• Natural Product Sciences
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• 제14권3호
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• pp.147-151
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• 2008

A high performance liquid chromatographic (HPLC) method for the simultaneous determination of marker constituents, baicalin and glycyrrhizin was established for the quality control of traditional herbal medicinal preparation, Eul-Ja-Tang (EJT). Separation and quantification were successfully achieved with a Waters XTerra RP18 column ($5{\mu}m$, 4.6 mm I.D. ${\times}$ 150 mm) 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 250 nm. The presence of baicalin and glycyrrhizin in this decoction was ascertained by retention time, spiking with each authentic standard and UV spectrum. Both baicalin and glycyrrhizin showed good linearity ($r^2$ > 0.999) in a relatively wide concentration ranges. The R.S.D. for intra-day and inter-day precision was less than 5% and the limits of detection (LOD) were about 30 ng. The mean recovery of each compound was 99.5 - 101.2% with R.S.D. values less than 4.0%. This method was successfully applied to the determination of contents of baicalin and glycyrrhizin in three commercial products of EJT, which resulted in the difference in the contents of these compounds. These results suggest that the developed HPLC method is simple, effective and could be readily utilized as a quality control method for commercial EJT products.

• 생약학회지
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• 제47권4호
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• pp.366-373
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• 2016

Bulhwangeumjeonggisan (BHGJGS) is a traditional herbal formulation generally used in the treatment of cold and gastritis. BHGJGS consists of eight herbal plants; Atractylodis Rhizoma, Magnoliae Cortex, Citri Pericarpium, Glycyrrhizae Radix, Agastachis Herba, Pinelliae Rhizoma, Zingiberis Rhizoma and Zizyphi Fructus. Complete standardization of this formulation has not been done yet. So, a simple and accurate method was developed and validated using Ultra Performance Liquid Chromatography (UPLC) with Diode Array Detector (DAD) for the standardization of BHGJGS. UPLC conditions were optimized using a c18 RP-Amide column with mobile phase; 0.1% phosphate buffer and acetonitrile, detection wavelength; 210 and 325 nm. The linearities of calibration curves were acceptable ($R^2$>0.9994), and the limit of detection and quantification were within the ranges of 0.011-0.091 and $0.034-0.277{\mu}g/ml$ respectively. The relative standard deviation (RSD) of intra- and inter-day precisions were under 3.61%. The RSD of repeatability was under 0.68 %. The results of recovery test were 94.4-107.9%, and the RSD were under 4.6%. The developed method was used to find the contents of standard constituents in BHGJGS mix extract powder, and two commercial formulation (A and B). The data show that the developed method was specific, sensitive, accurate, and precise for analysis of BHGJGS components.

• 한국의학물리학회지:의학물리
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• 제28권4호
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• pp.164-170
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• 2017

One of the methods to consider the effect of respiratory motion of a tumor target in radiotherapy is to establish a treatment plan with the internal target volume (ITV) created based on an accurate analysis of the target motion displacement. When this method is applied to intensity modulated radiotherapy (IMRT), it is expected to yield a different treatment dose distribution under the motion condition according to the IMRT method. In this study, we prepared ITV-based IMRT plans with conventional IMRT using fixed gantry angle beams, RapidArc using volumetric modulated arc therapy, and tomotherapy using helical therapy. Then, the variation in dose distribution caused by the target motion was analyzed by the dose measurement in the actual motion condition. A delivery quality assurance plan was prepared for the established IMRT plan and the dose distribution in the actual motion condition was measured and analyzed using a two-dimensional diode detector placed on a moving phantom capable of simulating breathing movements. The dose measurement was performed considering only a uniform target shape and motion in the superior-inferior (SI) direction. In this condition, it was confirmed that the error of the dose distribution due to the target motion is minimum in tomotherapy. This is thought to be due to the characteristic of tomotherapy that treats the target sequentially by dividing it into several slices. When the target shape is uniform and the main target motion direction is SI, it is considered that tomotherapy for the ITV-based IMRT method has a characteristic which can reduce the dose difference compared with the plan dose under the target motion condition.

