured. Water soaked in her hair is the very holy water poured by the Buddha whenever he made a merit in his former lives. The amount of water flowed from the hair of the earth goddess representing the amount of his merit making was so huge and enough to defeat the Mara's army and for the Buddha to reach the Enlightenment. This legend explains the significance of the notion of merit in the Theravada Buddhist countries such as Thailand and Myanmar where the water pouring rites still take place and the images of the hair wringing earth goddess are commonly discovered. Fourthly, the first image of the earth goddess as the witness of merits for the Buddha appeared in some Gandharan Buddhist sculptures in the form of devotional gesture with her both hands pressed together and the upper half of her body above the ground. The appearance is in accordance with the description of her in the Lalitavistara canon. In later periods, the form changed into various types and finally the image of the earth goddess wringing her hair appeared in Southeast Asia around 11 century C.E. Some researchers argue this image form of the earth goddess shows the influence from China or India. However, the arguments are considered to be hypothetical as they have no strong evidence to prove. Finally, the modern image of earth goddess shows richer and more dynamic expression compared with its predecessors. Especially, outstanding is the standing earth goddess images found in the scene of the victory over Mara in many temple murals of central region of Thailand. The earth goddess in her voluptuous body shape is usually depicted as wringing her hair with her arms wide open in a posture of S curve. This appearance strongly reminds us of the postures of Salabhanjika and Tribhanga originated from Indian art. The adoption presumably has been made to signify her fertile and affluent characteristics.

Measurement uncertainty analysis of radiophotoluminescent glass dosimeter reader system based on GD-352M for estimation of protection quantity

  • Kim, Jae Seok;Park, Byeong Ryong;Yoo, Jaeryong;Ha, Wi-Ho;Jang, Seongjae;Jang, Won Il;Cho, Gyu Seok;Kim, Hyun;Chang, Insu;Kim, Yong Kyun
    • Nuclear Engineering and Technology
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    • v.54 no.2
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    • pp.479-485
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    • 2022
  • At the Korea Institute of Radiological and Medical Sciences, physical human phantoms were developed to evaluate various radiation protection quantities, based on the mesh-type reference computational phantoms of the International Commission on Radiological Protection. The physical human phantoms were fabricated such that a radiophotoluminescent glass dosimeter (RPLGD) with a Tin filter, namely GD-352M, could be inserted into them. A Tin filter is used to eliminate the overestimated signals in low-energy photons below 100 keV. The measurement uncertainty of the RPLGD reader system based on GD-352M should be analyzed for obtaining reliable protection quantities before using it for practical applications. Generally, the measurement uncertainty of RPLGD systems without Tin filters is analyzed for quality assurance of radiotherapy units using a high-energy photon beam. However, in this study, the measurement uncertainty of GD-352M was analyzed for evaluating the protection quantities. The measurement uncertainty factors in the RPLGD include the reference irradiation, regression curve, reproducibility, uniformity, energy dependence, and angular dependence, as described by the International Organization for Standardization (ISO). These factors were calculated using the Guide to the Expression of Uncertainty in Measurement method, applying ISO/ASTM standards 51261(2013), 51707(2015), and SS-ISO 22127(2019). The measurement uncertainties of the RPLGD reader system with a coverage factor of k = 2 were calculated to be 9.26% from 0.005 to 1 Gy and 8.16% from 1 to 10 Gy. A blind test was conducted to validate the RPLGD reader system, which demonstrated that the readout doses included blind doses of 0.1, 1, 2, and 5 Gy. Overall, the En values were considered satisfactory.

THE ANTICANCER EFFECT OF PACLITAXEL($Taxol^{(R)}$) IN ORAL SQUAMOUS CELL CARCINOMA XENOGRAFT (이종 이식된 구강편평세포 암종에서 Paclitaxel ($Taxol^{(R)}$)의 항암 효과)

