Inhibitory Potential of Red Cabbage against Digestive Enzymes

Jul 28, 2017 - Among the varieties, the highest inhibitory activity against α-glucosidase (IC50 = 3.87 ± 0.12 mg dry weight (DW) of cabbage/mL) and ...
0 downloads 9 Views 519KB Size
Subscriber access provided by UNIV OF NEWCASTLE

Article

Inhibitory potential of red cabbage against digestive enzymes linked to obesity and type 2 diabetes Anna Podsedek, Iwona Majewska, and Alicja Kucharska J. Agric. Food Chem., Just Accepted Manuscript • DOI: 10.1021/acs.jafc.7b02499 • Publication Date (Web): 28 Jul 2017 Downloaded from http://pubs.acs.org on July 29, 2017

Just Accepted “Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted online prior to technical editing, formatting for publication and author proofing. The American Chemical Society provides “Just Accepted” as a free service to the research community to expedite the dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been fully peer reviewed, but should not be considered the official version of record. They are accessible to all readers and citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors or consequences arising from the use of information contained in these “Just Accepted” manuscripts.

Journal of Agricultural and Food Chemistry is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.

Page 1 of 28

Journal of Agricultural and Food Chemistry

Inhibitory potential of red cabbage against digestive enzymes linked to obesity and type 2 diabetes Anna Podsędeka*, Iwona Majewskaa, Alicja Z. Kucharskab a

Institute of Technical Biochemistry, Faculty of Biotechnology and Food Sciences,

Lodz University of Technology, Stefanowskiego 4/10, 90-924 Łódź, Poland b

Department of Fruit, Vegetable and Plant Nutraceutical Technology, Wroclaw University of Environmental

and Life Sciences, Chełmońskiego 37, 51-630 Wrocław, Poland

*

Corresponding author: Anna Podsędek

Institute of Technical Biochemistry, Faculty of Biotechnology and Food Sciences, Lodz University of Technology, Stefanowskiego 4/10, 90-924 Łódź, Poland Fax number: 48 42 6366618 Phone: 48 42 6313435 E-mail: [email protected]

1

ACS Paragon Plus Environment

Journal of Agricultural and Food Chemistry

Page 2 of 28

1

ABSTRACT

2

Assays of the inhibitory potential against enzymes involved in carbohydrate and lipid

3

digestion (α-amylase, α-glucosidase and lipase) as well as the total contents of phenolics and

4

anthocyanins, anthocyanin profile, and antioxidant capacity revealed significant differences (p

5

< 0.05) between five varieties of red cabbage. Among the varieties, the highest inhibitory

6

activity against α-glucosidase (IC50 = 3.87±0.12 mg DW of cabbage/mL) and lipase

7

(IC50=1.57±0.06 mg DW/mL) was exhibited by Koda variety, which showed the highest

8

antioxidant capacity in ABTS (TEAC = 141±4.71 µmol/g DW) and FRAP (TEAC = 125±

9

1.94 µmol/g DW) assays, and had the highest total phenolics level (19.6±0.48 mg/g DW). The

10

highest total anthocyanin content (12.0±0.16 mg/g DW) and inhibitory activity against α-

11

amylase (IC50 = 69.0±3.65 mg DW of cabbage/mL) was shown by the Kissendrup variety.

12

The anthocyanin profiles of these two varieties were characterized by the highest percentages

13

of diacylated cyanidin derivatives. There was no correlation between the contents of phenolic

14

compounds and lipase inhibitory activity, but inhibition of α-amylase was correlated with

15

concentrations of monoacylated and diacylated anthocyanins, while inhibition of α-

16

glucosidase increased with total phenolics and diacylated anthocyanins levels.

17 18

KEYWORDS: red cabbage, α-amylase, α-glucosidase, lipase, antioxidant capacity

19

20

INTRODUCTION

21

The increasing prevalence of obesity is accompanied by a growing prevalence of type 2

22

diabetes and is associated in part with major worldwide changes in caloric intake and diet

23

composition. One of strategies aiming at prevention and treatment of obesity and type 2 2

ACS Paragon Plus Environment

Page 3 of 28

Journal of Agricultural and Food Chemistry

24

diabetes relies on application of nutrient digestion and absorption inhibitors. Suppression of

25

activity of pancreatic lipase, α-glucosidase and α-amylase is positively associated with the

26

reduction of fat and sugar absorption from the gastrointestinal tract. Plant derived foods,

27

especially fruits and vegetables, can be a dietary source of polyphenolic inhibitors of digestive

28

enzymes involved in lipid and carbohydrate metabolism.1-3

29

Numerous fruits have been screened in vitro as the source of pancreatic lipase, α-

30

glucosidase and α-amylase inhibitors,2-7 while there are only a few reports on the occurrence

31

of such inhibitors in vegetables.5,8-10 The results of these studies suggest the positive

32

correlation between the inhibitory activity of vegetables and polyphenol and anthocyanin

33

contents. It was found that an anthocyanin extract and a polyphenol extract from red cabbage

34

inhibited α-glucosidase and lipase, respectively.5,8 Therefore, we decided to determine the

35

inhibitory activity of five red cabbage varieties against digestive enzymes degrading dietary

36

sugars and lipids.

