• Journal of Ginseng Research
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• 제45권5호
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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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• 제7권1호
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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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• 제6권2호
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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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• 제19권5호
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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.

• 대한화장품학회지
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• 제46권1호
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• pp.49-55
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• 2020

본 연구는 아위느타리버섯(Pleurotus eryngii var. ferulae)에 함유되어 있는 항산화 물질인 에르고티오네인의 정확한 함량을 분석할 수 있는 최적 분석법을 확립하고 아위느타리버섯 3종(맥송, 비산 2호, 백황)의 에르고티오네인 함량 및 항산화 능력을 검증하고자 수행되었다. 서로 다른 네 가지의 HPLC 분석 조건을 비교하여 아위느타리버섯 열수 추출물의 에르고티오네인 함량을 정량할 수 있는 최적 조건(shodex HILIC column, 35 ℃, 1.0 mL/min)을 확립하였으며 이를 이용하여 3종류의 아위느타리버섯 열수 추출물에 포함된 에르고티오네인 함량을 결정하였다(맥송: 3.04 ± 0.02 mg/g, 비산 2호: 3.15 ± 0.05 mg/g, 백황: 1.13 ± 0.07 mg/g). 또한, 3종류의 아위느타리버섯 열수추출물의 DPPH 및 ABTS 라디컬 소거활성을 측정하였으며, 총 페놀 함량 및 총 플라보노이드 함량을 측정하였다. 결론적으로 본 연구를 통하여 아위느타리버섯 열수 추출물로부터 에르고티오네인 함량을 가장 정확하게 측정할 수 있는 분석법을 확립하였으며, 맥송 및 비산 2호 아위느타리버섯 열수추출물 소재는 높은 에르고티오네인 함량과, 높은 항산화 활성을 보유하고 있는 것으로 밝혀져 향후 항산화 효능을 보유한 기능성 화장품 개발에 이용될 수 있을 것으로 판단된다.

• Mass Spectrometry Letters
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• 제11권1호
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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.

• 한국물환경학회지
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• 제32권6호
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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.

• 한국식품영양과학회지
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• 제37권5호
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• pp.612-617
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• 2008

우리나라 남해안 연안에 서식하는 복섬(Takifugu niphobles) 간장의 TTX유도체들을 활성탄 칼럼을 이용하여 부분 정제하고, Hydrophilic Interaction Liquid Chromatography(HILIC)를 사용한 LC/MS(SIM mode)로 분석하였으며, pH와 가열 조건에 따른 독성 변화를 조사하고 복섬 fillet으로 복국 조리시 독성의 분포를 조사하였다. 복섬 간장의 독성분은 LC/MS에 의하여 7개의 성분이 분석되었으며, 각 성분의 함량과 조성은 5,6,11-trideoxyTTX(34.0%, 1,029.6nmol/g), 6,11-dideoxyTTX(29.3%, 887.6 nmol/g), TTX (22.1%, 667.8 nmol/g), 4,9-anhydroTTX(11.2%, 339.3nmol/g), 11-deoxyTTX＋5-deoxyTTX(2.6%, 78.6 nmol/g), 4-epiTTX(0.8%, 23.6 nmol/g), 5,6,11-trideoxyTTX(34.0%), 6,11-dideoxyTTX(29.3%), TTX(22.1%), 4,9-anhydroTTX (11.2%), 4-epiTTX(0.8%)이었다. 복섬 간장 추출물 희석액의 독성은 pH에 따라 크게 변하여 pH 3에서 8.4 MU/mL로 최고의 독성을 나타내었고, 알칼리로 갈수록 독성이 감소하여 pH 10에서는 pH 3의 1/7(1.4 MU/mL)을 나타내었다. 각각의 pH(pH 5, 7, 9)에서 $80^{\circ}C,\;100^{\circ}C,\;115^{\circ}C$를 유지하며 가열 시 온도는 높을수록, 시간은 길수록 독성은 감소하였다. 특히, 산성이나 중성에서는 독성이 완만하게 감소하는 경향을 보였으나, 알칼리 영역인 pH 9에서는 가열 10분 후에 $80^{\circ}C$의 경우라도 최초 독성(79 MU/mL)이 1/2 이하로 급격히 감소하였으며, $115^{\circ}C$에서는 완전 소멸하였다. 복섬 개체별 fillet 중의 총 독량은 $43.2{\sim}106.7$ MU로 개체에 따라서 2.5배까지 독량의 차이를 나타내었으며, 복섬 가식부에 3배량의 물을 가하여 10분간 끓일 경우 총 독량의 $64{\sim}78%$는 국물 중으로 용출되어 나와 육에 남아있는 것보다 독성이 강하였다.