Park, Youngchun;Lim, Jinsook;Ko, Younghuyn;Kwon, Kyechul;Koo, Sunhoe;Kim, Jimyung
The Korean Journal of Blood Transfusion
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v.23
no.2
/
pp.127-135
/
2012
Background: Despite modern advances in laboratory automated medicine, work-process in the blood bank is still handled manually. Several automated immunohematological instruments have been developed and are available in the market. The IH-1000 (Bio-Rad Laboratories, Hercules, CA, USA), a fully automated instrument for immunohematology, was recently introduced. In this study, we evaluated the performance of the IH-1000 for ABO/Rh typing and irregular antibody screening. Methods: In October 2011, a total of 373 blood samples for ABO/Rh typing and 303 cases for unexpected antibody screening were collected. The IH-1000 was compared to the manual tube and slide methods for ABO/Rh typing and to the microcolumn agglutination method (DiaMed-ID system) for antibody screening. Results: For ABO/Rh typing, concordance rate was 100%. For unexpected antibody screening, positive results for both column agglutination and IH-1000 were observed in 10 cases (four cases of anti-E and c, three of anti-E, one of anti-D, one of anti-M, and one of anti-Xg) and negative results for both were observed in 289 cases. The concordance rate between IH-1000 and column agglutination was 98.7%. Sensitivity and specificity were 90.9% and 99.3%, respectively. Conclusion: The automated IH-1000 showed good correlation with the manual tube and slide methods and the microcolumn agglutination method for ABO-RhD typing and irregular antibody screening. The IH-1000 can be used for routine pre-transfusion testing in the blood bank.
Se-Yun Lee;Hee-Sun Jeong;Kyu-Heon Kim;Mi-Young Lee;Jung-Ho Choi;Jeong-Sun Ahn;Kwang-Il Kwon;Hye-Young Lee
Journal of Food Hygiene and Safety
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v.39
no.2
/
pp.72-82
/
2024
This study aimed to develop a scientifically and systematically standardized xylooligosaccharide analytical method that can be applied to products with various formulations. The analysis method was conducted using HPLC with Cadenza C18 column, involving pre-column derivatization with 1-phenyl-3-methyl-5-pyrazoline (PMP) and UV detection at 254 nm. The xylooligosaccharide content was analyzed by converting xylooligosaccharide into xylose through acid hydrolysis. The pre-treated methods were compared and evaluated by varying sonication time, acid hydrolysis time, and concentration. Optimal equipment conditions were achieved with a mobile phase consisting of 20 mM potassium phosphate buffer (pH 6)-acetonitrile (78:22, v/v) through isocratic elution at a flow rate of 0.5 mL/min (254 nm). Furthermore, we validated the advanced standardized analysis method to support the suitability of the proposed analytical procedure such as specificity, linearity, detection limits (LOD), quantitative limits (LOQ), accuracy, and precision. The standardized analysis method is now in use for monitoring relevant health-functional food products available in the market. Our results have demonstrated that the standardized analysis method is expected to enhance the reliability of quality control for healthy functional foods containing xylooligosaccharide.
Lee, Ji Hyun;Cho, So-Hyun;Park, Han Na;Park, Hyoung Joon;Kim, Nam Sook;Park, Sung Kwan;Kang, Hoil
Analytical Science and Technology
/
v.31
no.4
/
pp.171-178
/
2018
Recently, for successful lactation, many breastfeeding mothers seek various products, including herbal medicine, dietary supplements, and prescribed medicines, to improve milk production. As demand for galactogogues grows, it is highly possible that pharmaceutical galactogogues may be adulterated with illegal products to maximize their efficacy. For continuous control and supervision of illegal products, we developed and validated a simple and sensitive LC-MS/MS method capable of simultaneously determining five galactogogues. Chromatographic separation was conducted using an Agilent Poroshell $120SB-C_{18}$ column with a mobile phase consisting of 20 mM ammonium formate (pH 5.4) and 100 % acetonitrile. The total run time was 13 min per analyte. The proposed method was performed according to the guidelines of the International Conference of Harmonization and it produced reliable results. This method showed high sensitivity and specificity, with a limit of detection (LOD) and limit of quantitation (LOQ) of 0.01-0.82 ng/mL and 0.02-2.45 ng/mL, respectively, for the solid- and liquid-type samples. Specificity was evaluated by analyzing matrix-blank samples spiked with the target compounds at LOQ levels, which provided a good separation of all peaks without interference. Additionally, the repeatability and intermediate precision were typically <15 %, whereas the recovery was 80-120 % of the values obtained using blank samples. Thus, we concluded that this method could be used for the identification and quantification of galactogogues in food or herbal products.
