• Bulletin of the Korean Chemical Society
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• 제31권5호
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• pp.1159-1164
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• 2010

Several triterpenoid saponins from root of Pulsatilla koreana Nakai (Ranunculaceae) were studied and their biological activities were reported. It is difficult to analyze triterpenoid saponins using HPLC-UV due to the lack of chromophores. So, evaporative light scattering detection (ELSD) is used as a valuable alternative to UV detection. More recently, a charged aerosol detection (CAD) has been developed to improve the sensitivity and reproducibility of ELSD. In this study, we developed and validated a novel method of high performance liquid chromatography coupled with a charged aerosol detector for the simultaneous determination of four triterpenoid saponins: pulsatilloside E, pulsatilla saponin H, anemoside B4 and cussosaponin C. Analytes were separated by the Supelco Ascentis$^{(R)}$ Express C18 column (4.6 mm ${\times}$ 150 mm, 2.7 ${\mu}m$) with gradient elution of methanol and water at a flow rate of 0.8 mL/min at $30^{\circ}C$. We examined various factors that could affect the sensitivity of the detectors, including various concentrations of additives, the pH of the mobile phase, and the CAD range. Linear calibration curves were obtained within the concentration ranges of 2 - 200 ${\mu}g$/mL for pulsatilloside E, anemoside $B_4$ and cussosaponin C, and 5 - 500 ${\mu}g$/mL for pulsatilla saponin H with correlation coefficient ($R^2$) greater than 0.995. The limit of detection (LOD) and quantification (LOQ) were 0.04 - 0.2 and 2 - 5 ${\mu}g$/mL, respectively. The validity of the developed HPLC-CAD method was confirmed by satisfactory values of linearity, intra- and inter-day accuracy and precision. This method could be successfully applied to quality evaluation, quality control and monitoring of Pulsatilla koreana.

• 한국동물위생학회지
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• 제28권3호
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• pp.203-213
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• 2005

Oxytetracycline, tetracycline, chlortetracycline and doxycycline in honey were separated by solid phase extraction (SPE) and determined with high performance liquid chromatography (HPLC) with UV/Visible detector. Analysis was carried out using following conditions: XTerra $C_8$ column $(3.9\times150mm\;i.d. 5{\mu}m)$, mobile phase composed of 0.01M oxalic acid : methanol : acetonitrile (820 : 80 : 100, v/v/v), isocratic pump at a flow rate of 0.9 ml/min. and $50{\mu}l$ of injection volume, UV/Visible detector with wavelength of 360nm. The calibration curves of four tetracyclines showed linearity $(\gamma^2>0.999)$ at concentration range of $100\~1,000 ng/ml$. The recoveries in fortified honey represented more than $70\%$ with low coefficient of variation $(<10\%)$ for concentration range of four tetracyclines. The detection limits for oxytetracycline, tetracycline, chlortetracycline and doxycycline were 13.8, 14.6, 26.2 and 24.9ng/g in acacia honey. respectively. We also monitored tetracyclines residue in domestic honey [n : 38, acacia (20), wild flower (18) ] and foreign honey [n=22, legally distributed (13), illegally distributed (9)] using modified Charm II screening and HPLC confirmation methods. Seven of the 60 samples $(11.7\%)$ were suspect positive using modified Charm II screening test. Chlortetracycline residue was found in one foreign honey (illegally distributed) tested at concentrations of 0.22 ppm. Conclusively, for more effective control of tetracyclines used in beekeeping should be further survey for residues in honey and also national guidelines (maximum residue limit : MRL) and methods should be obligatory.

• 분석과학
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• 제17권2호
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• pp.98-107
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• 2004

본 연구에서는 $^1H$ NMR 분광분석법 및 HPLC를 이용하여 자동차 부품에 사용된 아크릴 코팅의 성분 및 함량을 분석하였다. $^1H$ NMR 분석 결과 아크릴 코팅은 88.40 wt%의 PMMA, 7.05 wt%의 MMA 및 2.36 wt%의 allyl methacrylate를 포함하고 있다. 또한, 고분자 첨가제로 벤조트리아졸 (Hisorb 328) 및 oxanilide 광안정제, BHT 및 DMP가 $^1H$ NMR 스펙트럼에서 확인되었고 정량적으로 측정되었다. 그러나, Bondapak C18 칼럼, 메탄올 이동상, PDA 검출기를 사용한 역상 HPLC 분석을 통해서는 두개의 광안정제만이 확인되었다. 두 광안정제의 함량은 UV-Vis 흡광광도계를 이용한 정량분석법을 통해 측정되고 NMR 분석 결과와 비교하였다. $^1H$ NMR 스펙트럼으로부터 얻은 분석 정보들이 HPLC에서 얻는 것들 보다 우수하였다.

