• 한국응용과학기술학회지
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• 제33권3호
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• pp.459-465
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• 2016

This study investigated reusability of costly guard column by ultrasonic. It also investigated various effects that affect to guard column generation by ultrasonic. When investigated 30 KHz of frequency, area of ascorbic acid is 73.0% compared to unused guard column. As a result of investigation of effect of pH, guard column by ultrasonic is effective at alkali area. As a result of investigation of solvent effect, when ethanol is used, generation rate is 81.9% as of peak area compared to the case of analysis in un used column. From the result, it indicates that regenerated guard by ultrasonic is reusable.

• 환경위생공학
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• 제21권4호
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• pp.29-38
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• 2006

This study included the development of analytical method for determining perchlorate in water sample. The analytical condition was referred in EPA 314.0 method which use ion chromatography and the concentrator column was replaced by the guard column. Concentrating 10mL raw or treated water sample on to AGl6 guard column made it possible to get the LOD(Limit of Detection) of $0.73\;{\mu}g/L$. The total run time was 11 minutes and during run time next sample could be concentrated on AGl6 guard column. Compared to the Concentration method which needed manual operation, the Direct Injection method could screen the many water samples. The LOD of the Direct Injection method was higher and the sensitivity was lower than that of the Concentration method. The RSDs(Relative Standard Deviations) were lower than 2.5 % for peak height and 0.7 % for retention time in pre-concentration methods. This method Showed good reproducibility and reliability and it was thought the deviations of recovery value could be reduced by considering column capacity and making water sample homogeneous. Matrix Elimination could be done using the pre-concentration method if perchlorate were in complex matrix of sample.

• FOCUS: LIFE SCIENCE
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• 제1호
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• pp.5.1-5.3
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• 2024

The document discusses the regeneration of Partisil/Partisphere ion-exchange columns in chromatography. It mentions that column efficiency can diminish with use due to the accumulation of sample and/or mobile phase impurities at the head of the column. This can lead to a change in back pressure, lower column efficiency, and sometimes a change in selectivity. The document outlines a procedure that may restore column performance. The document also provides everyday practices to enhance the lifetime of a column. These include using only high-purity HPLC solvents and buffers, using freshly prepared mobile phases and buffers, filtering mobile phases to remove particulates, using appropriate sample clean-up procedures, using a guard column or pre-column filter, and working within the pressure and flow rate limitations of the column. For the regeneration of Partisil/Partisphere SAX, SCX, WAX, and WCX columns, the document suggests passing 20 column volumes of various mobile phases through the column. These include a buffer wash, distilled water, an acid wash, a chelating wash, a methanol wash, and a buffer for separation. The document emphasizes that not all of these wash steps are required for every column clean-up and that some chromatographers require only a combination of certain steps.

• FOCUS: LIFE SCIENCE
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• 제1호
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• pp.1.1-1.4
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• 2024

This article provides comprehensive guidance on the maintenance, cleaning, regeneration, and storage of silica-based HPLC (High-Performance Liquid Chromatography) columns. The general considerations emphasize the importance of using in-line filters and guard cartridges to protect columns from blockage and irreversible sample adsorption. While these measures help, contamination by strongly adsorbed sample components can still occur over time, leading to an increase in back pressure, loss of efficiency, and other issues. To maximize column lifetime, especially with UHPLC (Ultra-High Performance Liquid Chromatography) columns, it is advisable to use ultra-pure solvents, freshly prepared aqueous mobile phases, and to filter all samples, standards, and mobile phases. Additionally, an in-line filter system and sample clean-up on dirty samples are recommended. However, in cases of irreversible compound adsorption or column voiding, regeneration may not be possible. The document also provides specific recommendations for column cleaning procedures, including the flushing procedures for various types of columns such as reversed phase, unbonded silica, bonded normal phase, anion exchange, cation exchange, and size exclusion columns for proteins. The flushing procedures involve using specific solvents in a series to clean and regenerate the columns. It is emphasized that the flow rate during flushing should not exceed the specified limit for the particular column, and the last solvent used should be compatible with the mobile phase. Furthermore, the article outlines the storage conditions for silica based HPLC columns, highlighting the impact of storage conditions on the column's lifetime. It is recommended to flush all buffers, salts, and ion-pairing reagents from the column before storage. The storage solvent should ideally match the one used in the initial column test chromatogram provided by the manufacturer, and column end plugs should be fitted to prevent solvent evaporation and drying out of the packing bed.