37

Red cabbage (Brassica oleracea var. capitata rubra) is a rich dietary source of phenolic

38

compounds, especially structurally differentiated acylated anthocyanins.11-14 This widely

39

cultivated in Europe, North America, China and Japan vegetable demonstrated the potential

40

therapeutic effect in diabetic and obese rats.15,16 To the best of our knowledge there are no

41

literature reports comparing the anti-obesity and anti-diabetic effects, antioxidant capacity

42

and anthocyanin profiles of different red cabbage varieties. The objective of this study was to

43

determine: inhibitory activities of five varieties of red cabbage against digestive enzymes such

44

as: porcine pancreatic lipase type II, intestinal rat α-glucosidase and α-amylase from porcine

45

pancreas type VI-B, antioxidant capacities (radical-scavenging capacity (ABTS assay) and

46

ferric-reducing antioxidant power (FRAP assay), the contents of total phenols (using Folin-

47

Ciocalteu reagent) and total anthocyanins (by pH-differential assay) as well as anthocyanins

48

profiles (by UPLC-PDA-MS technique). 3

ACS Paragon Plus Environment

Journal of Agricultural and Food Chemistry

49

Page 4 of 28

MATERIALS AND METHODS

50

Chemicals. Gallic acid, cyanidin-3-glucoside, rat intestinal α-glucosidase (EC 3.2.1.20), α-

51

amylase (EC 3.2.11) from porcine pancreas type VI-B, lipase (EC 3.1.1.3) from porcine

52

pancreas type II, 4-methylumbelliferyl α-D-glucopyranoside (4-MUG), 4-methylumbelliferyl

53

oleate (4-MUO), TRIS-base, potassium persulfate, 2,2’-azinobis(3-ethyl-benzothiazoline-6-

54

sulphonic

55

tetramethychroman-2-carboxylic acid (Trolox), acarbose, orlistat and acetonitrile were

56

obtained from Sigma. Cyanidin 3-glucoside was purchased from Extrasynthese (Genay,

57

France). Methanol and sodium carbonate were purchased from Chempur. Folin-Ciocalteu

58

reagent, potato starch, hydrochloric acid, iodine, potassium iodide, sodium chloride and

59

calcium chloride of analytical grade were purchased from POCH (Gliwice, Poland). Ultra

60

purity water was prepared in the laboratory using a SimplicityTM Water Purification System

61

(Millipore, Marlborough, MA, USA).

acid)

(ABTS),

2,4,6-tris-2-pyridyl-s-triazine

(TPTZ),

6-hydroxy-2,5,7,8-

62

Plant Material. Koda and Kissendrup red cabbage (Brassica oleracea var capitata rubra)

63

varieties were purchased from a farm of the PlantiCo Horticulture Breeding and Seed

64

Production (Golębiew, Poland) while varieties Haco, Kalibos and Langedijker were obtained

65

from commercial gardens near Łódź (central region of Poland). The edible parts of raw red

66

cabbage were chopped, lyophilised (Alpha 1-2 LDplus, Christ, Osterode, Germany), finely

67

ground using a coffee grinder and applied for preparation of crude extracts.

68

Extraction Procedure. Samples of lyophilised cabbage (2 g) were extracted twice with 50

69

mL of 70% methanol (v/v) for 15 min at room temperature using a magnetic stirrer and the

70

suspensions were centrifuged at 4000 rpm for 15 min.17 The two supernatants were combined

71

and concentrated using a rotary evaporator (Büchi Labortechnik AG, Switzerland) at 40 °C to

72

the volume of 20 mL. Thus, 1 mL of crude extract was equivalent to 0.1 g of lyophilised red

73

cabbage. 4

ACS Paragon Plus Environment

Page 5 of 28

Journal of Agricultural and Food Chemistry

74

Lipase Inhibition Assay. The activity of lipase was determined in the absence (control)

75

and presence of the inhibitor by measuring the release of 4-methylumbelliferone from 4-

76

MUO, using the fluorimetric method, as described previously.6 Reaction mixtures containing

77

4-MUO as substrate, the lipase from porcine pancreas type II, crude extract and Tris buffer

78

(pH 7.4) were incubated at 37 °C for 20 min. The controls contained the buffer instead of the

79

extract. Orlistat was used as positive control. The amount of 4-methylumbelliferone released

80

by lipase was measured with a microplate reader (SynergyTM2, BioTek Instruments Inc.) at

81

an excitation wavelength of 360 nm and at an emission wavelength of 460 nm. The

82

percentage inhibition (%) was calculated according to the following formula: ℎ % = 100 ∗ 1 −

 − 

  − 

83

Where FA, FC, FB and FD were the values of fluorescence of the sample, control, blank sample

84

and blank control, respectively.

85

α-Glucosidase Inhibition Assay. The inhibition of α-glucosidase activity was determined

86

by measuring the amount of 4-methylumbelliferone hydrolyzed from a fluorogenic substrate

87

(4-MUG). The α-glucosidase solution was prepared as described previously by Podsędek et

88

al.6 Aliquots (20 µL) of the red cabbage extracts were combined with 40 µL of freshly

89

prepared enzyme solution and 40 µL of 0.5 mM 4-MUG solution in the Tris buffer (pH 6.9).

90

The mixtures were incubated at 37 °C for 20 min and the fluorescence (FA) (excitation at 390

91

nm and emission at 410 nm) was measured using a SynergyTM2 (BioTek Instruments Inc.).

