• Korean Journal of Pharmacognosy
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• v.45 no.1
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• pp.84-87
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• 2014

A high performance liquid chromatography (HPLC) method for the quantitation of ellagic acid in Nymphaea tetragona was developed for the quality control of functional cosmetic ingredient, the extract of N. tetragona. Separation and quantitation were successfully achieved with a Kromasil C18 column ($5{\mu}m$, $250mm{\times}4.6mm$, i.d.) by isocratic elution of a mixture of acetonitrile containing 0.1% trifluoroacetic acid and water containing 0.03% phosphoric acid at a flow rate of 1.0 ml/min. The UV detector was used for the detection and the wavelength for quantitation was set at 254 nm. The presence of ellagic acid in the extract was determined by comparison of retention time and spiking with authentic standard. Analytical results showed good linearity ($R^2=0.99996$) in relatively wide concentration ranges. The R.S.D. for precision test was less than 3.0%. Recovery of the compound was 98.55~101.72% with R.S.D values less than 4.0%. In conclusion, this method has been successfully applied to the determination of ellagic acid in N. tetragona.

• Journal of Pharmaceutical Investigation
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• v.39 no.1
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• pp.23-27
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• 2009

A high performance liquid chromatographic (HPLC) method for the simultaneous determination of marker constituents, paeoniflorin and glycyrrhizin was established for the quality control of traditional herbal medicinal preparation, Sayuk-san (SYS). Separation and quantification were successfully achieved with a Waters XTerra RP18 column ($5{\mu}m$, $4.6mm\;I.D.{\times}150mm$) 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 paeoniflorin and glycyrrhizin in this decoction was ascertained by retention time, spiking with each authentic standard and UV spectrum. All two compounds showed good linearity ($r^2$>0.996) in a relatively wide concentration ranges. The R.S.D. for intra-day and inter-day precision was less than 7.3% and the limits of detection (LOD) were less than 55.7 ng. The mean recovery of each compound was $102.3{\sim}111.1%$ with R.S.D. values less than 4.6%. This method was successfully applied to the determination of contents of paeoniflorin and glycyrrhizin in three commercial products of SYS. These results suggest that the developed HPLC method is simple, effective and could be readily utilized as a quality control method for commercial SYS products.

• Journal of Pharmaceutical Investigation
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• v.35 no.6
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• pp.431-436
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• 2005

A high-performance liquid chromatographic method was validated for quantitation of nadolol in human plasma. Nadolol and internal standard, pindolol, were extracted with tert-butyl methyl ether after addition of 10 M sodium hydroxide solution. The analytes were separated on a reverse phased C18 column using a mobile phase consisting of 0.05 M ammonium phosphate monobasic buffer, acetonitrile and methanol (81: 17:2 v/v/v) and detected using a fluorescence detector (excitation wavelength 230 nm, emission wavelength 294 nm). The method was specific and sensitive enough to detect as low as 3 ng/mL of nadolol in human plasma. Linear calibration range was 3-150 ng/mL with correlation coefficient greater than 0.999. The overall accuracy was in the range of 96.8 to 103% and precision C.V.(%) 7.30 to 12.2%. The recovery was approximately 100% and stability was confirmed during storage and sample preparation. The present HPLC method was successfully applied to study bioavailability after oral administration of 80 mg of nadolol in healthy Korean subjects. The mean $AUC_{t}$ was $1968{\pm}397\;ng{\cdot}hr/mL$ and $C_{max}$ of $186{\pm}79.3\;ng/mL$ was reached at $3.5{\pm}0.76\;hr$. The mean $t_{1/2}$ of nadolol was $17.3{\pm}2.59\;hr$.

• Korean Journal of Oriental Medicine
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• v.15 no.1
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• pp.85-89
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• 2009

