• Title/Summary/Keyword: Serum samples

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The Effects of Endogenous and Exogenous Estrogen on Risk Factors of Coronary Heart Disease -Part I : On Serum Lipid Levels- (내인성 및 외인성 Estrogen이 관상심장질환 위험인자에 미치는 영향 -제1보 : 내인성 및 외인성 Estrogen이 혈액 지질 농도에 미치는 영향-)

  • 박유신
    • Journal of Nutrition and Health
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    • v.30 no.3
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    • pp.307-317
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    • 1997
  • To determine the effects of endogenous and exogenous strogen on serum lipid levels, twenty nonsmoking healthy Korean women were participated in this experiment for 12 weeks. They were assigned to three groups : (1) eight women aged 22 to 30(yr) for the premenopausal(Pre) group, (2) eight, aged 49 to 60(yr) for the postmenoparusal(Pst) group, (3) four, aged 23 to 30(yr) for the oral contraceptive(OC) group which used triphasic OC formulation. Fasting blood samples representing every phase of the hormonal levels were obtained from the subjects of the Pre and the OC group. From the subjects of the Pst group, fasting blood samples were obtained once per three weeks for 12 weeks. All the serum data were adjusted for dietary effects, exercise, personality type and body mass index(BMI) by using analysis of covariation(ANCOVA). Serum lipid levels of the three groups were significantly different. While serum levels of triglycerides(TG)(p<0.0001), low density lipoprotein-chloesterol(LDL-C)/high density liporotein-cholesterol(HDL-C) ratio (LDC-C/HDL-C)(P<0.01) and total cholesterol (TC)/HDL-C ratio (TC/HDL-C)(P<0.001) were significatnly high in the Pst group, serum HDL-C(P0.001) level was significantly high in the Pre group. The OC group showed significantly low serum TC(P<0.0001) and LDL-C(P<0.0001) levels. There was no signidicant difference in the fluictuation of serum lipid levels during the menstrual cycle of the Pre group. However, in the OC group, serum TG level was significantly increased at phase 2(P<0.05) where exogenous estrogen administration was highest. Even though other serum lipid levels of the OC group were not significantly fluctuated according to the exogenous estrogen administration, there was a trend of increased levels of serum TC, LDL-C, LDL-C/HDL-C and TC/HDL-C and decreased level of HDL-C during the menstruation period. Also, serum TC level was high(P<0.005) and serum TG level was low (P<0.005) at the baseline of the OC group compared with the periods of OC administration. When screening and counseling the female population at risk for coronary heart disease(CHD), the result of this study suggest that in may be desirable to divide the population into several groups according to their personal physiological characteristics, such as age, OC administration, menstrual cycle and menopause, as well as general risk factors for CHD.

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Detecting Potassium Imbalance: Whole Blood vs. Serum (전혈과 혈청에서의 칼륨 이상소견 검사의 차이)

  • Cho, Young-Duck;Choi, Sung-Hyuk;Yoon, Young-Hoon;Park, Sang-Min;Kim, Jung-Youn;Lim, Chae-Seung
    • The Korean Journal of Blood Transfusion
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    • v.23 no.2
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    • pp.162-168
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    • 2012
  • Background: Potassium, the most common cation in the intracellular space, plays a critical role in our physiology. Potassium imbalance may cause life-threatening problems, ranging from general weakness to cardiac arrest due to ventricular fibrillation. For emergency physicians, detection of such derangement within a short period of time is of critical importance. In this study, we wanted to determine whether analysis of whole blood samples can be used as a screening tool for potassium imbalance by comparative analysis of whole blood and serum samples. Methods: Two samples were drawn from 227 patients. The whole blood sample was taken from the radial artery and contained in a commercially available arterial blood collection syringe with a lithium-heparin coating. The serum sample was contained in a commercially available vacuum bottle in a non-additive silicone coated tube and transported to the laboratory. The study population was divided into three groups, patients with normal whole blood potassium, patients with decreased whole blood potassium, and patients with elevated whole blood potassium. Potassium levels for each group were coupled with serum potassium levels and compared. Results: No significant difference in potassium values was observed between whole blood and serum samples (P<0.05). Strong associations were observed among the three groups (normal range, hypokalemia, and hyperkalemia group). Compared to the normal group (r=0.851), the hyperkalemia group showed a stronger association between variables (r=0.897), and the hypokalemia group showed a weaker association (r=0.760). Their correlation coefficients were highly significant (P<0.05). Conclusion: Our study illustrates that point-of-care testing using whole blood with whole blood can be a reliable screening tool when treating patients with suspicious potassium abnormality, especially in hyperkalemia patients.