92

Blank and α-glucosidase controls were also prepared. The α-glucosidase control (FC)

93

contained the buffer (20 µL), substrate solution (40 µL) and enzyme (40 µL). Blank (FB)

94

contained the cabbage extract (20 µL), buffer (20 µL) and enzyme (40 µL) while the blank to

95

control (FD) contained the buffer and enzyme solution. Acarbose was used as positive control.

5

ACS Paragon Plus Environment

Journal of Agricultural and Food Chemistry

96 97

Page 6 of 28

The percentage α-glucosidase activity inhibition was calculated according to the following equation: ℎ % = 100 ∗ 1 −

 − 

  − 

98

Where: FA,FB, FC and FD were the values of fluorescence of the sample, blank sample, control

99

and blank control, respectively.

100

α-Amylase Inhibition Assay. The inhibiting effect of the red cabbage crude extracts on α-

101

amylase activity was assayed as previously described.6 Before this assay the red cabbage

102

extracts were concentrated tenfold. The reaction mixture contained 1% potato starch gel (the

103

substrate), cabbage extract, phosphate buffer (pH 6.9) and α-amylase from porcine pancreas

104

type VI-B. Acarbose was used as positive control. After incubation at 37 °C for 10 min, the

105

reaction was stopped by the addition of HCl. After that, I2 in KI was added and the absorbance

106

was read at 600 nm (using a SynergyTM2, BioTek Instruments Inc.). The percentage inhibition

107

of α-amylase activity was calculated as follows: ℎ % = 100 ∗ 1 −

 − 

  − 

108

Where: AA and AB were the values of absorbance of mixtures containing crude extract and

109

starch with or without amylase, respectively. AC and AD were the values of absorbance of

110

mixtures containing starch and amylase or only starch.

111

IC50 Values for Enzyme Inhibition. Inhibitory activities of crude extracts of red cabbage

112

against the lipase, α-glucosidase and α-amylase were expressed as IC50 values. IC50 is defined

113

as the concentration of the dry weight (DW) of red cabbage per mL of reaction mixture under

114

assay conditions that halves the enzyme activity. The IC50 values were obtained from the

115

least-squares regression line of the plots presenting the logarithm of the sample concentration

116

(log) vs the percentage enzyme inhibition (%). 6

ACS Paragon Plus Environment

Page 7 of 28

Journal of Agricultural and Food Chemistry

117

Antioxidant Capacity Assays. The ABTS•+ and FRAP assays were carried out according

118

to Re et al.18 and Benzie & Strain.19 The procedures were the same as described elsewhere.6

119

The antioxidant capacity was expressed as µmol of Trolox/g DW of red cabbage (TEAC).

120

The linearity range for ABTS assay was determined as 2-17 µM Trolox (R2 = 0.9981), and for

121

FRAP assy as 0.5-15 µM Trolox (R2 = 0.9986).

122

Total Phenolics Assay. Total phenolics content (TPC) assay was based on reaction with

123

the Folin-Ciocalteu reagent and absorbance measurements at 760 nm as described

124

previously.6 TPC was expressed as mg of gallic acid equivalents (GAE)/g DW of red

125

cabbage. The calibration curve ranged from 0.5-4 µg gallic acid/mL (R2 = 0.9981).

126

Total Anthocyanins Assay. Total anthocyanin content (TAC) was determined by the pH-

127

differential method according to Nicoue et al.20 The absorbance of diluted crude extracts from

128

red cabbage varieties was measured at 530 and 700 nm. TAC was expressed as mg of

129

cyanidin-3-glucoside equivalents (CGE)/g DW of red cabbage.

130

Identification and Quantification of Individual Anthocyanins. Anthocyanins were

131

identified using an Acquity UPLC system coupled with a quadruple-time of flight (Q-TOF)

132

MS instrument (Waters Corp., Milford, MA, USA), equipped with an electrospray ionization

133

(ESI) source. Anthocyanins were separated on an Acquity TMBEH C18 column (100 x 2.1

134

mm, 1.7 µm; Waters) operating at 30 °C. The mobile phase was a mixture of 0.1% formic

135

acid (A) and acetonitrile (B). The gradient program was as follows: initial conditions 99%

136

(A), 12 min 65% (A), 12.5 min 100% (B), 13.5 min 99% (A). The flow rate was 0.42 mL/min

137

and the injection volume was 5 µL. The major operating parameters for the Q-TOF-MS were

138

set as follows: capillary voltage of 2.0 kV, cone voltage of 40 V, cone gas flow of 1 L/h,

139

collision energy of 28-30 eV, source temperature of 100 °C, desolvation temperature of 250

140

°C, argon - collision gas, desolvation gas (nitrogen) at a flow rate of 600 L/h, data acquisition 7

ACS Paragon Plus Environment

Journal of Agricultural and Food Chemistry

Page 8 of 28

141

range of m/z 100-2000 Da, and the positive ionization mode. The data were collected by

142

Mass-LynxTM V 4.1 software. Anthocyanins were identified basing on the comparison of

143

their UV-Vis spectra and MS/MS fragmentation spectra with the previously published data.11-

144

13

145

Anthocyanins were quantified according to Mizgier et al.12 using a Dionex HPLC system

