表七、

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捌、表

表一、SDR-E 及其區分物之產率及乾重

Table 1. Yield and dry weight of SDR-E and its fractions

Yield⁽³⁾ Dry weight Fractions⁽¹⁾⁽²⁾

(%) (g)

SDR-E 100.0 180.7

SDR-E-H₂O 49.3 89.2

SDR-E-EA 43.4 78.5

SDR-E-EA(S) 25.9 46.8

SDR-E-EA(L) 17.6 31.7

(1) SDR: sorghum distilled residue

SDR-E: 95% ethanolic extract of SDR SDR-E-EA: ethyl acetate extract of SDR-E SDR-E-H2O: deionized water extract of SDR-E

(2) Concentration of fractions were 0.5 mg/mL

(3) Yield (%) = (dry weight of SDR-E fraction / SDR dry weight) × 100%

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表二、SDR-E 及其區分物之抗氧化能力

Table 2. DPPH scavenging, 15-LOX inhibition and antioxidative activity of SDR-E and its fractions

Concentration DPPH scavenging 15-LOX Antioxydative⁽¹⁾ Fractions⁽²⁾

(μg/mL) efficiecy (%) inhibition (%) activity (%) SDR-E 500 8.03 ± 1.96 ^c 31.13 ± 4.63 ^b 38.73 ± 3.63 ^b SDR-E-EA(S) 500 44.57 ± 0.29 ^a 71.29 ± 4.96 ^a 71.48 ± 4.41 ^a SDR-E-EA(L) 500 10.86 ± 0.59 ^c 29.35 ± 4.61 ^bc 21.41 ± 2.79 ^c SDR-E-H2O 500 15.99 ± 2.21 ^b 23.26 ± 4.21 ^c 43.10 ± 4.30 ^b

Trolox 25 41.45 ± 0.90 ─ 94.18 ± 1.39

NDGA 3.125 ─ 83.87 ± 2.30 ─

Values were mean ± SD (n = 3).

Letters in the same column with different means difference significantly (p < 0.05).

(1) Antioxidative activity: inhibitory ability of hemoglobin-induced linolenic acid oxidation.

(2) SDR: sorghum distilled residue

SDR-E: 95% ethanolic extract of SDR SDR-E-EA: ethyl acetate extract of SDR-E SDR-E-H₂O: d.d.H₂O extract of SDR-E

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表三、SDR-E 及其區分物之總多酚含量及類黃酮含量

Table 3. Total phenols and flavonoids contents of SDR-E and its fractions

Gallic acid equivalent Quercetin equivalent Fractions

(mg/g extract⁽¹⁾) (mg/g extract⁽¹⁾) SDR-E 69.55 ± 5.06 ^b 1.69 ± 0.08 ^b SDR-E-H₂O 39.55 ± 2.34 ^c 1.38 ± 0.00 ^c SDR-E-EA(S) 153.28 ± 2.80 ^a 8.44 ± 0.43 ^a SDR-E-EA(L) 38.21 ± 1.29 ^c 1.11 ± 0.06 ^bc Values were mean ± SD (n = 3).

Letters in the same column with different means difference significantly (p < 0.05).

(1) The extract means each fraction weight.

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表四、SDR-E-EA(S)管柱層析區分物之乾重及回收率

Table 4. Dry weight and yield of SDR-E-EA(S) and its fractions by the silica gel column chromatography

Solvent system (%) Dry weight Yield⁽¹⁾ Fractions

Hexane Ethyl acetate Acetone Methanol H₂O (g) (%)

SDR-E-EA(S) 1.6615 25.89

SDR-E-EA(S)-I 90 10 0.3490 5.44

SDR-E-EA(S)-II 70 30 0.2267 3.53

SDR-E-EA(S)-III 50 50 0.2447 3.81

SDR-E-EA(S)-IV 100 0.1485 2.31

SDR-E-EA(S)-V 100 0.2330 3.63

SDR-E-EA(S)-VI 100 0.2505 3.90

SDR-E-EA(S)-VII 100 0.1909 2.98

(1) Yield (%) = (dry weight of SDR-E-EA(S) fraction / SDR-E dry weight) × 100%

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表五、SDR-E-EA(S)管柱層析區分物之抑制脂氧合酶活性半抑制濃度

Table 5. Inhibitory effects of lipoxygenase activity by SDR-E-EA(S) of chromatography on the silica gel column

Fractions IC₅₀ (μg/mL)

SDR-E-EA(S)-I 91.741 ± 3.47 ^b SDR-E-EA(S)-II 19.048 ± 1.79 ^a SDR-E-EA(S)-III 17.987 ± 1.24 ^a SDR-E-EA(S)-IV 14.351 ± 1.98 ^a SDR-E-EA(S)-V 12.881 ± 1.37 ^a SDR-E-EA(S)-VI 20.073 ± 2.03 ^a SDR-E-EA(S)-VII 144.495 ± 7.85 ^c Values were mean ± SD (n = 3).

