• 약학회지
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• 제41권4호
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• pp.484-491
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• 1997

The effect of prolonged treatment with Mori Folium column fraction(MFCF) on intestinal ${\alpha}$-glycosidase activity has been studied in mice made hyperglycemic and hyperinsulinemic by feeding a high carbohydrate-containing diet. Mice were treated for 10 week with or without MFCF, added to the high carbohydrate-containing diet at 50mg/100g food. While MFCF had no effect on body weight, it prevented the rise in glycemia and insulinemia. Maltase, sucrase and lactase activities were measured in intestinal homogenates of proximal, middle and distal segments of jejunoileum. Following 10 week of MFCF administration, MFCF significantly induced all three enzyme activities especially at middle and distal segments. The mechanisms responsible for these changes and their potential biochemical implications remain to be determined.

• 대한수의학회지
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• 제40권1호
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• pp.72-80
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• 2000

Lipopolysaccharides (LPS) of Pasteurella multocida (P multocida) and several Gram-negative bacterial pathogens were analyzed by methanolysis, trifluoroacetylation and gas chromatography (GC) on a fused-silica capillary column. The GC analysis indicated that LPS prepared from a strain of P multocida by phenol-water (PW) or trichloroacetic acid (TCA) extraction were quite different in chemical composition. However, LPS prepared from Salmonella enteritidis by the two extraction methods were very similar. PW-LPS extracts from different Pasteurella strains of a serotype had essentially identical GC patterns. Endotoxic LPS extracted from 16 different serotypes of P multocida by PW or by phenol-chloroform-petroleum ether procedures yielded chromatograms indicating similar composition of the fatty acid moieties but minor differences in carbohydrate content. When the chemical composition of endotoxic LPS extracted from several Gram-negative bacteria (P multocida, Pasteurella hacmolytica, Haemophilus somnus, Actinobacillus ligniersii, Brucella abortus, Treponema hyodysenteriae, Escherichia coli, Bacteriodes fragilis, Salmonella abortus equi and Salmonella enteritidis) were examined, each bacteria showed a unique GC pattern. The carbohydrate constituents in LPS of various Gram-negative bacteria were quite variable not only in the O-specific polysaccharides but also in the core polysaccharides. The LPS of closely related bacteria shared more fatty acid constituents with each other than with unrelated bacteria.

• Archives of Pharmacal Research
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• 제21권5호
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• pp.521-526
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• 1998

The physicochmical properties of recombinant hepatitis B surface antigen (r-HBsAg), which was expressed in C127 mammalian cell were studied. Using roller bottle culture in DMEM supplemented with fetal bovine serum, 10-15 mg/L of r-HBsAg was produced with about 31% of purification yield. The purity of r-HBsAg by HPLC was 99.8% and electron microscopic examination showed homogeneous spherical particle with 22 nm in diameter, a morphological characteristic of HBsAg. The density of r-HBsAg by CsCI density gradient method was 1.19g/ml and the isoelectric point by Mono $P^{TM}$ HR 5/20 column was 4.6. The analysis of subunit protein pattern using SDS-PAGE followed by scanning densitometry gave 81.3% of S protein and 18.7% of pre-S protein. fluorophore-assisted-carbohydrate-electrophoresis analysis showed the relative amount of carbohydrate to protein was 1.7% and it smajr component was N-acetyl glucosamine, which was about 39% of total carbohydrate. The relative amount of lipid to protein determined by vanillin phosphoric acid method was 32.5% and its major component was phospholipid, which was about 70% of total lipid. The physicochemical properties of C127 mammalian cell-derved r-HBsAg are similar to those of p-HBsAg, suggesting that the r-HBsAg can be used in developing a new preventive vaccine against hepatitis B.

• Applied Biological Chemistry
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• 제36권4호
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• pp.310-314
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• 1993

HPLC를 이용하여 fructose oligomer인 fructo올리고당(GF2-GF7) 및 inulo올리고당(F2-F4)의 분리, 정량을 실시하였다. TSK-gel amide 80 column과 acetonitrile-water(65:35; v/v) 용매를 사용하여 각 표준당들을 효과적으로 분리할 수 있었으며 이들의 retention time은 분석시마다 거의 변하지않아 재현성이 있었다. 각 당의 농도에 따른 peak면적을 이용하여 표준곡선을 작성한 결과 넓은 당량의 범위에 걸쳐 결정계수가 0.9884이상의 값을 보여 본 HPLC방법에 의한 정량법이 타당한 것으로 나타났다. 또한 기울기는 비슷하나 y축 절편 값이 당마다 크게 상이하여 몇가지 표준당들을 기준으로 하여 모든 당을 일률적으로 정량하는 것은 적합하지 않으며 각 당에 대한 표준곡선을 하나씩 작성하여 정량하여야함을 알 수 있었다.