146

(Germering, Germany) equipped with an Ultimate 3000 diode array detector. The detector

147

was coupled with a LPG-3400A pump, an EWPS-3000SI autosampler, a column thermostat

148

TCC-3000SD and Chromeleonv. 6.8 software. A Cadenza Imtakt column C5-C18 (75 x 4.6

149

mm) equipped with a guard column were used. The mobile phase was a mixture of solvent A

150

(4.5% formic acid, v/v) and solvent B (acetonitrile). The elution system was as follows: 0–1

151

min 5% B; 20 min 25% B; 21 min 100% B; 26 min 100% B; 27 min 5% B. The flow rate of

152

the mobile phase was 1.0 mL/min and the injection volume was 20 µL. The column operated

153

at 30 °C and the separated compounds were monitored at 520 nm. All samples were analysed

154

in duplicate. The results were calculated as mg of cyanidin-3-glucoside equivalents (CGE)/g

155

DW of red cabbage. The linearity range for this assay was determined as 10-75 µg/mL (R2 =

156

0.9995).

157

Statistical Analysis. Unless otherwise stated, data were expressed as the means ± standard

158

deviations of triplicate measurements. The results were analyzed by means of a one-way

159

analysis of variance (ANOVA). A Tukey’s post hoc test was used to determine differences

160

between the means at the significance level p

326

Kissendrup > Langedijker > Haco > Kalibos, regardless of the assay method. These data were

327

consistent with the previous results and confirmed the conclusion that the antioxidant

328

potential of red cabbage is variety-dependent.13,17,35 For comparison, Wiczkowski et al.13

329

reported that among five red cabbage varieties, the highest antioxidant capacity against

330

superoxide anion radical, ABTS●+ cation radical and peroxyl radical was exhibited by the

331

Langedijker Polona variety, followed by the Langedijker Dauer 2, Kissendrup and Koda ones.

332

A few authors reported that acylation of anthocyanins increased their antioxidant activity.

333

This relationship was observed for delphinidin37, pelargonidin38 and for cyanidin32

334

derivatives. According to the latter authors, the diacylated form of cyanidin-3-diglucoside-5-

335

glucoside showed the stronger antioxidant activity against ABTS●+cation radical and peroxyl

336

radical in comparison to the monoacylated and not acylated forms. Among the five red

337

cabbage varieties tested in our study, the percentage of diacylated cyanidin-3-diglucoside-515

ACS Paragon Plus Environment

Journal of Agricultural and Food Chemistry

Page 16 of 28

338

glucoside among total anthocyanins was the highest in the var. Kissendrup (24.74%) and

339

Koda (23.44%), which showed the highest antioxidant potential.

340

Pearson Correlation Coefficient Analysis. The Pearson correlation analysis was

341

conducted to assess the interplay between the content of phenolic substances and both enzyme

342

inhibition potential and antioxidant capacity. The data presented in Table 4, suggest the

343

moderate correlation (the values of Pearson coefficient range from -0.413 to -0.655) between

344

the contents of total phenolics and total anthocyanins and the degree of both α-amylase and

345

α-glucosidase inhibition, which in turn suggests the synergistic or antagonistic effect between

346

structurally different phenolic compounds. Therefore, the inhibitory effects of red cabbage

347

against the aforementioned digestive enzymes cannot be predicted based on the high phenolic

348

and anthocyanin levels. Also our previous study6 showed no correlation between the total

349

phenolics content in fruit extracts and inhibition of pancreatic lipase, α-glucosidase or α-

350

amylase (r < -0.530). Moreover, some other compounds present in the crude extracts may

351

influence the inhibitory effect of phenolics. The solvent used for extraction (70% methanol) in

352

this study is nonselective and dissolves not only phenolic compounds but also large amounts

353

of sugars, organic acids and proteins.

354

The data of this study showed that the inhibitory activities of the red cabbage extracts

355

toward α-amylase were correlated with mono- and diacylated anthocyanins (r > - 0.70). In

356

case of α-glucosidase, the strongest correlation was observed between the IC50 values and the

357

total diacylated anthocyanins. Sreerama et al. suggested that the inhibition of α-glucosidase

358

might be caused mainly by the occurrence of anthocyanins in methanolic extracts of different

359

beans.26 Gironés-Vilaplana et al. showed that the inhibition of α-glucosidase was correlated

360

with both the total contents of anthocyanins and non-red polyphenols in Latin-American

361

fruits.4 16

ACS Paragon Plus Environment

Page 17 of 28

Journal of Agricultural and Food Chemistry

362

The inhibition of pancreatic lipase was negatively correlated with the contents of

363

anthocyanins, with an exception of diacylated anthocyanins. The lack of correlation between

364

inhibition of lipase activity and total anthocyanin content was also reported for the black and

365

red beans.26 However, the inhibition of lipase by fruits4 and anthocyanin-containing extracts

366

from fruits, vegetables, legumes and cereals5 was positively correlated with total anthocyanins

367

contents.

