參考文獻 - 氧化鐵奈米粒子萃取之兒茶素

1. 廖慶樑。2000。茶葉中兒茶素的用途與萃取製程。食品資訊，2000，

6，22-25。

2. 原征彥。1998。兒茶素(catechin)類的生理活性作用。茶多元酚在食品/保健食品產業之製造及應用研討會。

3. Hertog, M.G.L.; Hollman, P.C.H.; Van de Putte, B. J. Agric. Food.

Chem. 1993, 41, 1242.

4. Serafini, M.; Ghiselli, A.; Ferro-Luzzi, A. Eur. J. Clin. Nutr. 1996, 50, 28.

5. Gao, G.; Sofic, E.; Prior, R. J. Agric. Food. Chem. 1996, 44, 3426.

6. Richelle, M.; Tavazzi, I.; Offord, E. J. Agric. Food. Chem. 2001, 49, 3438.

11. Caffeine Content of Food and Drugs. (1996, December 1), Nutrition Action Health Newsletter. Center For Science in the Public Interest.

From the World Wide Web :

http://cspinet.org/nah/caffeine/caffeine_content.htm

12. Erowid. (2006, July 7), Caffeine Content of Beverages, Food, &

Medications. The Vaults of Erowid. From the Word Wide Web:

http://www.erowid.org/chemicals/caffeine/caffeine_info1.shtml 13. Bolton, S.; Gary Null, M.S.; Orthomolecular Psychiatry. 1981, 10, 3,

202.

14. Caffeine-related disorders. Encyclopedia of Mental Disorders. From the World Wide Web:

http://www.minddisorders.com/Br-Del/Caffeine-related-disorders.html 15. Kamijo, Y.; Soma, K.; Asari, Y.; Ohwada, T.; Vet. Hum. Toxicol. 1999,

41, 381.

16. Cedars-Sinai. Gastroesophageal Reflux Disease. (GERD) From the World Wide Web: http://www.csmc.edu/pf_5543.html

17. Goto, T.; Yoshida, Y.; Kiso, M.; Nagashima, H. J.Chromatogr. A. 1996, 749, 295.

18. Price, W. E.; Spitzer, J. C. Food. Chem. 1993, 47, 271.

19. 阮逸明。1991。茶葉可溶分及主要化學成分萃取之研究。台灣茶葉研究彙報。10，89。

20. Lee, B. L.; Ong, C. N. J.Chromatogr. A. 2000, 881, 439.

21. Weiss, D. J.; Anderton, C. R. J.Chromatogr. A. 2003, 1101, 173.

22. 王煌忠；張慶源；邱國隆；陳英玲；張傑明。1999。超臨界二氧化碳萃取茶葉精油及濃縮研究。興大工程學刊。10，3，1。

23. 吳長煜；邱國隆；張慶源；張傑明。1999。二氧化碳萃取烏龍茶葉之兒茶酚類機能性成分。食品科學，26，614。

24. Varike, M.; Koel, M. J.Chromatogr. A. 2003, 990, 225.

25. 呂宗昕。2003。圖解奈米科技與光觸媒。商周。100。

26. Rajh, T.; Chen, L.X.; Lukas, K.; Liu, T. Thurnauer, M.C.; Tiede, D.M.

J. Phys. Chem. B 2002, 106, 10543.

27. Lee, K.H.; Chiang, C.K.; Lin, Z.H.; Chang, H.T. Rapid. Commun.

Mass Spectrom. 2007, 21, 2023.

28. 陳學森。茶葉茶多酚類物質的測定。From the World Wide Web：

http://202.194.137.16/yyzwyz/ziyuan/shiyan/12.pdf

29. Suntornsuk, L.; Kasemsook, S.; Wongyai, S. Electrophoresis. 2003, 24,

30. Volpi, N. Electrophoresis. 2004, 25, 1872.

31. Xu, X.; Qi, X.; Wang, W.; Chen, G. J. Sep. Sci. 2005, 28, 647.

32. Heiger, D. N. High performance capillary electrophoresis-An

introduction, Hewleet-Packard Company, 1992.

