• Journal of Ginseng Research
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• v.45 no.5
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• pp.539-545
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• 2021

Background: Red ginseng polysaccharides (RGPs) have been acknowledged for their outstanding immunomodulation and anti-tumor activities. However, their studies are still limited by the complexity of their structural features, the absence of purification and enrichment methods, and the rarity of the analytical instruments that apply to the analysis of such macromolecules. Thus, this study is an attempt to establish a new mass spectrometry (MS)-based analysis procedure for RGPs. Methods: Saponin pre-excluded powder of RG (RG-SPEP, 10 mg) was treated with 200 µL of distilled water and centrifuged for 5 h at 1000 rpm and 85 ℃. Ethanol-based precipitation and centrifugation were applied to obtain RGPs from the heated extracts. Further, endo-carbohydrase treatments were performed to produce specific saccharide fragments. Solid-phase extraction (SPE) processes were implemented to purify and enrich the enzyme-treated RGPs, while matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) MS was employed for the partial structural analysis of the obtained RGPs. Results: Utilizing cellulase, porous graphitized carbon (PGC), hydrophilic interaction chromatography (HILIC), and MALDI-TOF/TOF MS, the neutral and acidic RGPs were qualitatively analyzed. Hex_n and Hex_n-18 (cellulose analogs) were determined to be novel neutral RGPs. Additionally, the [Unknown + Hex_n] species were also determined as new acidic RGPs. Furthermore, HexA_n (H) was determined as another form of the acidic RGPs. Conclusion: Compared to the previous methods of analysis, these unprecedented applications of HILIC-SPE and MALDI-TOF/TOF MS to analyze RGPs proved to be fairly effective for fractionating and detecting neutral and acidic components. This new procedure exhibits great potential as a specific tool for searching and determining various polysaccharides in many herbal medicines.

• Mass Spectrometry Letters
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• v.7 no.1
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• pp.12-15
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• 2016

Results of free and bound myo-inositol in infant formula (IF) are presented. Inositol was analyzed by HILIC ultra-performance liquid chromatography coupled with mass spectrometer. The levels of free myo-inositol in 27 Australian and 4 EU originated IF samples were 300-600 mg/kg of powder or 1.6-3.1 mg/100 kJ. The amount of bound inositol in lipid fraction of IF was, on average, 10% of free myo-inositol.

• Mass Spectrometry Letters
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• v.6 no.2
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• pp.31-37
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• 2015

Phosphorylation upon protein is well known to a key regulator that implicates in modulating many cellular processes like growth, migration, and differentiation. Up to date, grafting of multidimensional separation techniques onto advanced mass spectrometry (MS) has emerged as a promising tool for figuring out the biological functions of phosphorylation in a cell. However, advanced MS-based phosphoproteomics is still challenging, due to its intrinsic issues, i.e., low stoichiometry, less susceptibility in positive ion mode, and low abundance in biological sample. To overcome these bottlenecks, diverse techniques (e.g., SCX, HILIC, ERLIC, IMAC, TiO₂, etc.) are continuously developed for on-/off-line enrichment of phosphorylated protein (or peptide) from biological samples, thereby helping qualitative/quantitative determination of phosphorylated protein and its phosphorylated sites. In this review, we introduce to the overall views of enrichment tools that are universally used to selectively isolate targeted phosphorylated protein (or peptide) from ordinary ones before MS-based phospoproteomic analysis.

• Archives of Pharmacal Research
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• v.19 no.5
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• pp.337-341
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• 1996

Propolid (bee-glue), known as a folk medicine, is a lipo;hilic material found in honeybee hives. In the present study on the anti-inflammatory effect of Korean propolis, it was extracted with ethanol, and used as a test material. The $LD_{50}$ value with the oral administration of ethanolic extract of Korean propolis (EEKP) was higher than 2g/kg in mice. The oral administration of the propolis extract (100mg/kg) significantly inhibited the development of hind paw edema induced by carrageenin in rats. the oral pretreatment of the propolis extract markedly inhibited the increase in vascular permeability and the number of writhing induced by acetic acetic acid in mice. Propolis extract, 50 and 100 mg/kg p.o. per day for 7 days, produced a significant inhibitory effect on granuloma and exudate formation in rats. This inhibitory effect was enhanced with the concomitant use of prednisolone (2.5 mg/kg). These results suggest that Korean propolis apparently has a strong anti-inflammatory activity.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.46 no.1
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• pp.49-55
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• 2020

