• Applied Biological Chemistry
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• 제10권
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• pp.1-13
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• 1968

phytin은 phytic acid의 금속염(金屬鹽)(주(主)로 Ca 와 Mg)임으로 그중(中)의 P,Ca 및 Mg를 정량(定量)하면 순도(純度)를 알 수 있고, 또 분자식(分子式)을 추정(推定)할 수 있다. 저자(著者)는 phytin 중(中)의 P,Ca 및 Mg를 정량분석(定量分析)하는 새로운 방법(방법)으로 서 phytin을 건식(乾式) 분해(分解)하고 ion 교환수지(交換樹脂)로 처리한 다음 Chelate 법(法)으로 정량(定量)하는 방법(方法)을 확정(確定)켰으며 그 결과(結果)를 요약(要約)하면 다음과 같다. 1) phytin 분석(分析)의 전처리과정(前處理課程)으로서는 phytin을 conc. $HNO_3$로 적시면서 $550{\sim}660^{\circ}C$에서 회화(灰化)하는 건식분해법(乾式分解法)을 썼다. 이 방법(方法)은 습식분해법(濕式分解法)보다 분석결과(分析結果)가 정확(正確)하다. 2) phytin을 건식분해(乾式分解)한 시료(試料)를 가지고 종래법(從來法)과 새로운 분석법(分析法) (본법(本法))에 의하여 P,Ca 및 Mg를 정량(定量)하였으며, 본법(本法)은 다음고 같다. phytin 회분(灰分 HCl 용액(溶液)을 양(陽) ion 교환수지(交換樹脂)로 처리하여 양(陽) ion 구분(區分)과 음(陰) ion 분리(分離)하고 양(陽) ion 구분(區分)의 일부(一部)를 pH 7.0로 한다음 완충액(緩衝液)($NH_3-NH_4Cl$으로 pH 10으로 하고 BT 지시약(指示藥)을 써서 표준(標準) EDTA 용액(溶液적정(滴定)하여 Ca와 Mg의 합계치(合計値)를 얻었다. 또 양(陽) ion 구분(區分)의 일부(一部)를 pH 7.0로 하고 표준(標準) EDTA 용액(溶液)을 소량(少量)넣고 8N-KOH로 pH $12{\sim}13$으로 하고 N-N 희석분말(稀釋粉末)을 지시약(指示藥) 으로써 표준(標準) EDTA 용액(溶液)으로 적정(滴定)하여 Ca 치(値)를 얻었다. Ca와 Mg의 합계결정치(合計決定値)와 Ca 적정치(滴定値) 차(差)로 Mg 치(値)를 얻었다. 음(陰) ion 구분(區分)으로부터 상법(常法)에 의하여 $MgNH_4PO_4$의 침전(沈澱)을 만들어서 HCl에 녹키고 일정량(一定量)의 표준(標準) EDTA 용액(溶液)을 넣어 pH 7.0로 한다음 완충액(緩衝液)으로 pH 10으로 하고 BT 지시약(指示藥)을 써서 표준(標準) Mg $SO_4$용액(溶液)으로 적정(滴定)하여 P 치(値)를 얻었다. 본법(本法)으로 Na-phytate를 분석(分析)한 결과(結果) Na-phytate의 분자식(分子式)을 $C_6H_6O_{24}P_6Mg_4CaNa_2{\cdot}5H_2O$라고 하였을 때의 이론치(理論値)에 비(比)하여 P가 98.9% Cark 97.1%, Mg가 99.1%이고 통계처리(統計處理)한 결과분석치(結果分析値)와 이론치(理論値)는 잘 일치(一致)된다. 그러나 종래법(從來法)에 의(依)한 분석치(分析値)는 이론치(理論値)에 비(比)하여 P가 92.40%, Cark 86.80%, Mg가 93.80%로서 이론치(理論値)와 일치(一致)하지 않는다. 3) Na-phytate를 전분(澱粉)과 일정(一定)한 비(比)로 혼합(混合)하고 본법(本法)으로 P,Ca 및 Mc를 정량(定量)한 결과(結果) 이들의 회수율(回收率)은 거의 100%이었다. 4) 본분석법(本分析法)의 정확성(正確性)을 재확인(再確認)하기 위하여 phytic acid 수용액(水溶液)에 $CaCl_2$수용액(水溶液)을 phytic acid 1M:$CaCl_25M:McCl_220M$의 비(比)로 반응(反應)서키어서 Ca 1 원자(原子), Mg 4원자함유(原子含有)된 Na-phytate를 합성(合成)하였으며 이것의 P,Ca 및 Mg 분석치(分析値)와 의(依한) 조제(調製) Naphytate의 분석치(分析値)와 일치(一致)되었다. 이상(以上)과 같이 phytin 시료(試料)를 건식분해(乾式分解)하고 ion 교환수지(交換樹脂)로 처리(處理)한 다음 Chelate 법(法)으로 P,Ca 및 Mg를 정량(定量)하는 본법(本法)은 정확(正確)하고 신속(迅速)한 phytin의 새 분석방법(分析方法)이라고 사료(思料)되는 바이다.