33. Pomponio, R.; Gotti, R.; Santagati, N. A.; Cavrini, V. J.Chromatogr. A.

2003, 990, 215.

34. Kodama, S.; Yamamoto, A.; Matsunaga, A.; Yanai, H. Electrophoresis.

2004, 25, 2892.

35. Huang, H. Y.; Lien, W. C. Electrophoresis. 2005, 26, 3134.

36. Gotti, R.; Furlanetto, S.; Pinzauti, S.; Cavrini, V. J.Chromatogr. A.

2006, 1112, 345.

37. Turney, K.; Drake, T. J.; Smith, J. E.; Tan, W.; Harrison, W. W. Rapid.

Commun. Mass Spectrom. 2004, 18, 2367.

38. Massart, R. Magnetic fluids and process for obtaining them. US Patent 4’329’241, 1982.

39. Morin, Ph.; Villard, F.; Dreux, M. J.Chromatogr. 1993, 628, 153.

40. Morin, Ph.; Villard, F.; Dreux, M. J.Chromatogr. 1993, 628, 161.

41. Harris, D. C.; Quantitative Chemical Analysis 7^th

Edition, W. H.

Freeman and company, 2007.

42. Magnetite. Mindat.org. (2001)。From the World Wide Web：

http://www.mindat.org/min-2538.html

43. Sillén, L.G.; Martell, A.E. Stability Constants of Metal-Ion Complexes, The Chemical Society (London), 1964 and 1971.

Table 3. Effect of various pH value on migration time and resolution of catechins

Migration time (min) Resolution

pH Caffeine (-)-EGC (-)-EC (-)-EGCG EGC / EC EGCG / ECG

7.0 4.29 5.78 5.78 6.88 – –

7.5 4.95 6.91 6.98 8.44 0.35 0.50

8.0 5.51 8.13 8.28 10.11 0.51 0.64

8.5 7.74 13.10 13.57 16.71 1.30 1.60

9.0 10.65 20.73 21.59 26.01 1.70 1.73

Table 4. Effect of various methanol concentration on migration time and resolution of catechins

Methanol Migration time (min) Resolution

concentration Caffeine (-)-EGC (-)-EC (-)-EGCG (-)-ECG EGC / EC EGCG / ECG

5% 6.21 10.03 10.31 13.33 13.84 0.97 1.15

10% 7.46 12.49 12.90 17.24 18.03 1.16 1.33

15% 9.13 15.87 16.51 22.90 24.23 1.34 1.58

20% 10.77 19.71 20.69 30.12 32.43 1.59 1.88

Table 5. Effect of various ethanol concentration on migration time and resolution of the catechins

Migration time (min) Resolution

Ethanol Caffeine (-)-EGC (-)-EC (-)-EGCG (-)-ECG EGC / EC EGCG / ECG

5% 6.37 10.24 10.53 13.47 13.98 0.99 1.22

10% 7.78 12.82 13.23 17.03 17.72 1.17 1.34

15% 9.84 16.90 17.55 23.06 24.15 1.42 1.58

20% 12.66 22.60 23.65 31.57 33.45 1.67 1.81

Table 6. Effect of various voltage on migration time and resolution of catechins

Migration time (min) Resolution

Voltage (kV) Caffeine (-)-EGC (-)-EC (-)-EGCG (-)-ECG EGC / EC EGCG / ECG

+15 19.95 33.50 34.72 44.73 46.87 1.58 1.71

+20 13.97 23.24 24.08 30.74 32.16 1.44 1.60

+25 9.84 16.90 17.55 23.06 24.15 1.42 1.57

+30 7.96 13.41 13.84 17.49 18.17 1.26 1.38

Table 7. Optimum conditions for capillary electrophoresis experiment

Experiment conditions

Buffer 360 mM borate with 200 mM tris (pH8) Organic modifier 15% ethanol

Applied voltage +30 kV

Table 8. Migration time, LOD, LOQ, linear range and R square of the caffeine and catechins

Migration Time (min)

LOD (μM)

