• 약학회지
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• 제57권3호
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• pp.187-193
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• 2013

Bojungikgi-tang has been widely used for enhancement of physical fitness in Korea. The convenient, simple, and accurate high-performance liquid chromatography (HPLC) method was established for simultaneous determination of four marker compounds, liquiritin, nodakenin, hesperidin, and glycyrrhizin in Bojungikgi-tang (Buzhongyiqi-tang in Chinese, Hochuekkito in Japanese), a traditional Korean herbal prescription. The column for optimizing HPLC separation was used a Gemini $C_{18}$ column at column oven temperature of $40^{\circ}C$ with 1.0% (v/v) aqueous acetic acid (A) and 1.0% (v/v) acetic acid in acetonitirle (B) by gradient flow. The flow rate was 1.0 ml/min and the detector was a photodiode array (PDA) set at 254 nm, 280 nm, and 335 nm. Calibration curves of four components were acquired with $r^2$ values ${\geq}0.9999$. The recoveries were found to range 92.11~105.68% with relative standard deviations (RSDs, %) value less than 2.50%. The RSD values of intraand inter-day precision were 0.07~2.50% and 0.16~1.99%, respectively. The contents of liquiritin, nodakenin, hesperidin and glycyrrhizin in Bojungikgi-tang were 3.85~3.92 mg/g, 2.27~2.32 mg/g, 4.14~4.19 mg/g, and 3.39~3.45 mg/g, respectively. The established simultaneous analysis method will be effective for quality control of Bojungikgi-tang.

• Bulletin of the Korean Chemical Society
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• 제32권8호
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• pp.2666-2670
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• 2011

High-performance liquid chromatography (HPLC) and capillary electrophoresis (CE) were used to identify five active components in the modified herbal decoction Bo-Yang-Hwan-O-Tang (mBHT), i.e., amygdalin, decursin, paeoniflorin, salvianolic acid B, and calycosin-7-O-${\beta}$-D-glycoside. These components were identified by comparing their retention times and mass spectra with those of reference compounds. The conditions of both analytical methods were optimized and validated. Sufficient separation of target analytes was achieved using a buffer consisting of 40 mM sodium borate and 60 mM sodium dodecylsulfate (SDS) containing 10% methanol (pH 9.5) at 250 nm for CE analysis and gradient elution with a water-methanol mobile phase and ultraviolet (UV) photodiode array detector (DAD) at 250 nm for HPLC analysis. The mBHT components were determined within 65 min by HPLC and 16 min by CE. All calibration curves showed high linearity (R > 0.999) within the ranges tested. Intra-day and inter-day precision were less than 1.6% and 1.8% for HPLC and 2.5% and 4.8% for CE, respectively. The accuracy of the methods ranged from 98.8% to 102.3% for HPLC and from 95.9% to 108.2% for CE.

• 한국한의학연구원논문집
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• 제15권1호
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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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• 제25권5호
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• pp.694-698
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• 2004

A new method for the simultaneous determination of seven transition metal ions in water and food by microcolumn high-performance liquid chromatography has been developed. The lead, cadmium, mercury, nickel, cobalt, silver and tin ions were pre-column derivatized with tetra-(4-aminophenyl)-porphyrin ($T_4$-APP) to form the colored chelates which were then enriched by solid phase extraction with $C_{18}$ cartridge. The enrichment factor of 50 was achieved by eluted the retained chelates from the cartridge with tetrahydrofuran (THF). The chelates were separated on a ZORBAX Stable Bound microcolumn ($2.0{\times}50\;mm,\;1.8\;{\mu}m$)with methanol-tetrahydrofuran (95 : 5, v/v, containing 0.05 mol/L pyrrolidine-acetic acid buffer salt, pH = 10.0) as mobile phase at a flow rate of 0.5 mL/min and detected with a photodiode array detector from 350-600 nm. The seven chelates were separated completely within 2.0 min. The detection limits of lead, cadmium, mercury, nickel, cobalt, silver and tin are 4 ng/L, 3 ng/L, 6 ng/L, 5 ng/L, 5 ng/L, 6 ng/L, 4 ng/L respectively in the original samples. This method was applied to the determination of the seven transition metal in water and food samples with good results.

• Bulletin of the Korean Chemical Society
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• 제35권7호
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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.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권1호
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• pp.78-84
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• 2005

The purpose of this study was to optimize the solid state cultivation of Monascus ruber on sterile rice. A single-level-multiple-factor and a single-factor-multiple-level experimental design were employed to determine the optimal medium constituents and to optimize carbon and nitrogen source concentrations for lovastatin production. Simultaneous quantitative analyses of the ${\beta}$-hydroxyacid form and ${\beta}$-hydroxylactone for of lovastatin were performed by the high performance liquid chromatography (HPLC) method with a UV photodiode-array (PDA) detector. The total lovastatin yield ($4{\sim}6\;mg/g$, average of five repeats) was achieved by adding soybean powder, glycerol, sodium nitrate, and acetic acid at optimized levels after 14 days of fermentation. The maximal yield of lovastatin under the optimal composition of the medium increased by almost 2 times the yield observed prior to optimization. The experimental results also indicated that the ${\beta}$-hydroxylactone form of lovastatin (LFL) and the ${\beta}$-hydroxyacid form of lovastatin (AFL) simultaneously existed in solid state cultures of Monascus ruber. while the latter was the dominant form in the middle-late stage of continued fermentation. These results indicate that optimized culture conditions can be used for industrial production of lovastatin to obtain high yields.

