• Mass Spectrometry Letters
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• 제7권4호
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• pp.111-115
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• 2016

Glycerol was identified and isolated from endogenous interferences during analysis of human urine using high-resolution mass spectrometry (HRMS) for doping control. Urinary sample preparation was simple; the samples were diluted with an organic solvent and then analyzed using a liquid chromatography-mass spectrometry ("dilute and shoot" method). Although the interfering ion peaks were observed at the similar retention time of glycerol, the inference could be identified by isolation with HRMS and further investigation. Thus, creatinine was identified as the endogenous interference for glycerol analysis and it also caused ion suppression resulting in the decrease of glycerol signal. This study reports the first identification and efficient isolation of endogenous interferences in human urine for "dilute and shoot" method. The information about ion suppression could be novel to prevent overestimation or a false result for antidoping analysis.

• Mass Spectrometry Letters
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• 제7권3호
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• pp.55-63
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• 2016

Growth hormone (GH)-releasing peptides (GHRPs) and GH secretagogues (GHSs) are listed in the World Anti-Doping Agency (WADA) Prohibited List. In the present study, we developed and validated a method for the simultaneous analysis of seven GHRPs (alexamorelin, GHRP-1, -2, -4, -5, -6, and hexarelin) and three GHSs (anamorelin, ibutamoren, and ipamorelin) in human urine. Method validation was performed at minimum required performance levels specified by WADA technical documents (2 ng/mL) for all substances, and the method was validated with regard to selectivity (no interference), linearity (R2 > 0.9986), matrix effects (50.0%-141.2%), recovery (10.4%-100.8%), and intra- (2.8%-16.5%) and inter-day (7.0%-22.6%) precisions. The limits of detection for screening and confirmation were 0.05-0.5 ng/mL and 0.05-1 ng/mL, respectively.

• 한국컴퓨터정보학회논문지
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• 제28권5호
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• pp.171-177
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• 2023

본 연구는 다양한 이론적 접근을 통해 진행되어왔던 선행연구들을 바탕으로 국내 엘리트 선수들의 도핑 의도에 영향을 미치는 요인들을 파악하기 위해 단계적 회귀분석을 활용하여 운동경력 및 도핑교육 경험과 같은 인구통계학적 요인들과 통제적 동기, 도핑방지에 대한 태도 및 행동통제인식 요인과 도핑 의도의 관계를 규명함으로써 도핑방지에 중요한 정보를 제공하는데 목적이 있다. 연구의 목적을 달성하기 위해 SPSS 27.0 ver을 사용하여 분석하였다. 상관분석과 단계선택 회귀분석 사용하여 도출된 연구결과는 다음과 같다. 운동경력, 도핑 교육 경험 유·무, 통제적 동기, 도핑방지에 대한 태도 및 행동통제인식 요인 모두 도핑의도에 유의한 영향을 미치는 것으로 확인되었으며 이 중 영향력이 가장 큰 변인들을 각 순서대로 투입하여 유의한 영향이 있는지 검증하였다. 검증 결과 통제 동기가 가장 큰 영향을 미치는 것으로 나타났다. 다음으로는 도핑에 대한 행동통제인식, 도핑 교육 경험 유·무, 도핑에 대한 태도, 운동경력 순서대로 영향을 미치는 것을 확인하였다.

• 분석과학
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• 제8권4호
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• pp.881-886
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• 1995

Several forms of gangliosides have been separated from various types of biological matrices using cyclodextrin-modified capillary zone electrophoresis (CZE). Quantitative analysis of phospholipids from biological fluids was achieved by micellar electrokinetic capillary chromatography (MECC) using 35mM sodium dodecyl sulfate. Phosphorylation, one of the most important post-translational modifications of proteins at serine, threonine and tyrosine residues in small peptides were identified and quantitative analyses of phosphopeptides were performed. Seven different neuropeptides which are relative the pain reachanism in the vertebrate central nervons system were also separated by CZE.

• Mass Spectrometry Letters
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• 제11권4호
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• pp.118-124
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• 2020

An analytical method was developed for hypoxia-inducible factor (HIF) stabilizers based on QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) sample preparation and liquid chromatography-high resolution mass spectrometry analysis. HIF stabilizers potentially enhance the performance of athletes, and hence, they have been prohibited. However, the analysis of urinary HIF stabilizers is not easy owing to their unique structure and characteristics. Hence, we developed the QuEChERS preparation technique for a complementary method and optimized the pH, volume of extraction solvent, and number of extractions. We found that double extraction with 1% of formic acid in acetonitrile provided the highest recovery of HIF stabilizers. Moreover, the composition of the mobile phase was also optimized for better separation of molidustat and IOX4. The developed method was validated in terms of its precision, detection limit, matrix effect, and recovery for ISO accreditation. To the best of our knowledge, this is the first demonstration of the application of the QuEChERS method, which is suitable as a complementary analytical method, in antidoping.

• 대한화학회지
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• 제35권6호
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• pp.659-666
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• 1991

질소를 함유한 흥분제와 마약성 진통제 18종류의 약물들을 gas chromatography-nitrogen phosphorus detector(GC-NPD)를 사용하여 human urine으로부터 동시에 신속하게 분석할 수 있는 최적조건을 찾기 위하여 pH 변화와 추출용매 변화에 따른 회수율을 조사하였다. pH 8.5에서 에테르를 추출용매로 사용하였을 때 가장 적은 방해영향과 가장 좋은 회수율을 나타냈다. NPD에 대한 각 약물들의 상대 감응인자를 구하였고, 이 상대감응인자는 약물이 가지고 있는 질소원자의 갯수가 증가할수록 작은 값을 나타냈다. 생체시료 중의 약물들을 신속하게 검정하기 위하여 내부표준물질인 diphenylamine에 대한 relative retention time(RRT)을 작성하였다. 상대머무름 시간은 0.1% 이하의 정밀도를 나타냈다.

