• Journal of Ginseng Research
• /
• v.40 no.4
• /
• pp.382-394
• /
• 2016

Background: Ginsenosides are the characteristic and principal components which manifest a variety of the biological and pharmacological activities of the roots and rhizomes of Panax ginseng (GRR). This study was carried out to qualitatively and quantitatively determine the ginsenosides in the cultivated and forest GRR. Methods: A rapid and sensitive ultra-high-performance liquid chromatography coupled with diode-array detector and quadrupole/time of flight tandem mass spectrometry (UPLC-DAD-QTOF-MS/MS) was applied to the qualitative analysis of ginsenosides and a 4000 QTRAP triple quadrupole tandem mass spectrometer (HPLC-ESI-MS) was applied to quantitative analysis of 19 ginsenosides. Results: In the qualitative analysis, all ingredients were separated in 10 min. A total of 131 ginsenosides were detected in cultivated and forest GRR. The method for the quantitative determination was validated for linearity, precision, and limits of detection and quantification. 19 representative ginsenosides were quantitated. The total content of all 19 ginsenosides in the forest GRR were much higher than those in the cultivated GRR, and were increased with the growing ages. Conclusion: This newly developed analysis method could be applied to the quality assessment of GRR as well as the distinction between cultivated and forest GRR.

• Food Science and Biotechnology
• /
• v.18 no.3
• /
• pp.641-648
• /
• 2009

A one-step simultaneous immunchromatographic (OS-ICG) assay using colloidal gold-monoclonal antibody (gold-MAb) conjugates was developed for the rapid multianalysis of aflatoxin B1 (AFB1) and ochratoxin A (OTA) in feed samples. Visual detection limits for AFB1 and OTA were 0.5 and 2.5 ng/mL, respectively, and the results were obtained within 15 min. Matrix interference from the feed extracts was efficiently reduced by appropriate dilution with buffer. Cut-off values of the OS-ICG assay for the feed spiked with AFB1/OTA mixtures (5/5, 10/10, 25/25, 50/55, 100/100 ${\mu}g/kg$) were 10 and 50 ${\mu}g/kg$ for AFB1 and OTA. The comparative analyses of 65 feed samples by OS-ICG, enzyme-linked immunosorbent assay (ELISA), and high performance liquid chromatography (HPLC) showed good agreement. In this study, we confirmed that simultaneous analysis based on immunoassay is possible and it can be used as an on-site multianalysis of AFB1 and OTA in feed, food, and agricultural products.

• Journal of Microbiology and Biotechnology
• /
• v.23 no.1
• /
• pp.69-75
• /
• 2013

Enrofloxacin is a fluoroquinolone antibiotic approved for the treatment of infections in animals. Because of the side effects to consumers of animal products, the maximum residue limits (MRLs) of enrofloxacin in animal tissues for consumption are regulated. In this study, a monoclonal antibody (mAb) against enrofloxacin was prepared and characterized for the development of a direct competitive enzyme-linked immunosorbent assay (ELISA). The obtained mAb, Enro44, was highly specific for enrofloxacin and had a 50% inhibition concentration ($IC_{50}$) of 1.99 ng/ml in a competitive ELISA, and the limit of detection (LOD) was 0.50 ng/ml. The cross-reactivity of the mAb with other quinolones and fluoroquinolones was lower than 0.01%. The subclass of the mAb Enro44 was identified as IgG1. The antigen (Ag)-captured direct competitive ELISA using the mAb Enro44 was tested on different spiked samples, including chicken muscle, cattle milk, and cattle urine, and the assay demonstrated recoveries of 82-112%, 80-125%, and 78-124%, respectively. Furthermore, the quantitation of enrofloxacin obtained from the ELISA and from high-performance liquid chromatography (HPLC) was in good agreement, with the linear regression coefficient between 0.933 and 1.056. The cDNAs encoding a heavy-chain Fd fragment (VH and CH1) and a light chain of the mAb Enro44 were cloned and sequenced. Taken together, the results obtained reveal a potential use of this mAb in an ELISA for the detection of enrofloxacin in food samples. The information of amino acid sequence of this mAb will be useful for further modification and production of the mAb in a bioreactor.

