• Analytical Science and Technology
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• v.25 no.4
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• pp.250-256
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• 2012

Two marker compounds of Achyranthis Radix, ecdysterone and inokosterone, were analyzed by HPLC on an ODS column ($250{\times}4.6$ mm, 5 ${\mu}m$) with a mobile phase of 15% acetonitrile containing 0.08% formic acid at a flow rate of 1.0 mL/min and a detection wavelength of UV 254 nm. The method was validated by ICH guideline and applied to the monitoring of marker compounds in 93 samples of Achyranthis Radix collected at various areas in Korea and China. The new content criteria of ecdysterone and inokosterone, established using linear regression method were 0.033% and 0.020%, respectively. When the new content criteria were applied to the quality control test of commercial Achyranthis Radix, 95.4% of total samples including 100% of Korean and 92.6% of Chinese samples were passed the test. Application of new content criteria could protect the Korean products and decrease the distribution of Chinese products with lower quality.

• Korean Journal of Pharmacognosy
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• v.43 no.4
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• pp.279-285
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• 2012

The Mume Fructus (MF) has been used for relieves cough, arrests arrest chronic diarrhea, treat fluid depletion, and treat ascariasis in Korea. In this study, a high-performance liquid chromatography (HPLC) method was established for simultaneous determination of six main components of MF. Additionally, we were investigated the anti-inflammatory and anti-allergic effects of MF extract on lipopolysaccharide (LPS)-treated RAW264.7 cells and tumor necrosis factor (TNF)-${\alpha}$/interferon (IFN)-${\gamma}$-treated HaCaT cells. The analytical column for separation was used a Gemini $C_{18}$ column maintained at $40^{\circ}C$. The mobile phase consisted of 1.0% (v/v) acetic acid in water (A) and 1.0% (v/v) acetic acid in acetonitrile (B). The flow rate was 1.0 mL/min and the detector was a photodiode array (PDA) set at 280 nm and 320 nm. We evaluated the inhibitory effect of MF extract on the production of inflammatory markers, nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) in LPS-stimulated RAW264.7 cells and thymus- and activation-regulated chemokine (TARC/CCL17) in TNF-${\alpha}$/IFN-${\gamma}$-treated HaCaT cells, respectively. We confirmed the genes expression related with TARC, macrophage-derived chemokine (MDC/CCL22) and regulated on activation, normal T cell expressed and secreted (RANTES/CCL5) in HaCaT keratinocyte cells by MF extract. The contents of the five compounds in MF were 0.22-1.01 mg/g. Also, the MF extract show inhibition of about 78% and 75% on NO and $PGE_2$ production at the concentration 1000 mg/mL in RAW264.7 cells. MF extract suppressed the hTARC level and genes expression such as TARC, MDC, and RANTES on TNF-${\alpha}$/IFN-${\gamma}$-treated HaCaT cells.

• Journal of Food Hygiene and Safety
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• v.23 no.4
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• pp.319-323
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• 2008

Analytical method for determination of florfenicol was developed for estimate veterinary drug residue of unestablished MRLs in meat. The method was validated in correspondence with the CODEX guideline for florfenicol residue in meat. The samples mixed with sodium sulfate were extracted with ethyl acetate. After clean-up, the residue was dissolved in mobile phase and analyzed using high-performance liquid chromatography with fluorescence detector. The calibration curve showed good linearity($r^2=0.9997$) within the concentration range of $0.05{\sim}1.0\;mg/kg$. The limit of detection(LOD) and limit of quantification(LOQ) were validated at 0.012 and 0.039 mg/kg, respectively. The recoveries in fortified meat ranged from 85.6 to 95.6%($1.1{\sim}5.3%$ RSD) at the 0.05 to 0.4 spiking levels. We monitored 150 samples of meats that were purchased in Korea(Seoul, Busan, Daegu, Daejeon and Gwangju). Among tested samples, florfenicol was detected in 1 of pig at the level of 0.040 mg/kg, and below LOQ in 1 of cattle, 2 of pig and 2 of chicken. The residues of florfenicol in the tested samples were within the MRLs.

• Korean Journal of Food Science and Technology
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• v.35 no.6
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• pp.1028-1032
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• 2003

To assess accurate methods for measuring sulfur dioxide residue in pickles, the acid distillation HPLC-UV method and Monnier Willams modified method were examined. By the acid distillation HPLC-UV method, sulfites released from pickles by acid distillation were absorbed in 1% triethanolamine solution and detected as sulfite ion by HPLC with UV monitoring at 240nm. An anion exchange column was employed with 1.8mM $Ma_2CO_3-1.7mM\;NaHCO_3$ solution as a mobile phase, $84.0{\sim}91.7%$ of sulfite added to pickled radish were recovered. Total sulfite levels from 48 kinds of pickles analyzed by acid distillation HPLC-UV was compared with those analyzed by the Monnier Williams modified method. The Monnier Williams modified method showed higher levels of sulfur dioxide than the acid distillation HPLC-UV method due to the presence of volatile acids in pickles. The concentration of sulfur dioxide was in the range of $N.D{\sim}173.05ppm$ in pickled radish and over 30ppm of sulfur dioxide from 3 samples by the acid distillation-HPLC-UV method.