• 한국식품과학회지
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• 제28권1호
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• pp.99-104
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• 1996

탄수화물을 갖고 있지 않은 caseinomacropeptide (CMP)를 감성유청분말로부터 12% TCA법에 의해 분리한 결과 100 g의 감성유청분말로 부터 2.7 g을 얻을 수 있었다. 분리된 CMP는 전기영동과 아미노산 조성을 분석한 결과 매우 순도가 높은 CMP로 나타나 12% TCA침전법은 감성유청분말로 부터 CMP를 분리하는 효과적인 방법임을 알 수 있었다. 트립신을 pore glass beads에 carbodiimide (EDC) 방법으로 amide결합에 의해 고정화시켰으며 고정화된 트립신의 양은 1 g glass beads당 20 mg이었다. 수용성 트립신에 의한 CMP 가수분해는 24시간이 지나면 거의 완료되었으며 고정화 트립신은 24시간이 지난 후에도 가수분해가 진행되고 있었다. 가수분해물을 Bio-Gel P4 column에 의해 분획한 후 혈소판 응집 저해작용을 측정한 결과 고정화 트립신에 의해 24시간 가수분해시킨 CMP 분해물이 가장 높은 혈소판 응집 억제능을 보였다.

• 대한수의학회지
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• 제36권3호
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• pp.521-529
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• 1996

In an effort to obtain basic data of carbohydrate metabolism during spermiogenesis of the sexually-matured Jindo dog, the glycogen distribution in the testis was investigated by light and transmission electron microscopy. Periodic acid thiocarbohydrazide silver proteinate physical development(PA-TCH-SP-PD) staining method provided better results in the detection of glycogen granules from Sertoli cells and germ cells than the periodic acid schiff(PAS) staining method did. Pre-treatment of the tissue sections with ${\alpha}$-amylase elicited a significant decrease in PA-TCH-SP-PD stained granules, which suggested that the stained granules were of glycogen origin. High concentration of the glycogen granules were observed in the Sertoli cells, especially in its column, sheet-like processes, club-like processes, and tubular processes. The glycogen granules were unevenly distributed in some Sertoli cell columns. These results strongly indicated that the Sertoli cells of Jindo dogs showed vigorous activity of carbohydrate metabolism.

• 한국식품영양과학회지
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• 제11권3호
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• pp.81-86
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• 1982

인삼(人蔘)사포닌중의 각(各) ginsenosides를 효과적(效果的)으로 분리(分離) 정량(定量)하기 위하여 carbohydrate analysis column을 사용(使用)한 HPLC로 전형적(典型的)인 용매(溶媒) system인 acetonitrile/water의 혼합비율(混合比率)을 80/20에서 94/6까지 조정(調整)하여 retention time과 분리능(分離能)과의 관계(關係)를 비교실험(比較試驗)하였고 또 n-butanol의 첨가효과(添加效果)도 조사(調査)하였다. 기본구조(基本構造)가 다른 diol saponin과 triol saponin을 같은 mobile phase로 만족하게 분리정량(分離定量)하기는 어렵다. 따라서 diol saponin은 acetonitrile/water system(80/20), trol saponin은 acetonitrile/water/n-butanol system(86/14/10)을 mobile phase로 하여 분석(分析)함이 효과적(效果的)이었다. 이 방법(方法)에 따라 백삼(白蔘)과 홍삼(紅蔘)을 정량(定量)한 결과(結果), diol saponin 함량(含量)은 큰 차이(差異)가 없으나 triol saponin 함량(含量)은 홍삼(紅蔘)이 백삼(白蔘)보다 증가(增加)하였다. 특(特)히 PT/PD ratin가 백삼(白蔘)은 0.401인데 비(比)해 홍삼(紅蔘)은 0.561로서 홍삼(紅蔘)이 백삼(白蔘)보다 약 1.4배(倍)나 높았다. 이것이 홍삼(紅蔘)의 생화학적약리효능(生化學的藥理效能)과 깊은 관계(關係)가 있다고 판단(判斷)되며 열처리(熱處理)에 의한 제조공정(製造工程)과 관계(關係)가 있다.

