• Korean Journal of Pharmacognosy
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• v.36 no.3 s.142
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• pp.177-185
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• 2005

For the purpose of the quality control of Poria cocos, three major compounds were isolated and identified as pachymic acid (1), $3{\beta}-hydroxylanosta-7,9(11),24-trien-21-oic$ acid (2), and dehydroeburicoic acid (3). The optimal extraction conditions for the quantification of pachymic acid its analogues were the 3 hours of reflux with 15g of P. cocos in 100ml 95% ethanol. HPLC conditions were as follows: Column; ZORBAX Eclipse XDB C18 $(4.6{\times}250\;mm,\;Agillent)$, mobile phase; 1% HOAc in 70% $MeOH{\rightarrow}1%$ HOAc in 100% MeOH for 25 min, then 1% HOAc in 100% MeOH for 15 min, detector, ELSD, flow rate; 1ml/min. The mean contents of 1,2, and 3 in Poria cocos cultivated at 18 different site were $0.65{\pm}0.19,\;0.88{\pm}0.72,\;and\;0.84{\pm}0.54\;mg/g$, respectively, and values might be the guide line for the quality control of P. cocos.

• Food Science and Biotechnology
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• v.14 no.3
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• pp.317-322
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• 2005

A simple and convenient method of immobilizing dextransucrase via an affinity interaction is described, along with the use of this system to synthesize leucrose. Dextransucrase was produced in sucrose-free medium by fermenting a constitutive mutant of Leuconostoc mesenteroides NRRL B-512F and was separated using an ultrafiltration membrane. The purified enzyme was free of dextran polymer, which previously was always found with the sucrose-induced enzyme. Therefore, it was possible to immobilize the enzyme on dextran-based resins using an affinity interaction. Sephadex G-200 was the best resin for immobilizing the dextransucrase and gave a fast flow rate through the packed column. The immobilized dextransucrase retained more than 80% of its specific activity after immobilization ($K_m\;=\;18.1\;mM$ and $k_{cat}\;=\;450\;sec^{-1}$ vs. 13.1 mM and $640\;sec^{-1}$, respectively, for the free enzyme). The immobilized dextransucrase showed improved stability over a pH range of 4.0 to 6.5 and at moderately high temperatures over $40^{\circ}C$. When immobilized dextransucrase was used to synthesize leucrose via the transfer reaction with sucrose and fructose, about 74% of the sucrose was converted into leucrose after one day, and the half-life of the enzyme activity was 15 days. Regeneration of the resin by supplementation with dextransucrase enabled the recovery of the initial activity of the system, but both the reaction and the flow rate were lower, probably owing to the accumulation of dextran inside the resin.

• Analytical Science and Technology
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• v.34 no.5
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• pp.212-224
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• 2021

The intent of the present work was to develop a simple, sensitive, accurate, precise, rapid and economical UV- spectrophotometric and reverse phase high pressure liquid chromatographic method for the simultaneous estimation of Spironolactone and Furosemide in bulk and combined tablet dosage forms. UV-Spectrophotometry was carried out by simultaneous equation method using 0.02 M potassium dihydrogen phosphate buffer pH 3.5: Acetonitrile (50:50) v/v as a solvent. The linearity range was 2-14 ㎍ mL^-1 for Spironolactone and Furosemide with a correlation coefficient > 0.99. The chromatographic separation was achieved on 250 mm × 4.6 mm, hypersil BDS C18 column with particle size 5 ㎛, by using an isocratic mixture of 0.02 M potassium dihydrogen phosphate buffer pH 3.5: Acetonitrile: tert butyl methyl ether (49:50:1) v/v/v as a solvent at a flow rate of 1 mL min^-1 and UV detection was carried out at 254 nm. The retention time were observed to be 3.666 and 6.661 minutes for Furosemide and Spironolactone respectively. The two developed methods were validated according to the ICH guidelines for accuracy, precision, linearity, LOD, LOQ and were found to be within the limits. It can be concluded that these two methods could be successfully used for the simultaneous estimation of Spironolactone and Furosemide in bulk and combined tablet dosage forms.

