• Journal of Applied Biological Chemistry
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• v.62 no.2
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• pp.129-135
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• 2019

Salvia miltiorrhiza Radix is cultivated in Korea and China and is traditionally used to treat cardiovascular diseases. In this study, we developed and validated a quantitative analysis method for S. miltiorrhiza Radix using high-performance liquid chromatography (HPLC). Identification was performed using ultra performance liquid chromatography-tandem mass spectrometry. For quantitative analysis, we used seven marker compounds. Separation conditions for HPLC were optimized using an ODS column with gradient conditions of 1% formic acid in distilled water and 1% formic acid in acetonitrile, with a flow rate of 0.8 mL/min and a detection wavelength of 280 nm. This method showed good linearity ($R^2=0.9998$), precision (relative standard deviation ${\leq}3.3%$), accuracy (recovery of 94.16-102.89%), limit of detection ($7.53{\mu}g/mL$), and limit of quantification ($23.71{\mu}g/mL$). This approach successfully quantified marker compounds in S. miltiorrhiza Radix. The individual marker compounds were identified by comparing the molecular masses and retention times with does standard compounds. Marker compound contents of S. miltiorrhiza Radix were investigated with different cultivation regions. Seven marker compounds were detected and quantified in all samples. Among them, salvianolic acid B showed the highest contents and it ranged from 4.13 to 7.15%. The salvianolic acid B content (7.15%) of marker compound was the highest in Bonghwa, and the tanshinone IIA content (1.90%) was the highest in Pohang. The results of marker compounds and developed method were intended to provide a favorable reference for the study of S. miltiorrhiza Radix from different regions of Korea.

• The Korea Journal of Herbology
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• v.30 no.4
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• pp.11-20
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• 2015

Objectives : Ongyeong-tang (OGT) is a traditional herbal formula used to cure gynaecological disorders. OGT consists of 12 herbal medicines containing various bioactive components. Therefore, the development of suitable analytical method for the marker compounds is necessary for the quality control of OGT. Methods : Determination of the 18 marker compounds in OGT preparations was quantitatively performed by high-performance liquid chromatography-photodiode array detection analysis. The marker compounds were separated on a reversed-phase C ₁₈ column and the analytical method was successfully validated, which was applied to compare OGT extracts from laboratory preparation and commercial OGT granules. Results : Limit of detection and limit of quantification values were in the ranges of $0.001-0.016{\mu}g/mL$ and $0.003-0.047{\mu}g/mL$, respectively. Precision was 0.03-3.71 % within a day and 0.03-3.81 % over four consecutive days. Recovery of marker compounds ranged from 90.63-108.26 %, with relative standard deviation (RSD) values < 4.0 %. Reproducibility was < 2.5 % of the RSD value. The 18 marker compounds were stable within 16 h at $10^{\circ}C$, with the RSD value < 3.5 %. Quantitative analysis results showed that the quantities of the 18 marker compounds varied among OGT samples. Pearson coefficient evaluation and principal component analysis demonstrated that an OGT water extract produced by a laboratory method clearly differed from commercial OGT granules. Conclusions : The developed analytical method was simple, precise, and reliable. Therefore, it can be used for the quality assessment of OGT preparations.

• The Korea Journal of Herbology
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• v.31 no.3
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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.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2023.04a
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• pp.62-62
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• 2023

Zanthoxylum piperitum ("chopi" in Korean) has been used as traditional medicinal plants with high anti-inflammatory, antioxidant, and antifungal activities. The aims of the study were to identify marker compounds and to investigate metabolites variation of chopi according to different parts and harvest times. Every month from June to September, chopi were harvested with three different parts: leaves, leaf-twig mixtures, twigs. Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), two main marker compounds (quercitrin and quercetin-3-O-glucoside) were characterized in 70% ethanol extracts of chopi. Quantification of the two marker compounds were subsequently conducted by high performance liquid chromatography (HPLC), representing that contents of these compounds were higher in leaves and leaf-twig mixtures rather than twigs. For the comprehensive analysis of metabolites associated with production of marker compounds, 35 primary metabolites were identified using gas chromatography-mass spectrometry (GC-MS). Multivariate analysis results represented that plant parts were main contributors to the separation of chopi. However, significant differences were not observed between leaves and leaf-twig mixtures samples. The partial least square (PLS) predictive model revealed that monosaccharides (fructose, galactose, glucose, mannose, xylose) and branched-chain amino acids (isoleucine, valine, leucine) were important determinants for the production of marker compounds together with alanine, inositol, GABA, and theronic acid. This study could be extended to stabilize and utilize chopi as an industrial material, as well as to find good candidates with various nutritional traits.