• Korean journal of food and cookery science
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• v.28 no.6
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• pp.805-811
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• 2012

This study was carried out in order to optimize the manufacturing condition of fried garlic flakes as well as to investigate the physicochemical properties of the flakes. Fried garlic flake samples were prepared as follows: garlic was sliced by a thickness of 1.5 mm, 2.0 mm, 2.5 mm, which were measured by a thickness gage. The samples were fried in vegetable oil under different temperatures of $140{\sim}150^{\circ}C$, $160{\sim}170^{\circ}C$ and $180{\sim}185^{\circ}C$. The compression strength depending on the height (h) was measured in order to find the thickness effect by the rheometer (force control: 50 N, h: 3.25 mm). Moreover, the sample with 1.5 mm thickness showed crisp phenomena of the split compared with the crush shape of the 2.0 mm and 2.5 mm thick samples. The result of strength for time dependence showed a sample with a thickness of 1.5 mm, which was measured 5~9 times more than the 2.0 mm and 2.5 mm thick samples. We thought the reason that the 1.5 mm sample had less response power equivalent to compression force than the other samples. Alliin has been found to affect the immune responses in the blood, it is a derivative of the amino acid cysteine and is also quite heat stable. The LC system with a UV detection at 210 nm consists of a separation on a Zorbax TMS column and isocratic elution with water and ACN as a mobile phase. The alliin contents of raw and fried garlic flake under $140{\sim}150^{\circ}C$, $160{\sim}170^{\circ}C$ and $180{\sim}185^{\circ}C$ were 18.10 mg/mL, 14.0 mg/mL, 11.6 mg/mL and 11.1 mg/mL, respectively. The decrement of alliin content under different temperature was a small quantity hence, we confirmed that the increasing manufacturing temperature was not affected by the alliin content. Examining for the particle structure of fried garlic flakes by a polarization microscope, the color of the sample treated at $160{\sim}170^{\circ}C$ was pure yellow. Furder, the fiber shaped particle, which has an effect on the tough texture, almost did not appear compared to the different temperature conditions. Finally, the sensory test for the preference of fried garlic flake under different conditions was carried out and the scores for various sensory characteristics were surveyed. According to the physicochemical measurements and sensory evaluation, we confirmed that the optimum manufacturing condition of fried garlic flake was 1.5 mm thick at a temperature of $160{\sim}170^{\circ}C$.

• Korean Journal of Veterinary Research
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• v.40 no.4
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• pp.741-746
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• 2000

We have developed a rapid, simple and precise high-performance liquid chromatographic (HPLC) method using an UV detection system for the determination of new anti-HIV candidates, nineteen KR-V compounds, in rat plasma. We used a analytical columnn of $C_{18}$ ($5{\mu}m$, $250{\times}2.0mm$ I.D.) and a mobile phase of water and ACN mixture (40/60, v/v). Under these conditions, all the KR-V compounds were readily separated from plasma with retention times of 4-12 min. The limits of quantitation for the 19 KR-V compounds were 15-30 ng/ml. The recoveries from the plasma were higher than 85% (C.V.<10%) with exception of KR-V 2, 7 and 15. The compounds KR-V 2, 7 and 15, containing ester moieties, were found to be unstable in plasma. This result suggests that esters, like KR -V 2, 7 and 15, should be excluded from future structure design studies of anti-HIV KR-V agents. In conclusion, the current HPLC method is a valuable analytical tool for investigating the pharmacokinetics of the KR-V series in rats.

• Analytical Science and Technology
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• v.18 no.4
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• pp.271-277
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• 2005

Urea in blood has been measured as an effective marker for diagnosis of renal function. Urea which is e end-product of nitrogen containing metabolites such as proteins is filtered through glomeruli of kidneys and then excreted as urine. If the renal function is deteriorated, the urea concentration in blood will be increased, from which the healthiness of renal function is judged. In order to improve the confidence of diagnosis results, the results must keep traceability chain to certified reference materials, which was certified by primary reference method. In this study, we proposed isotope dilution-liquid chromatography/mass spectrometry (ID-LC/MS) as a candidate primary method, in which $15^N_2$-urea is used as an internal reference material. The developed method is highly accurate in principle and is convenient as it does not require cumbersome derivatization. 0.1 mmol/L ammonium chloride was selected as a mobile phase for HPLC because it provided low interference in MS analysis of relatively low molecular weighted urea. HPLC and MS were connected with an electrospray ionization (ESI) interface of positive mode, which provided high sensitivity and reproducibility. The developed method was validated with internationally recognized reference materials, and we have obtained satisfactory results in an international ring trial. The expanded uncertainty calculated according to ISO guide was 1.8% at 95% confidence interval. The developed method is being used as a primary reference measurement method such as for certification of serum certified reference materials (CRMs).