Development and Validation of an HPLC Method for the Pharmacokinetic Study of Fexofenadine in Human (테르페나딘 체내동태 연구를 위한 혈청 중 펙소페나딘의 HPLC 정량법 개발 및 검증)

  • Cho, Hye-Young;Kang, Hyun-Ah;Kim, Yoon-Gyoon;Choi, Hoo-Kyun;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.35 no.6
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    • pp.437-443
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    • 2005
  • A rapid, selective and sensitive reversed-phase HPLC method for the determination of a major metabolite of terfenadine, fexofenadine, in human serum was developed, validated, and applied to the pharmacokinetic study of terfenadine. Fexofenadine and internal standard, haloperidol were extracted from human serum by liquid-liquid extraction with acetonitrile and analyzed on a $Symmetry^{TM}$ C8 column with the mobile phase of 1% triethylamine phosphate (pH 3.7)-acetonitrile (67:33, v/v, adjusted to pH 5.6 with triethylamine). Detection wavelength of 230 nm for excitation, 280 nm for emission and flow rate of 1.0 mL/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^{3}$ factorial design using a fixed fexofenadine concentration (50 ng/mL) with respect to its peak area and retention time. In addition, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of 10-500 ng/mL with correlation coefficients greater than 0.999. The lower limit of quantification using 0.5 mL of serum was 10 ng/mL, which was sensitive enough for the pharmacokinetic studies of terfenadine. The overall accuracy of the quality control samples ranged from 95.70 to 114.58% for fexofenadine with overall precision (% C.V.) being 3.53-14.39%. The relative mean recovery of fexofenadine for human serum was 90.17%. Stability studies (freeze-thaw, short-term, extracted serum sample and stock solution) showed that fexofenadine was stable during storage, or during the assay procedure in human serum. However, the storage at $-70^{\circ}C$ for 4 weeks showed that fexofenadine was not stable. The peak area and retention time of fexofenadine were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of fexofenadine in human serum samples for the pharmacokinetic studies of orally administered Tafedine tablet (60 mg as terfenadine) at three different laboratories, demonstrating the suitability of the method.

Development and Validation of an HPLC Method for the Pharmacokinetic Study of Pentoxifylline in Human Serum (체내동태 연구를 위한 혈청 중 펜톡시필린의 HPLC 정량법 개발 및 검증)

  • Cho, Hea-Young;Kang, Hyun-Ah;Yoo, Hee-Doo;Lee, Hwa-Jeong;Moon, Jai-Dong;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.36 no.2
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    • pp.89-95
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    • 2006
  • A selective and sensitive reversed-phase HPLC method for the determination of pentoxifylline in human serum was developed, validated, and applied to the pharmacokinetic study of pentoxifylline. Pentoxifylline and internal standard, chloramphenicol, were extracted from the serum by liquid-liquid extraction with dichloromethane and analyzed on a Luna CI8(2) column with the mobile phase of acetonitrile-0.034 M phosphoric acid (25:75, v/v, adjusted to pH 4.0 with 10 M NaOH). Detection wavelength of 273 nm and flow rate of 0.8 mL/min were used. This method showed linear response over the concentration range of 10-500 ng/mL with correlation coefficients greater than 0.999. The lower limit of quantification using 0.5 mL of the serum was 10 ng/mL, which was sensitive enough for pharmacokinetic studies of pentoxifylline. The overall accuracy of the quality control samples ranged from 89.3 to 92.7% for pentoxifylline with overall precision (% C.V.) being 4.1-9.2%. The relative mean recovery of pentoxifylline for human serum was 105.8%. Stability (stock solution, short and long-term) studies showed that pentoxifylline was not stable during storage. But three freeze-thaw cycles and extracted serum samples were stable. This method showed good ruggedness (within 15% C.V.) and was successfully applied for the analysis of pentoxifylline in human serum samples for the pharmacokinetic studies of orally administered $Trental^{\circledR}$ tablet (400 mg pentoxifylline), demonstrating the suitability of the method.