• Korean Journal of Pharmacognosy
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• v.48 no.4
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• pp.320-328
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• 2017

Yongdamsagan-tang has been used to treat the urinary disorders, acute- and chronic-urethritis, and cystitis in Korea. In this study, an ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS/MS) method was established for simultaneous analysis of the 20 bioactive marker compounds, geniposidic acid, chlorogenic acid, geniposide, liquiritin apioside, acteoside, calceolarioside B, liquiritin, nodakenin, baicalin, liquiritigenin, wogonoside, baicalein, glycyrrhizin, wogonin, glycyrrhizin, wogonin, saikosaponin A, decursin, decursinol angelate, alisol B, alisol B acetate, and pachymic acid in traditional herbal formula, Yongdamsagan-tang. Chromatographic separations of all marker compounds were conducted using a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$) at $45^{\circ}C$ using a mobile phase of 0.1% (v/v) formic acid in water and acetonitrile with gradient elution. The MS analysis was performed using a Waters ACQUITY TQD LC-MS/MS coupled with an electrospray ionization source in the positive and negative modes. The flow rate was 0.3 mL/min and injection volume was $2.0{\mu}L$. The correlation coefficient of 20 marker compounds in the test ranges was 0.9943-1.0000. The limits of detection and quantification values of the all marker components were 0.11-6.66 and 0.34-19.99 ng/mL, respectively. As a result of the analysis using the optimized LC-ESI-MS/MS method, three compounds, geniposidic acid (from Plantaginis Semen), alisol B (from Alismatis Rhizoma), and pachymic acid (from Poria Sclerotium), were not detected in this sample. While the amounts of the 17 compounds except for the geniposidic acid, alisol B, and pachymic acid were $0.04-548.13{\mu}g/g$ in Yongdamsagan-tang sample. Among these compounds, baicalin, bioactive marker compound of Scutellariae Radix, was detected at the highest amount as a $548.13{\mu}g/g$.

• Korean Journal of Pharmacognosy
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• v.45 no.3
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• pp.249-255
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• 2014

This study compared Bojungikgi-tang (BIT) and Yukmijihwang-tang (YJT) decoctions produced using the extractors made of various materials, such as glass, earthenware, and stainless steel, for 60, 90, and 120 min. The extraction yield was measured and the amounts of marker compounds in decoctions were determined by quantitative analysis using high performance liquid chromatography. The extraction yields of BIT decoctions produced by three extractors were not significantly different, whereas the extraction yield of YJT produced using glass extractor was significantly higher than that of YJT produced using earthenware extractor after 90 min. The amounts of most marker compounds were highest in BIT decoctions produced using glass extractor. However, variations of the amounts of marker compounds were observed in YJT decoctions and the volatile compound was not easily vaporized in YJT decoction produced using earthenware extractor. This study suggests that the extractor made of various materials may affect the extraction efficiency and the contents of marker compounds, especially in the decoction produced using earthenware extractor, although the results were dependent on the characteristics of herbal formula.

• Journal of Food Hygiene and Safety
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• v.37 no.1
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• pp.29-37
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• 2022

This study will evaluate the effect of fermented Morinda citrifolia L. extracts and its marker compounds to provide baseline data for utilizing Morinda citrifolia L. as functional health products. Morinda citrifolia L. and six marker compounds were processed on RAW 246.7 macrophage to test for XTT Cytotoxicity, measure Nitric Oxide and Cyokine formation, and analyze the expression of immune marker genes. Furthermore, LPS and fermented red ginseng extract, a common functional ingredient, are used as positive controls. Our results showed that fermented Morinda citrifolia L and six bioactive compounds did not have any cytotoxic effect in all treatment concentrations and groups. Among six bioactive compounds, SCP and ASE confirmed the formation of NO. In addition, the ASE treatment group showed increased formation of IL-6 and IL-1β and the expression of iNOS and TNF-α. Also, fermented Morinda citrifolia L extract activated the macrophage by enhancing the production of nitric oxide (NO), interleukin (IL)-6, and IL-1β, and the expression of COX2 compared to Morinda citrifolia L. extracts. The result of the study showed that Fermented Morinda citrifolia L. (Noni) and marker compound enhance the innate immunity activity and suggested that the bioactive compound could be applied as a marker compound. Thus, Fermented Morinda citrifolia L. (Noni) could be used as functional food material to develop immunity-enhancing products, and highly functional marker compounds can be utilized as the effective components.