LOQ

(μM) Linear range (μM) Slope Intercept R

Caffeine 7.96 75 90 90 – 500 108.2 7061.4 0.9993

(-)-EGC 13.41 0.9 5 5 – 500 998.7 2918.4 0.9994

(-)-EC 13.84 1.6 5 5 – 500 881.3 2776.9 0.9904

(-)-EGCG 17.49 0.9 2.5 2.5 – 500 2830.6 8128.2 0.9992

(-)-ECG 18.17 1 5 5 – 500 2711.7 10857 0.9988

Table 9. Adsorption ratio of the (-)-EC in various adsorption time

^a)

Adsorption time (min)

Concentration after absorption (M)

Absorption ratio (%)

5 1.1×10^-5 89.01

10 <5×10^-6 > 95 15 <5×10^-6 > 95

Formate 1.85 3.61 3.95 5.4 25 1

Acetate 3.38 7.1 9.7 25 0

Oxalate 7.54 14.59 20 ─ 0.5

Tartrate 6.49 11.87 9.48 20 0.1

Citrate 11.04 21.2 20 0.1

a) The overall (cumulative) formation constant, β

, is the equilibrium constant for the reaction

M + nL ML

: β

=[ML

]/([M][L]

ⁿ

). β

is related to stepwise formation constants (K

) by β

=K

K

...K

.

Table 11. Recovery of (-)-EC desorbed in various pH of tartrate buffer

^a)

Tartrate buffer pH value

Concentration after

desorption (M) Recovery (%) 3 4.36×10^-5 43.56 3.4 4.37×10^-5 43.74

4 4.77×10^-5 47.67 5.3 5.16×10^-5 51.62

11.2 0 0 a) Original concentration: 10^-4 M

Table 12. Recovery of (-)-EC desorbed in various tartrate buffer

concentration^a)

Tartrate buffer concentration (mM)

Concentration

after desorption(M) Recovery (%) 10 4.29×10^-5 42.91 20 4.31×10^-5 43.10 40 4.41×10^-5 44.14 60 4.57×10^-5 45.72 80 5.15×10^-5 51.58

100 5.16×10^-5 51.62

a) Original concentration: 10^-4 M

Table 13. Recovery of (-)-EC in various desorbed time

^a)

Reaction time (min)

(-)-EC

concentration (M) Recovery (%)

5 4.52×10^-5 42.55

10 5.21×10^-5 52.08 20 5.16×10^-5 51.61 40 4.99×10^-5 49.93 60 5.19×10^-5 51.99 a) Original concentration: 10^-4 M

Table 14. Recovery of (-)-EC in various desorbed times

^a)

Desorption times

Concentration

after desorption (M) Recovery (%) 1^st 5.21×10^-5 52.08 2^nd 8.81×10^-6 8.81 3^rd < 5×10^-6 < 5 4^th < 5×10^-6 < 5 a) Original concentration: 10^-4 M

Table 15. Recovery of (-)-EC desorbed in tartrate buffer with various organic solvent

^a)

10% organic solvent

Concentration 1

^st

desorption (M)

Concentration 2

^nd

desorption (M)

1

^st

recovery (%)

2

^nd

recovery (%)

1

^st

+2

^nd

recovery (%)

none 5.21×10

^-5

8.81×10

49.90 5.27 55.17

a) Original concentration 10

^-4

M

Table 17. Adsorption ratio of the real sample in various MNPs concentration MNPs

concentration Catechin concentration (μM) Adsorption ratio (%)

(pmole) (-)-EGC (-)-EC (-)-EGCG (-)-ECG (-)-EGC (-)-EC (-)-EGCG (-)-ECG 0.096 30.11 5.84 7.26 <5 62.7 63.5 84.1 >38.8 0.192 27.93 5.35 2.72 N.D. 65.4 66.5 94.0 100 0.480 8.31 5.07 N.D. N.D. 90.0 68.3 100 100 0.768 2.56 <5 N.D. N.D. 96.9 >68.8 100 100 0.960 <5 N.D. N.D. N.D. >93.9 100 100 100 1.152 N.D.