Development and Validation of HPLC Method for Pharmacokinetic Study of Promethazine in Human (염산프로메타진 체내동태 연구를 위한 혈청 중 프로메타진의 HPLC 정량법 개발 및 검증)

  • Cho, Hae-Young;Kang, Hyun-Ah;Lee, Hwa-Jeong;Choi, Hoo-Kyun;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.36 no.1
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    • pp.23-29
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    • 2006
  • A rapid, selective and sensitive reversed-phase HPLC method for the determination of promethazine in human serum was developed, validated, and applied to the pharmacokinetic study of promethazine. Promethazine and internal standard, chlorpromazine, were extracted from human serum by liquid-liquid extraction with n-hexane containing 0.8% isopropanol and analyzed on a Capcell Pak CN column with the mobile phase of acetonitrile-0.2 M potassium dihydrogen phosphate (42:58, v/v, adjusted to pH 6.0 with 1 M NaOH). Detection wavelength of 251 nm and flow rate of 0.9 mL/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^{3}$ factorial design using a fixed promethazine concentration (10 ng/mL) with respect to its peak area and retention time. In addition, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of 1-40 ng/mL with correlation coefficients greater than 0.999. The lower limit of quantification using 1 mL of serum was 1 ng/mL, which was sensitive enough for pharmacokinetic studies. The overall accuracy of the quality control samples ranged from 96.15 to 105.40% for promethazine with overall precision (% C.V.) being 6.70-11.22%. The relative mean recovery of promethazine for human serum was 63.54%. Stability (freeze-thaw and short-term) studies showed that promethazine was stable during storage, or during the assay procedure in human serum. However, the storage at $-80^{\circ}C$ for 4 weeks showed that promethazine was not stable. Extracted serum sample and stock solution were not allowed to stand at ambient temperature for 12 hr prior to injection. The peak area and retention time of promethazine were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of promethazine in human serum samples for the pharmacokinetic studies of orally administered Himazin tablet (25 mg as promethazine hydrochloride) at three different laboratories, demonstrating the suitability of the method.

Isolation and characterization of Brucella abortus isolates from wildlife species in South Korea

  • Truong, Quang Lam;Kim, Kiju;Kim, Jong-Taek;Her, Moon;Jung, Suk-Chan;Hahn, Tae-Wook
    • Korean Journal of Veterinary Research
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    • v.56 no.3
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    • pp.147-153
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    • 2016
  • A total of 782 blood and 465 tissue samples from 1,039 wild animals and 127 dairy goats were collected from January 2011 to December 2013 in 10 provinces of South Korea and tested for the presence of brucellosis. The Rose Bengal test revealed that 8.0% (52/650) of the serum samples were seropositive, while 4.2% (33/782) of the serum samples were positive for Brucella antibodies by competitive enzyme-linked immunosorbent assay. Of the 650 sera examined, only 16 (2.5%) were positive by both serological tests. Direct polymerase chain reaction (PCR) assay using B4/B5 primers for Brucella abortus (BCSP31) revealed the prevalence of Brucella to be 26.5% (129/487) in blood samples and 21% (98/465) in tissue samples while, 16S rRNA PCR detected Brucella DNA in 6.8% (33/487) and 2.6% (12/465) in blood and tissue samples, respectively. Of PCR-positive samples, only 6.2% (30/487) of blood samples and 2.4% (11/465) of tissue samples were found to be positive by both BCSP31 and 16S rRNA PCRs. However, Brucella strains were isolated by blood culture from only two out of 487 blood samples (0.4%). This characterization and identification of pathogenic Brucella isolates is the first to clearly indicate that the organisms were Brucella abortus biovar 1.

