• 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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• 제9권4호
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• pp.95-99
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• 2018

An innovative, simple, and rapid assay method based on liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) was developed and validated for the simultaneous determination of eight statin drugs in human urine. A simple sample clean-up procedure using the "dilute and shoot" (DAS) approach enabled a fast and reliable analysis. The influence of the dilution factor was investigated to ensure detectability and reduce the matrix effect. Chromatographic separation was performed on a Phenomenex Kinetex C18 column ($50{\times}3.0mm$ i.d., $2.6{\mu}m$) using an elution gradient of mobile phase A composed of 0.1% acetic acid, and mobile phase B composed of acetonitrile, at a flow rate of 0.35 mL/min. Quantitation was performed on a triple quadrupole mass spectrometer operated in multiple reaction monitoring (MRM) mode using electrospray ionization in positive ion mode. The total chromatographic run time was 15 min. The method was validated for selectivity, sensitivity, recovery, linearity, accuracy, precision, and stability. The present method was successfully applied to the analysis of Rosuvastatin in urine samples after oral administration to healthy human subjects.

• 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.

• Mass Spectrometry Letters
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• 제12권1호
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• pp.1-10
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• 2021

A simple, specific, and economical LC-MS/MS method was investigated for the screening of 43 prescribed antihypertensive and related drugs in human urine. The urine samples were simply prepared by diluting and mixing with internal standard before directly introduced to the LC-MS/MS system, which is fast, straightforward, and cost-effective. Fractional factorial, Box-Behnken, and I-optimal design were applied to screen and optimize the mass spectrometric and chromatographic factors. The analysis was carried out on a triple quadrupole mass spectrometer system utilizing multiple reaction monitoring with positive and negative electrospray ionization method. Chromatographic separation was performed on a Thermo Scientific Accucore RP-MS column (50 × 3.0 mm ID., 2.6 ㎛) using two separate gradient elution programs established with the same mobile phases. Chromatographic separation was performed within 12 min. The optimal method was validated based on FDA guideline. The results indicated that the assay was specific, reproducible, and sensitive with the limit of detection from 0.1 to 50.0 ㎍/L. The method was linear for all analytes with coefficient of determination ranging from 0.9870 to 0.9981. The intra-assay precision was from 1.44 to 19.87% and the inter-assay precision was between 2.69 and 18.54% with the recovery rate ranges from 84.54 to 119.78% for all drugs measured. All analytes in urine samples were stable for 24 h at 25℃, and for 2 weeks at -60℃. The developed method improves on currently existing methods by including larger number of cardiovascular medications and better sensitivity of 12 analytes.

• 약학회지
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• 제60권3호
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• pp.112-117
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• 2016

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the determination of caffeine in forensic aqueous sample. The centrifuged sample ($100{\mu}l$) was diluted 50-fold with distilled water. The diluted sample ($400{\mu}l$) was then diluted further with $200{\mu}l$ of 0.1% formic acid solution and $400{\mu}l$ of acetonitrile containing 500 ng of caffeine-(3-methyl-$^{13}C_3$) prior to LC-MS/MS analysis. The mobile phase was composed of 0.1% formic acid in distilled water (A) and acetonitrile (B). Chromatographic separation was performed by using a Zorbax SB-C18 ($100mm{\times}2.1mm$ i.d., $3.5{\mu}m$) column and caffeine was eluted within 1.1 min. Linear least-squares regression with a 1/x weighting factor was used to generate a calibration curve with the coefficients of determination ($r^2=0.9983$). The lower limit of quantification was $25ng/ml$ for the analyte. The process efficiency was 98.6~100.1%. Intra- and inter-day precisions were not more than 2.1% and 1.7%, while intra- and inter-day accuracies were ranged from -6.8 to 4.5%, respectively. The suitability of the method was examined by analyzing unknown forensic aqueous samples.

• Journal of Forest and Environmental Science
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• 제35권4호
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• pp.223-231
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• 2019

Castanopsis indica (Sil Batna) is an ecologically valuable multipurpose indigenous tree species of Bangladesh. Considering its high value but poor natural regeneration due to seed dormancy, the authors conducted an experiment at the Institute of Forestry and Environmental Sciences Chittagong University (IFESCU) during 2012-2013 to find out effective pre-sowing treatments. Ten pre-sowing seed treatments were applied on mature, even-sized seeds namely-control; sandpaper rubbing; nail clipping; immersion in normal water (at room temperature: 24℃) for 24 hrs., 48 hrs. and 72 hrs.; immersion in hot water (80℃) for 1 minute; soaking in 10% dilute H₂SO₄; soaking in 10% HCl for 5 minutes; and sowing in propagator house. Seeds sown after sandpaper rubbing at the distal end revealed best performances by providing highest germination percent (66.7%), germination energy (30%), germination index (0.17), germination rate (0.0145), germination value (30%) and plant percent (66.7%) within shortest period (38 days). The treatment also produced most vigor seedlings with 20.9 cm shoot height, 15 node number, and largest leaves (11.1 cm×2.9 cm). Hence, it is recommended to adopt sandpaper rubbing method for maximum germination and quality seedlings.