^a)

N.D. N.D. N.D. 100 100 100 100

1.440 N.D. N.D. N.D. N.D. 100 100 100 100

a)Not detected

Table 18. Recovery of real sample A with optimum extract method

Recovery (%)

Desorption

times (-)-EGC (-)-EC (-)-EGCG (-)-ECG

1^st 6.8% 79.0% 0% 0%

2^nd 2.6% 7.0% 0% 0%

Reaction formula :

Fig. 4. Scheme of Magnetic nano-particles synthesis.

shake Catechin Solution

CE-UV

Tartarate buffer Solution absorption

desorption MNPs

Solution

shake Static 20min

Static 10min

Fig. 5. Scheme of sample preparation process.

Fig. 6. UV absorbance of catechins and caffeine. Samples: 5×10

^-5

M(A) caffeine; (B) (-)-EGCG; (C) (-)-ECG; (D) (-)-EGC;

(E) (-)-EC.

200 220 240 260 280 300

Absorbance (a. u .)

W avelength (nm )

240 260 280 300

0.00 0.05 0.10 0.15

(C)

Abs

Wavelength (nm)

(D)

(E)

(A)

(B)

O OH

OH

O

OH

B OH HO

Fig. 7. The probable structure of the borate - catechin complex.

0 2 4 6 8 10

(2)

(3) (4) (1) (5)

(A)

(B)

mV

(C)

(D)

Time (min)

Fig. 8. Electropherograms of the catechins and caffeine in various borate

buffer concentration. Samples: 5×10^-5 M of (1)caffeine; (2) (-)-EGC; (3) (-)-EC; (4) (-)-EGCG; and (5) (-)-ECG. Separation electrolyte: (A)60 mM; (B)70 mM; (C)80 mM; and (D)90 mM, pH9 borate buffer. Total length of capillary 65 cm (53 cm effective length); Voltage, +15 kV; UV detection at 275 nm.

0 2 4 6 8 10 12 14 (3) (4) (5)

(2)

(1)

(A)

(B)

(C) (D)

Time (min)

Fig. 9. Electropherograms of the catechins and caffeine in various

tris-borate buffer concentration. Samples: 5×10^-5 M of (1)caffeine; (2) (-)-EGC; (3) (-)-EC; (4) (-)-EGCG; and (5) (-)-ECG. Separation electrolyte: (A)200 mM; (B)150 mM;

(C)100 mM; and (D)50 mM, pH8 Tris-borate buffer. Total length of capillary 65 cm (53 cm effective length); Voltage, +25 kV; UV detection at 275 nm.

0 5 10 15 20 25 30

Fig. 10. Electropherograms of the catechins and caffeine

in various tris-borate buffer pH value. Separation electrolyte: (A)pH 7;

(B)pH 7.5; (C)pH 8; (D)pH 8.5; and(E)pH 9, 200 mM tris-borate buffer. Total length of capillary 65 cm (53 cm effective length);

Voltage: +25 kV; UV detection at 275 nm; Samples: 5×10^-5M of (1)caffeine; (2) (-)-EGC; (3) (-)-EC; (4) (-)-EGCG; and (5) (-)-ECG.

0 5 10 15 20 25 30 35

(2) (3)(4) (5)

(1)

(A)

(3)(4) (5)

mV (1) (2) (B)

(3) (4) (5)

(1) (2)

(C)

(3) (4) (5)

(1) (2)

(D)

(3) (4) (5)

(2)

(E)

(1)

Time (min)

Fig. 11. Electropherograms of the catechins and caffeine

in various methanol concentration. Separation electrolyte: pH 8, 200 mM tris-borate buffer with (A)0%; (B)5%; (C)10%; (D)15% and (E)20% methanol. Total length of capillary 65 cm (53 cm effective length); Voltage: +25 kV; UV detection at 275 nm;

Samples: 5×10^-5M of (1)caffeine; (2) (-)-EGC; (3) (-)-EC; (4) (-)-EGCG; and (5) (-)-ECG.