Ahn, Eun Jeong;Lee, Hwa Shim;Kim, Byung Joo;Lee, Gae Ho
Analytical Science and Technology
/
v.28
no.6
/
pp.460-466
/
2015
An isotope dilution liquid chromatography tandem mass spectrometry was developed as a primary method for the quantitative analysis of cholesterol in infant formula. Cholesterol-d4 was used as an internal standard and spiked into the infant formula sample. In order to release cholesterol out of cholesteryl ester, which is cholesterol bound to fatty acids in infant formula, saponification was carried out. Saponification conditions were optimized with heating temperature, reaction time and the concentration of KOH. The optimum conditions were as follows; heating temperature was 70 ℃, reaction time was 180 min and the concentration of KOH was 0.8 mL of 8 M KOH for about 0.1 g infant formula sample. Extraction of cholesterol out of sample solution was carried out with hexane uisng liquid-liquid extraction. Chromatographic analysis was carried out using Phenomenex Kinetex C18 column. Mobile phase was 0.1% acetic acid in methanol/water (v/v, 99/1) and flow rate was 0.3 mL/min. Cholesterol and cholesterol-d4 were monitored at mass transfer m/z 369/259 and 373/263 respectively. Reproducibility of the method was evaluated to be 0.23% of the measurement result. The expanded uncertainty of the measurement result of cholesterol in infant formula was approximately 1.9% at a 95% confidence level. NIST standard reference material having certified values of cholesterol in infant formula, was analyzed in order to verify this method. The ID-LC/MS/MS results were well agreed with the certified values of NIST SRM within the uncertainty.
Piscicidal substance produced by Streptomyces sp. isolated from soil was toxic against various kinds of fish. After extraction with CH$Cl_3$ from the culture medium, the substance was purified by avicel column chromatography. In order to test toxicity, various kinds of fish were subjected to the acqueous solution of 100 us of the substance per liter of water. Generally, the substance was toxic to most fish, but Macropodus chinenes and Misgurnus mizolepis are resistant to the substance than Gobius similis and Pseudorasbora parva. The substance was stable at pH range, 3.0 to 7.0, but labile at alkaline pH, and to heat as well. Succinic dehydrogenase on most of tissue cell of Cyprinus carpio was inhibited by this substance strongly, but spinal cord was not inhibited. By addition of Cu and Pb salts to the culture medium, piscicidal substance producibility was activated.