• Natural Product Sciences
• /
• v.16 no.2
• /
• pp.116-122
• /
• 2010

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 five representative metabolites of the iridoid and phenolic classes from Rehmannia glutinosa. The optimal chromatographic conditions were obtained on an ODS column (5 mm, $4.6{\times}250\;mm$) with the column temperature at $25^{\circ}C$. The mobile phase was composed of water and acetonitrile using a gradient elution with the flow rate 0.3 mL/min. Detection wavelength was set at 205 nm. All calibration curves showed good linear regression ($r^2$ > 0.997) within test ranges. Limits of detection (LOD) and quantitation (LOQ) values were lower than 0.123 and $0.373\;{\mu}g/mL$, respectively. The developed method provided satisfactory precision and accuracy with overall intra-day and inter-day variations of 0.09 - 0.76% and 0.16 - 1.41%, respectively, and the overall recoveries of 99.03 - 102.67% for all of the compounds analyzed. In addition, effectiveness of diverse extraction methods was compared to each other for the development of standard analytic method. The verified method was successfully applied to the quantitative determination of five representative metabolites in twenty-one commercial Rehmannia glutinosa samples from different markets in Korea and China. The analytical results showed that the contents of the five analytes vary significantly with sources.

• Natural Product Sciences
• /
• v.16 no.1
• /
• pp.26-31
• /
• 2010

High performance liquid chromatographic method with diode-array detection (HPLC-DAD) has been performed for the simultaneous determination of four marker constituents, paeoniflorin, trans-cinnamic acid, schisandrin and glycyrrhizin in traditional herbal medicinal preparation, So-Cheong-Ryong-Tang (SCRT). The presence of paeoniflorin, trans-cinnamic acid, schisandrin and glycyrrhizin in this decoction was ascertained by retention time, spiking with each authentic standard, UV spectrum and ESI mass spectrum. All four compounds showed good linearity ($r^2$ > 0.998) in a relatively wide concentration ranges. The RSD for intra-day and inter-day precision was less than 3% and the limits of detection (LOD) were less than 30 ng. The mean recovery of each compound was 94.1-113.0% with RSD values less than 3.0%. These results suggest that the developed HPLC method is simple, effective and could be readily utilized as a quality control method for commercial SCRT products.

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

• Journal of Environmental Health Sciences
• /
• v.47 no.5
• /
• pp.496-503
• /
• 2021

Background: Considering the expenses of and difficulties in arsenic speciation by high performance liquid chromatography-inductively coupled plasma-mass spectrometry (HPLC-ICP-MS), alternative measurement methods should be useful, especially for large-scale research and projects. Objectives: A measurement method was developed for arsenic speciation using HPLC-atomic fluorescence spectrometry (HPLC-AFS) as an alternative to HPLC-ICP-MS. Methods: Total arsenic and toxic arsenic species in some seafoods were determined by atomic absorption spectrometry coupled with hydride vapor generation (AAS-HVG) and HPLC-AFS, respectively. Recovery rate of arsenic species in seafood was evaluated by ultra sonication, microwave and enzyme (pepsin) for the optimal extraction method. Results: Limits of detection of HPLC-AFS for As³⁺, dimethylarsinate (DMA), monomethylarsonate (MMA) and As⁵⁺ were 0.39, 0.53, 0.60 and 0.64 ㎍/L, respectively. The average accuracy ranged from 97.5 to 108.7%, and the coefficient of variation was in the range of 1.2~16.7%. As³⁺, DMA, MMA and As⁵⁺ were detected in kelp, the sum of toxic arsenic in kelp was 40.4 mg/kg. As³⁺, DMA, MMA and As⁵⁺ were not detected in shrimp and squid, but total arsenic (iAS and oAS) content in shrimp and squid analyzed by AAS-HVG were 18.1 and 24.7 mg/kg, respectively. Conclusions: HPLC-AFS was recommendable for the quantitative analysis method of arsenic species. As toxic arsenic species are detected in seaweeds, further researches are needed for the contribution degree of seafood in arsenic exposure.