Letters in the same column with different means difference significantly (p < 0.05).

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表六、SDR-E-EA(S)管柱層析區分物之總多酚含量及類黃酮含量

Table 6. Total phenols and flavonoids contents of SDR-E-EA(S) and its fractions by the silica gel column chromatography

Gallic acid equivalent Quercetin equivalent Fractions

(mg/g extract⁽¹⁾) (mg/g extract⁽¹⁾) SDR-E-EA(S) 153.28 ± 2.80 ^c 8.44 ± 0.43 ^e SDR-E-EA(S)-I 28.75 ± 1.15 ^f 0.75 ± 0.03 ^h SDR-E-EA(S)-II 107.68 ± 2.43 ^e 4.10 ± 0.03 ^f SDR-E-EA(S)-III 133.28 ± 2.12 ^d 25.36 ± 0.50 ^a SDR-E-EA(S)-IV 246.35 ± 4.74 ^a 19.49 ± 0.22 ^b SDR-E-EA(S)-V 188.88 ± 3.17 ^b 17.41 ± 0.15 ^c SDR-E-EA(S)-VI 152.75 ± 4.55 ^c 10.25 ± 0.11 ^d SDR-E-EA(S)-VII 25.15 ± 0.83 ^f 2.78 ± 0.13 ^g Values were mean ± SD (n = 3).

Letters in the same column with different means difference significantly (p < 0.05).

(1) The extract means each fraction weight.

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表七、SDR-E-EA(S)-V 管柱層析之各區分乾重、回收率及 DPPH 自由基清 除能力

Table 7. Dry weight, yield and DPPH scavenging efficiency of SDR-E-EA(S)-V and its fractions by chromatography on silica gel column

Dry weight Yield⁽²⁾ DPPH scavenging Fractions⁽¹⁾

(mg) (%) efficiency (%) SDR-E-EA(S)-V 2300 3.6306 62.35 ± 2.67 ^bc SDR-E-EA(S)-V-A 28.5 0.0450 80.52 ± 0.74 ^a

SDR-E-EA(S)-V-B 253.6 0.4002 49.66 ± 7.45 ^ef SDR-E-EA(S)-V-C 143.0 0.2257 28.57 ± 4.92 ^gh SDR-E-EA(S)-V-D 145.1 0.2290 28.32 ± 1.62 ^gh SDR-E-EA(S)-V-E 289.9 0.4575 20.04 ± 6.45 ^ij SDR-E-EA(S)-V-F 207.8 0.3280 30.24 ± 2.43 ^gh SDR-E-EA(S)-V-G 114.6 0.1809 59.49 ± 5.01 ^cd SDR-E-EA(S)-V-H 79.0 0.1247 53.31 ± 1.50 ^de SDR-E-EA(S)-V-I 32.4 0.0511 46.57 ± 5.79 ^ef SDR-E-EA(S)-V-J 37.8 0.0597 23.25 ± 6.55 ^hij SDR-E-EA(S)-V-K 50.6 0.0799 42.42 ± 7.15 ^f SDR-E-EA(S)-V-L 23.9 0.0377 46.75 ± 6.08 ^ef SDR-E-EA(S)-V-M 14.9 0.0235 68.98 ± 4.45 ^b SDR-E-EA(S)-V-N 280.7 0.4430 76.69 ± 5.94 ^a SDR-E-EA(S)-V-O 223.8 0.3532 54.17 ± 1.13 ^de SDR-E-EA(S)-V-P 68.6 0.1083 27.33 ± 1.26 ^ghi SDR-E-EA(S)-V-Q 28.4 0.0448 32.47 ± 0.37 ^g SDR-E-EA(S)-V-R 39.8 0.0628 16.88 ± 4.35 ^j SDR-E-EA(S)-V-S 49.3 0.0778 9.40 ± 1.02 ^k SDR-E-EA(S)-V-T 51.8 0.0818 48.61 ± 1.94 ^ef SDR-E-EA(S)-V-U 78.4 0.1237 19.79 ± 2.33 ^ij Values were mean ± SD (n = 3).

