• Food Science and Biotechnology
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• v.15 no.5
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• pp.778-780
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• 2006

High performance liquid chromatography (HPLC) was used for the analysis of the lignans eleutheroside B and eleutheroside E in the fruits of Acanthopanax species. A reverse-phase system using a gradient of $H_2O$ and acetonitrile as the mobile phase was developed and detection was at 210 nm. The analysis was successfully carried out within 20 min. The content of eleutheroside B and eleutheroside E in Acanthopanax species was measured in the fruits of A. senticosus (0.58 and $1.66\;{\mu}g/mg$, respectively), A. sessiliflorus (1.15 and $8.49\;{\mu}g/mg$, respectively), A. koreanum (2.16 and $1.80\;{\mu}g/mg$, respectively), and A. divaricatus (1.06 and $7.08\;{\mu}g/mg$, respectively).

• Journal of Pharmaceutical Investigation
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• v.22 no.1
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• pp.63-68
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• 1992

An HPLC procedure with UV detection has been developed for the quantitation of flurbiprofen released into isopropyl myristate used as the receptor phase in an in vitro membraneless drug diffusion cell. The drug and the internal standard (oxaprozin) were extracted from isopropyl myristate with a mixture of dimethylsulfoxide:methanol:water (2:1:1) and quantitated using a reverse phase $C_{18}$ column. The chromatograms were completely free from interfering peaks, and the relative retention times of flurbiprofen and the internal standard were 4.9 and 6.8 min, respectively. Calibration plots were linear over the concentration range of $1-200\;{\mu}g/ml$ of flurbiprofen with correlation coefficients, all higher than 0.99. The mean intra-day precision and accuracy among three replicate sets of the assay in a day were 4.26 and 4.52%, respectively, whereas the mean inter-day precision and accuracy were 3.35 and 3.64%, respectively. The mean recovery of the drug was 92.5% over the calibration range. The method was simple, reliable and accurate for the quantitation of flurbiprofen in unpurified isopropyl myristate.

• Korean Journal of Veterinary Service
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• v.26 no.2
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• pp.157-162
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• 2003

This stud was attempted to detect six benzimidazoles[thiabendazole(TBZ), oxfendazole(OFZ), mebendazole(MBZ), flubendazole(FLBZ), albendazole(ABZ), and febendazole(FBZ)] in eggs using high performance liquid chromatography(HPLC) with photodiode array detector(DAD) simultaneously. The eluates were determined by HPLC on a waters X-Terra$^{TM}\;C_{18}$ reverse-phase column($4.6{\times}250nm,\;5{\mu}m$) with DAD at 295nm. The mobile phase was 0.04M ammonium phosphate(pH 7.5)/ACN(62.28 v/v) run isocratically. The calibration curves were linear(r>0.999) for six benzimidazoles. Average recovery rate from eggs were 94.54% for TBZ, 97.71% for OFZ, 88.82% for MBZ, 93.71% for FLBZ, 78.61% for ABZ, and 75.09% for FBZ, respectively. The limits of detection and quantification were 2.27ppb, 3.88 for TBZ, 9.34ppb, 15.61ppb for OFZ, 28.53ppb, 45.15ppb for MBZ, 27.39ppb, 40.95ppb for FBZ, 8.61ppb, 13.95ppb for ABZ and 12.79ppb, 22.34ppb for FBZ, respectively.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.2
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• pp.300-306
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• 2000

In the present studies, we assessed the stability of active components and toxicological safety of irradiated Angelica gigas Nakai(Danggui). In order to confirm the stability of active components in the ${\gamma}$-irradiated roots of Danggui, the quantitative analysis of decursin and decursinol angelate of ${\gamma}$-irradiated sample was carried out by high performance liquid chromatographic (HPLC) methods using reverse phase columns and normal phase columns. From the root of Danggui, decursin and decursinol angelate were isolated by a silica gel column chromatography(toluene : ether (1 : 1), Hexane : EtOAc(15 : 1)). And then the structures were confirmed in the 1H and 13C-NMR analysis. The HPLC chromatograms of decursin and decursinol angelate in ${\gamma}$-irradiated Danggui were similar with those of non-irradiated sample. In the examination of in vitro genotoxicity of the water extract from ${\gamma}$-irradiated Danggui using Salmonella reversion assay(Ames test) and micronucleus test in Chinese hamster ovary (CHO) cells, mutagenicity was not exhibited in the two assays with or without metabolic activation. These resutls suggest that active components in the ${\gamma}$-irradiated Danggui should be stable and that the safety of ${\gamma}$-irradiated Danggui could be revealed in further test in vivo.