• Analytical Science and Technology
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• v.32 no.3
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• pp.96-104
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• 2019

Agastache rugosa, known as Korean mint, is a medicinal plant with many beneficial health effects. In this study, a simple and reliable HPLC-UV method was proposed for the quantification of rosmarinic acid (RA) in the aqueous extracts of A. rugosa. RA was selected as a quantification marker due to its easiness in procurement and analysis. The developed method involved chromatographic separation on a $C_{18}$ column ($250{\times}4.6mm$, $5{\mu}m$) at room temperature. The mobile phase consisted of water and acetonitrile both containing 2 % acetic acid and was run at a flow rate of $1mL\;min^{-1}$. The method was validated for specificity, linearity, precision, and accuracy. It was specific to RA and linear in the range of $50-300{\mu}g\;mL^{-1}$ ($r^2=0.9994$). Intra-day, inter-day, and inter-analyst precisions were ${\leq}0.91%\;RSD$, ${\leq}1.40%\;RSD$, and 1.94 % RSD, respectively. Accuracy was 93.3-95.9 % (${\leq}1.21%\;RSD$). The method could be applied to three batches of bulk samples and three batches of lab scale samples, which were found to be $0.64({\pm}0.04)mg\;g^{-1}$ and $0.48({\pm}0.02)mg\;g^{-1}$ for the dried raw materials of A. rugosa. The results show that the proposed method can be used as a readily applicable method for QC of health functional foods containing the aqueous extracts of A. rugosa.

• Korean Journal of Food Science and Technology
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• v.51 no.1
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• pp.12-17
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• 2019

Dichloroacetic acid (DCAA) and trichloroacetic acid (TCAA) in fresh-cut salads available from the market were determined by gas chromatography-mass spectrometry (GC-MS). The target compounds in 3 g of acidified homogenates were extracted with 20 mL of methyl t-butyl ether (MTBE). The extract was concentrated to 1 mL and heated for 1 h at $55^{\circ}C$. The analytes were separated using a DB-1701 column and detected with a mass spectrometer. The method detection limit was approximately $6{\mu}g/kg$, and both analytical accuracy and precision were found to be satisfactory. The linearity of the calibration curves expressed as the coefficients of determination was >0.996. The analysis of seven samples using the established method showed that the four samples contained considerable amounts of analytes ($25.4-31.2{\mu}g/kg$ of DCAA and $18.8-46.1{\mu}g/kg$ of TCAA). These results raised a concern about the impact of fresh-cut salad consumption on human health.

• Journal of Applied Biological Chemistry
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• v.62 no.3
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• pp.239-246
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• 2019

In this study, an evaluation of the anti-inflammatory effect of ferulic acid isolated from Tetragonia tetragonioides in lipopolysaccharide (LPS) simulated RAW 264.7 cells was made. The chemical structure of the active compound was elucidated by $^1H$-NMR, $^{13}C$-NMR, and FAB-MS, and was confirmed to be ferulic acid. Ferulic acid was purified via open column chromatography with Sephadex LH-20 and MCI gel CHP-20. To test the anti-inflammatory effect of ferulic acid, LPS-stimulated RAW 264.7 cells were treated in subsequent experiments with different concentrations of ferulic acid (5, 10, and $25{\mu}g/mL$) and the levels of inflammatory cytokines and enzymes were also measured by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. Cell viability was above 95% at acid concentrations ranging from $5-25{\mu}g/mL$. The results showed that 30% of the production of nitric oxide and 66% of prostaglandin $E_2$ were inhibited by $25{\mu}g/mL$ of ferulic acid, it also inhibited the protein expression of both inducible nitric oxide synthase and cyclooxygenase-2 by 70%. Additionally, it inhibited the production of the pro-inflammatory cytokines, tumor necrosis factor-${\alpha}$, interleukin-6, and interleukin-$1{\beta}$ by 40, 75, and 77%, respectively. According to these results, the anti-inflammatory activity of ferulic acid was demonstrated via his implication in the inhibition of the expression and secretion of inflammatory substances in LPS-stimulated RAW 264.7 cells. Therefore, we concluded that ferulic acid can be used as a functional additive having anti-inflammatory activity.

