• Natural Product Sciences
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• 제21권1호
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• pp.20-24
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• 2015

Cyanidin-3-glucoside (C3G) and cyanidin-3-rutinoside (C3R) were isolated by high-performance countercurrent chromatography (HPCCC) using a two-phase solvent system composed of tert-butyl methyl ether/n-butanol/acetonitrile/water/trifluoroacetic acid (1 : 3 : 1 : 5 : 0.01, v/v) to give pure C3G (34.1 mg) and C3R (14.3 mg) from 1.5 g crude mulberry fruit extract. Using the pure C3G and C3R, a reliable high-performance liquid chromatography (HPLC) method was developed and validated to determine the C3G and C3R contents in mulberry fruit. C3G and C3R were separated simultaneously using an Eclipse XDB-C18 column ($4.6{\times}250mm$ I.D., $5{\mu}m$) coupled with a photodiode array detector (PDA). The gradient elution of the mobile phase consisting of acetonitrile (0.5% formic acid) and water (0.5% formic acid) was applied (1.0 mL/min), and the detection wavelength was 520 nm. The calibration curves of C3G and C3R showed good linearity (both with $r^2=0.9996$) in the concentration range $15.625-500{\mu}g/mL$, and the relative standard deviations (RSD%) of intra- and inter-day variability were in the ranges 2.1 - 8.2% and 4.1 - 17.1%, respectively. The accuracies were ranged 96.5 - 102.6% for C3G and C3R, respectively. The developed HPLC method was used to determine the contents of C3G and C3R in newly harvested mulberry from eight different provinces of Korea.

• Journal of Pharmaceutical Investigation
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• 제35권3호
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• pp.137-142
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• 2005

A rapid, selective and sensitive reversed-phase HPLC method for the determination of glipizide in human serum was validated and applied to the pharmacokinetic study of glipizide. Glipizide and internal standard, tolbutamide, were extracted from human serum by liquid-liquid extraction with benzene and analyzed on a Nova Pak $C_{18}\;60{\AA}$ column with the mobile phase of acetonitrile-potassium dihydrogen phosphate (10 mM, pH 3.5) (4:6, v/v). Detection wavelength of 275 nm and flow rate of 0.7 ml/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^3$ factorial design using a fixed glipizide concentration (500 ng/ ml) with respect to its peak area and retention time. And also, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of 10-1000 ng/ml with correlation coefficient greater than 0.999. The lower limit of quantitation using 0.5 ml of serum was 10.0 ng/ml, which was sensitive enough for pharmacokinetic studies. The overall accuracy of the quality control samples ranged from 82.6 to 105.0% for glipizide with overall precision (% C.V.) being 1.13-13.20%. The percent recovery for human serum was in the range of 85.2 93.5%. Stability studies showed that glipizide was stable during storage, or during the assay procedure in human serum. The peak area and retention time of glipizide were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of glipizide in human serum samples for the pharmacokinetic studies at three different laboratories, demonstrating the suitability of the method.

• Journal of Pharmaceutical Investigation
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• 제35권6호
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• pp.437-443
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• 2005

A rapid, selective and sensitive reversed-phase HPLC method for the determination of a major metabolite of terfenadine, fexofenadine, in human serum was developed, validated, and applied to the pharmacokinetic study of terfenadine. Fexofenadine and internal standard, haloperidol were extracted from human serum by liquid-liquid extraction with acetonitrile and analyzed on a $Symmetry^{TM}$ C8 column with the mobile phase of 1% triethylamine phosphate (pH 3.7)-acetonitrile (67:33, v/v, adjusted to pH 5.6 with triethylamine). Detection wavelength of 230 nm for excitation, 280 nm for emission and flow rate of 1.0 mL/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^{3}$ factorial design using a fixed fexofenadine concentration (50 ng/mL) with respect to its peak area and retention time. In addition, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of 10-500 ng/mL with correlation coefficients greater than 0.999. The lower limit of quantification using 0.5 mL of serum was 10 ng/mL, which was sensitive enough for the pharmacokinetic studies of terfenadine. The overall accuracy of the quality control samples ranged from 95.70 to 114.58% for fexofenadine with overall precision (% C.V.) being 3.53-14.39%. The relative mean recovery of fexofenadine for human serum was 90.17%. Stability studies (freeze-thaw, short-term, extracted serum sample and stock solution) showed that fexofenadine was stable during storage, or during the assay procedure in human serum. However, the storage at $-70^{\circ}C$ for 4 weeks showed that fexofenadine was not stable. The peak area and retention time of fexofenadine were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of fexofenadine in human serum samples for the pharmacokinetic studies of orally administered Tafedine tablet (60 mg as terfenadine) at three different laboratories, demonstrating the suitability of the method.