  • Kim, Ki-Hwan;Kim, Chul-Hwan;Han, Se-Jin;Lee, Jae-Hoon
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.2
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    • pp.95-110
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    • 2006
  • The treatment for oral and maxillofacial carcinoma with chemotherapeutic agents is evaluated by many effective methods to reduce the tumor mass and cancer cell proliferation. However these chemotherapy have many serious side effects, such as bone marrow suppression, renal toxicity, G-I troubles. Therefore a possible approach to develop a clinically applicable chemotherapeutic agent is to screen anticancer activity of Taxol which is known to have very little side effect and have been used to breast cancer and ovarian carcinoma. Taxol is a new anti-microtubular anti-cancer agent extracted from the bark of the Pacific yew, Taxus brevifolia. Paclitaxel(Taxol) acts by promoting tubulin polymerization and over stabilizing microtubules agianst depolymerization. Despite the constant improvements of methods of the cancer treatment especially chemotherapy, the rate of cancer metastasis and recurrent are not decreased. Thus the investigation of new drug which have very little side effect and a possible clinically application continues to be a high priority. Considering that the Taxol have shown very effective chemotherapeutic agent with relatively low toxicity in many solid tumors, it deserves to evaluate its efficacy in oral squamous cell carcinoma. In this study, to investigate the in-vivo and in-vitro anti-cancer efficacy of Taxol in oral squamous cell carcinoma and lastly, the potency of Paclitaxel in the clinical application for oral cancer was evaluated. In vivo study, after HN22 cell line were xenografted in nude mice, the growth of tumor mass was observed, 3 mg/Kg taxol was injected intraperitoneally into nude mice containing tumor mass. The methods of these study were measurement of total volume of tumor mass, histopathologic study, immunohistochemical study, drug resistance assay, growth curve, MTT assay, flow cytometry, cDNA microarray in vivo and in vitro. The results were obtained as following. 1. The visual inspection of the experimental group showed that the volume of the tumor mass was slightly decreased but no significant difference with control group. 2. Ki-67 index was decreased at weeks 4 in experimental group. 3. Microscopic view of the xenografted tumor mass showed well differentiated squamous cell carcinoma and after Taxol injection, some necrotic tissue was seen weeks 4. 4. The growth curve of the tumor cells were decreased after 1day Taxol treatment. 5. According to the MTT assay, HN22 cell line showed relative drug resistancy above $5\;{\mu}g/ml$ concentrations of Taxol. 6. In drug resistance assay, the decrease of cell counts was seen relatively according to concentration. 7. In Flow cytometry, G2M phase cell arrests were seen in low concentration of the Taxol, while S phase cell arrests were seen in high concentration of the Taxol. 8. Using cDNA microarray technique, variable gene expression of ANGPTL4, TXNRD1, FAS, RRAGA, CTGF, CYCLINEA, P19, DUSP5, CEBPG, BTG1 were detacted in the oral squamous cell carcinoma cell after taxol treatment. In this study paclitaxel is effective against oral squamous cell carcinoma cell lines in vitro, but week effect was observed in vivo. So we need continuous study about anticancer effect of taxol in vivo in oral squamous cell carcinoma.

Quantifying Uncertainty of Calcium Determination in Infant Formula by AAS and ICP-AES (AAS 및 ICP-AES에 의한 조제분유 중 칼슘 함량 분석의 측정불확도 산정)

  • Jun, Jang-Young;Kwak, Byung-Man;Ahn, Jang-Hyuk;Kong, Un-Young
    • Korean Journal of Food Science and Technology
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    • v.36 no.5
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    • pp.701-710
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    • 2004
  • Uncertainty was quantified to evaluate calcium determination result in infant formula with AAS (Atomic Absorption Spectrometry) and ICP-AES (Inductively Coupled Plasma-Atomic Emission Spectrometry). Uncertainty sources in measurand, such as sample weight, final volume of sample, sample dilution and the instrumental result were identified and used as parameters for combined standard uncertainty based on the GUM (Guide to the expression of uncertainty in measurement) and Draft EURACHEM/CITAC Guide. Uncertainty components of each sources in measurand were identified as resolution, reproducibility and stability of chemical balance, standard material purity, standard material molecular weight, standard solution concentration, standard solution dilution factor, sample dilution factor, calibration curve, recovery, instrumental precision, reproducibility, and stability, Each uncertainty components were evaluated by uncertainty types and included to calculate combined uncertainty. The kinds of uncertainty sources and components in the analytical method by AAS and ICP-AES were same except sample dilution factor for AAS. The analytical results and combined standard uncertainties of calcium content were estimated within the certification range $(367{\pm}20\;mg/100g)$ of CRM (Certified Reference Material) and were not significantly different between method by AAS followed by ashing and method by ICP-AES followed by acid digestion as $359.52{\pm}23.61\;mg/100g\;and\;354.75{\pm}16.16\;mg/100g$, respectively. Identifying uncertainty sources related with precision, repeatability, stability, and maintaining proper instrumental conditions as well as personal proficiency was needed to reduce analytical error.