• 한국식품과학회지
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• 제35권3호
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• pp.519-526
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• 2003

본 연구에서는 한국인에게 적합한 비만 개선책을 찾고자 지방 흡수억제 기천뿐만 아니라 과잉의 탄수화물 흡수억제가 함께 작용하는 분화억제에 대한 지방흡수 억제 물질과 율무 열수에서 ${\alpha}$-amylase 활성 저해에 대한 탄수화물 흡수 억제물질이 첨가된 비만 개선 식이조성물이 동물 실험 시 체중 증가가 억제됨을 확인하였다. 호박의 열수 추출물을 ethyl acetate로 추출하여 췌장 lipase 활성 저해를 확인하였으며, 호박의 ethyl acetate 추출물을 농축하여 silica gel column chromatography와 C-18 column chromatography를 이용하여 활성 저해 물질을 분리하였다. 또한 율무 열수는 chloroform으로 추출하여 호박 열수와 같은 방법으로 분리하여 ${\alpha}$-amylase 활성 저해를 확인하였다. 3T3-L1 분화억제 확인 실험에서는 3T3-L1에 분화 유도물질만 처리한 경우 지방 세포로 분화되어 세포내에 지방축적이 일어났지만, 분화 유도물질과 호박 추출물을 동시처리 결과 지방 세포로의 분화가 호박 추출물 $120\;{\mu}g/mL$ 처리 시약 5% 이하로 일어났으며, 세포내에 지방축적도 거의 일어나지 않았다. 동물실험에서 고지방 식이와 비만개선용 식이조성물을 동시에 섭취한 군의 체중 증가가 고지방 식이만 섭취한 군보다 13% 정도 감소함을 확인할 수 있었고, 비만개선용 식이 조성물을 8주간 경구 투여한 뒤 간 기능 지표와 혈액학적 지표 및 흰쥐의 행동학적 소견에서 차이를 나타내지 않아 비만개선용 식이조성물이 경구 투여에 의한 안전성이 있는 것으로 확인되었다. 이들 결과를 종합하면, 호박의 췌장 lipase 활성 저해물질과 율무의 ${\alpha}$-amylase 활성 저해물질을 첨가한 비만개선용 식이조성물이 동물 실험에서 체중 증가를 효과적으로 억제할 수 있음을 관찰하였으며, 이는 비만 환자들의 다이어트 식이로 유용할 것으로 사료되었다.

• Journal of Ginseng Research
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• 제35권1호
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• pp.31-38
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• 2011

In order to distinguish the cultivation area of Panax ginseng, principal component analysis (PCA) using quantitative and qualitative data acquired from HPLC was carried out. A new HPLC method coupled with evaporative light scattering detection (HPLC-ELSD) was developed for the simultaneous quantification of ten major ginsenosides, namely $Rh_1$, $Rg_2$, $Rg_3$, $Rg_1$, Rf, Re, Rd, $Rb_2$, Rc, and $Rb_1$ in the root of P. ginseng C. A. Meyer. Simultaneous separations of these ten ginsenosides were achieved on a carbohydrate analytical column. The mobile phase consisted of acetonitrile-water-isopropanol, and acetonitrile-water-isopropanol using a gradient elution. Distinct differences in qualitative and quantitative characteristics for ginsenosides were found between the ginseng roots produced in two different Korean cultivation areas, Ganghwa and Punggi. The ginsenoside profiles obtained via HPLC analysis were subjected to PCA. PCA score plots using two principal components (PCs) showed good separation for the ginseng roots cultivated in Ganghwa and Punggi. PC1 influenced the separation, capturing 43.6% of the variance, while PC2 affected differentiation, explaining 18.0% of the variance. The highest contribution components were ginsenoside $Rg_3$ for PC1 and ginsenoside Rf for PC2. Particularly, the PCA score plot for the small ginseng roots of six-year old, each of which was light than 147 g fresh weight, showed more distinct discrimination. PC1 influenced the separation between different sample sets, capturing 51.8% of the variance, while PC2 affected differentiation, also explaining 28.0% of the variance. The highest contribution component was ginsenoside Rf for PC1 and ginsenoside $Rg_2$ for PC2. In conclusion, the HPLC-ELSD method using a carbohydrate column allowed for the simultaneous quantification of ten major ginsenosides, and PCA analysis of the ginsenoside peaks shown on the HPLC chromatogram would be a very acceptable strategy for discrimination of the cultivation area of ginseng roots.

• 대한의생명과학회지
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• 제18권2호
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• pp.96-103
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• 2012

Kv 4.2 ion channel protein has an ability to open at subthreshold membrane potentials and to recover quickly from inactivation. That is very important for neuronal signal transmission in vertebrate brain. In order to purify Kv 4.2 protein, the novel purification methods were experimented. The purification procedure utilized chromatography on DE-52 ion exchange column and affinity chromatography on a WGA-Sepharose 4B, and Kv 4.2 affinity column chromatography. It was found that 0.5% (wt./vol.) Triton X-100 detergent in lysis buffer worked well for Kv 4.2 protein solubilization from rat brain membrane. Protein quantitative determination was conducted by BCA method at 562 nm for each purification step to avoid determination interference of protein at 280 nm by detergent. The confirmation of Kv 4.2 existence and amount is performed using by SDS-PAGE/immunoblotting or 96-well dot blotting. The Kv 4.2 without interacting protein that contains carbohydrate, was purified from novel biochemical 3-steps purification method for further research.