Evaluation of Enzyme-linked Immunosorbent Assay in Serological Diagnosis of Human Neurocysticercosis using paired Samples of Serum and Cerebrospinal Fluid (면역효소측정법을 이용한 뇌낭미충증의 혈청학적 진단의 평가)

  • 조승열;김석일
    • Parasites, Hosts and Diseases
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    • v.24 no.1
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    • pp.25-41
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    • 1986
  • The applicability of micro-ELISA was evaluated in human neuro-cysticercosis using paired samples of serum and CSF. A total of 355 cases who were mostly neurologic patients was subjected. Cystic fluid of C. cellulosae was used as antigen in protein concentration of $2.5{\;}{\mu}g/ml$. Serum was diluted to 1 : 100 and CSF was undiluted in the assay for the specific IgG antibody level. The differential criterion of the positive reaction was the abs. of o. 18 in both samples. The results were summarized as follows: 1. The overall sensitivity of the micro-ELISA in 71 confirmed neurocysticercosis was 90.1% ; the sensitivity by serum was 77.5% and that by CSF was 83.1%. CSF was a more sensitive and valuable material. Most of the false negative cases of neuro-cysticercosis showed far lower level of abs. rather than marginal. 2. The overall specificity of the micro-ELISA in 52 confirmed other neurologic diseases was 88.5%; the specificities by serum and by CSF were 94.2% respectively. Cases of other neurologic diseases did not show false positive reactions in both samples. 3. When serum was assayed, taeniasis(2/18), sparganosis(2/20), paragonimiasis(1/56), clonorchiasis(1/15) and fascioliasis(1/1) cases showed cross reactions. When CSF was assayed, 2 ot 10 neuro-sparganosis showed cross reactions while none of 9 neuro-paragonimiasis showed it. Out of 71 confirmed neuro-cysticercosis cases, 6 and 11 showed cross reactions by serum and CSF to crude extract antigen of sparganum; but no case did show it to crude extract antigen of Paragonimus westermani. 4. Ventricular CSF showed low or negative levels of IgG antibody than lumbar CSF unless the lesion was at the lateral ventricle itself. 5. Out of 4 racemose cysticercosis cases, 3 showed positive reaction in serum while all of 3 examined CSF were positive. The above results indicated that the serological test for detecting the specific IgG antibody by micro-ELISA using paired samples of serum and CSF was very helpful for clinical differentiation of neuro-cysticercosis from neurologic diseases of other causes.

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A Compact Optical System using LED and CMOS Image Sensor for Liver Function Analysis (LED와 CMOS 이미지 센서 기반 간 기능 분석용 소형 광학장치)

  • Kim, Chul;Lim, Chang-Jin;Nam, Myung-Hyun;Kim, Dong-Sik;Seo, Sung-Kyu;Pak, Jung-Ho
    • The Transactions of The Korean Institute of Electrical Engineers
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    • v.61 no.2
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    • pp.270-275
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    • 2012
  • This paper presents a portable and compact optical device which can conveniently be used to perform a functional analysis of human liver function. The proposed system employed red/green LEDs, as a light source, and CMOS image sensor, which is commonly used in cellular phones. With this system, several blood serum samples have been evaluated for liver functional analysis by measuring light absorption level through the blood serum samples depending on aspartate aminotransferase (AST), alanine aminotransferase (ALT), and total bilirubin concentration. The light absorption through the blood serum samples containing AST, ALT, or total bilirubin can provide their concentrations. The green light absorption is more sensitive to the concentration of AST or ALT, and the red light absorption is more sensitive to the total bilirubuin concentration. Additional calibration steps were performed by using a MATLAB program in order to eliminate the light scattering effects from the extraneous particles existing in each blood serum sample. From the blind test, three standard light intensity curves through each enzyme have been obtained and the enzyme concentration values have been compared to those obtained from a commercially available biochemistry analyzer (Toshiba 200 FR). The average percent difference in the obtained concentrations from two systems for AST, ALT, and total bilirubin concentration came out to be 7.79%, 7.98%. and 7.56%, respectively, with the adjusted coefficient of determination (R2) higher than 0.98. This system can possibly lead to a low-cost and simple system that can be used as a point-of-care (POC) system in a condition without advanced equipments.