To elucidate pathogenesis of bovine leukemia virus(BLV) in Korean native goats, the goats experimentally infected with BLV were studied especially for the aspects of infectivity and hematological changes. The experimental goats were examined for 27 months by agar-gel immunodiffusion(AGID) test and syncytium formation assay. During this period, changes of total leucocyte, absolute Iymphocyte and atypical Iymphocyte were examined, and the distribution of surface immunoglobulin ( sIg ) -bearing cells and rosette forming cell (RFC) in the peripheral Iymphocyte were also investigated. By indirect immunofluorescence (IFA) and complement dependent antibody cytotoxicity (CDAC) assay using monoclonal antibody(Mab) against bovine leukosis tumor-associated anti-gen(BL-TAA), changes of BL-TAA positive Iymphocyte in peripheral blood were measured. The results obtained through the experiment were summarized as follows. 1. Antibody titers were measured by AGID using gP51 and P24 antigens. The animals were serologically converted at 2 months post-inoculation(pi) in gP51 antigen, whereas sero-converted at 4 months pi in P24 antigen. In comparison with antibody titers for gP51, P24 antigen showed lower titers throughout the trial period. 2. The peripheral lymphocytes from all of the infected goats, as co-cultivated with F8l cells manifested syncytial formation at 4 months pi. 3. On counting total leucocyte, Iymphocyte and atypical Iymphocyte, two out of four infected goats showed normal distribution, while No 2 of the remaining two revealed temporal and No 3, Persistant increasing number of the cells. 4. The optimal condition of rosette formation of the peripheral Iymphocyte of normal Korean native goats was shown in the sheep erythrocyte treated with 0.1M AET for 30 nun at $37^{\circ}C$. When the Iymphocytes were treated in nylon wool column, the number of sIg-bear-ing cell were increased in the nylon wool adherent cells, but RFC was increased in the non-adherent cells. Of the infected goats, No 2 and No 3 showed significantly increasing number of sIg-bearing cells at 18 months pi. 5. The Iymphocytes of No 2 and No 3 goats reacted positively in IFA using Mab against BL-TAA at 12 months pi and 18 months pi, respectively. In CDAC test, all of four infected goats revealed positive reaction at 24 months pi. The higher positive rates were observed in No 2 and No 3 as compared with the remainders.
Journal of the Korean Society of Food Science and Nutrition
/
v.45
no.8
/
pp.1208-1213
/
2016
Advanced glycation end product (AGE) formation and reactive oxygen species are potential therapeutic targets for the prevention of diabetic nephropathy and other pathogenic complications. Activity-guided isolation of an ethylacetate-soluble portion of 80% methanolic extract from fruits of Sorbus commixta of the Ulleung Island origin using AGE formation inhibition assay led to the isolation and identification of three caffeoylquinic acid derivatives of a previously known structure, 3-O-caffeoylquinic acid (neochlorogenic acid; 1), 4-O-caffeoylquinic acid (cryptochlorogenic acid; 2), and 5-O-caffeoylquinic acid (chlorogenic acid; 3). The structures of these compounds were confirmed by interpretation of nuclear magnetic resonance and mass spectrometry data. Among the isolates, the major metabolite, neochlorogenic acid (1) showed the most potent inhibitory effect against AGE formation with an $IC_{50}$ value of $167.5{\pm}3.5{\mu}M$. Furthermore, all isolated chlorogenic acid isomers were evaluated for their radical scavenging activity against peroxynitrite, and structurally related isomers 1, 2, and 3 exhibited potent inhibitory effects in this radical scavenging assay. This result suggests that the monocaffeoyl quinic acid derivatives isolated from S. commixta might be beneficial for the regulation of diabetic complications and related diseases.
In order to elucidate the influence of intestinal and hepatic first-pass effect on the pharmacokinetics of triflusal, the biotransformation of triflusal in the gastrointestinal tract and liver was designed. Moreover, we tried to establish an HPLC method applicable for bioassay and available to pharmacokinetics, not only with the simultaneous determination of triflusal and its active metabolite, 2-hydroxy-4-trifluoromethyl benzoic acid (HTB), but also with improving sensitivity. After the administration of triflusal (10 mg/kg) and HTB (10 mg/kg) into femoral vein, portal vein (only triflusal) and oral route (only triflusal), pharmacokinetic parameters were investigated from the plasma concentration-time profiles of triflusal and HTB in rats. An HPLC method was developed for the simultaneous determination of triflusal and HTB in rat plasma, urine and bile. The HPLC analysis was carried out using a C18 column and acetonitrile-methanol-water (25:10:65, v/v/v) as the mobile phase and UV detection at 234 nm. Furosemide was used as the internal standard. The calibration curves were linear over the concentration range $0.05-5.0\;{\mu}g/ml$ for triflusal and $0.2-200.0\;{\mu}g/ml$ for HTB with correlation coefficients greater than 0.999 and with intra-day or inter-day coefficients of variation not exceeding 10.0%. This assay procedure was applied to the study of metabolite pharmacokinetics of triflusal and HTB in rats. It was supposed that triflusal was almost metabolized in vivo because urinary and biliary excreted amounts of triflusal could be ignored as it was lower than 1.2% of the administered dose. According to the gastrointestinal and hepatic biotransformation pathways of triflusal, it was found that triflusal was hydrolyzed by about 5% in intestine and metabolized by about 53% in liver, and that the bioavailability of triflusal after oral administration of triflusal was 0.44, and also that the fraction of total elimination rate of triflusal which formed HTB in liver $(F_{mi},\;%)$ was about 98%. These results showed that triflusal was almost metabolized in liver, and the total elimination of triflusal in the body was dependent to the formation rate of HTB from triflusal in liver.