• 분석과학
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• 제19권5호
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• pp.383-388
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• 2006

간단한 후세척의 이온 크로마토그래피를 이용하여 형석 광화대 지역의 광산 폐수에서 플루오라이드 이온을 빠르고 정확하게 정량하는 방법을 확립하였다. 시료중의 높은 농도의 황산 이온은 전치 컬럼(pre-column)에 머물러있고 머무름 시간이 적은 플루오라이드 이온은 제시된 이온 크로마토그래프 시스템과 스위칭 기술을 이용하여 분리 시스템에 주입되었다. 사전 농축(pre-concentration) 컬럼은 다량의 황산 이온을 보유할 수 있는 이온 교환 용량이 큰 분석용 컬럼(AS 9 HC)을 사용하였다. 분리용 컬럼은 플루오라이드 이온의 감도를 증가시키기 위하여 틈새 부피(void volumn)와 시스템 압력을 감소시킬 수 있도록 보호 컬럼(AG 14)을 사용하였다. 첫 번째 전기전도도 검출기로는 플루오라이드 이온의 회수율을, 두 번째 전기전도도 검출기로는 제거되는 황산 이온 피크를 관찰함으로써 결정된 최적의 10-포트주입기의 스위칭 시간은 4.3 분 이었다. 시료 주입량이 $25{\mu}L$ 일때, $500mg\;L^{-1}$ 의 높은 황산 이온이 포함된 용액에서 플루오라이드 이온의 검출한계(S/N = 3)는 $2.4{\mu}g/L$였다.

• 해양환경안전학회지
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• 제13권1호
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• pp.61-67
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• 2007

본 논문에서는 어장에 침입하는 도적을 감시하기위한 어장보호 시스템의 구현절차와 구현결과를 기술하였다. 이 시스템은 사용자의 요구에 따라 저가부터 고가 및 단순 기능부터 복합 기능 등 다양한 형태로 구성할 수 있도록 설계하였다. 육상으로부터 0.5 마일 이내에 총 50개(가로 10열, 세로 5열)의 케이지가 밀집된 전라남도 진도군 소재 전복 양식장에서 구축한 시스템을 현장 실험하였다. 그 결과, 구축한 시스템이 양식장 전체는 물론 단위 케이지까지 세밀하게 감시할 수 있음을 확인하였다.

• 대한본초학회지
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• 제22권4호
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• pp.261-270
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• 2007

Objective : The 5-HMF was not index material suitable to do the quality control of Rehmanniae Radix Preparata. In this study, We estimated the changes of oligosaccharide contents in Rehmanniae Radix Preparata using high-performance anion-exchange chromatography with pulsed amperometric detection(HPAEC-PAD). Methods : The analysis of oligosaccharide was conducted by HPAEC-PAD with Carbopac PA1, $250{\times}4mm$, 5um, and Carbopac PA1 guard column. Column temperature was kept at $30^{\circ}C$. Elution was carried out at 1000 ${\mu}l/min$ with 70mM NaOH and the injection volume was $10{\mu}l$. Each component was detected by PAD. Results : Nine constituents were found from merchandising Rehmanniae Radix Preparata(MR), while seven constituents were found in various processed Rehmanniae Radix Preparata. Not all constituents were defined but stachyose and raffinose were found in all cases. And The most common constituents of Rehmanniae radix was stachyose. In the course of processing, most of stachyose and raffinose were decreased. Stachyose was decreased slowly in the course of processing with rice wine(RR), amomi and rice wine(AR), and crataegi and rice wine(CR). However stachyose was decreased rapidly in the course of processing with fresh rehmannia juice(FR). The method with crataegi and rice wine(CR) showed the smallest decrease of stachyose. And processing method with crataegi and rice wine(CR) showed the most abundant amount for stachyose after the nineth processing. Conclusion : The changes of oligosaccharides in the course of processing were a very important direct barometers to do the quality control and set up a standard of Rehmanniae Radix Preparata.

• 한국산업보건학회지
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• 제6권1호
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• pp.28-37
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• 1996

Benzidine, an aromatic amine used primarily in the manufacture of azo dyes, is recognized as a urinary bladder carcinogen in humans. In rats, mice, and hamsters, chronic exposure to benzidine resulted in tumors of the liver. The present study was undertaken to suggest analyzing the metabolites of benzidine with the optimal condition, identify the metabolites of benzidine, and observe time variance of the metabolites in the isolated perfusated rat liver. N-acetylbenzidine was synthesized by acetylation of benzidine with acetic anhydride and separated by thin layer chromatography(TLC) and high performance liquid chromatography(HPLC). To analysis benzidine and the metabolites of benzidine, HPLC operating condition has been optimized by means of preliminary experiment. The mobile phase consisted of acetonitrile(37%) in phosphate buffer, flow rate maintained at 1.0 ml/min. Optimal detective conditions were electrochemicaldetector(ECD) at 0.75 V for benzidine and N-acetylbenzidine and ultravioletdetector(UVD) at 287 nm for N,N'-diacetylbenzidine. The separation system was composed of a guard column and a separation column(Polymer C18, $4.6{\times}250cm$) at a temparature of $40^{\circ}C$. The perfusion system was equilibrated for 30 minutes before addition of benzidine to the perfusate. Samples of the perfusate were collected at time intervals(0, 10, 20, 30, 60, 90, 120 min) during the 2 hour perfusion. Before analyzing samples by HPLC/ECD/UVD, samples had been treated with sep-pak. Samples of perfusate analyzed by HPLC/ECD/UVD and the metabolites of benzidine in the isolated perfused rat liver were N-acetylbenzidine and N,N'-diacetylbenzidine. Benzidine metabolized over 60% during the initial 30 minutes of perfusion, extensively by 1 hour, and was undetectable in the perfusate. N-acetylbenzidine increased by 30 minutes of perfusion, declined. N,N'-diacetylbenzidine increased the 0-90 minutes period, remained constant during the 90-120 minutes period.