• 대한한의학방제학회지
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• 제20권2호
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• pp.37-46
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• 2012

Objectives : Oyaksungi-san(Wuyaoshunqisan) has been used for treatment of stroke and rheumatoid arthritis in Korea. In this study, a simple and accurate high-performance liquid chromatography(HPLC) method was established for simultaneous determination of six main components, liquiritin, ferulic acid, naringin, hesperidin, neohesperidin, and glycyrrhizin in Oyaksungi-san, a traditional Korean herbal prescription. Methods : The analytical column for separation of six constituents was used a Gemini $C_{18}$ column maintained at $40^{\circ}C$. The mobile phase consisted of two solvent systems, 1.0% (v/v) acetic acid in $H_2O$ (A) and 1.0% (v/v) acetic acid in acetonitrile (B) by gradient flow. The flow rate was 1.0 mL/min and the detector was a photodiode array (PDA) set at 254 nm for glycyrrhizin, 280 nm for liquiritin, naringin, hesperidin, and neohesperidin, and 320 nm for ferulic acid. Results : Calibration curves were acquired with $r^2$ values ${\geq}0.9998$. The results of recovery test were 91.58%-105.90% with a relative standard deviations (RSDs, %) value less than 2.0%. The values of RSD for intra- and inter-day precision were 0.03%-1.72% and 0.03%-1.63%, respectively. The contents of the six compounds in Oyaksungi-san were 0.33-9.30 mg/g. Conclusions : The newly established HPLC method will be helpful to improve quality control of Oyaksungi-san.

• Journal of Applied Biological Chemistry
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• 제50권4호
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• pp.244-248
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• 2007

The contents of total phenol and total flavonoid of artichoke (Cynara scolymus L.) were measured. The antioxidant activity of the artichoke was evaluated based on its potential as a scavenging the ABTS radical. These results showed the antioxidant activity of artichoke has a close relationship with the total flavonoid content. The compound showing antioxidant activity was isolated from the artichoke by repeated column chromatography and recrystallization. Based on the spectrometric studies, the compound was identified as 1,3-dicaffeoylquinic acid, known as cynarin. The content of cynarin from heads and leafs of the artichoke determined by $C_{18}$ reversed phase HPLC (high-performance liquid chromatography) coupled with photodiode array detector was 10.15 and 0.67 mg/g, respectively. This compound showed potent antioxidant activities against DPPH and ABTS radicals ($EC_{50}$ = 14.09 and 28.85 ${\mu}M$, respectively).

• 한국한의학연구원논문집
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• 제15권2호
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• pp.119-124
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• 2009

The purpose of this study was investigation of quantitative analysis of marker substances in Paeonia lactiflora extracts by solid fermentation. High performance liquid chromatography (HPLC) for the determination of albiflorin and paeoniflorin in P. lactiflora extracts by solid fermentation, the separation 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 (230nm). The flow rate was 1.0 ml/min. Retention time of albiflorin and paeoniflorin was about 28.88, 31.92 min and linearity of calibration was showed good result(r2 = 0.9998, 0.9996), respectively. Content of albiflorin was $0.090\;{\pm}\;0.03%$ in P. lactiflora extract(control), $0.102\;{\pm}\;0.00%$ in P. lactiflora extract fermented with Paecilomyces japonica, $0.056\;{\pm}\;0.01%$ in P. lactiflora extract fermented with Ganoderma lucidum, $0.093\;{\pm}\;0.00%$ in P. lactiflora extract fermented with honey and $0.046\;{\pm}\;0.00%$ in P. lactiflora extract fermented with Nuruk. Content of paeoniflorin was $4.506\;{\pm}\;0.13%$ in control, $2.599\;{\pm}\;0.04%$ in P. lactiflora extract fermented with Paecilomyces japonica, $1.222\;{\pm}\;0.03%$ in P. lactiflora extract fermented with Ganoderma lucidum, $2.750\;{\pm}\;0.05%$ in P. lactiflora extract fermented with honey and $0.847\;{\pm}\;0.00%$ in P. lactiflora extract fermented with Nuruk, respectively. Content of the marker substances did not increase in all fermentation experiment group.

• 한국한의학연구원논문집
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• 제16권1호
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• pp.173-178
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• 2010

The purpose of this study was investigation of quantitative analysis of marker substances in solid fermented Angelicae Gigantis Radix by High performance liquid chromatography(HPLC). HPLC was performed for determination of nodakenin and decursin in solid fermented Angelicae Gigantis Radix extract, the separation 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 (330 nm). The flow rate was 1.0 ml/min. Retention time of nodakenin and decursin was about 11.47, 46.79 min and linearity of calibration was showed good result(r2=0.9999, 0.9999), respectively. Content of nodakenin was $0.76\;{\pm}\;0.02%$ in control, $0.31\;{\pm}\;0.00%$ in Angelicae Gigantis Radix extract fermented with Paecilomyces japonica(SDT)(p<0.01), $0.51\;{\pm}\;0.02%$ in Angelicae Gigantis Radix extract fermented with Ganoderma lucidum(SYT)(p<0.01), $0.82\;{\pm}\;0.03%$ in Angelicae Gigantis Radix extract fermented with honey(SST)(p<0.05) and $0.88\;{\pm}\;0.01%$ in Angelicae Gigantis Radix extract fermented with Nuruk(SNT)(p<0.01). Content of decursin was $4.50\;{\pm}\;0.08%$ in control, $2.90\;{\pm}\;0.05%$ in Angelicae Gigantis Radix extract fermented with Paecilomyces japonica(SDT)(p<0.01), $2.65\;{\pm}\;0.08%$ in Angelicae Gigantis Radix extract fermented with Ganoderma lucidum(SYT)(p<0.01), $4.46\;{\pm}\;0.11%$ in Angelicae Gigantis Radix extract fermented with honey(SST) and $4.73\;{\pm}\;0.04%$ in Angelicae Gigantis Radix extract fermented with Nuruk(SNT)(p<0.05), respectively.