• Natural Product Sciences
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• 제17권4호
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• pp.328-336
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• 2011

A high-performance liquid chromatography (HPLC) with diode array detector (DAD) method was established for the discrimination of a folk medicine Forsythia suspensa and Forsythia viridissima. Five and three representative metabolites of the lignan and phenolic glycoside classes were selected for the analysis from F. suspensa and F. viridissima, respectively. The optimal chromatographic conditions were obtained on an ODS column (5 ${\mu}m$, $4.6{\times}250$ mm) with the column temperature at $40^{\circ}C$. The mobile phase was composed of methanol and 0.3% acetic acid using an isocratic elution with the flow rate 1 mL/min. Detection wavelength was set at 280 nm. All calibration curves showed good linear regression ($r^2$ > 0.996) within test ranges. Limits of detection (LOD) and limits of quantitation (LOQ) values were lower than 0.096 and 0.291 ${\mu}g/mL$, respectively. The developed method provided satisfactory precision and accuracy with overall intra-day and inter-day variations of 0.07-0.63% and 0.14-0.62%, respectively, and the overall recoveries of 97.79-102.46% for all of the compounds analyzed. In addition, effectiveness of diverse extraction methods was compared to each other for the development of standard analytical method. The verified method was successfully applied to the quantitative determination of representative metabolites in fifty-three commercial F. suspensa samples and fifteen commercial F. viridissima samples from diverse sources. The overall analytical results showed the unequivocal differences in bioactive constituents between F. suspensa and F. viridissima.

• ALGAE
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• 제27권4호
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• pp.303-313
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• 2012

Cyanobacteria are well known for their production of a multitude of highly toxic and / or allelopathic compounds. Among the photosynthetic microorganisms, cyanobacteria, belonging to the genus Nostoc are regarded as good candidate for producing biologically active secondary metabolites which are highly toxic to humans and other animals. Since so many reports have been published on the poisoning of different animals from drinking water contaminated with cyanobacteria toxins, it might be assumed that bioactive compounds are found only in aquatic species causes toxicity. However, the discovery of several dead dogs, mice, ducks, and fish around paddy fields, prompted us to study the toxic compounds in a strain of Nostoc which is most abundant in the paddy fields of Iran, using polymerase chain reaction and liquid chromatography coupled with a diode array detector and mass spectrophotometer. Results of molecular analysis demonstrated that the ASN_M strain contains the nosF gene. Also, the result of ion chromatograms and $MS^2$ fragmentation patterns showed that while there were three different peptidic compound classes (anabaenopeptin, cryptophycin, and nostocyclopeptides), there were no signs of the presence of anatoxin-a, homoanatoxin-a, hassallidin or microcystins. Moreover, a remarkable antifungal activity was identified in the methanolic extracts. Based on the results, this study suggests that three diverse groups of potentially bioactive compounds might account for the death of these animals. This case is the first documented incident of toxicity from aquatic cyanobacteria related intoxication in dogs, mice, and aquatic organisms in Iran.

• Journal of Pharmaceutical Investigation
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• 제39권1호
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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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• 제38권5호
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• pp.343-348
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• 2008

The bioequivalence of two cyclosporine A (CyA) 100 mg soft capsules (Chong Kun Dang's $Cipol-N^{(R)}$ as the test drug; Korea Novartis' Sandimmun $Neoral^{(R)}$ as the reference drug) was assessed in healthy male Korean volunteers after oral administration of 200 mg CyA according to a randomized crossover design. The whole blood samples were analyzed for the determination of parent CyA in the blood by using a validated HPLC/diode array detector method. The mean $AUC_t$ values for reference and test products were $4095.3{\pm}1397.2$ and $3958.3{\pm}1138.2\;ng{\cdot}hr/mL$, respectively. The mean $C_{max}$ values were $1135.9{\pm}293.2\;ng/mL$ for the reference product, and $985.0{\pm}207.9\;ng/mL$ for the test product. $T_{max}$ was $1.6{\pm}0.4\;hr$ for the reference and $1.8{\pm}0.5\;hr$ for the test product. The differences of $AUC_t$, $C_{max}$ and $T_{max}$ were -3.35, -13.28 and +10.63%, respectively. The point estimates and 90% confidence intervals for $AUC_t$ and $C_{max}$ were 0.981 (0.9171 to 1.0514) and 0.876 (0.8229 to 0.9336), respectively. Based on the pharmacokinetic and statistical data, we conclude that these two products are bioequivalent and can be considered interchangeable in the medical practice.