• Food Science of Animal Resources
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• v.39 no.4
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• pp.601-609
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• 2019

Bifidobacterium longum KACC 91563 secretes family 5 extracellular solute-binding protein via extracellular vesicle. In our previous work, it was demonstrated that the protein effectively alleviated food allergy symptoms via mast cell specific apoptosis, and it has revealed a therapeutic potential of this protein in allergy treatment. In the present study, we cloned the gene encoding extracellular solute-binding protein of the strain into the histidine-tagged pET-28a(+) vector and transformed the resulting plasmid into the Escherichia coli strain BL21 (DE3). The histidine-tagged extracellular solute-binding protein expressed in the transformed cells was purified using Ni-NTA affinity column. To enhance the efficiency of the protein purification, three parameters were optimized; the host bacterial strain, the culturing and induction temperature, and the purification protocol. After the process, two liters of transformed culture produced 7.15 mg of the recombinant proteins. This is the first study describing the production of extracellular solute-binding protein of probiotic bacteria. Establishment of large-scale production strategy for the protein will further contribute to the development of functional foods and potential alternative treatments for allergies.

• Journal of Korean Neurosurgical Society
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• v.62 no.6
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• pp.671-680
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• 2019

Objective : We introduce innovative method of cervical column reconstruction and performed the reconstruction with a flanged titanium mesh cage (TMC) instead of a plate after anterior corpectomy for cervical spondylotic myelopathy (CSM) and an ossified posterior longitudinal ligament (OPLL). Methods : Fifty patients with CSM or OPLL who underwent anterior cervical reconstruction with a flanged TMC were investigated retrospectively. Odom's criteria were used to assess the clinical outcomes. The radiographic evaluation included TMC subsidence, fusion status, and interbody height. Thirty-eight patients underwent single-level and 12 patients underwent two-level corpectomy with a mean follow-up period of 16.8 months. Results : In all, 19 patients (38%) had excellent outcomes and 25 patients (50%) had good outcomes. Two patients (4%) in whom C5 palsy occurred were categorized as poor. The fusion rate at the last follow-up was 98%, and the severe subsidence rate was 34%. No differences in subsidence were observed among Odom's criteria or between the single-level and two-level corpectomy groups. Conclusion : The satisfactory outcomes in this study indicate that the flanged TMC is an effective graft for cervical reconstruction.

• Korean Journal of Pharmacognosy
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• v.52 no.3
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• pp.186-191
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• 2021

We developed a method to identify and quantify 6-hydroxyluteolin 7-O-glucoside in the powder of Salvia plebeia (PS) using high-performance liquid chromatography coupled with diode array detector (HPLC/DAD) and equipped with reverse-phase INNO C18 column. The analytical method was optimized and validated using novel parameters. The obtained values for the limits of detection and quantification were 3.60 and 10.90 ㎍/mL, respectively. Calibration curve showed good linearity in the concentration range tested (0.00625-0.1 mg/mL, r² = 1.0000), high accuracy (96.2-101.4%), and precision values (RSD ≤ 0.27%). Our analysis support the use of our method for accurately identifying and quantifying 6-hydroxyluteolin 7-O-glucoside from PS in routine analyses and large-scale extraction processes for content determination.

• Natural Product Sciences
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• v.27 no.1
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• pp.54-59
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• 2021

Bergenia ciliata (Family: Saxifragaceae) is a folklore remedy for the treatment of various ailments in Asian countries. Bergenin (1) has been isolated as an active constituent in many studies, however, the amount of bergenin has not been determined in all parts of the plant. A simple RP-HPLC method was developed to determine the amount of bergenin in methanol extracts of leaves, rhizomes and roots of the plant. Separation was achieved on an Agilent Eclipse XDB-C18 column maintained at 25 ℃ using isocratic solvent system (water: methanol: acetic acid; 62.5:37:0.5 v/v/v) adjusted at pH 2 0 at a flow rate of 1.0 mL/min. and detected at 275 nm. Correlation coefficient (0.9952) showed linearity of concentration (5-200 ㎍/mL) and response. The values of LOD (0.00947 ㎍/mL) and LOQ (0.02869 ㎍/mL) indicated that method was sensitive. The recovery of bergenin was 99.99-100% indicating accuracy of method. The methanol extract of rhizomes contained higher amount of bergenin (19.4%) than roots (9.2%) and leaves (6.9%). It is concluded that methanol extract of rhizomes is a better source of bergenin than other parts of the plant. The findings are useful for standardization of bergenin containing extracts and herbal preparations.