This study was conducted to establish optimal high pressure liquid chromatography (HPLC) conditions for estimation of the ergothioneine contents in the three kinds of water extracts of Pleurotus eryngii var. ferulae (Meaksong, Beesan No. 2, Baekwhang). By comparing the four different HPLC conditions, optimun condition for quantifying the contents of ergotioneine was established (shodex HILIC column, 35 ℃, 1.0 mL/min). By this method, the contents of ergothioneine in Meakong (3.04 ± 0.02 mg/g), Beesan No. 2 (3.15 ± 0.05 mg/g) and Baewhang (1.13 ± 0.07 mg/g) were estimated. DPPH and ABTS radical scavenger activities of these three kinds of Pleurotus eryngii var. ferulae were estimated and the contents of total phenol and flavonoid were also estimated. Taken together, this study establish an optimun HPLC condition for determining the ergothioneine contents in water extracts of Pleurotus eryngii var. ferulae. Furthermore water extracts of Maesong and Beesan No. 2 showed relatively high contents of ergothioneine and antioxidant activity, suggesting that these materials could be used as potential antioxidant in developing functional cosmetics.

• Mass Spectrometry Letters
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• v.11 no.1
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• pp.10-14
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• 2020

Riboflavin is a water-soluble vitamin, which serves as a precursor to flavin mononucleotide and flavin adenine dinucleotide. This study aimed to develop a simple and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis for the quantification of riboflavin in the Beagle dog plasma. This method utilized simple protein precipitation with acetonitrile and ¹³C₄, ¹⁵N₂-riboflavin was used as an internal standard (IS). For chromatographic separation, a hydrophilic interaction liquid chromatography (HILIC) column was used with gradient elution. The mobile phase consisted of 0.1% (v/v) aqueous formic acid with 10 mM ammonium formate and acetonitrile with 0.1% (v/v) formic acid. Since riboflavin is an endogenous compound, 4% bovine serum albumin in phosphate buffered saline was used as a surrogate matrix to prepare the calibration curve. The quantification limit for riboflavin in the Beagle dog plasma was 5 ng/mL. The method was fully validated for its specificity, sensitivity, accuracy and precision, recovery, and stability according to the US FDA guidance. The developed LC-MS/MS method may be useful for the in vivo pharmacokinetic studies of riboflavin.

• Journal of Korean Society on Water Environment
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• v.32 no.6
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• pp.649-669
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• 2016

This study explored the analytical conditions for 21 veterinary antibiotics which have been popularly sold in South Korea in 2014 but have not yet been targeted in EPA method 1694. Most of the selected antibiotics were separated by a reverse-phase C18 column with a combination of (buffered) water and organic polar solvent, which was commonly methanol and acetonitrile in the gradient elution mode. Volatile additives such as formic acid, ammonium acetate and ammonium formate were usually added to the mobile phases to minimize asymmetrical and tailing of antibiotics' peaks and to increase their ionization in mass spectrometry. The analytical methods of aminoglycoside antibiotics were distinct from those of the other antibiotics in terms of adoption of ion-pair chromatography (IPC) and hydrophilic interaction liquid chromatography (HILIC) capable of retaining and separating extremely polar compounds due to their hydrophilicity. Trifluoroacetic acid or heptafluorobutyric acid was frequently added to the mobile phase as an ion-pair reagent for the IPC. Tandem mass spectrometry was numerously applied to the detection of antibiotics using positive electrospray ionization (ESI) and the selected reaction monitoring (SRM) mode. All reviewed analytical methods had been/were validated by evaluating recovery, limits of detection and quantification, decision limit or detection capability of the methods.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.5
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• pp.612-617
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• 2008

Tetrodotoxin (TTX) analogues were first determined from the liver extracts of puffer fish, Takifugu niphobles, by LC/MS with Hydrophilic Interaction Liquid Chromatography (HILIC). In total, 7 TTX analogues were detected within 20 minutes as follows; 5,6,11-trideoxyTTX (34.0%, 1,029.6 nmol/g), 6,11-dideoxyTTX (29.3%, 887.6 nmol/g), TTX (22.1%, 667.8 nmol/g), 4,9-anhydro-TTX (11.2%, 339.3 nmol/g), 11-deoxyTTX＋5-deoxyTTX (2.6%, 78.6 nmol/g), and 4-epiTTX (0.8%, 23.6 nmol/g). Mouse toxicity of diluted liver extracts showed the highest toxicity at pH 3 (8.7 MU/mL) and decreased, as increasing pH, to 1.4 MU/mL at pH 10. At acidic (pH 5) and neutral conditions (pH 7), mouse toxicity of liver extracts (79 MU/mL) decreased slowly, as increasing temperature from $80^{\circ}C$ to $115^{\circ}C$, and time until 1 hour; in contrast, at the akaline condition (pH 9), the toxicity decreased rapidly to the more than half within 10 minutes. Individual toxicity of the fillet of T. niphobles were between $43.2{\sim}106.7$ MU, and $64{\sim}78%$ of its toxicity was eluted to soup when boiled with 3 volumes of water during 10 minutes.