Letters in the same column with different means difference significantly (p <

0.05).

(1)Concentration of fractions were 1.0 mg/mL

(2)Yield (%) = (dry weight of SDR-E-EA(S)-V fraction / SDR-E dry weight)

× 100%

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表八、SDR-E-EA(S)-V 管柱層析之區分物清除 DPPH 自由基之半抑制濃度

Table 8. EC50 values for DPPH scavenging efficiency of SDR-E-EA(S)-V and its fractions by the silica gel column chromatography

Fractions EC50 (μg/mL)

SDR-E-EA(S)-V 725.15 ± 38.72 ^e SDR-E-EA(S)-V-A 291.83 ± 14.08 ^a SDR-E-EA(S)-V-G 651.82 ± 17.57 ^c SDR-E-EA(S)-V-M 663.94 ± 8.20 ^d SDR-E-EA(S)-V-N 581.11 ± 5.39 ^b

Trolox 30.09 ± 1.24

Values were mean ± SD (n = 3).

Letters in the same column with different means difference significantly (p < 0.05).

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表九、SDR-E-EA(S)-V 管柱層析區分物之抑制脂氧合酶活性半抑制濃度

Table 9. Inhibitory effect of lipoxygenase activity by SDR-E-EA(S)-V of chromatography on silica gel column

Fractions IC50 (μg/mL)

SDR-E-EA(S)-V 12.88 ± 1.37 ^b SDR-E-EA(S)-V-A 8.94 ± 0.49 ^a SDR-E-EA(S)-V-G 14.63 ± 2.61 ^b SDR-E-EA(S)-V-M 14.94 ± 3.28 ^b SDR-E-EA(S)-V-N 11.40 ± 1.78 ^ab

NDGA 0.12 ± 0.03 Values were mean ± SD (n = 3).

Letters in the same column with different means difference significantly (p < 0.05).

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表十、SDR-E-EA(S)-V 管柱層析區分物之總多酚含量及類黃酮含量

Table 10. Total phenols and flavonoids contents of SDR-E-EA(S)-V and its fractions by the silica gel column chromatography

Gallic acid equivalent Quercetin equivalent Fractions

(mg/g extract⁽¹⁾) (mg/g extract⁽¹⁾) SDR-E-EA(S)-V 188.88 ± 3.17 ^c 17.41 ± 0.15 ^e SDR-E-EA(S)-V-A 282.56 ± 6.97 ^b 55.46 ± 0.80 ^a SDR-E-EA(S)-V-G 275.89 ± 4.69 ^b 23.36 ± 0.46 ^d SDR-E-EA(S)-V-M 173.49 ± 2.31 ^d 32.84 ± 0.61 ^b SDR-E-EA(S)-V-N 306.29 ± 5.21 ^a 27.54 ± 0.23 ^c Values were mean ± SD (n = 3).

Letters in the same column with different means difference significantly (p < 0.05).

(1) The extract means each fraction weight.

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表十一、高粱酒糟乙醇萃取物區分物之 TLC 呈色反應 Table 11. Characteristic tests for the SDR-E fractions

＋: detectable

－: non-detectable

呈色劑 物質 SDR-E SDR-E-EA(S) SDR-E-EA(L) SDR-E-H₂O

Ninhydrin

＋ － ＋ ＋

Vanillin- sulfuric acid

高級醇、固醇類、

酚類

＋ ＋ ＋ －

Sulfuric acid

＋ ＋ ＋ －

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表十二、類黃酮化合物對不同試劑的呈色反應

Table 12. Coloring reaction of dye reagents on flavonoids

氨蒸氣 三氯化鋁

試劑化合物

可見光 紫外光 可見光 紫外光

氫氧化鈉 鎂+鹽酸 硫酸

硼氫化鉀 + 鹽酸

三氯化銻

黃酮 黃 亮黃、黃綠 淺黃 綠、黃、褐 黃 橙 深黃→橙

常有特殊螢光 黃 黃

黃酮醇 黃 亮黃、黃

綠、綠 黃 黃、綠 淺黃→橙

(空氣氧化褐變) 紅 深黃→橙

常有特殊螢光 黃 黃

雙氫黃酮 — —

淺黃、黃綠 — 綠黃、

藍-白

黃→橙、

深紅(冷)、紫(熱) 紫紅 橙→深紅 桃紅 黃

異黃酮 — 與未薰前

同 — 黃 黃 — 黃 黃 —

查耳酮 黃、橙紅、

橙、粉紅

橙、紅、

紫、黑

黃、橙、

黃綠 橙→紅 橙→紅 — 橙紅、洋紅 — 紫紅、橙

(中國醫學科學院藥物研究所，1978)