• Korean Journal of Breeding Science
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• v.42 no.2
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• pp.129-138
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• 2010

Total phenolic content, total anthocyanin content and antioxidant activity were analyzed from rice samples collected in Korea, Japan and China. The results showed that the total phenolic content and free-radical scavenging activity differed significantly in these rice lines. The correlation between content and activity was subsequently investigated. The results showed that in black rice, anthocyanin was the major phenolic component and that both phenolic content and anthocyanin content were closely correlated with free-radical scavenging activity. Reverse Phase High Performance Liquid Chromatography (RP-HPLC) data showed that cyanidin-3-O-glucoside and peonidin-3-O-glucoside composed about 90% of the total anthocyanin content in black rice and in red rice. In the red rice extract, the total phenolic content produced a high correlation coefficient with antioxidant activity but correlated very poorly with the total anthocyanin content. The $OD_{458}$ and the $OD_{500}$ values which represent the proanthocyanidin content of the rice extract, produced high correlation coefficients with antioxidant activity and total phenolic content. These results suggest that the $OD_{458}$ and the $OD_{500}$ values can be used to evaluate the quality of red rice. In addition, based on the data obtained, a competitive accumulation model of anthocyanin and proanthocyanidin in black and red rice was proposed.

• Analytical Science and Technology
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• v.20 no.5
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• pp.424-433
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• 2007

An analytical method for the determination of thyroid hormones in plasma samples has been studied by solid-phase extraction and high-performance liquid chromatography/diode array detector (DAD)/electrospray ionization (ESI)-mass spectrometer. Seven thyroid hormones were successfully separated by gradient elution on the reverse phase Hypersil ODS column (4.6 mm I.D., 250 mm length, particle size $5{\mu}m$) with ammonium formate buffer and acetonitrile. In addition, these compounds were confirmed by UV spectra and ESI-mass Spectra. The extraction recoveries of thyroid hormones in the plasma sample (at pH 3) were in the range of 74.5-115.7 % with solid-phase extraction by C18, followed by elution with 4 mL of methanol. The calibration curves showed good linearity with the correlation coefficients ($r^2$) varying from 0.9939 to 0.9978 and the detection limits of all analytes were obtained in the range of 20-50 ng/mL (38.1-162.8 pmol/mL). As a result, thyroxine was found in the range of 50.98-112.97 ng/mL in normal plasma samples.

• Biomolecules & Therapeutics
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• v.27 no.2
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• pp.193-200
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• 2019

Ceramide metabolism is known to be an essential etiology for various diseases, such as atopic dermatitis and Gaucher disease. Glucosylceramide synthase (GCS) is a key enzyme for the synthesis of glucosylceramide (GlcCer), which is a main ceramide metabolism pathway in mammalian cells. In this article, we developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to determine GCS activity using synthetic non-natural sphingolipid C8-ceramide as a substrate. The reaction products, C8-GlcCer for GCS, could be separated on a C18 column by reverse-phase high-performance liquid chromatography (HPLC). Quantification was conducted using the multiple reaction monitoring (MRM) mode to monitor the precursor-to-product ion transitions of m/z $588.6{\rightarrow}264.4$ for C8-GlcCer at positive ionization mode. The calibration curve was established over the range of 0.625-160 ng/mL, and the correlation coefficient was larger than 0.999. This method was successfully applied to detect GCS in the human hepatocellular carcinoma cell line (HepG2 cells) and mouse peripheral blood mononuclear cells. We also evaluated the inhibition degree of a known GCS inhibitor 1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) on GCS enzymatic activity and proved that this method could be successfully applied to GCS inhibitor screening of preventive and therapeutic drugs for ceramide metabolism diseases, such as atopic dermatitis and Gaucher disease.

• Applied Biological Chemistry
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• v.40 no.3
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• pp.178-183
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• 1997

A protease was purified from Bacillus subtilis CCKS-111 by ammonium sulfate treatment, DEAE-cellulose ion-exchange chromatography, Sephadex G-100 gel filtration and high performance liquid chromatography (HPLC). The specific activity of the purified enzyme was 24.3 unit/mg protein and the purification fold of enzyme was 50.6. Molecular weight of the purified enzyme estimated about 28,000 by HPLC gel filtration. The amino acid residues of this enzyme were 251.3 except threonine, serine and glycine. This result was similar to Bacillus subtilis subtilisin DY. From the first N-terminal amino acid to the 32th amino acid, the amino acid sequence was estimated after RP-HPLC elution. N-terminal and the 32th amino acids were alanine and aspartic acid. Alanine, serine, glycine and arginine were four major acids in the enzyme.

• Journal of Nutrition and Health
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• v.24 no.3
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• pp.199-205
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• 1991

Retinoic acid. the active metabolite of vitamin A. was detected in the human liver for the first time using a new method. A rapid and sensitive technique has been developed using gradient-elution. reverse-phase high performance liquid chromatography. This assay. with simultaneous multiwavelength detection at 294nm, 325nm and 450nm after saponifcation of liver samples. allows us seperation and quantitation of vitamin E, retinoic acid, total retinoids and various carotenoids in one small sample. The proportion of retinoic acid to total retinoids in human liver appears to be quite $consistent(2.4\pm0.2%$ ). With low vitamin A storage in liver, detection at another wavelenth 354nm would increase the sensitivity for retinoic acid of small quantity This method of analysis could be used for other tissues like red blood cells, plasma or serum, also. Hepatic retinoic acid level with total retinoids and carotenoids would serve a better indicator of functional vitamin A nutriture especially for those with disease requiring needle biopsy of liver.

• Mass Spectrometry Letters
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• v.8 no.2
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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.