• Natural Product Sciences
• /
• v.25 no.3
• /
• pp.238-243
• /
• 2019

In this study, the marker compounds of Curcumae Rhizoma (CR) were simultaneously quantified by high-performance liquid chromatography equipped with a photodiode array detector and the anti-inflammatory effects of CR extract and marker compounds in human benign prostatic hyperplasia epithelial-1 (BPH-1) cell lines were investigated. The marker components (4S,5S)-(+)-germacrone-4,5-epoxide, furanodienone, and germacrone, were separated on Gemini $C_{18}$ columns ($250mm{\times}4.6mm$, $5{\mu}m$) at $40^{\circ}C$ by using a gradient of two mobile phases eluting at 1.0 mL/min. Prostaglandin $E_2$ ($PGE_2$) levels in Human BPH-1 cells were determined with an ELISA kit. The coefficients of determination in a calibration curve of each analyte were all 0.9997. The limits of detection and quantification of the three compounds were $0.10-0.32{\mu}g/mL$ and $0.30-0.98{\mu}g/mL$, respectively. The content of three compounds, (4S,5S)-(+)-germacrone-4,5-epoxide, furanodienone, and germacrone, in the CR sample were found to be 5.79 - 5.92 mg/g, 4.72 - 4.86 mg/g, and 1.06 - 1.09 mg/g, respectively. Regarding pharmacological activity against benign prostatic hyperplasia, CR and its components significantly suppressed $PGE_2$ levels of BPH-1 cells. The established analysis method will help to improve quality assessment of CR samples and related products. In addition, CR and its components exhibit antiinflammatory activity in BPH-1 cells, suggesting the inhibitory efficacy of these compounds against the pathogenesis of BPH.

• Geomechanics and Engineering
• /
• v.18 no.2
• /
• pp.179-188
• /
• 2019

This paper presents an experimental study to evaluate the effect of palm oil on the selected basic physical-chemical and geotechnical properties of kaolin. The experimental findings are further compared with literature outcomes investigating similar properties of fine grained soils subjected to contamination by different types of oils. To this end, palm oil was mixed with oven dried kaolin samples-aiding oil's interaction (coating) with dry particles first, in anticipation to emphasize the effect of oil on the properties of kaolin, which would be difficult to achieve otherwise. Oil content was limited to 40% by dry weight of kaolin, supplemented at intervals of 10% from clean kaolin samples. Observations highlight physical particle-to-particle bonding resulting in the formation of pseudo-silt sized clusters due to palm oil's interaction as evinced in the particle size distribution and SEM micrographs. These clusters, aided by water repellency property of the oil coating the kaolin particles, was analyzed to show notable variations in kaolin's consistency-measured as liquid and plastic limits. Furthermore, results from compaction tests indicates contribution of oil's viscosity on the compaction behavior of kaolin - showing decrease in the maximum dry unit weight (${\gamma}_{d,max}$) and optimum moisture content ($w_{opt}$) values with increasing oil contents, while their decrease rates were directly and inversely proportional in ${\gamma}_{d,max}$ and $w_{opt}$ values with oil contents respectively. Comparative study in similar terms, also validates this lower and higher decrease rates in ${\gamma}_{d,max}$ and $w_{opt}$ values of the fine grained soils respectively, when subjected to contamination by oil with higher viscosity.

• Natural Product Sciences
• /
• v.25 no.1
• /
• pp.49-58
• /
• 2019

Eleven steroid hormones (SHs: androstene-3,17-dione, estrone, ${\beta}$-estradiol, ${\alpha}$-estradiol, testosterone, dehydroepiandrosterone, $17{\acute{a}}$-hydroxyprogesterone, medroxyprogesterone, megestrol acetate, progesterone, and androsterone) were detected from New Zealand deer (Cervus elaphus var. scoticus) velvet antler (NZA, 鹿茸 ). A method for the quantification of eleven SHs was established by using ultraperformance liquid chromatography (UPLC)-MS/MS. The linearities ($R^2$ > 0.991), limits of quantification (LOQ values, 0.3 ng/mL to 23.1 ng/mL), intraday and interday precisions (relative standard deviation: RSD < 2.43%), and recovery rates (97.3% to 104.6%) for all eleven SHs were determined. In addition, a method for the quantification of three 7-oxycholesterols (7-O-CSs: 7-ketocholesterol, $7{\alpha}$-hydroxycholesterol, and $7{\beta}$-hydroxycholesterol) in the NZA was established by using an HPLC-photodiode array (PDA) method. The linearities ($R^2$ > 0.999), LOQ values (30 ng/mL to 350 ng/mL), intraday and interday precisions (RSD < 1.93%), and recovery rates (97.2% to 103.5%) for the three 7-O-CSs were determined. These quantitative methods are accurate, precise, and reproducible. As a result, it is suggested that the five steroid compounds of androstene-3,17-dione, androsterone, 7-ketocholesterol, $7{\alpha}$-hydroxycholesterol, and $7{\beta}$-hydroxycholesterol could be marker steroids of NZA. These methods can be applied to quantify or standardize the marker steroids present in NZA.