368

The data presented in Table 4 (the values of Pearson correlation coefficient of r ≥ 0.72)

369

provide evidence of the highly positive correlation between the contents of total phenolics,

370

total anthocyanins, not acylated, monoacylated and diacylated pigments contents, and

371

antioxidant capacities (ABTS scavenging and FRAP assays). The high correlation between

372

the results of ABTS scavenging and FRAP assays and the contents of total phenols is not

373

surprising because the latter compounds were assayed using the Folin-Ciocialteu reagent,

374

which is also applied to determine the reducing capacity. Also other authors observed the

375

positive correlation between the levels of total phenolics and total anthocyanins, and the

376

antioxidant potential of plant derived food.4,9,13,17,25

377

To summarize, the inhibitory activities against α-glucosidase, α-amylase and lipase as well

378

as the high antioxidant activities of five red cabbage varieties suggest that this vegetable may

379

be used to prevent and treat diabetes and obesity. Our results showed that the biological

380

activities and levels of phenolic compounds were variety-dependent. These results may help

381

to exploit the potential of red cabbage as a healthy diet component for obese and diabetic

382

people as well as a source of nutraceuticals. However, further studies are necessary to

383

determine the effects of red cabbage phenolics in more complex systems and in various food

384

matrices, both in the in vitro and in vivo digestion processes.

385

Notes

386

The authors declare no competing financial interest. 17

ACS Paragon Plus Environment

Journal of Agricultural and Food Chemistry

387

Page 18 of 28

REFERENCES

388

(1)

Fu, C.;

Jiang, Y.; Guo, J.; Su, Z. Natural products with anti-obesity effects and

389

different mechanisms of action. J.Agric. Food Chem., 2016, 64, 9571-9585.

390

(2)

391

Silveira, D. α-Amylase inhibitors: A review of raw material and isolated compounds from

392

plant source. J. Pharmacol. Pharm. Sci., 2012, 15, 141-183.

393

(3)

394

A natural approach to treat diabetes. Pharmacogn. Rev., 2011, 5(9), 19-29.

395

(4)

396

Moreno, A. A. Evaluation of Latin-American fruits rich in phytochemicals with biological

397

effects. J. Funct. Foods, 2014, 7, 599-608.

398

(5)

399

the anthocyanin profile and in vitro pancreatic lipase inhibition by anthocyanin-containing

400

extracts of fruits, vegetables, legumes and cereals. J. Sci. Food Agric., 2016, 96, 4713-4723.

401

(6)

402

inhibitory effect on digestive enzymes and antioxidant potential of commonly consumed

403

fruits. J. Agric. Food Chem., 2014, 62, 4610-4617.

404

(7)

405

of polyphenolic extracts from selected edible plants as α-amylase, α-glucosidase and PTP1B

406

inhibitors, and β pancreatic cells cytoprotective agents – A comparative study. Curr. Top.

407

Med. Chem., 2015, 15, 2431-2444.

408

(8)

409

α-glucosidase in an immobilized enzyme assay system. Food Sci. Technol. Res., 2006, 12,

410

275-277.

De Sales, P. M.; de Souza, P. M.; Simeoni, L. A.; de Oliveira Magalhães, P.;

Kumar, S.; Narwal, S.; Kumar, V.; Prakash, O. α-Glucosidase inhibitors from plants:

Gironés-Vilaplana, A.; Baenas, N.; Villaño, D.; Speisky, H.; Garcia-Viguera, C.;

Fabroni, S.; Ballistreri, G.; Amenta, M.; Romeo, F. V.; Rapisarda, P. Screening of

Podsędek, A.; Majewska, I.; Redzynia, M.; Sosnowska, D.; Koziołkiewicz, M. In vitro

Zakłos-Szyda, M.; Majewska, I.; Redzynia, M.; Koziołkiewicz, M. Antidiabetic effect

Kawada, Y.; Miura, M.; Gomyo, T. Inhibitory effect of vegetables, fruits and herbs on

18

ACS Paragon Plus Environment

Page 19 of 28

Journal of Agricultural and Food Chemistry

411

(9)

Mai, T. T.; Thu, N. N.; Tien, P. G.; Chuyen, N. V. Alpha-glucosidase inhibitory and

412

antioxidant activities of Vietnamese edible plants and their relationships with polyphenol

413

contents. J. Nutr. Sci. Vitaminol., 2007, 53, 267-276.

414

(10) Matsui, T.; Ebuchi, S.; Kobayaschi, M.; Fukui, K.; Sugita, K.; Terahara, N.;

415

Matsumoto, K. Anti-hyperglycemic effect of diacylated anthocyanin from Ipomea batatas

416

cultivar Ayamurasaki can be achieved by through the α-glucosidase inhibitory action. J.

417

Agric. Food Chem., 2002, 50, 7244-7248.

418

(11) Charron, C. S.; Clevidence, B. A.; Britz, S. J.; Novotny, S. J. Effect of dose size on

419

bioavailability of acylated and nonacylated anthocyanins from red cabbage (Brassica

420

oleracea L. var. capitata). J. Agric. Food Chem., 2007, 55, 5354-5362.

421

(12) Mizgier, P.; Kucharska, A. Z.; Sokół-Łętowska, A.; Kolniak-Ostek, J.; Kidoń, M.;

422

Fecka, I. Characterization of phenolic compounds and antioxidant and anti-inflamatory

423

properties of red cabbage and purple carrot extracts. J. Funct. Foods, 2016, 21, 133-146.