• 분석과학
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• 제20권5호
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• pp.424-433
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• 2007

혈장시료 중의 갑상선 호르몬을 고체상 추출법을 이용하여 추출한 후 HPLC/DAD/ESI-MS (high-performance liquid chromatograph/diode array detector/electrospray ionization-mass spectrometer)를 사용하여 분석하였다. 7종의 thyroid hormones의 HPLC 분리조건은 Hypersil ODS 컬럼(4.6 mm I.D., 250 mm length, particle size $5{\mu}m$)을 사용하고 ammonium formate buffer와 아세토나이트릴을 이동상으로 하여 기울기 용리한 결과 완전 분리가 가능하였으며, UV spectra 및 질량스펙트럼을 확인할 수 있었다. 고체상추출법에 의한 전처리 최적 조건을 조사한 결과 시료를 pH 3으로 한 후 C18 고체상을 사용하여 4 mL의 메탄올로 용리할 경우 회수율이 74.5-115.7%로 나타났다. HPLC/ESI-MS를 이용하여 20-500 ng/mL범위에서 검량선을 작성한 결과 $r^2$값은 0.9939-0.9978으로 나타났으며 검출한계는 20-50 ng/mL (38.1-162.8 pmol/mL)로 계산되었다. 정상인의 혈장에서 thyroxine의 정량이 가능하였으며 그 결과 50.98-112.97 ng/mL로 검출되었다.

• Biomolecules & Therapeutics
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• 제16권2호
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• pp.168-172
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• 2008

The present study is to determine of sensitive nicorandil analysis method using HPLC and measure the pharmacokinetics parameters (bioavailability, $C_{max}$, $T_{max}$, Ke, $T_{1/2}$) of nicorandil (5 mg, Tab; Choongwae Pharma Corporation). Plasma (500 ul) was mixed with furosemide (internal standard, 500 ug/ml). Detection wavelength was 256 nm. The mixture of 0.01 M ammonium acetate and acetonitrile 80:20 (v/v) was used mobile phase. The HPLC separation was accomplished on ODC reverse HPLC column. The nicorandil was analyzed by a HPLC system, which consists of CAPCELL PAK C18 column (5 ${\mu}$m, 4.6 × 150 mm) and a chromatography data analysis S/W, using a isocratic mobile phase (mixture of 0.01 M ammonium acetate and acetonitrile 80:20 ) at 1.0 ml/min. Its sensitivity, selectivity, accuracy and precision must be adequate for the bioavailabilty study of nicorandil, and the linearity ($r^2$ ≥ 0.9994) of nicorandil was also proved in the range of 0.05 ug/ml . 3 ug/ml. The pharmacokinetic parameters of nicorandil (5 mg) tablets were measured as the follow. AUC: 0.19 ug/ml·hr, $C_{max}$: 0.14 ug/ml, $t_{max}$: 0.58 hr, Ke: 0.11 hr., $t_{1/2\beta}$: 6.76 hrs. This method is simple and sensitive HPLC method using UV detector for determination of nicorandil in human plasma.

• 한국어병학회지
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• 제20권2호
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• pp.139-145
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• 2007