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表十三、類黃酮化合物紫外光吸光範圍

Table 13. The UV absorbance extent of flavonoids

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玖、圖

Gallic acid concentration (μg/mL)

0 20 40 60 80 100 120

Absorbance 675 nm

0.0 0.1 0.2 0.3 0.4 0.5

Y = 0.005X + 0.0128 r

= 0.9991

圖一、沒食子酸之標準檢量曲線 Figure 1. Standard curve of gallic acid

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Quercetin concentration (μg/mL)

0 10 20 30 40 50 60

Absor bance 430 nm

0.0 0.2 0.4 0.6 0.8 1.0 1.2

Y = 0.0201X － 0.0117 r

= 0.9995

圖二、Quercetin 之標準檢量曲線 Figure 2. Standard curve of quercetin

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Concentration (μg/mL)

0 500 1000 1500 2000

DPPH scavenging efficiency (%)

0 20 40 60

80 SDR-E-EA(S)-I

SDR-E-EA(S)-II SDR-E-EA(S)-III SDR-E-EA(S)-IV SDR-E-EA(S)-V SDR-E-EA(S)-VI SDR-E-EA(S)-VII

圖三、SDR-E-EA(S)管柱層析區分物在不同濃度下 DPPH 自由基清除能力 Figure 3. Concentration effect of SDR-E-EA(S) and its fractions by the silica

gel column chromatography on DPPH scavenging efficiency

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SDR-E-EA(S) I II III IV V VI VII Trolox

A ntio xi da tiv e a ct iv ity (% )

0 20 40 60 80 100

bc

e cde cd

ab

a

de

bc 25 μg/mL

圖四、SDR-E-EA(S)及其管柱層析區分物之抑制血色素誘導亞麻油酸氧化作

用

Figure 4. The inhibitory effect of SDR-E-EA(S) and its fractions by the silica gel column chromatography to hemoglobin-induced linoleic acid oxidation

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Concentration (μg/mL)

0 50 100 150 200 250

LOX inhibition (%)

10 20 30 40 50 60 70 80 90

SDR-E-EA(S)-I SDR-E-EA(S)-II SDR-E-EA(S)-III SDR-E-EA(S)-IV SDR-E-EA(S)-V SDR-E-EA(S)-VI SDR-E-EA(S)-VII

圖五、SDR-E-EA(S)管柱層析之區分物在不同濃度下脂氧合酶活性的抑制率

Figure 5. Inhibitory effects of SDR-E-EA(S) and its fractions on the level of lipoxygenase activity

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Concentration (μg/mL)

0 200 400 600 800 1000 1200

DPPH Scavenging efficiency (%)

10 20 30 40 50 60 70 80 90

SDR-E-EA(S)-V-A SDR-E-EA(S)-V-G SDR-E-EA(S)-V-M SDR-E-EA(S)-V-N

圖六、SDR-E-EA(S)-V 管柱層析區分物在不同濃度下 DPPH 自由基清除能 力

Figure 6. Concentration effect of SDR-E-EA(S)-V fractions by the silica gel column chromatography on DPPH scavenging efficiency

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SDR-E-EA(S)-V A G M N Trolox

A ntio xi da tiv e acti v ity ( % )

0 20 40 60 80 100 120

c

a a

a b

12.5 μg/mL

圖七、SDR-E-EA(S)-V 及其管柱層析區分物之抑制血色素誘導亞麻油酸氧 化作用

Figure 7. The inhibitory effect of SDR-E-EA(S)-V and its fractions by the silica gel column chromatography to hemoglobin-induced linoleic acid oxidation

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Concentration (μg/mL)

0 10 20 30 40 50

LOX inhibition (%)

0 20 40 60 80

SDR-E-EA(S)-V-A SDR-E-EA(S)-V-G SDR-E-EA(S)-V-M SDR-E-EA(S)-V-N

圖八、SDR-E-EA(S)-V 管柱層析區分物在不同濃度下脂氧合酶活性的抑制 率

Figure 8. Inhibitory effects of SDR-E-EA(S)-V and its fractions on the level of lipoxygenase activity

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(A) ninhydrin (B) 10% sulfuric acid and (C) vanillin-sulfuric acid

圖九、SDR-E 及其區分物之薄層層析圖譜

Figure 9. TLC chromatograms of SDR-E fractions sprayed with (A) ninhydrin, (B) 10% sulfuric acid and (C) vanillin-sulfuric acid