0 10 20 30

Fig. 12. Electropherograms of the catechins and caffeine in

various ethanol concentration. Separation electrolyte: pH 8, 200 mM tris-borate buffer with (A)0%; (B)5%; (C)10%; (D)15% and (E)20% ethanol. Total length of capillary 65 cm (53 cm effective length); Voltage: +25 kV; UV detection at 275 nm;

Samples: 5×10^-5M of (1)caffeine; (2) (-)-EGC; (3) (-)-EC; (4) (-)-EGCG; and (5) (-)-ECG.

10 20 30 40 50

(4) (5) (2)(3)

(1)

(A)

(4) (5) (2)(3)

mV (1)

(B)

(2)(3)

(4) (5) (1)

(C)

(2) (3)

(4) (5)

(1)

(D)

Time (min)

Fig. 13. Electropherograms of the catechins and caffeine in various

voltage. Separation electrolyte: pH 8, 200 mM tris-borate buffer with 15% ethanol. Total length of capillary 65 cm (53 cm effective length); Voltage. (A)+15 kV; (B)+20 kV; (C) +25 kV;

(D)+30 kV; UV detection at 275 nm; Samples: 5×10^-5M of (1)caffeine; (2) (-)-EGC; (3) (-)-EC; (4) (-)-EGCG; and (5) (-)-ECG.

0 5 10 15 20

(4) (5) (3)

(2)

(1)

Time (min)

Fig. 14. Electropherograms of the catechins and caffeine under optimum

conditions. Sample: 10^-4M of (1) caffeine; (2) (-)-EGC; (3) (-)-EC; (4) (-)-EGCG; and (5) (-)-ECG . Separation electrolyte:

200 mM Tris-borate buffer containing 15% ethanol. Total length of capillary 65 cm (53 cm effective length); Voltage, +30 kV; UV detection at 275 nm.

0 100 200 300 400 500

(A)

(B)

Peak Area (a.u.)

(C) (D) (E)

Concentration (μM)

Fig. 15. The calibration curves of the catechins and caffeine. (A)

(-)-EGCG; (B) (-)-ECG; (C) (-)-EGC; (D) (-)-EC; and (E) caffeine . Separation electrolyte: 200 mM tris-borate buffer containing 15% ethanol. Total length of capillary 65 cm (53 cm effective length); Voltage, +30 kV; UV detection at 275 nm.

Fig. 16. TEM of the magnetic nano-particles.

240 260 280 300 320

Absorption (a.u.)

(A)

(B)

Wavelength (nm)

Fig. 17. UV absorbance of 5×10

^-5M (-)-EC. (A) Without; (B)After MNPs adsorption.

5 10

(A)

(B)

(C)

(D) (E) (F)

Time (min)

Fig. 18. Electropherograms of (-)-EC absorbed by various MNPs

concentration. (A)0; (B)0.032; (C)0.16; (D)0.32; (E)0.64, and (F)0.98 pmole . Separation electrolyte as Fig. 15.

0 5 10

(A)

(B) mV

(C)

(D) (E) (F) (G) (H) (I)

Time (min)

Fig. 19. Electropherograms of caffeine absorbed by various MNPs

concentration. (A)0; (B)0.032; (C)0.16; (D)0.32 (E)0.64; and (F)0.98; (G)1.92; (H)2.56; (I)3.2 pmole. Separation electrolyte as

Fig. 15.

0.0 0.5 1.0 1.5 0

20 40 60 80 100

Absorption %

0.64 pmole

EC Caffeine EGC ECG EGCG

pmole

Fig.20. Adsorption ratio of 10

^-4 M catechins and caffeine adsorbed by various mole of MNPs.

5 10

(A)

mV (B)

(C) (D) (E) (F)

Time (min)

Fig. 21. Electropherograms of (-)-EC after various adsorption time. (A)0;

(B)5; (C)10; (D)15; (E)20, and (F)25 min . Sample: 10^-4M (-)-EC. Separation electrolyte as Fig. 15.