인체에 널리 사용되고 있는 Aspirin (ASA)을 양식 뱀장어 (Anguilla japonica) 약욕 또는 경구 투여한 다음 혈장 중의 ASA와 salicylic acid (SA)의 양을 HPLC로 측정하였다. 뱀장어 혈장은 0.2 M HCl과 0.2 M orthophosphoric acid로 산성화시킨 다음 acetonitrile과 혼합하여 ASA와 SA를 추출하였다. 2약제의 정량은 Novapak C18과 UV detector (237 nm)가 장착된 HPLC로 측정하였다. 이 때 이동상은 740 ㎖의 증류수, 900 ㎕의 orthophosphoric acid와 180㎖의 acetonitrile을 사용하였다. ASA, SA 및 내부 표준물질로 사용한 2-methylbenzoic acid (MBA)의 retention time은 각각 4.8분, 8.4분 및 11.4분이였으며, 측정한계 농도는 ASA가 0.05 ㎍/㎖, SA는 0.01 ㎍/㎖였다. 혈장으로 부터의 평균회수율은 ASA가 70.8-99.6%, SA는 95.2-100.3%였다. 뱀장어에 ASA를 약욕 (20 ppm) 또는 경구투여 (50 ㎎/kg BW) 한 다음 채취한 혈장을 시료로 이 방법으로 ASA와 SA의 양을 측정한 결과 단지 SA만 검출되어졌고, ASA는 검출되지 않았다. 이는 ASA가 혈장내에서 신속히 SA로 분해되기 때문으로 판명되었다. 또 ASA에 약욕시킨 경우에는 약욕 후 3시간후에 혈장내의 SA양이 최고치에 도달하였으며, 경구투여 한 경우에는 7일후에 최고치에 도달하였다. 한편 ASA를 투여한 다음 ASA 무첨가 수조에 수용한 결과 2투여 경로 모두 48시간 이후에는 SA가 0.02-0.03 ㎍/㎖이 검출되어 잔류의 문제도 거의 없었다. HPLC를 이용한 혈장내의 ASA와 SA의 검출법은 신속하며 정확한 방법으로 뱀장어 이외의 어류에도 활용 가능할 것으로 생각된다.

• 약학회지
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• 제31권2호
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• pp.105-111
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• 1987

A high performance liquid chromatographic method was developed for the simultaneous determination of free and glycine conjugated bile acids. Free and glycine conjugated bile acids were extracted from bear gall bladder by methanol and from serum using a Sep-pak $C_{18}$ catridge. The extracted bile acids were labeled with 2-bromoacetyltriphenylene in acetonitrite using 18-crown-6-ether as a catalyst. Derivatized bile acids were separated from the individual bile acids on a reversedphase column (Chemcosorb 5-ODS-H) using acetonitrile-methanol-water(10:50:30) as a mobile phase and monitored by an UV-detector at 254nm. Linearities of calibration curve were obtained between 4 ng and 24 ng, and recoveries from bear gall bladder sample were higher than 94%.

• 한국식품위생안전성학회지
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• 제10권3호
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• pp.169-174
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• 1995

Piperine, component of pure ground black pepper, has strong stimulative and hot. Analytical method for piperine was developed by high performance liquid chromatography. Analytical conditions are as follows, mobile phase is 70% methanol, detector UV 343 nm (0.05 AuFs), column is Novapak 5 C18 (15 cm $\times$ 4.6mm), flow rate is 1.0ml/min, chart speed is 0.25 cm/min and injection volume is 20 ul. Analytical results are as follows that relative standard deviation is 1.15%, calibration curve is y=170473.1x-7848.5 (R2=0.999) that shows good linearity. Standard solution of piperine is stable up to 10 hr and content of piperine in pureground black pepper is 4.97$\pm$0.86% Retention time of piperine in HPLC method is about 7 min. Therefore, the developed HPLC method including simple pretreatment of sample will be contribute to quality mangement.

• Asian-Australasian Journal of Animal Sciences
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• 제16권12호
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• pp.1799-1803
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• 2003

The possibility of lycopene affecting egg yolk pigmentation was studied with lycopene diets containing 0, 4, 8, and $12{\mu}g/g$ meal, respectively. The addition of lycopene above $4{\mu}g/g$ meal significantly improved yolk color after four days of supplementation. The transfer of lycopene into egg yolk was confirmed by thin layer chromatography, and high-performance liquid chromatography-mass spectrometry (HPLC-MS). The deposition rate of lycopene into egg yolk was approximately 2%, which was quantitatively determined using a HPLC with a UV detector. The result indicates that lycopene is a good candidate for egg yolk pigmentation and for making functional eggs.

• 대한화장품학회지
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• 제24권3호
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• pp.123-128
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• 1998

Dihydroxyacetone (DHA) has been used as a self tanning agent and many emulsion formulations containing DHA have been studied. In an emulsion, many factors which have negative effect on DHP and the resultant DHA decomposition can destabilize the emulsion base. In this study, two kinds of emulsion with 5％ DHA were prepared, O/W type emulsion and Multilamellavesicle (MLV) type emulsion to compare the stabilization effects of both emulsions on the DHA. The OHA concentration was analyzed quantitatively by high performance liquid Chromatography (HPLC), also the pH and viscosity of both emulsions were measured for stability. This process was carried out over 4 months. For HPLC, a bondaclone $C_{18}$ column with a mobile phase of distilled water and UV detector were used. The results of these experiment showed that DHA is more stable in an MLV emulsion than it is in an O/W type emulsion.