• Mass Spectrometry Letters
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• 제8권2호
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• pp.39-43
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• 2017

Meldonium is a drug for treating ischemia by expanding the arteries but it can also enhance the performance of sports players. The World Anti-Doping Agency (WADA) has included it in the list of prohibited substances since 2016. Meldonium is one of the challenging substances for anti-doping testing because it is difficult to recover by general liquid-liquid or solid phase extraction due to its permanent charge and high polarity. Therefore, high-performance liquid chromatography (HPLC) is currently used by injecting a diluted urine sample (known as the "dilute-and-shoot" strategy). There is no loss of target compounds in the extraction/cleanup procedure but its high matrix effect could interfere in their separation or detection from the endogenous urinary compounds. We report a single method using high-resolution mass spectrometry that can be used for both screening and confirmation, which follows the "dilute-and-shoot" strategy. In this method, the endogenous compounds' interfering peaks in the mass spectrum are separated at a high resolution of FWHM 140,000, and the results are suitable for substance detection following the WADA guidelines. The interferences in the obtained mass spectrum of the urine matrix are identified as acetylcholine, lysine, and glutamine by further analysis and database searching. Validation of the method is performed in routine anti-doping testing, and the limit of detection is 50 ng/mL. This method uses simple sample preparation and a general reverse phase HPLC column, and it can be easily applied to other substances.

• Bulletin of the Korean Chemical Society
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• 제30권1호
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• pp.137-144
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• 2009

In this study, quantitative and pattern recognition analyses for the quality evaluation of Herba Epimedii using HPLC was developed. For quantitative analysis, five major bioactive constituents, hyperin, epimedin A, epimedin B, epimedin C, and icariin were determined. Analysis was carried out on Capcell pak $C_{18}$ column ($250{\time}4.6$ mm, 5 ${\mu}m$) with a mobile phase of mixture of acetonitrile and 0.1% formic acid, using UV detection at 270 nm. The linear behavior was observed over the investigated concentration range (2-50 ${\mu}g/mL;\;r_2\;>$ 0.99) for all analytes. The intraand inter-day precisions were lower than 4.3% (as a relative standard deviation, RSD) and accuracies between 95.1% and 104.4%. The HPLC analytical method for pattern recognition analysis was validated by repeated analysis of one reference sample. The RSD of intra- and inter-day variation of relative retention time (RRT) and relative peak area (RPA) of the 12 selected common peaks were below 0.8% and 4.7%, respectively. The developed methods were applied to analysis of twenty Herba Epimedii extract samples. Contents of hyperin, epimedin A, epimedin B, epimedin C, and icariin were calculated to be 0$\sim$0.79, 0.69$\sim$1.91, 0.93$\sim$9.58, 0.65$\sim$3.05, and 2.43$\sim$11.8 mg/g dried plant. Principal component analysis (PCA) showed that most samples were clustered together with the reference samples but several apart from the main cluster in the PC score plot, indicating differences in overall chemical composition between two clusters. The present study suggests that quantitative determination of marker compounds combined with pattern-recognition method can provide a comprehensive approach for the quality assessment of herbal medicines.

• Mass Spectrometry Letters
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• 제3권2호
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• pp.50-53
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• 2012

After success of sildenafil for the treatment of erectile dysfunction, a large number of its analogues have been approved from FDA. Recently, the illegal dietary supplements which include sildenafil, vardenafil, tadalafil, or analogues of these drugs as ingredient have been widely distributed. Therefore, the determination of the residue of synthetic phosphodiesterase- 5 (PDE-5) inhibitors in dietary supplements is highly required due to indiscriminate and unintentional overdose caused nausea, chest pains, fainting and irregular heartbeat. In this paper, we report a rapid and sensitive analytical method for the simultaneous determination of nine phosphodiesterase-5 inhibitors by liquid chromatography-high resolution mass spectrometry. The present method was found to be accurate and reproducible with 40 ${\mu}g$/g of the limit of quantification for the nine PDE-5 inhibitors. The developed method can be successfully applied to the analysis of the seven illegal dietary supplements.

• Mass Spectrometry Letters
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• 제4권3호
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• pp.47-50
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• 2013

An ultra-fast generic LC-MS/MS method was developed for high-throughput quantification of discovery pharmacokinetic (PK) samples and its reliability was verified. The method involves a simple protein precipitation for sample preparation and the analysis by ultra-fast generic LC-MS/MS with the ballistic gradient program and selected reaction monitoring (SRM) mode. Approximately 290 new chemical entities (NCEs) (over 10,000 samples) from 5 therapeutic programs were analyzed. The calibration curves showed good linearity in the concentration range of 1, 2 or 5 to 2000 ng/mL. No significant ion suppression was observed in the elution region of all the NCEs. When approximately 300 plasma samples were continuously analyzed, the peak area of internal standard was constant and reproducible. In the repeated analysis of samples, the plasma concentrations and the area under the curve (AUC) were consistent with the results from the first analysis. These results showed that the present ultra-fast generic LC-MS/MS method is reliable in terms of selectivity, sensitivity, and reproducibility and could be useful for high-throughput quantification and other bioanalysis in drug discovery.