The aim of this study was to study the quantitative analysis of glycyrrhizic acid in Glycyrrhizae Radix extract fermented with Paecilomyces japonica, Ganoderma lucidum, honey or Nuruk. The amounts of dry on loss were measured and the quantitative analysis of glycyrrhizic acid was performed by high performance liquid chromatographic (HPLC). HPLC method was performed on C18 column ($250\;mm\;{\times}\;4.6\;mm$, $5\;{\mu}m$, RS tech) using gradient solvent mixtures of water-acetonitrile with photodiode array detector (254 nm). The flow rate was $1.0\;m{\ell}/min$. Retention time of glycyrrhizic acid was about 23.96 min and linearity of calibration was $R^2$=0.9998. Contents of glycyrrhizic acid in Glycyrrhizae Radix extract (control) was $5.048\;{\pm}\;0.14$; Contents of glycyrrhizic acid in Glycyrrhizae Radix extract fermented with Paecilomyces japonica (SDT) was $1.975\;{\pm}\;0.07$; Contents of glycyrrhizic acid in Glycyrrhizae Radix extract fermented with Ganoderma lucidum (SYT) was $2.676 \;{\pm}\;0.07$; Contents of glycyrrhizic acid in Glycyrrhizae Radix extract fermented with honey (SST) was $5.191\;{\pm}\;0.06$; Contents of glycyrrhizic acid in Glycyrrhizae Radix extract fermented with Nuruk (SNT) was $5.305\;{\pm}\;0.34$, respectively. Contents of glycyrrhizic acid in SDT and SYT were decreased but that in SST and SNT was increased when compared to control.

• Bulletin of the Korean Chemical Society
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• v.35 no.7
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• pp.1956-1964
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• 2014

To provide the scientific corroboration of the traditional uses of Akebia quinata (Thunb.) Decne., a detailed analytical examination of A. quinata stems was carried out using a reversed-phase high performance liquid chromatography (RP-HPLC) method coupled to photodiode array detector (PDA) for the simultaneous determination of four phenolic substances; cuneataside D (1), 2-(3,4-dihydroxyphenyl)ethyl-O-${\beta}$-D-glucopyranoside (2), 3-caffeoylquinic acid (3) and calceolarioside B (4). Particular attention was focused on the main compound, 3-caffeoylquinic acid (3), which has a range of biological functions. In addition, 2-(3,4-dihydroxyphenyl)ethyl-O-${\beta}$-D-glucopyranoside (2) was considered as a discernible marker of A. quinata from its easy confuse plants. The contents of compounds 2 and 3 ranged from 0.72 to 2.68 mg/g and from 1.66 to 5.64 mg/g, respectively. The validation data indicated that this HPLC/PDA assay was used successfully to quantify the four phenolic compounds in A. quinata from different locations using relatively simple conditions and procedures. The pattern-recognition analysis data from 53 samples classified them into two groups, allowing discrimination between A. quinata and comparable herbs. The results suggest that the established HPLC/PDA method is suitable for quantitation and pattern-recognition analyses for a quality evaluation of this medicinal herb.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.39 no.2
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• pp.59-65
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• 2006

Fluoroquinolones are the most common group of antibacterial agents currently used in the Korean aquaculture industry, and use of these agents has been increasing steadily. High performance liquid chromatography (HPLC) with fluorescence detection was used for the simultaneous determination of five fluoroquinolones in fish and shellfish: ofloxacin (OFL), pefloxacin (PEF), norfloxacin (NOR), ciprofloxacin (CIP), and enrofloxacin (ENRO). Fish and shellfish muscle was homogenized, and protein, lipid, and low molecular weight pigments were then excluded from the homogenate. The final eluates were analyzed by HPLC equipped with a Shiseido UG-120 type C18 reverse-phase column ($4.6{\times}250 mm$, $5{\mu}m$) and a fluorescence detector (excitation at 280 nm, emission at 450 nm). The mobile phase was 0.1 M phosphoric acid and acetonitrile solution (91:9, v/v) and tetrahydrofuran (THF) was added to it at a rate of 5 mL per a liter of the mobile phase. Adequate chromatography separation was obtained using the above method. Average recoveries of fortified samples at levels from 0.05 to 0.5 mg/kg were $72.3{\pm}2.5-84.5{\pm}1.2%$ for OFL, $82.7{\pm}3.3- 109.3{\pm}7.5%$ for NOR, $85.3{\pm}6.6-116.0{\pm}7.9%$ for PEF, $76.0{\pm}4.3-109.3{\pm}12.4%$ for CIP, and $78.7{\pm}5.9-100.0{\pm}9.8%$ for ENRO. The limit of detection of OFL was $5{\mu}g/L$, the others were $1{\mu}g/L$. We concluded that the new analytical method was suitable for the determination of fluoroquinolones in fish and shellfish.