• Journal of Pharmaceutical Investigation
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• 제35권4호
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• pp.265-271
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• 2005

A rapid, selective and sensitive reversed-phase HPLC method for the determination of etodolac in human serum was developed, validated, and applied to the pharmacokinetic study of etodolac. Etodolac and internal standard, ibuprofen were extracted from human serum by liquid-liquid extraction with hexane/isopropanol (95:5, v/v) and analyzed on a Luna C18(2) column with the mobile phase of 1% aqueous acetic acid-acetonitrile (4:6, v/v). Detection wavelength of 227 nm and flow rate of 1.0 mL/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^3$ factorial design using a fixed etodolac concentration $(1\;{\mu}g/mL)$ with respect to its peak area and retention time. And also, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of $0.05-40\;{\mu}g/mL$ with correlation coefficients greater than 0.999. The lower limit of quantification using 0.5 mL of serum was 0.05 ${\mu}g/mL$, which was sensitive enough for pharmacokinetic studies. The overall accuracy of the quality control samples ranged from 92.00 to 110.00% for etodolac with overall precision (% C.V.) being 1.08-10.11%. The percent recovery for human serum was in the range of 76.73-115.30%. Stability studies showed that etodolac was stable during storage, or during the assay procedure in human serum. The peak area and retention time of etodolac were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of etodolac in human serum samples for the pharmacokinetic studies of orally administered Lodin XL tablet (400 mg as etodolac) at three different laboratories, demonstrating the suitability of the method.

• Journal of Pharmaceutical Investigation
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• 제35권6호
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• pp.423-429
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• 2005

A selective and sensitive reversed-phase HPLC method for the determination of fenoprofen in human serum was developed, validated, and applied to the pharmacokinetic study of fenoprofen calcium. Fenoprofen and internal standard, ketoprofen, were extracted from human serum by liquid-liquid extraction with diethyl ether and analyzed on a Luna C18(2) column with the mobile phase of acetonitrile-3 mM potassium dihydrogen phosphate (32:68, v/v, adjusted to pH 6.6 with phosphoric acid). Detection wavelength of 272 nm and flow rate of 0.25 mL/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^{3}$ factorial design using a fixed fenoprofen concentration $(2\;{\mu}g/mL)$ with respect to its peak area and retention time. And also, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of $0.05-100\;{\mu}g/mL$ with correlation coefficients greater than 0.999. The lower limit of quantification using 1 mL of serum was $0.05\;{\mu}g/mL$, which was sensitive enough for pharmacokinetic studies. The overall accuracy of the quality control samples ranged from 92.27 to 109.20% for fenoprofen with overall precision (% C.V.) being 5.51-11.71 %. The relative mean recovery of fenoprofen for human serum was 81.7%. Stability (freeze-thaw, short and long-term) studies showed that fenoprofen was not stable during storage. But, extracted serum sample and stock solution were allowed to stand at ambient temperature for 12 hr prior to injection without affecting the quantification. The peak area and retention time of fenoprofen were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of fenoprofen in human serum samples for the pharmacokinetic studies of orally administered Fenopron tablet (600 mg as fenoprofen) at three different laboratories, demonstrating the suitability of the method.