424

(13) Wiczkowski, W.; Topolska, J.; Honke, J. Anthocyanins profile and antioxidant

425

capacity of red cabbage are influenced by genotype and vegetation period. J. Funct. Foods,

426

2014, 7, 201-211.

427

(14) Wu, X.; Beecher, G. R.; Holden, J. M.; Haytowitz, D.; Gebhardt, S. E.; Prior, R. L.

428

Concentrations of anthocyanins in common foods in the United States and estimation of

429

normal consumption. J. Agric. Food Chem., 2006, 54, 4069-4075.

430

(15) Farag, M. A.; Ammar, N. M.; Kholeif, T. E.; Metwally, N. S.; El-Sheikh, N. M.;

431

Wessjohan, L. A.; Abdet-Hamid, A. Z. Rats’ urinary metabolomes reveal the potential roles

432

of functional foods and exercise in obesity management. Food Funct., 2017, 8, 985-996.

433

(16) Kataya, H. A.; Hamza, A. A. Red cabbage (Brasica oleracea) ameliorates diabetic

434

nephrophaty in rats. J. Evidence-Based Complementary Altern. Med., 2008, 5, 281-287.

19

ACS Paragon Plus Environment

Journal of Agricultural and Food Chemistry

Page 20 of 28

435

(17) Podsędek, A.; Sosnowska, D.; Redzynia, M.; Anders, B. Antioxidant capacity and

436

content of Brassica oleracea dietary antioxidants. Int. J. Food Sci. Technol., 2006, 41 (Supp

437

1), 49-58.

438

(18) Re, R.; Pellergini, N.; Protegente, A.; Pannal, A.; Yang, M.; Rice-Evans, C.

439

Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free

440

Radical Biol. Med., 1999, 26, 1231-1237.

441

(19) Benzie, J. F. F.; Strain, J. J. The ferric reducing ability of plasma (FRAP) as

442

measurement of „antioxidant power“: the FRAP assay. Anal. Biochem., 1996, 239, 70-76.

443

(20) Nicoue, E. E.;

444

Quebec: Extraction and identification. J. Agric. Food Chem., 2007, 55, 5626-5635.

445

(21) Wu, X.; Beecher, G. R.; Holden, J. M.; Haytowitz, D.; Gebhardt, S. E.; Prior, R. L.

446

Lipophilic and hydrophilic antioxidant capacities of common foods in the United States. J.

447

Agric. Food Chem., 2004, 52, 4026-4037.

448

(22) Ahmadiani, N.; Robbins, R. J.; Collins, T. M.; Giusti, M. M. Anthocyanins contents,

449

profiles, and color characteristic of red cabbage extracts from different cultivars and

450

maturity stages. J. Agric. Food Chem., 2014, 62, 7524-7531.

451

(23) Podsędek, A.; Redzynia, M.; Klewicka, E.; Koziołkiewicz, M. Matrix effects on the

452

stability and antioxidant activity of red cabbage anthocyanins under simulated

453

gastrointestinal digestion. BioMed Res. Int., 2014, ID 365738, 11 pages.

454

(24) Birari, R. B.; Bhutani, K. K. Pancreatic lipase inhibitors from natural sources:

455

unexplored potential. Drug Discovery Today, 2007, 12, 879–889.

456

(25) Zhang, B.; Deng, Z.; Ramdath, D. D.; Tang, Y.; Chen, P. X.; Liu, R.; Liu, R., Q.;

457

Tsao, R. Phenolic profiles of 20 Canadian lentil cultivars and their contribution to

458

antioxidant activity and inhibitory effects on α-glucosidase and pancreatic lipase. Food

459

Chem., 2015, 172, 862-872.

Savard, S.;

Belkacemi, K. Anthocyanins in wild blueberries of

20

ACS Paragon Plus Environment

Page 21 of 28

Journal of Agricultural and Food Chemistry

460

(26) Sreerama, Y.N.; Takahashi, Y.; Yamaki, K. Phenolic antioxidants in some Vigna

461

species of legumes and their distinct inhibitory effects on α-glucosidase and pancreatic

462

lipase activities. J. Food Sci., 2012, 77, C927-C933.

463

(27) Esatbeyoglu, T.; Rodrigez-Werner, M.; Schlösser, A.; Winterhalter, P.; Rimbach, G.

464

Fractionation, enzyme inhibitory and cellular antioxidant activity of bioactives from purple

465

sweet potato (Ipomoea batatas). Food Chem., 2017, 221, 447-456.

466

(28) Li, Y.; Wen, S.; Kota, B. P.; Peng, G.; Li, G. Q.; Yamahara, J.; Roufogalis, B. D.

467

Punica granatum flower extract, a potent alpha-glucosidase inhibitor, improves postprandial

468

hyperglycemia in Zucker diabetic fatty rats. J. Ethnopharmacol., 2005, 99, 239–244.

469

(29) Kwon, Y.I.; Apostolidis, E.; Shetty, K. Inhibitory potential of wine and tea against α-

470

amylase and α-glucosidase for management of hyperglycemia linked to type 2 diabetes. J.

471

Food Biochem., 2008, 32, 15-31.

472

(30) Matsui, T.; Ueda, T.; Oki, T.; Sugita, K.; Terahara, N.; Matsumoto, K. α-Glucosidase

473

inhibitory action of natural acylated anthocyanins. 1. Survey of natural pigments with potent

474

inhibitory activity. J. Agric. Food Chem., 2001, 49, 1948-1951.