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(A) sulfuric acid

(B) vanillin-sulfuric acid

圖十、SDR-E-EA(S)及其管柱層析區分物之薄層層析圖譜

Figure 10. TLC chromatograms of SDR-E-EA(S) and its fractions sprayed with (A) sulfuric acid and (B) vanillin-sulfuric acid

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(A) Standard

(B) SDR-E-EA(S)

(C) SDR-E-EA(S)-I

(D) SDR-E-EA(S)-II

(E) SDR-E-EA(S)-III

圖十一、SDR-E-EA(S)及其管柱層析區分物之 HPLC 圖譜

Figure 11. HPLC chromatograms of (A) mixed standard of phenolic acid (B) SDR-E-EA(S), (C) SDR-E-EA(S)-I, (D) SDR-E-EA(S)-II and (E) SDR-E-EA(S)-III

Vanillic acid

p-Coumaric acid

Ferulic acid

p-Coumaric acid

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(A) Standard

(B) SDR-E-EA(S)-IV

(C) SDR-E-EA(S)-V

(D) SDR-E-EA(S)-VI

(E) SDR-E-EA(S)-VII

圖十二、SDR-E-EA(S)管柱層析區分物之 HPLC 圖譜

Figure 12. HPLC chromatograms of (A) mixed standard of phenolic acid (B) SDR-E-EA(S)-IV, (C) SDR-E-EA(S)-V, (D) SDR-E-EA(S)-VI and (E) SDR-E-EA(S)-VII

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(A) Vanillic acid (B)

p-Coumaric

圖十三、SDR-E-EA(S)-II 與酚酸標準品之紫外光圖譜

Figure 13. UV spectrum of the SDR-E-EA(S)-II compared with (A) vanillic acid, (B) p-coumaric acid and (C) ferulic acid

(A) Vanillic acid

(B) p-Coumaric acid (C) Ferulic acid

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Concentration (μg/mL)

0 100 200 300 400 500

LOX inhibition (%)

0 20 40 60 80

Sinapic acid Vanillic acid Ferulic acid p-Coumaric acid Genistein

圖十四、酚酸與異黃酮標準品在不同濃度下抑制脂氧合酶活性之抑制率 Figure 14. Inhibitory effect of standards on the level of lipoxygenase activity

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圖十五、SDR-E-EA(S)-V 及其管柱層析區分物之薄層層析圖譜

Figure 15. TLC chromatograms of SDR-E-EA(S)-V and its fractions sprayed with 10% sulfuric acid

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圖十六、SDR-E-EA(S)-V 及其管柱層析區分物之薄層層析圖譜

Figure 16. TLC chromatograms of SDR-E-EA(S)-V and its fractions sprayed with vanillin-sulfuric acid

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(A) SDR-E-EA(S)-V

(B) SDR-E-EA(S)-V-A

(C) SDR-E-EA(S)-V-G

(D) SDR-E-EA(S)-V-M

(E) SDR-E-EA(S)-V-N

圖十七、SDR-E-EA(S)-V 及其管柱層析區分物之 HPLC 圖譜

Figure 17. HPLC chromatograms of (A) SDR-E-EA(S)-V, (B) SDR-E-EA(S)-V-A, (C) SDR-E-EA(S)-V-G, (D) SDR-E-EA(S)-V-M and (E) SDR-E-EA(S)-V-N

genistein genistein

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(A) mixed isoflavone standard

(B) SDR-E-EA(S)-V-M with standard

(C) SDR-E-EA(S)-V-N with standard

圖十八、SDR-E-EA(S)-V-M 及 SDR-E-EA(S)-V-N 添加 genistin、daidzein 及 genistein 之 HPLC 圖譜

Figure 18. HPLC chromatograms of (A) mixed isoflavone standard, (B) SDR-E-EA(S)-V-M with standard, (C) SDR-E-EA(S)-V-N with standard

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(A) SDR-E-EA(S)-V-A

(B) SDR-E-EA(S)-V-A-G

(C) SDR-E-EA(S)-V-A-M

(D) SDR-E-EA(S)-V-A-N

圖十九、SDR-E-EA(S)-V 管柱層析區分物之紫外光圖譜

Figure 19. UV spectrum of (A) SDR-E-EA(S)-V-A, (B) SDR-E-EA(S)-V-G, (C) SDR-E-EA(S)-V-M, (D) SDR-E-EA(S)-V-N