• Natural Product Sciences
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• v.23 no.2
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• pp.75-83
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• 2017

The optimization and microwave assisted extraction of stem bark of Terminalia arjuna, quantitative estimation of the marker compounds arjunic acid and arjunolic acid using HPTLC and the evaluation of free radical scavenging activity has been performed in this study. The central composite design was used for optimization and the values of parameters for optimized batch of microwave assisted extraction were 1000 W (Power), 3 minutes (Time) and 1/120 (Solid/solvent ratio). The solvent system to carry out the HPTLC was toluene: acetic acid: ethyl acetate (5: 5: 0.5) and quantitative estimation was done using standard equations obtained from the marker compounds. The in-vitro free radical scavenging activity was performed spectrophotometrically using ascorbic acid as standard. The value of estimated percentage yield of arjunic acid and arjunolic acid was 1.42% and 1.52% which upon experimentation was obtained as 1.38% and 1.51% respectively. The DPPH assay of the different batches of microwave assisted extraction and marker compounds taken suggested that the marker compounds arjunic acid and the arjunolic acid were responsible for the free radical scavenging activity as the batch having the maximum percentage yield of the marker compounds showed best free radical scavenging effect as compared to standard ascorbic acid. The $IC_{50}$ value of the optimized batch was found to be 24.72 while that of the standard ascorbic acid was 29.83. Hence, the yield of arjunic acid and arjunolic acid has direct correlation with the free radical scavenging activity of stem bark extract of Terminalia arjuna and have potential to serve as active lead compounds for free radical scavenging activity.

• Natural Product Sciences
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• v.17 no.1
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• pp.58-63
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• 2011

Insampaedok-san is a traditional medicine used for the treatment of colds. We investigated several compounds in Insampeadok-san, and tested their neuroprotective and anti-oxidative activities after fermentation with Lactobacillus. The amounts of four marker compounds (ferulic acid, hesperidin, 6-gingerol and glycyrrhizin) and unidentified compounds in Insampaedok-san (IS) and fermented Insampaedok-san (FIS) were measured and compared by an established HPLC-DAD method. Neuroprotective activity of IS and FIS extracts was evaluated and compared after glutamate-induced neurotoxicity in HT22 cells. Anti-oxidative activity of IS and FIS was also compared in DPPH free radical, hydroxyl radical and hydrogen peroxide scavenging activity assays. Contents of two compounds, ferulic acid and glycyrrhizin were decreased while 6-gingerol was increased by fermentation. FIS showed more potent neuroprotective activity than IS. DPPH, hydroxyl radical and hydrogen peroxide scavenging was slightly increased by FIS when compared to IS. In conclusion, fermentation with Lactobacillus can vary the amounts of the marker compounds in IS and improve neuroprotective and anti-oxidative activities of IS.

• Journal of Ginseng Research
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• v.26 no.3
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• pp.165-169
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• 2002

Experiments were carried out to select marker for rusty tolerance ginseng root using high rusty degree lines(HRL) and low rusty degree lines (LRL) in ginseng plant. A strong positive correlation was detected between degree of rusty-root in 4-year-root and that in 6-year-root. The contents of phenolic compounds among samples were not different in stele and branch & fine roots. The contents of phenolic compounds of rusty-roots was higher than that of healthy-roots in cortex, but those of high 겨sty degree lines (HRL) were not different compared with low rusty degree lines (LRL) in cortex using same rusty-degree samples. These suggest that phenolic compounds in cortex tissue were not adequate as a marker to select rusty tolerance ginseng roots. The contents of phenolic compounds of rusty-roots were higher than that of healthy-roots in epidermis, and those of HRL were higher than LRL in epidermis using same rusty-degree samples. These suggested that the contents of phenolic compounds in epidermis tissue might be a potent marker to select rusty tolerance ginseng roots.