• Mass Spectrometry Letters
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• v.8 no.3
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• pp.59-64
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• 2017

In clinical diagnosis, it's well known that the abnormal level of uric acid (UA) in human body is implicated in diverse human diseases, for instance, chronic heart failure, gouty arthritis, diabetes, and so on. As a primary method, an isotope dilution mass spectrometry (IDMS) has been used to obtain the accurate quantity of UA in blood or serum and also develop the certificated reference material (CRM) so as to provide a SI-traceability to clinical laboratories. Due to the low solubility of UA in water, an ammonium hydroxide ($NH_4OH$) has been considered as a promising solvent to increase the solubility of UA that enables the preparation of both UA and its isotope standard solution for next IDMS-based absolute quantification. But, because of using this $NH_4OH$ solvent, it gives rise to the unwanted degradation of UA. In this study, we sought to optimize condition for the stability of UA in $NH_4OH$ solution by varying the mole ratios of UA to $NH_4OH$, followed by ID-LC-MRM analysis. In addition, we also inspected minutely the effect of the storage temperatures. Additionally, we also performed the quantitative analysis of UA in the KRISS serum certificated reference material (CRM, 111-01-02A) with diverse mixing ratios of UA to $NH_4OH$ and then compared those values to its certification value. Based on our experiments, adjusting the mole ratio of 1/2 ($UA/NH_4OH$) with the storage temperature of $-20^{\circ}C$ is an effective way to secure both the solubility and stability of UA in $NH_4OH$ solution for next IDMS-based quantification of UA in serum.

• Analytical Science and Technology
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• v.18 no.4
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• pp.271-277
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• 2005

Urea in blood has been measured as an effective marker for diagnosis of renal function. Urea which is e end-product of nitrogen containing metabolites such as proteins is filtered through glomeruli of kidneys and then excreted as urine. If the renal function is deteriorated, the urea concentration in blood will be increased, from which the healthiness of renal function is judged. In order to improve the confidence of diagnosis results, the results must keep traceability chain to certified reference materials, which was certified by primary reference method. In this study, we proposed isotope dilution-liquid chromatography/mass spectrometry (ID-LC/MS) as a candidate primary method, in which $15^N_2$-urea is used as an internal reference material. The developed method is highly accurate in principle and is convenient as it does not require cumbersome derivatization. 0.1 mmol/L ammonium chloride was selected as a mobile phase for HPLC because it provided low interference in MS analysis of relatively low molecular weighted urea. HPLC and MS were connected with an electrospray ionization (ESI) interface of positive mode, which provided high sensitivity and reproducibility. The developed method was validated with internationally recognized reference materials, and we have obtained satisfactory results in an international ring trial. The expanded uncertainty calculated according to ISO guide was 1.8% at 95% confidence interval. The developed method is being used as a primary reference measurement method such as for certification of serum certified reference materials (CRMs).

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.6
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• pp.894-899
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• 2007

An experiment was conducted using a [6, 6-$^2H$]glucose isotope dilution method to determine the effects of plantain (Plantago lanceolata L.) on plasma glucose metabolism in sheep taken from a thermoneutral environment and exposed to a hot environment. The sheep were fed either mixed hay (MH) of orchardgrass (Dactylis glomerata L.) and reed canarygrass (Phalaris arundinacea L.) at a 60:40 ratio or MH and plantain (PL) at a 9:1 ratio in a crossover design for each 23-day period. In both dietary treatments the metabolizable energy (ME) and crude protein intake were designed to be isoenergetic and isoproteinous at around maintenance level. The sheep were taken from a thermoneutral environment ($20^{\circ}C$, 70% RH) and exposed to a hot environment ($28-30^{\circ}C$, 70% RH) for 5 days. The isotope dilution method using a single injection of [6, 6-$^2H$]glucose was performed on the $18^{th}$ day of the thermoneutral environment and on the $5^{th}$ day of heat exposure. Plasma glucose pool size was numerically lower (p = 0.26) during heat exposure on both dietary treatments, and numerically higher (p = 0.13) on the MH diet irrespective of environmental temperature. Plasma NEFA concentration (p = 0.01) and glucose turnover rate (p = 0.03) were decreased during heat exposure, but remained similar between diets. It could be concluded that, although no positive impact of plantain on glucose metabolism was found under the present experimental conditions (plantain constituted only 10% of basal diet), plantain herb is an alternative to MH for rearing sheep in both thermoneutral and hot environments.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.4
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• pp.523-529
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• 2016