• 약학회지
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• 제51권2호
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• pp.96-102
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• 2007

Measurement of globotriaosylceramide (Gb3, ceramide trihexoside) in urine has clinical importance for monitoring after enzyme replacement therapy in Fabry disease patients. The disease is an X-linked lipid storage disorder that results from a deficiency of the enzyme ${\alpha}$-galactosidase A (${\alpha}$-Gal A). The lack of ${\alpha}$-Gal A causes an intracellular accumulation of glycosphingolipids, mainly Gb3. A simple, rapid, and highly sensitive analytical method for Gb3 in urine was developed without labor-extensive pre-treatment by electrospray ionization MS/MS (ESI-MS/MS). Only simple 5-fold dilution of urine is necessary for the extraction and isolation of Gb3 in urine. Gb3 in diluted urine was dissolved in dioxane containing C17:0 Gb3 as an internal standard. After centrifugation it was directly injected and analyzed through guard column by in combination with multiple reaction monitoring mode of ESI-MS/MS. Eight isoforms of Gb3 were completely resolved from urine matrix. C24:0 Gb3 occupied 50% of total Gb3 as a major component in urine. Linear relationship for Gb3 isoforms was found in the range of 0.005${\sim}$5.0 ${\mu}$g/ml. The limit of detection (S/N=5) was 0.005 ${\mu}$g/ml and limit of quantification was 0.05 ${\mu}$g/ml for C24:0 Gb3 with acceptable precision and accuracy. Correlation coefficient of calibration curves for 8 Gb3 isoforms ranged from 0.9598 to 0.9975. This method could be useful for rapid and sensitive 1st line Fabry disease screening, monitoring and/or diagnostic tool for Fabry disease.

• Journal of Genetic Medicine
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• 제4권1호
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• pp.45-52
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• 2007

목 적 : 패브리병은 X-linked 지질 축적 질환으로 ${\alpha}$-galactosidase A (${\alpha}$-Gal A)의 결손으로 인해 스핑고당지질인 Gb3의 세포내 축적을 일으키는 병이다. 혈장 중 Gb3 측정은 패브리병 환자의 효소대체요법 후의 모니터링이나 진단에 임상적 의의가 크므로 ESI-MS/MS를 이용한 시료 전처리를 위한 노동력이 덜 들면서 간단, 신속, 고감도로 정량할 수 있는 혈장 중 Gb3분석법을 개발하고자 하였다. 방 법 : 혈장을 디옥산으로 50배 희석하여 vortex-mix 및 원심분리를 거쳐 Gb3의 추출 및 분리를 수행한다. 이 때 내부표준액인 C17:0 Gb3를 혈장에 처음부터 첨가한다. 희석과 원심분리된 혈장은 가드컬럼을 통하여 ESI-MS/MS의 다중성분 모니터링 모드에서 분석하여 내부표준액에 대한 8종 Gb3 isoform의 피크면적비를 이용하여 정량한다. 결 과 : 혈장의 바탕성분 하에서 8종의 Gb3 isoform이 완전히 잘 분리됨을 확인할 수 있었다. 혈장 중의 8종의 Gb3 isoform 중 50% 이상 차지하는 종류는 C16:0 Gb3 임이 확인되었다. Gb3 isoform이 직선성을 이루는 농도 범위는 0.001-1.0 ${\mu}g$/mL이었다. 검출한계(S/N=3)는 C16:0 Gb3의 경우 0.001 ${\mu}g$/mL 이었고 정량한계는 0.01 ${\mu}g$/mL 이었으며 회수율의 일내재현성(정확도 87-108%와 정밀도 7% 이하)과 일간재현성(정확도 87-110%와 정밀도 13% 이하)은 매우 양호 하였다. 결 론 : 본 연구에서 개발된 Gb3 분석법은 신속, 정확, 간편하게 패브리병의 1차 스크리닝이나 효소대체요법 전후의 모니터링 및 진단에 유용하게 적용될 수 있을 것이다.