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3.3. CUPRAC Assay (Spectrophotometry)

XL Xican Li
HX Hong Xie

Last updated date: Jan 25, 2023 Views: 485 Forks: 0

original An abbreviated version of this protocol was published in Molecules in Mar, 2018
Effect of Double Bond Position on 2-Phenyl-benzofuran Antioxidants: A Comparative Study of Moracin C and Iso-Moracin C
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总还原能力-Cu

Reducing power assay-Cu

 

1  溶液的配制

 1.1  0.01mol/L CuSO4: 8mg CuSO4+5mL蒸馏水(可测40个数据) (CuSO4:MW=160) 。

 1.2  7.5 mmol/L新铜试剂Neocuproine: 8.1 mg + 5mL无水乙醇 (可测40个数据) (MW=217.3) 

 1.3  0.1mol/L乙酸铵缓冲液（pH = 7.5）：0.385g + 50mL 蒸馏水 (MW= 77.08) 。

   

2  实验方法

 

 

3  计算公式

   相对总还原百分率 = （A- A0）/Amax- A0）

   式中，A0为样品浓度为0时的A450nm值；Amax为在一次测量内最大的A450nm值；A为样品的A450nm值。

 

[附]参考数据

 

 

 

 

BHA 1 mg/mL

体积uL

最终浓度ug/mL

A450nm

Percentage

Mean

SD

0

0

0.027

 

0.03167

 

0.034

 

0.034

 

2

2

0.187

15.21881123

16.91704768

1.550156014

0.208

17.27629001

0.218

18.2560418

4

4

0.436

39.61463096

38.50424559

1.595923151

0.432

39.22273024

0.406

36.67537557

6

6

0.603

55.97648596

55.58458524

0.353254534

0.596

55.2906597

0.598

55.48661006

8

8

0.806

75.86544742

68.58262573

9.870887919

0.772

72.53429131

0.617

57.34813847

10

10

0.931

88.11234487

87.36120183

0.910343557

0.913

86.34879164

0.926

87.62246897

12

12

1.044

99.18354017

100

1.499799151

1.07

101.7308948

1.043

99.08556499

 

 

1.05233

（MAX）

100

Bai/uL

 

 

 

 

 

2

2

0.199

16.39451339

18.58262573

2.42838817

 

 

0.217

18.15806662

 

 

 

 

0.248

21.19529719

 

 

4

4

0.477

43.63161332

42.74983671

1.527281901

 

 

0.45

40.98628347

 

 

 

 

0.477

43.63161332

 

 

6

6

0.704

65.8719791

64.72893534

1.019758001

 

 

0.684

63.91247551

 

 

 

 

0.689

64.4023514

 

 

8

8

0.81

76.25734814

75.21227956

1.982232646

 

 

0.812

76.4532985

 

 

 

 

0.776

72.92619203

 

 

10

10

0.9

85.0751143

84.42194644

1.945574266

 

 

0.909

85.95689092

 

 

 

 

0.871

82.2338341

 

 

12

12

0.975

92.42325278

95.10124102

2.91356067

 

 

1.034

98.20378837

 

 

 

 

0.998

94.67668191

 

 

Trolox/uL

 

 

 

 

 

2

2

0.116

8.262573481

8.229915088

1.029128103

 

 

0.105

7.184846506

 

 

 

 

0.126

9.242325278

 

 

4

4

0.242

20.60744611

18.61528413

1.731510926

 

 

0.21

17.47224037

 

 

 

 

0.213

17.7661659

 

 

6

6

0.304

26.68190725

24.85303723

1.862387646

 

 

0.266

22.95885042

 

 

 

 

0.286

24.91835402

 

 

8

8

0.427

38.73285434

36.51208361

2.254138985

 

 

0.381

34.22599608

 

 

 

 

0.405

36.57740039

 

 

10

10

0.49

44.90529066

44.7093403

2.064465138

 

 

0.508

46.66884389

 

 

 

 

0.466

42.55388635

 

 

12

12

0.618

57.44611365

54.83344219

2.809571027

 

 

0.595

55.19268452

 

 

 

 

0.561

51.86152841

 

 

 

 

 

 

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How to cite：
Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
  1. Li, X and Xie, H(2023). 3.3. CUPRAC Assay (Spectrophotometry). Bio-protocol Preprint. bio-protocol.org/prep2131.
  2. Li, X., Xie, H. and Chen, D.(2018). Effect of Double Bond Position on 2-Phenyl-benzofuran Antioxidants: A Comparative Study of Moracin C and Iso-Moracin C. Molecules 23(4). DOI: 10.3390/molecules23040754

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