• Natural Product Sciences
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• v.19 no.1
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• pp.20-27
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• 2013

A simple high performance liquid chromatography - diode array detector (HPLC-DAD) method has been developed and validated for simultaneous determination of the six components (cinnamic acid, cinnamaldehyde, eugenol, atractylenolide I, atractylenolide III, and ergosterol) in Oryeongsan. In addition, identification of six marker compounds was conducted by a LC-MS/MS analysis. The six compounds in Oryeongsan were separated on Shishedo $C_{18}$ column (5 ${\mu}m$, $4.6{\times}250$ mm) at a column temperature of $30^{\circ}C$. The mobile phase was a mixture of 0.1% trifluoroacetic acid (TFA) water and acetonitrile employing gradient elution at a flow rate of 1.0 mL/min. The detection wavelength was set at 205, 250, 280, and 330 nm. The developed method had good linearity ($R^2$ > 0.9997) and the limit of detection (LOD) and limit of quantification (LOQ) were observed within the ranges 0.01~0.15 and 0.05~0.45 ${\mu}g/mL$, respectively. The relative standard deviation (RSD) values of intra- and inter-day testing were indicated that less than 3% and 90.31~103.31% of accuracy. The results of recovery test were 90.56~106.72% and RSD range was measured from 0.84 to 2.95%. In conclusion, this HPLC-DAD method has been successfully applied to the simultaneous determination of six compounds in Oryeongsan samples.

• Journal of Mushroom
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• v.11 no.2
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• pp.107-110
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• 2013

This study was carried out to optimize the conditions for a chemotaxonomic classification of Ganoderma species. The mycelia of Ganoderma species were extracted with 100% MeOH, and the concentrated extracts were further purified and partitioned with column chromatography (HP20) and n-BuOH, respectively. From the result of high-performance liquid chromatography (HPLC), the constituents of the samples were efficiently separated with a Chemco Pak $C_{18}$ column ($250mm{\times}4.6mm$) by linear gradient elution using water and acetonitrile as mobile phase components at a flow rate of 0.8 ml/min and detector wavelength at 210 nm. However, the samples without purification or partition were not detected the characteristic peaks. This profile could be used to classify and identify the various Ganoderma species.

• YAKHAK HOEJI
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• v.54 no.3
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• pp.157-163
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• 2010

A high-performance liquid chromatography (HPLC) with DAD detector and electrospray ionization mass spectrometry (ESI-MS) was established for the simultaneous determination of coniferin (1), loganic acid (2), demethylsecologanol (3), sweroside (4) and loganin (5) from caulis Lonicera joponica. The optimal chromatographic conditions were obtained on an ODS column ($5{\mu}m$, $4.6{\times}150mm$) with the column temperature $35^{\circ}C$. The mobile phase was composed of (A) water with 0.1% formic acid and (B) methanol with 0.1% formic acid using a gradient elution, the flow rate was 0.3 ml/min. Detection wavelength was set at 254 nm. All calibration curves showed good linear regression ($r^2$>0.998) within test ranges. The developed method provided satisfactory precision and accuracy with overall intra-day and interday variations of 0.16~3.28% and 0.14~1.99%, respectively, and the overall recoveries of 99.39~105.89% for the five compounds analyzed. The verified method was successfully applied to quantitative determination of the two types (phenolic compounds and iridoids) of bioactive compounds in 24 commercial caulis L. japonica samples from different markets in Korea and China. The analytical results demonstrated that the contents of the five analytes vary significantly with sources.

• YAKHAK HOEJI
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• v.52 no.6
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• pp.446-451
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• 2008

A high-performance liquid chromatography (HPLC) with diode array detector (DAD) and electrospray ionization mass spectrometry (ESI-MS) was established for the simultaneous determination of chlorogenic acid (1), sweroside (2), luteolin-7-O-glucoside (3), (E)-aldosecologanin (4) and 3,5-dicaffeoylquinic acid (5) from Lonicera joponica flower buds. The optimal chromatographic conditions were obtained on an ODS column (5 ${\mu}m$, 4.6${\times}$150 mm) with the column temperature $25^{\circ}C$. The mobile phase was composed of (A) water with 0.1% formic acid and (B) acetonitrile with 0.1% formic acid using a gradient elution, the flow rate was 0.3 ml/min. Detection wavelength was set at 250 nm. All calibration curves showed good linear regression ($r^2$>0.994) within test ranges. The developed method provided satisfactory precision and accuracy with overall intra-day and inter-day variations of 0.05${\sim}$1.95% and 0.15${\sim}$2.26%, respectively, and the overall recoveries of 97.71${\sim}$103.65% for the five compounds analyzed. The verified method was successfully applied to quantitative determination of the three types (phenolic compounds, iridoids and flavonoids) of bioactive compounds in 21 commercial L. japonica flower buds samples from different markets in Korea and China. The analytical results demonstrated that the contents of the five analytes vary significantly with sources.