• 대한수의학회지
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• 제36권3호
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• pp.577-598
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• 1996

This study was conducted to identify the effects of caffeine or combinations of caffeine and iron or vitamin E on the lipid and protein components or blood chemistry levels of the serum as well as the total homogenate, mitochondrial and microsomal fraction of the rat(Sprague-Dawley, female) liver. Chronic test were conducted to determine those effects. The chronic test was conducted by dividing rats into 5 groups according to the type of drugs and dosages administrated as follows; the control(group A), and group B was given 25mg/kg caffeine orally once daily for 30 days, group C was given 50mg/kg caffeine orally once daily for 30 days, group D was given 25mg/kg caffeine and orally ferric chloride once daily for 30 days and group E was given 25mg/kg caffeine and 25mg/kg vitamin E once daily for 30 days. The concentrations of glucose, urea nitrogen, uric acid, creatinine, total protein, albumin, A/G ratio, triglyceride, total cholesterol, HDL-cholesterol, free fatty acid, phospholipid as well as the activities of alanine aminotransferase(ALT), aspartate aminotransferase(AST) and alkaline phosphatase(ALP) were measured in the serum of each experimental groups. The concentrations of the carbonyl group and malondiaidehyde(MDA) and the patterns of the SDS-PAGE(Sodium Dodecyl Sulfate - Polyacrylamide Gel Electrophoresis) and fatty acid compositions in free fatty acids and phospholipids were analyzed to determine the oxidative damages and metabolic changes on the lipid and protein components in the serum, and total homogenate, mitochondrial and microsomal fractions of the rat liver. The results obtained from this study were summarized as follows; 1. Body weights of groups B, C, D and E were significantly decreased(p < 0.01) in comparison with that of the control in the chronic test. 2. The concentrations of serum glucose in groups B(124.5mg/dl), C(130.1mg/dl), D(122.1mg/dl), E(119.3mg/dl) were significantly higher(p < 0.01) in comparison to that of the control(101.5mg/dl). But, there were no significant differences in the concentrations of urea nitrogen, uric acid, creatinine, total protein, albumin and A/G ratio in comparison to that of the control. 3. The concentrations of total cholesterol and HDL-cholesterol in serum of groups B(69.6, 53.4mg/dl), C(73.0, 56.3mg/dl), D(68.9, 51.1mg/dl) and E(68.2, 51.3mg/dl) were significantly higher(p < 0.01) in comparison to that of the control(52.6, 38.8mg/dl). On the other hand, the concentrations of triglyceride in serum of groups B(45.0mg/dl), C(40.4mg/dl), D(33.8mg/dl) and E(47.2mg/dl) were significantly lower(p < 0.01) in comparison to that of the control(66.2mg/dl). There were no significant differences in the activities of ALT, AST and ALP in comparison to that of the control. 4. The concentrations of free fatty acid and phospholipid in serum of groups B(45.7, 154.4mg/dl), C(50.0, 167.2mg/dl), D(52.5, 148.4mg/dl) and E(41.1, 159.2mg/dl) were higher(p < 0.01) in comparison to that of the control(35.2, 125.3mg/dl). And the concentrations of the carbonyl group and malondialdehyde in serum of group D(1.82, 0.52nM/mg protein) were significantly higher(p < 0.01) in comparison to the control(1.53nM/mg protein). 5. The concentrations of carbonyl group in total homogenate, mitochondrial and microsomal fraction of group D(1.45, 0.94, 1.67nM/mg protein) were significantly higher (p < 0.01) in comparison to the control(1.16, 0.66, 1.27nM/mg protein). And the concentrations of malondialdehyde in the total homogenate, mitochondrial and microsomal fraction of group D(6.70, 6.10, 1.36nM/mg protein) were significantly higher(p < 0.01) in comparison to the control(5.17, 3.64, 0.68nM/mg protein). 