475

(31) Matsui, T.; Ueda, T.; Oki, T.; Sugita, K.; Terahara, N.; Matsumoto, K. α-Glucosidase

476

inhibitory action of natural acylated anthocyanins. 2. α-Glucosidase inhibition by isolated

477

acylated anthocyanins. J. Agric. Food Chem., 2001, 49, 1952-1956.

478

(32) Wiczkowski, W.; Szawara-Nowak, D.; Romaszko, J. The impact of red cabbage

479

fermentation on bioavailability of anthocyanins and antioxidant capacity of human plasma.

480

Food Chem., 2016, 190, 730-740.

481

(33) McDougall, G.J.; Fyffe, S.; Dobson, P.; Stewart, D. Anthocyanins from red cabbage –

482

stability to simulated gastrointestinal digestion. Phytochemistry, 2007, 68, 1285-1294.

483

(34) McMurray, F.; Patten, D. A.; Harper, M.-E. Reactive oxygen species and oxidative

484

stress in obesity – recent findings and empirical approaches. Obesity, 2016, 24, 2301-2310. 21

ACS Paragon Plus Environment

Journal of Agricultural and Food Chemistry

Page 22 of 28

485

(35) Wold, A. B.; Wicklund, T.; Hafner, K. Antioxidant activity in commonly grown and

486

consumed vegetables: a screening survey. J. Appl. Bot. Food Qual., 2006, 80, 111-115.

487

(36) Tan, J. B. L.; Lim, Y. Y. Critical analysis of current methods for assessing the in vitro

488

antioxidant and antibacterial activity of plant extracts. Food Chem., 2015, 172, 814-822.

489

(37) Azuma, K.; Ohyama, A.; Ippoushi, K.; Ichiyanagi, T.; Takeuchi, A.; Saito, T.;

490

Fukuoka, H. Structure and antioxidant activity of anthocyanins in many accessions of

491

eggplant and its related species. J. Agric. Food Chem., 2008, 56, 10154-10159.

492

(38) Matsufuji, H.; Kido, H.; Misawa, H.; Yaguchi,J.; Otsuki, T.; Chinol, M.; Takeda, M.;

493

Yamagata, K. Stability to light, and hydrogen peroxide at different pH values and DPPH

494

radical scavenging activity of acylated anthocyanins from red radish extract. J. Agric. Food

495

Chem., 2007, 55, 3692-3701.

496

(39) Wiczkowski, W.; Szawara-Nowak, D.; Topolska, J. Red cabbage anthocyanins:

497

Profile, isolation, identification, and antioxidant activity. Food Res. Int., 2013, 51, 303-309.

22

ACS Paragon Plus Environment

Page 23 of 28

Journal of Agricultural and Food Chemistry

Table 1 The total phenolics (TPC), total anthocyanins (TAC) and dry matter of the red cabbage TPC

TAC

(mg GAE/g DW)

(mg CGE/g DW)

Dry matter of lyophilised cabbage (%)

Haco

12.1±0.47b

6.38±0.27b

89.7±0.18d

Kalibos

10.1±0.10a

4.65±0.05a

84.8±0.17a

Kissendrup

18.7±0.22d

12.0±0.16d

90.1±0.20d

Koda

19.6±0.48e

10.4±0.16c

89.0±0.07c

Langedijker

16.3±0.59c

10.8±0.12c

86.8±0.25b

Red cabbage varieties

Values are expressed as mean ± standard deviation (n =3). Mean ± standard deviation in the same column with different letters denote statistically significant difference at p < 0.05. GAE, gallic acid equivalents; CGE, cyanidin-3-glucoside equivalents; DW, dry weight.

23

ACS Paragon Plus Environment

Journal of Agricultural and Food Chemistry

Page 24 of 28

Table 2 Profiles of anthocyanins in five red cabbage varieties Peak

Compound

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17

cyanidin 3-diglucoside-5-glucoside cyanidin 3,5-diglucoside cyanidin 3-(sinapoyl)-diglucoside-5-glucoside cyanidin 3-(sinapoyl)-triglucoside-5-glucoside cyanidin 3-(p-coumaroyl)-triglucoside-5-glucoside cyanidin 3-(feruloyl)-triglucoside-5-glucoside cyanidin 3-(sinapoyl)-diglucoside-5-glucoside cyanidin 3-(sinapoyl)-triglucoside-5-glucoside cyanidin 3-(sinapoyl)-diglucoside-5-glucoside cyanidin 3-(p-coumaroyl)(sinapoyl)-triglucoside-5-glucoside cyanidin 3-(feruloyl)(sinapoyl)-triglucoside-5-glucoside cyanidin 3-(caffeoyl)-diglucoside-5-glucoside cyanidin 3-(sinapoyl)(sinapoyl)-triglucoside-5-glucoside cyanidin 3-(p-coumaroyl)-diglucoside-5-glucoside cyanidin 3-(feruloyl)-diglucoside-5-glucoside cyanidin 3-(sinapoyl)-diglucoside-5-glucoside cyanidin 3-(sinapoyl)-glucoside-5-glucoside

[MS+H]+ (m/z) 773 611 979 1141 1081 1111 979 1141 979 1287 1317 935 1347 919 949 979 817