An isotope dilution method using $[U-^{13}C]glucose$ and $[1-^{13}C]leucine$ (Leu) was conducted to evaluate the effects of rice straw supplemented with urea and molasses (RSUM-diet) on plasma glucose and Leu turnover rates in sheep. Nitrogen (N) balance, rumen fermentation characteristics and blood metabolite concentrations were also determined. Four sheep were fed either mixed hay (MH-diet), or a RSUM-diet with a crossover design for two 21 days period. Feed allowance was computed on the basis of metabolizable energy at maintenance level. The isotope dilution method was performed as the primed-continuous infusion on day 21 of each dietary period. Nitrogen intake was lower (p = 0.01) for the RSUM-diet and N digestibility did not differ (p = 0.57) between diets. Concentrations of rumen total volatile fatty acids tended to be higher (p = 0.09) for the RSUM-diet than the MH-diet. Acetate concentration in the rumen did not differ (p = 0.38) between diets, whereas propionate concentration was higher (p = 0.01) for the RSUM-diet compared to the MH-diet. Turnover rates as well as concentrations of plasma glucose and Leu did not differ between diets. It can be concluded that kinetics of plasma glucose and Leu metabolism were comparable between the RSUM-diet and the MH-diet, and rumen fermentation characteristics were improved in sheep fed the RSUM-diet compared to the MH-diet.

• Bulletin of the Korean Chemical Society
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• v.34 no.6
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• pp.1698-1702
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• 2013

Glucose is a common medical analyte measuring in human serum or blood samples. The development of a primary method is necessary for the establishment of traceability in measurements. We have developed an isotope dilution liquid chromatography tandem mass spectrometry as a primary method for the measurement of glucose in human serum. Glucose and glucose-$^{13}C_6$ in sample were ionized in ESI negative mode and monitored at mass transfers of m/z 179/89 and 185/92 in MRM, respectively. Glucose was separated on $NH_2P$-50 2D column, and the mobile phase was 20 mM $NH_4OAc$ in 30% acetonitrile/70% water. Verification of this method was performed by the comparison with NIST SRMs. Our results agreed well with the SRM values. We have developed two levels of glucose serum certified reference material using this method and distributed them to the clinical laboratories in Korea as samples for proficiency testings. The expended uncertainty was about 1.2% on 95% confidence level. In proficiency testings, the results obtained from the clinical laboratories showed about 3.6% and 3.9% RSD to the certified values. Primary method can provide the traceability to the field laboratories through proficiency testings or certified reference materials.

• Nuclear Engineering and Technology
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• v.38 no.3
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• pp.301-310
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• 2006

Destructive methods were used for the burnup determination of U-Zr alloy nuclear fuel irradiated in the High-flux Advanced Neutron Application Reactor (HANARO) at KAERI. The dissolution rate of unirradiated U-Zr alloy fuel in $HNO_3$/HF mixtures was investigated for the experimental conditions of a different temperature, and initial concentrations of HF and $HNO_3$. The irradiated U-Zr alloy fuel specimen was dissolved in a mixed acid condition of 3 M HNO3 and 1 M HF at $90^{\circ}C$ for 8 hours under reflux. The total burnup was determined from measurement of the Nd isotope burnup monitors. The method includes U, Pu, $^{148}Nd,\;^P{145}Nd+^{146}Nd,\;^{144}Nd+^{143}Nd$ and total Nd isotopes determination by the isotope dilution mass spectrometric method (IDMS) using triple spikes $(^{233}U,\;^{242}Pu\;and\;^{150}Nd)$. The effective fission yield was calculated from the weighted fission yields averaged over the irradiation period. The results are compared with that obtained by the destructive -spectrometric measurement of the $^{137}Cs$ monitor.