Sample Preparation and Stability of Human Serum and Urine Based on HPLC-DAD for Metabonomics Studies

  • Liu, Yun;Sun, Xiaoming;Di, Duolong;Feng, Yuxiang;Jin, Fengling
    • Bulletin of the Korean Chemical Society
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    • v.33 no.7
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    • pp.2156-2162
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    • 2012
  • Many literatures focus on the biological relevance and the identification of biomarkers for disease activity assessment while less attention has been paid to the development of standard procedures for sample preparation and storage based on liquid chromatography technique. The influencing factors including protein precipitation, storage temperature, storage time, and reconstitution by ultra pure water were analyzed employing HPLC-DAD. The effects were investigated from five participants over three months by principal components analysis (PCA) and the values of percent changes (PC). The samples with protein precipitation might slow the rate of bacterial enzymatic conversion. After protein precipitation, the average PC of urine samples ($0.136{\pm}0.013$, n = 5) is relatively less than that of the serum samples ($0.173{\pm}0.026$, n = 5) for three months. Minimal effects on metabolic profiles of serum and urine (PC < 0.15) are reasonable for metabolomic studies after protein precipitation and storage at $-20^{\circ}C$ for two months.

Removing Lipemia in Serum/Plasma Samples: A Multicenter Study

  • Castro-Castro, Maria-Jose;Candas-Estebanez, Beatriz;Esteban-Salan, Margarita;Calmarza, Pilar;Arrobas-Velilla, Teresa;Romero-Roman, Carlos;Pocovi-Mieras, Miguel;Aguilar-Doreste, Jose-Angel;Commission on Lipoprotein and Vascular Diseases, Sociedad Espanola de Quimica Clinica
    • Annals of Laboratory Medicine
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    • v.38 no.6
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    • pp.518-523
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    • 2018
  • Background: Lipemia, a significant source of analytical errors in clinical laboratory settings, should be removed prior to measuring biochemical parameters. We investigated whether lipemia in serum/plasma samples can be removed using a method that is easier and more practicable than ultracentrifugation, the current reference method. Methods: Seven hospital laboratories in Spain participated in this study. We first compared the effectiveness of ultracentrifugation ($108,200{\times}g$) and high-speed centrifugation ($10,000{\times}g$ for 15 minutes) in removing lipemia. Second, we compared high-speed centrifugation with two liquid-liquid extraction methods-LipoClear (StatSpin, Norwood, USA), and 1,1,2-trichlorotrifluoroethane (Merck, Darmstadt, Germany). We assessed 14 biochemical parameters: serum/plasma concentrations of sodium ion, potassium ion, chloride ion, glucose, total protein, albumin, creatinine, urea, alkaline phosphatase, gamma-glutamyl transferase, alanine aminotransferase, aspartate-aminotransferase, calcium, and bilirubin. We analyzed whether the differences between lipemia removal methods exceeded the limit for clinically significant interference (LCSI). Results: When ultracentrifugation and high-speed centrifugation were compared, no parameter had a difference that exceeded the LCSI. When high-speed centrifugation was compared with the two liquid-liquid extraction methods, we found differences exceeding the LCSI in protein, calcium, and aspartate aminotransferase in the comparison with 1,1,2-trichlorotrifluoroethane, and in protein, albumin, and calcium in the comparison with LipoClear. Differences in other parameters did not exceed the LCSI. Conclusions: High-speed centrifugation ($10,000{\times}g$ for 15 minutes) can be used instead of ultracentrifugation to remove lipemia in serum/plasma samples. LipoClear and 1,1,2-trichlorotrifluoroethane are unsuitable as they interfere with the measurement of certain parameters.