18 19 20 21

cyanidin 3-(p-coumaroyl)(sinapoyl)-diglucoside-5-glucoside cyanidin 3-(p-hydroxyferuoyl)(sinapoyl)-triglucoside-5-glucoside cyanidin 3-(feruloyl)(sinapoyl)-diglucoside-5-glucoside cyanidin 3-(sinapoyl)(sinapoyl)-diglucoside-5-glucoside

1125 1171 1155 1185

Fragments (m/z) 611/449/287 449/287 817/449/287 979/449/287 919/449/287 949/787/449/287 817/449/287 979/817/449/287 817/449/287 1125/449/287 449/287 773/449/287 1023/773/449/287 757/449/287 787/449/287 817/449/287 655/449/287 963/449/287 1009/449/287 993/449/287 1025/449/287 total1

% of contribution in total anthocyanins content Haco Kalibos Kissendrup Koda 22.1 39.4 18.3 36.4 2.36 0.81 1.64 2.03 0.70 0.70 0.86 1.63 1.22 1.62 1.64 4.92 2.54 8.31 3.90 4.63

Langedijker 36.7 3.34 0.48 0.75 4.29

2.25

5.45

2.12

3.17

3.95

4.39 0.41 1.66 1.33 0.70 1.51 19.9

0.65 0.15 1.08 1.19 0.05 0.27 13.1

1.91 0.07 1.89 1.46 0.52 1.00 23.6

2.16 0.25 1.21 1.47 0.87 0.39 9.90

3.91 0.41 0.88 0.71 0.54 0.82 19.4

24.8

20.7

19.5

10.6

17.4

1.59

0

1.20

0

0

5.55 0.18 3.85 2.96 6.14±0.28b

1.88 0 2.05 2.59 4.64±0.06a

8.43 0 4.54 7.42 11.9±0.28d

5.47 0.08 6.38 8.44 10.1±0.21c

3.31 0.20 2.09 0.82 10.5±0.25c

Data represent mean values ± standard deviation (n=2). Means in line related to total anthocyanin content followed by different letters are significantly different (p < 0.05). 1 – total anthocyanin content was expressed as mg of cyanidin-3 glucoside equivalents per gram dry weight of red cabbage.

24

ACS Paragon Plus Environment

Page 25 of 28

Journal of Agricultural and Food Chemistry

Table 3 Inhibitory activities against pancreatic lipase, α-amylase, and α-glucosidase of five red cabbage varieties IC50 (mg DW/mL) Variety

pancreatic lipase

α-amylase

α-glucosidase

Haco

1.90±0.10b

87.1±0.33b

7.07±0.32d

Kalibos

1.79±0.11ab

119±7.49d

6.00±0.57c

Kissendrup

2.19±0.02c

69.0±3.65a

4.97±0.46b

Koda

1.57±0.06a

101±4.84c

3.87±0.12a

Langedijker

4.10±0.12d

107±3.13cd

7.06±0.17d

Values are expressed as mean ± standard deviation (n =3). Mean ± standard deviation in the same column with different letters denote statistically significant difference at p < 0.05. DW, dry weight of red cabbage.

Table 4 Pearson’s correlation coefficients (r) between total phenolics, total anthocyanins,nonmono- and diacylated anthocyanins and antioxidant capacities, and digestive enzymes inhibitory activities of five red cabbage varieties IC50 (mg DW of red cabbage/mL)

TEAC (µmol Trolox/g DW of red cabbage)

α-amylase

αpancreatic glucosidase lipase

ABTS

FRAP

total phenolics

-0.476

-0.655

0.129

0.990

0.991

total anthocyanins

-0,537

-0.413

0.393

0.971

0.969

nonacylated anthocyanins

0,253

-0.280

0.557

0.734

0.736

monoacylated anthocyanins -0,709

-0.105

0.487

0.760

0.754

diacylated anthocyanins

-0.726

-0.261

0.793

0.792

-0,782

25

ACS Paragon Plus Environment

Journal of Agricultural and Food Chemistry

15+16 8.40

Page 26 of 28

Range: 4e-1

21

9.37

14 3.0e-1

8.19

2.5e-1

1 3.59

AU

2.0e-1

18

1.5e-1

9.02

8

3+4

1.0e-1

20

6.41

10 11 13 12 7.41

7.04 7.26

4.82

5 6.11

5.0e-2

2

9

17 19 8.79

6.71

3.87

0.0 3.00

4.00

5.00

6.00

7.00

8.00

Time 10.00

9.00

Fig.1.

160 d

TEAC (µ µmol Trolox/g DW)

140

d c

d

d

120 100 80

c b

Haco b

a

Kalibos a

60

Kissendrup Koda

40

Langedijker 20 0 ABTS

FRAP Methods

Fig.2.

26

ACS Paragon Plus Environment

Page 27 of 28

Journal of Agricultural and Food Chemistry

List of figure captions Fig.1. UPLC-DAD chromatogram (520 nm) of anthocyanins from red cabbage var. Haco Fig.2. The dependence of TEAC value on the red cabbage variety and assay method

27

ACS Paragon Plus Environment

Journal of Agricultural and Food Chemistry

Page 28 of 28

TOC GRAPHIC

28

ACS Paragon Plus Environment