• Journal of the Korean Chemical Society
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• v.55 no.3
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• pp.472-477
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• 2011

The concentration of selenium in grain samples was determined using isotope dilution method in ORC-ICPMS. The experimental conditions were optimized to $H_2$ mode and the flow rate was $4.0\;mL\;min^{-1}$. ORC in $H_2$ mode proved to eliminate most of polyatomic interferences except $BrH^+$ when Br is present in sample matrix. Chemical removal of Br was very difficult and the mathematical correction was successfully employed. The fraction of $BrH^+$ generated from Br at the current experimental condition was 14.1%. The signal on m/z 82 was corrected and calculated for isotope dilution. The analytical reliability of the propose method was successfully evaluated by analyzing the certified standard reference material NIST SRM 1566 and 1567. The method was applied to real samples and the results are $0.034{\pm}0.001\;{\mu}g\;g^{-1}$ for white rice, $0.059{\pm}0.002_5\;{\mu}g\;g^{-1}$ for brown rice, $0.029{\pm}0.001_4\;{\mu}g\;g^{-1}$ for black rice, and $0.034{\pm}0.002\;{\mu}g \;g^{-1}$ for barley. The detection limits ($3\sigma$) for Se was $0.012\;ng\;g^{-1}$.

• Analytical Science and Technology
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• v.19 no.1
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• pp.1-8
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• 2006

The isotope dilution method was used for the determination of Br impurity in $1000{\mu}g/g$ Cl standard solution. Since relatively pure KCl salt was used for the preparation of the Cl standard solution, the Br impurity determination suffers from both spectral and non-spectral interferences due to the presence of a large amount of K and Cl matrices. AG2-X8 anion-exchange resin was employed to separate the Br analyte from the matrices, and RF power was raised to 1500 W and nebulizer gas flow rate was lowered to 0.77 L/min to reduce background from the $ArArH^+$ molecular ions. The Br impurity in the $1000{\mu}g/g$ Cl standard solution was determined to be 43.7 ng/g with the standard addition method. The analytical result was in good agreement with 41.2 ng/g (RSD 1.6%) determined by the isotope dilution method to lower uncertainty from poor reproducibility of the anion-exchange process.

• Nuclear Engineering and Technology
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• v.33 no.4
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• pp.375-385
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• 2001

The total burnup in the spent U$_3$Si/Al fuel samples from Hanaro reactor was determined by destructive methods using $^{148}$ Nd, the sum of $^{143}$ Nd and $^{144}$ Nd, the sum of $^{145}$ Nd and $^{146}$ Nd, and the sum of total Nd isotopes($^{143}$ Nd, $^{144}$ Nd, $^{145}$ Nd, $^{146}$ Nd, $^{148}$ Nd and $^{150}$ Nd) monitors. The fractional($^{235}$ U) turnup in the spent fuel samples was also determined by U and Pu mass spectrometric method. The samples were dissolved in a mixture of 4 M HCI and 10 M HNO$_3$ without any catalyst. The separation of U, Pu and Nd from the spiked and unspiked sample solutions was achieved by two sequential anion exchange separation methods. The isotope compositions of these elements, after their separation from the fuel samples were measured by mass spectrometry. The contents of the elements in the spent fuel samples were determined by isotope dilution mass spectrometric method(IDMS) using $^{233}$ U, $^{242}$ Pu and $^{150}$ Nd as spikes. The effective fission yield was calculated from the weighted fission yields averaged over the irradiation period. The difference between total turnup values determined by various Nd monitors were in the range of 1.8%.

• Analytical Science and Technology
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• v.25 no.1
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• pp.25-32
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• 2012

Isotope dilution mass spectrometry (ID-MS) based on liquid chromatography tandem mass spectrometry (LC/MS/MS) was developed for the accurate determination of sorbic acid in tea-drink. An isotope analogue of sorbic acid, $^{13}C_2$-sorbic acid, was obtained by custom synthesis. MS was operated in the negative mode with selected reaction monitoring (SRM) mode of $[M-H]^-$ ${\rightarrow}$ $[M-CO_2H]^-$ channel at m/z 111 ${\rightarrow}$ 67 for sorbic acid and at m/z 113 ${\rightarrow}$ 68 for its isotope analogue. Chromatographic separation was accomplished with a C18 column and an isocratic mobile phase of 55% of 50 mM ammonium acetate (pH 4.5) and 45% of methanol. Homogeneous reference materials were prepared for validation of this method, including repeatability and reproducibility tests, by fortifying tea-drink with sorbic acid in our laboratory. Repeatability and reproducibility studies showed that the ID-LC/MS method is a reliable and reproducible method which provides less than 3.8% of relative standard deviation (RSD) for the analysis of sorbic acid.