• Journal of Pharmaceutical Investigation
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• v.31 no.4
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• pp.265-271
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• 2001

In order to elucidate the influence of intestinal and hepatic first-pass effect on the pharmacokinetics of triflusal, the biotransformation of triflusal in the gastrointestinal tract and liver was designed. Moreover, we tried to establish an HPLC method applicable for bioassay and available to pharmacokinetics, not only with the simultaneous determination of triflusal and its active metabolite, 2-hydroxy-4-trifluoromethyl benzoic acid (HTB), but also with improving sensitivity. After the administration of triflusal (10 mg/kg) and HTB (10 mg/kg) into femoral vein, portal vein (only triflusal) and oral route (only triflusal), pharmacokinetic parameters were investigated from the plasma concentration-time profiles of triflusal and HTB in rats. An HPLC method was developed for the simultaneous determination of triflusal and HTB in rat plasma, urine and bile. The HPLC analysis was carried out using a C18 column and acetonitrile-methanol-water (25:10:65, v/v/v) as the mobile phase and UV detection at 234 nm. Furosemide was used as the internal standard. The calibration curves were linear over the concentration range $0.05-5.0\;{\mu}g/ml$ for triflusal and $0.2-200.0\;{\mu}g/ml$ for HTB with correlation coefficients greater than 0.999 and with intra-day or inter-day coefficients of variation not exceeding 10.0%. This assay procedure was applied to the study of metabolite pharmacokinetics of triflusal and HTB in rats. It was supposed that triflusal was almost metabolized in vivo because urinary and biliary excreted amounts of triflusal could be ignored as it was lower than 1.2% of the administered dose. According to the gastrointestinal and hepatic biotransformation pathways of triflusal, it was found that triflusal was hydrolyzed by about 5% in intestine and metabolized by about 53% in liver, and that the bioavailability of triflusal after oral administration of triflusal was 0.44, and also that the fraction of total elimination rate of triflusal which formed HTB in liver $(F_{mi},\;%)$ was about 98%. These results showed that triflusal was almost metabolized in liver, and the total elimination of triflusal in the body was dependent to the formation rate of HTB from triflusal in liver.

• Journal of Food Hygiene and Safety
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• v.30 no.2
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• pp.173-177
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• 2015

Rosa canina L. is health functional food materials that can help to temporarily relieve symptoms of arthritis. This study has been conducted to develop and validate analytical methods for hyperoside of Rosa canina L.. Methods based on HPLC with ultraviolet detection (UVD) were established through instrumental analytical conditions, and the examination of data, such as domestic and foreign reliable methods and journals. HPLC UVD analysis using Capcell Pak $C_{18}$ MG II column at 353 nm was determined on test through the column, mobile phase. The validation has been performed on the method to determine linearity, accuracy, limits of quantification (LOQ) and repeatability for hyperoside. The method showed high linearity in the calibration curve at a coefficient of correlation ($R^2$) of 0.999, and the LOQ was $0.393{\mu}g/mL$. Relative standard deviation (RSD) values of data from repeatability precision was between 0.6 and 2.6%. Recovery rate test at hyperoside scored between 98 and 99%. These results indicate that the established HPLC method is very useful for the determination of hyperoside in Rosa canina L. to develop a health functional material.

• Analytical Science and Technology
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• v.18 no.3
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• pp.250-256
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• 2005

A selective method of high performance liquid chromatography with UV detector has been applied to determine 4 tetracycline antibiotics in the animal food, simultaneously. The targets were chlortetracycline (CTC), doxycycline (DC), oxytetracycline (OTC), and tetracycline (TC) that are used routinely in veterinary medicine for prevention and control of disease. Food samples were beef, pork, chicken, milk, whole egg, flatfish (Limanda yokohamae), jacopever (Sebastes hubbsi), seabream (Chrysophrys major), eel (Anguilla japonica) and lobster (Hommarus americanus). After extracting food samples with 20% trichloroacetic acid and McIlvaine buffer, they were purified by a $C_18$ SPE cartridge with 0.01M methanolic oxalic acid solution. The concentrated residue was re-dissolved in methanol, filtered, cleaned up and analyzed on a $C_18$ column. The mobile phase was a mixture of 0.01M oxalic acid and acetonitrile with a gradient ratio from 85:15 to 60:40. The UV wavelength was 365 nm. The overall recoveries were ranged from 71% to 98% and the limit of detections were 0.022 for CTC, 0.012 for DC and OTC and 0.009 mg/kg for TC at signal/noise > 3, respectively. As results, CTC, DC and TC were not detected in all selected food samples, however, OTC was detected in meat and fishes. The determined level of OTC was 0.04 ppm for pork, 0.17 ppm for flatfish and 0.05 and 0.08 ppm for jacopever, that were within the Maximum Residue Limits (MRLs) in the food.