Journal of the Korean Society of Food Science and Nutrition
/
v.44
no.11
/
pp.1682-1686
/
2015
This study was conducted to establish an HPLC analysis method for determination of marker compounds as part of materials standardization for development of health functional food materials from pear pomace. The quantitative determination method of caffeic acid and chlorogenic acid as marker compounds of pear pomace extract (PPE) was optimized by HPLC analysis using a C18 column ($5{\times}250mm$, $5{\mu}m$) with a 0.2% elution gradient of acetic acid and methanol as the mobile phase at a flow rate of 0.8 mL/min and detection wavelength of 330 nm. The HPLC/UV method was applied successfully to the quantification of marker compounds in PPE after validation of the method with linearity, accuracy, and precision. The method showed high linearity of the calibration curve with a coefficient of correlation ($R^2$) of 0.9999, and limit of detection and limit of quantification were $1.14{\mu}g/mL$ (caffeic acid) and $1.61{\mu}g/mL$ (chlorogenic acid) as well as $4.9{\mu}g/mL$ (caffeic acid) and $4.9{\mu}g/mL$ (chlorogenic acid), respectively. Relative standard deviation values from intra- and inter-day precision were less than 3.1% (caffeic acid) and 4.0% (chlorogenic acid), respectively. Recovery rates of caffeic acid and chlorogenic acid at 12.5, 25, and $50{\mu}g/mL$ were 93.66~106.32% and 97.33~105.68%, respectively. An optimized method for extraction of caffeic acid and chlorogenic acid in PPE was established through diverse extraction conditions, and the validation indicated that the method is very useful for evaluation of marker compounds in PPE to develop a health functional food material.
Major egg white proteins and their hydrolysates serve as functional food ingredients that have certain metal-chelating properties. Employing egg white proteins and their hydrolysates to scavenge dietary oxalates is anticipated to have beneficial effect in the prevention of kidney stones. The objective of this study was to determine the biogenic oxalate-chelating activity of ovalbumin, ovomucin, and ovotransferrin and their hydrolysates. To prepare oxalate extracts, 30 mL of 0.25 N HCl was added to separately to 0.5 g of dried spinach and starfruit powders followed by boiling for 15 min, and after cooling, the addition of a further 20 mL of 0.25 N HCl. Having prepared these extracts, ovalbumin, ovomucin, and ovotransferrin and their hydrolysates were separately mixed with oxalate extracts and incubated at 3℃ for 24 h. Following centrifugation, supernatants were analyzed by HPLC using a reverse-phase C18 column coupled with a diode array detector. We found that all assessed proteins and their hydrolysates showed biogenic oxalate-chelating activity against the oxalates of spinach. In contrast, however, only ovalbumin, ovalbumin-hydrolysate, and ovomucin showed chelating activity (57.10%±8.84%, 85.44%±5.30%, 73.20%±4.13%, respectively) against the oxalates of starfruit (P<0.05). Overall, hydrolyzed ovalbumin was identified as the most effective chelator of the oxalates both spinach and starfruit. In this study, we thus established that the assessed egg white proteins and their hydrolysates have oxalate-chelating activity in vitro, thereby indicating that these compounds have potential utility as nutraceuticals for the chelation of dietary oxalate. However, further research will be necessary to verify their oxalate-chelating activities against different fruits and vegetables and under specific in vivo conditions and against purified oxalate.
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