O

Fig. 22. The structure of the (a)oxalate; (b)tartrate and probable structure

of the (c) iron - oxalate complex; (d) iron - tartrate complex.

6 8 10 12 14

(A)

mV (B)

(C) (D) (E) (F)

Time (min)

Fig. 23. Electropherograms of (-)-EC desorbed by various tartrate buffer

pH value. (A) 1×10^-4 M (-)-EC; (B) pH 3.0; (C) pH 3.4; (D) pH 4; (E) pH 5.3, and (F)pH 11.2 . Separation electrolyte as Fig.

15.

6 8 10 12 14

(A)

mV (B)

(C) (D) (E) (F) (G)

Time (min)

Fig. 24. Electropherograms of (-)-EC desorbed by various tartrate buffer

concentration. (A) 1×10^-4 M (-)-EC; (B) 10; (C) 20; (D) 40; (E) 60, (F)80; and (G)100 mM . Separation electrolyte as Fig. 15.

5 10

(A)

(C)

Time (min)

Fig. 28. Electropherograms of 50 mL 2 μM (-)-EC concentrated by MNPs.

(A) 2×10^-6 M (-)-EC; (B) 1^st desorption; and (C) 2^nd desorption.

Separation electrolyte as Fig. 15.

2 4 6 8 10 12 14

(A)

(4)

mV (2)

(3) (5) (1)

Time (min)

Fig. 33. Electropherograms of the real sample D diluted 10-fold. Sample:

(1) Caffeine; (2) (-)-EGC; (3) (-)-EC; (4) (-)-EGCG; and (5) (-)-ECG . Separation electrolyte as Fig. 15.

5 10 15 20

(2) (4)

(3) (5) (1)

Time (min)

Fig. 34. Electropherograms of the real sample E diluted 10-fold. Sample:

(1) Caffeine; (2) (-)-EGC; (3) (-)-EC; (4) (-)-EGCG; and (5) (-)-ECG . Separation electrolyte as Fig. 15.

5 10 15 20

(2) (4)

(3) (5)

(1)

Time (min)

Fig. 35. Electropherograms of the real sample F diluted 10-fold. Sample:

(1) Caffeine; (2) (-)-EGC; (3) (-)-EC; (4) (-)-EGCG; and (5) (-)-ECG . Separation electrolyte as Fig. 15.

5 10 15 20

(2) (4) mV

(3) (5) (1)

Time (min)

Fig. 36. Electropherograms of the real sample G diluted 10-fold. Sample:

(1) Caffeine; (2) (-)-EGC; (3) (-)-EC; (4) (-)-EGCG; and (5) (-)-ECG . Separation electrolyte as Fig. 15.

5 10 15 20

(2) (4)

5 10

(1)

(3) (5) (1)

(A)

(B)

Time (min)

Fig. 37. Electropherograms of the 10-fold diluted real sample (A) without;

and (B) after MNPs adsorption . Sample: (1) Caffeine; (2) (-)-EGC; (3) (-)-EC; (4) (-)-EGCG; and (5) (-)-ECG . Separation electrolyte as Fig 15.

5 10 15

(2)

(3) (4) (5)

(1)

(A)

(B) mV

(C) (D) (E) (F) (G)

Time (min)

Fig. 38. Electropherograms of the 10-fold diluted real sample A adsorbed

by various MNPs concentration. (A)0.096; (B)0.192; (C)0.48;

(D)0.768; (E)0.96; (F)1.152 and (G) 1.44 pmole. Sample: (1) Caffeine; (2) (-)-EGC; (3) (-)-EC; (4) (-)-EGCG; and (5) (-)-ECG . Separation electrolyte as Fig 15.

5 10 15

(A)

(B)

(C)

Time (min)

Fig. 39. Electropherograms of the real sample concentrated by MNPs. (A)

Original real sample concentration; (B) 1^st; and (C) 2^nd of 100 mM pH5.3 tartrate buffer with 10% ethanol desorption.

Separation electrolyte as Fig. 15.

在文檔中氧化鐵奈米粒子萃取之兒茶素 (頁 56-110)