• 분석과학
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• 제31권4호
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• pp.161-170
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• 2018

메스암페타민(methamphetamine, MA)과 암페타민(amphetamine, AP)은 전세계적으로 널리 퍼진합성 흥분제로 보건 및 사회 문제, 경제적 비용 발생 등의 다양한 문제를 일으킨다. 또한 이러한 마약은 중독 및 남용 가능성이 매우 높기 때문에 확산 방지를 위한 방안의 하나로 복용자의 생체시료에서 이들 마약을 검출하기 위한 분석법 개발이 필요하다. 따라서 본 연구에서는 액체크로마토그래피-질량분석법(liquid chromatography-tandem mass spectrometry, LC-MS/MS)를 이용하여 소변에서 MA와 4-하이드록시메스암페타민(4-hydroxymethamphetamine, 4HMA), AP, 4-하이드록시암페타민(4-hydroxyamphetamine, 4HA)을 동시에 분석할 수 있는 분석법을 개발하였고 유효성 평가를 수행하였다. 전처리 시간 단축을 위해 50000 g에서 3 min 동안 초고속원심분리 후 시료 희석 주입법을 이용하여 LC-MS/MS에 주입하였다. 분리관은 역상 C18 컬럼을 사용하였고, 정량분석을 위해 MRM (multiple reaction monitoring) 모드를 적용하였다. 가중계수 $1/x^2$에서 정량 범위는 MA는 10-2500 ng/mL, AP는 1.0-800 ng/mL, 4HA와 4HMA는 2.0-200 ng/mL로 설정하였고, 검량선의 직선성은 결정계수($r^2$)를 구하여 평가하였다. 초고속원심분리법의 효율성을 확인하기 위해 전처리 과정으로 시린지 필터(membrane-filtration)를 적용한 결과와 비교하였고, 그 결과 분석물질에 따라 6-15 % 성능이 우수한 것으로 나타났다. 일내(intra-day)와 일간(inter-day) 정밀도는 6.6 % 미만이었고, 정확도는 -14.9-11.3 % 였다. 최저정량한계(LLOQ)는 2.0 ng/mL (4HA 및 4HMA), 1.0 ng/mL (AP), 10 ng/mL (MA)로 확인되었다. 선택성, 검출한계, 희석무결성, 기질효과, 효율성, 안정성을 평가한 결과 만족스러운 측정값을 얻었다. 또한 개발된 분석법을 마약 복용자의 소변에 적용하여 분석법의 유용성을 확인하였다.

• 한국산림과학회지
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• 제1권1호
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• pp.12-21
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• 1962

리기다소나무와 젓나무의 야외삽목(野外揷木)이 수원농과대학(水原農科大學) 임업묘포(林業苗圃)에서 1956년(年) 실시(實施)되었는데 다음과 같은 사실(事實)이 확인(確認)되었다. 리기다소나무(pinus rigida)에 있어서는 총 1335본의 삽수가 3년생 및 10년생의 모수에서 얻어지고 표(表)에서 보이는바와 같은 처리(處理)를 받고 3월(月) 13일(日) 및 4월(月) 20일에 삽목(揷木)되었다. 이에 있어서는 (1) 유령목(幼齡木)일수록 발근(發根)이 높다는것과 50ppm의 IBA 또는 NAA가 가장 좋은 결과(34 per cent)를 보여주었다. (2) lammas shoot에서는 전연 발근이 되지 않았다. (3) 삽목시기(揷木時期)가 또한 발근율(發根率)에 영향을 주었다. (4) 발근기원(發根起元)은 삽수(揷穗)의 하단부위(下端部位) 또는 절구(切口)에 생긴 Callus 부터였다. 젓나무(Abies holophylla)에 있어서는 30년생의 모수에서 삽수(揷穗)가 채취(採取)되었고 3월(月) 23일(日)에 삽목(揷木)되었다. 이에 있어서는 (1) 수관(樹冠)의 상반부(上半部)에서 채취(採取)된 삽수는 전연 발근(發根)하지 않았다. (2) 수관(樹冠)의 하반부(下半部)에서 얻어진 삽수(揷穗)에 있어서는 20ppm IAA, 20ppm IBA, 2)ppm NAA, 20ppm thiamine 및 5 percent sucrose의 혼함용액이 가장 좋은 발근율(發根率)(22 per cent)를 보여주었다. (3) 모든 발근(發根)은 불규칙(不規則)한 parenchymatous 조직(組織)에서 유래했었다.