• Journal of Pharmaceutical Investigation
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• 제36권2호
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• pp.89-95
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• 2006

A selective and sensitive reversed-phase HPLC method for the determination of pentoxifylline in human serum was developed, validated, and applied to the pharmacokinetic study of pentoxifylline. Pentoxifylline and internal standard, chloramphenicol, were extracted from the serum by liquid-liquid extraction with dichloromethane and analyzed on a Luna CI8(2) column with the mobile phase of acetonitrile-0.034 M phosphoric acid (25:75, v/v, adjusted to pH 4.0 with 10 M NaOH). Detection wavelength of 273 nm and flow rate of 0.8 mL/min were used. This method showed linear response over the concentration range of 10-500 ng/mL with correlation coefficients greater than 0.999. The lower limit of quantification using 0.5 mL of the serum was 10 ng/mL, which was sensitive enough for pharmacokinetic studies of pentoxifylline. The overall accuracy of the quality control samples ranged from 89.3 to 92.7% for pentoxifylline with overall precision (% C.V.) being 4.1-9.2%. The relative mean recovery of pentoxifylline for human serum was 105.8%. Stability (stock solution, short and long-term) studies showed that pentoxifylline was not stable during storage. But three freeze-thaw cycles and extracted serum samples were stable. This method showed good ruggedness (within 15% C.V.) and was successfully applied for the analysis of pentoxifylline in human serum samples for the pharmacokinetic studies of orally administered $Trental^{\circledR}$ tablet (400 mg pentoxifylline), demonstrating the suitability of the method.

• 한국산업보건학회지
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• 제15권1호
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• pp.1-7
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• 2005

Although the Fourier Transform Infra-Red spectrophotometric Direct on Filter(FTIR-DOF) method is a useful analytical technique for quantifying quartz content in respirable dust samples, a number of analytical problems must be taken into consideration such as, to name only a few, inhomogeneous deposition of particles, level of environmental humidity, uneven surface of the filter, and interfering minerals in the sample. This study was designed to select the most suitable wavelength and proper filter material for the method, and to investigate effects of humidity and inhomogeneous deposition of particles on the filter. Samples of respirable dust, created in a dust chamber containing standard material of quartz, were collected using a cyclone equipped with a 25mm filter as a collection medium. The results were as follows; 1. Among seven (7) commercially available filters tested for the FTIR-DOF method, the DM 800 filter showed the best analytical performance having the lowest background absorbance bands and no overlapping peaks at 799, 779, and $695cm^{-1}$. 2. The variations of absorbance due to humidity ranged from 1.0% to 3.3% for $799cm^{-1}$, 1.0% to 3.3% for $779cm^{-1}$, and 8.9%~20.9% for $695cm^{-1}$ peaks, respectively. The $699cm^{-1}$ peak was proved to be most vulnerble to environmental humidity for quantitative analysis of quartz. 3. As for effects of inhomogeneous deposition of samples, the highest variation of absorbance of 10.9% ($13.5{\mu}g$) was observed when using the 695cm-1. The variations of absorbance from the other two peaks, 799 and $779cm^{-1}$, ranged from 1.2 to 3.2%, and 1.4 to 4.1%, respectively. Therefore, the $799cm^{-1}$ peak was considered to be most reliable for quantitative analysis of quartz. The results of this study suggest that, for quantitative analysis of quartz in the respirable dust samples, use of the $799cm^{-1}$ peak can minimize the influence of environmental humidity and inhomogeneous deposition of particles on the filter. The FTIR-DOF method, if adopted for routine analysis of quartz in the respirable dust samples, could save sample preparation time and efforts substantially and also could increase analytical throughputs. Since use of the $799cm^{-1}$ peak is prone to be affected by interferences in the sample, further research on minimizing the effects is needed.