6. As the analytical results of the fatty acid compositions of free fatty acid in serum, the proportions of stearic acid and arachidonic acid of groups B(16.52, 12.62%), C(17.52, 15.18%), D(19.73, 13.47%) and E(17.62, 13.28%) were significantly higher(p < 0.01) in comparison to the control(14.75, 7.88%), but the proportions of oleic acid and linoleic acid of groups B(12.97, 32.59%), C(10.88, 31.23%), D(12.37, 30.66%) and E(11.95, 32.41%) were significantly lower(p < 0.01) in comparison to the control(16.44, 35.12%). Otherwise, as the results of the fatty acid compositions of phospholipid in serum, the proportions of stearic acid and arachidonic acid of groups B(39.37, 16.39%), C(40.63, 17.83%), D(42.73, 15.39%) and E(39.16, 15.70%) were significantly higher(p < 0.01) in comparison to the control(37.74, 14.24%), but the proportions of oleic acid and linoleic acid of groups B(4.03, 14.38%), C(3.54, 12.38%), D(4.52, 11.68%) and E(4.29, 13.64%) were significantly lower(p < 0.01) in comparison to the control(5.53, 16.14%). 7. As the analytical results of the fatty acid compositions of free fatty acid in total homogenate, mitochondrial and microsomal fraction of liver, the proportions of oleic acid of groups B(7.8**, 8.73**, 6.88%) and C(6.89**, 7.75**, 6.58%) were lower(**:p < 0.01) in comparison to the control(8.67, 10.08, 7.81%), but the proportions of arachidonic acid of group C(22.62, 19.79, 23.71%) were significantly higher(p < 0.01) in comparison to the control(20.93, 18.47, 22.24%). And the proportions of palmitic acid of group D(25.95**, 26.16, 26.34**%) were significantly higher(**:p < 0.01) in comparison to the control(24.43, 25.42, 23.34%). In addition, the proportions of linoleic acid of group D(23.43, 25.02, 23.95%) were also significantly higher(p < 0.01) in comparison to the control(22.17, 23.75, 21.26%). The proportions of stearic acid of group D(19.87, 19.76**%) in mitochondrial and microsomal fraction were lower(**:p < 0.01) in comparison to the control(21.01, 24.18%), and the proportions of stearic acid of group E(16.71*, 19.65**%) in mitochondrial and microsomal fraction were significantly lower(**:p < 0.01, *:p < 0.05) in comparison to the control(21.01, 24.18%), and the proportions of linoleic acid of group E(25.04, 29.20, 26.48%) in total homogenate, mitochondria and microsome were significantly higher(p < 0.01) in comparison to the control(22.17, 23.75, 21.26%). 8. As the results of the fatty acid compositions of phospholipid in total homogenate, mitochondrial and microsomal fraction of liver, the proportions of palmitic acid of group D(17.58**, 18.78*, 18.23%**) were significantly higher(**:p < 0.01, *:p < 0.05) in comparison to the control(16.28, 17.22, 16.38%), and the proportions of stearic acid of group D(36.41, 37.23, 39.53%) were also significantly higher(p < 0.01) in comparison to the control(34.18, 34.16, 36.04%). But the proportions of oleic acid(3.41*, 3.11**, 3.12**%) and linoleic acid (18.03**, 15.79**, 14.74**%) of group D were significantly lower(**:p < 0.01, *:p < 0.05) in comparison to the control(oleic : 3.63, 3.72, 3.79%, linoleic : 20.03, 18.71, 18.48%). 9. In order to determine the oxidative damages to the protein in serum, mitochondrial and microsomal fraction of the rat liver, the patterns of the SDS-PAGE were identified, but the results of SDS-PAGE were not significantly different between the control and experimental groups.