• 약학회지
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• 제52권1호
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• pp.7-11
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• 2008

A high performance liquid chromatographic (HPLC) method for the determination of glycyrrhizic acid was developed for the quality control of traditional herbal medicinal preparation Bu-Zhong-Yi-Qi-Tang (BZYQT), which is well-known herbal medicine used as tonic. RP-HPLC analysis was carried out using Capcell pak $C_{18}$ MG column $(5\;{\mu},\;150{\times}4.6\;mm)$ and a mobile phase consisting of acetonitrile and water containing 0.03% phosphoric acid (pH 2.46) at a flow rate of 1.0 ml/min. The optimum wavelength for the detection of the glycyrrhizic acid was found at 250 nm using diode-array UV/VIS detector. The glycyrrhizic acid in BZYQT shows good linearity $(r^2>0.999)$ in the range of $15\;{\mu}g/ml$ to 500 ${\mu}g/ml$. The limit of detection (LOD) was less than 5 ng and R.S.D for intra-day and inter-day reproducibility was less than 7%. The mean recovery of the glycyrrhizic acid was $97.3{\sim}113.0%$. These results suggest that the developed HPLC method is simple and efficient, and could be contributed for the quality control of commercial BZYQT products.

• 대한본초학회지
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• 제31권3호
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• pp.29-35
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• 2016

Objectives : Ijung-tang (IJT) is a traditional herbal formula and has been used to treat digestive diseases such as abdominal pain, vomiting, and diarrhea. IJT consists of four herbal medicines, Ginseng radix, Atractylodis rhizoma alba, Zingiberis rhizoma, and Glycyrrhizae radix et rhizoma, containing various bioactive compounds. Quality assesment of IJT preparations was performed by analytical method for determining marker compounds.Methods : Determination of seven marker compounds in IJT preparations was quantitatively conducted by high-performance liquid chromatography equipped with a diode-array detector. The marker compounds were separated on a reversed-phase C18 column and the analytical method was successfully validated. Chemometric analysis was performed to compare IJT water extracts and commercial IJT granules.Results : Limit of detection and limit of quantification values were in the ranges of 0.093-2.649 μg/mL and 0.283-8.027 μg/mL, respectively. Precisions were 0.30-3.87% within a day and 0.23-2.35% over three consecutive days. Recoveries of the marker compounds ranged from 87.35-107.05%, with relative standard deviation (RSD) values < 6.15%. Repeatabilities were < 1.20% and < 1.71% of RSD value for retention time and absolute peak area, respectively. The results from quantitative analysis showed that the quantities of seven marker compounds of IJT samples varied, as were found in principal component analysis and hierarchical clustering analysis.Conclusions : The analytical method developed in the present study was precise and reliable to simultaneously determine marker compounds of IJT. Therefore, it can be used for the quality assessment of IJT preparations.

• Journal of Acupuncture Research
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• 제31권4호
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• pp.45-55
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• 2014

Objectives : The main aim of this study was to assess the comparative efficiency of two preparation types of Hwangryunhaedok decoction(HRHD-D) using distilled and mixed extraction by measuring the index components and indicators of antioxidant and anti-inflammatory effects. Methods : The antioxidant activity was assessed by comparing distilled and mixed extractions of HRHD-D using an ELISA reader. The anti-inflammatory effect was determined by measuring NO amounts in RAW 264.7 cells. The contents were analyzed with high performance liquid chromatography-diode array detector(HPLC-DAD). Results : The electron donating ability of mixed and distilled extractions obtained with 500 ppm DPPH(1,1-diphenyl-2-picrylhydrazyl assay) solution were 57.8 % and 4.2 %, respectively. The total phenolic content of mixed extraction was 6.9 times that of distilled extraction and total flavonoid content was 51.5 times higher. The anti-inflammatory effect was assessed by NO measurement, and was found to increase significantly dependent on concentration in all mixed extract concentrations(25, 50, 100, 200, $400{\mu}g/mL$), but the difference in distilled extraction by concentration was only significant at 200 and $400{\mu}g/mL$. The HPLC analysis results of mixed extract of HRHD-D showed detection of all four main active constituents of HRHD-D. However, they were not detected in the distilled extract of HRHD-D. Conclusions : Mixed extraction with distillation added to decoction of HRHD-D showed better efficacy in antioxidant and anti-inflammatory effects, and ingredient quantities compared to distilled extraction. Further stability and clinical efficacy studies for standardization of mixed extractions are required.