• 생약학회지
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• 제39권3호
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• pp.199-202
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• 2008

A high performance liquid chromatographic (HPLC) method for the simultaneous determination of hesperidin and glycyrrhizin was established for the quality control of traditional herbal medicinal preparation, Pyungwi-san (PWS). Separation and quantification were successfully achieved with a Waters XTerra RP18 column ($5{\mu}m$, 4.6 mm I.D. ${\times}$ 150 mm) by gradient elution of a mixture of acetonitrile and water containing 0.03% phosphoric acid (pH 2.03) at a flow rate of 1.0 ml/min. The diode-array UV/vis detector (DAD) was used for the detection and the wavelength for quantification was set at 230 nm. The presence of hesperidin and glycyrrhizin in this extract was ascertained by retention time, spiking with each authentic standard and UV spectrum. All four compounds showed good linearity $(r^2>0.995)$ in a relatively wide concentration ranges. The R.S.D. for intra-day and inter-day precision was less than 7.0% and the limits of detection (LOD) were less than 60 ng. The mean recovery of each compound was 99.0-105.6% with R.S.D. values less than 4.0%. This method was successfully applied to the determination of contents of hesperidin and glycyrrhizin in three commercial products of PWS. These results suggest that the developed HPLC method is simple, effective and could be readily utilized as a quality control method for commercial PWS products.

• 한국작물학회지
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• 제50권spc1호
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• pp.181-186
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• 2005

종실에 비해 다량의 rutin이 함유된 것으로 알려진 메밀 식물체의 최적 rutin함량 분석조건을 확립하기 위해 추출조건, HPLC 분석조건, 분석 재현성 및 최소검출한계를 검토한 결과 rutin은 미황색을 나타내는 폴리페놀 성분으로 에탄올에 용해 하여 최대 흡수파장을 측정하였을 때 359nm에서 흡수 극대파장을 나타내었다. 메밀 식물체 함유 rutin의 최적 HPLC분석 조건을 검토한 결과 컬럼은 Tosoh ODS 120T를 이용하고, 검출기 파장을 355nm, 이동상으로 A용매를 $2\%$ 초산함유-$45\%$ 아세토나이트릴로, B용매는 $2\%$ 초산함유-증류수로 하여 농도구배조건을 0분 : $50\%$ A용액, 0-18분 : $100\%$ A용액, 18-20분 : $50\%$ A용액, 20-22분 : $50\%$ A용액으로 조절하고, 분당 1mL를 용출 하였을 때 가장 우수한 분리가 유도되는 것을 확인할 수 있었다. 또한 HPLC 분석요인에 대한 재현성을 검정한 결과 머무름시간, peak면적 및 peak높이의 변이 계수가 모두 $0.6\%$미만을 나타내어 극히 높은 분석 재현성을 나타내는 것으로 조사되었으며, HPLC분석 시 최소검출한계는 20ng/mL수준이었다. 메밀 식물체 함유 rutin 분석을 위한 최적 추출조건을 검토한 결과 추출용매를 에탄올로 사용하고, $80^{\circ}C$ 조건에서 120분 이상의 환류추출을 시행하는 것이 가장 바람직한 추출법으로 조사되었다.

• KSBB Journal
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• 제23권1호
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• pp.83-89
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• 2008

음식물쓰레기를 처리하기 위한 3단계 메탄발효시스템으로부터 유출되는 음식물 발효 폐액은 고농도 유기성 폐수이다. 유기성 폐수는 고도처리 시스템에 의해 방류기준에 적합하게 처리되어져야만 한다. 본 연구에서는 유기성 폐수를 처리하기 위해 고효율 $UV/TiO_{2}$ 광촉매 산화공정의 최적 운전 조건을 조사하였다. 첫 번째 공정에서 폐수에 응집제인 $FeCl_{3}$를 전처리 하였으며, 응집을 위한 최적 pH와 응집제의 농도는 각각 pH 4와 2000 mg/L이었다. 이 공정을 통하여 최대 52.6%의 COD가 제거되었다. 두 번째는 $UV/TiO_{2}$ 광촉매 산화공정으로, 최적 운전 조건은 중심파장이 254 nm, 폐수 온도 및 pH가 각각 $40^{\circ}C$와 pH 8, 반응기 주입 공기량이 40 L/min인 것으로 조사되었다. 응집제를 이용한 전처리 공정과 광촉매 산화공정을 병합하여 최적조건에서 폐수를 처리할 경우 T-N과 COD의 제거율은 각각 69.7%와 70.9% 이었다.