Blackwell Science, LtdOxford, UKJFBCJournal of Food Biochemistry0145-8884Copyright 2005 by Food & Nutrition Press, Inc., Trumbull, Connecticut.

2005293278294Original Article ANTI-HYPERGLYCEMIC POTENTIAL OF

SELECTED FOODS P. MCCUE

ET AL.

ANTI-AMYLASE, ANTI-GLUCOSIDASE AND ANTI-ANGIOTENSIN I-

CONVERTING ENZYME POTENTIAL OF SELECTED FOODS

PATRICK MCCUE1, YOUNG-IN KWON2 and KALIDAS SHETTY2,3

1
Program in Molecular and Cellular Biology
University of Massachusetts
Amherst, MA 01003
2
Department of Food Science
University of Massachusetts
Amherst, MA 01003

Received for Publication May 15, 2004

Accepted for Publication December 9, 2004

ABSTRACT

a-Amylase and a-glucosidase have been targeted as potential avenues

for modulation of postprandial hyperglycemia through mild inhibition of the
enzymatic breakdown of complex carbohydrates to decrease meal-derived
glucose absorption. Water-soluble extracts with optimized phenolic content of
selected American and Asian foods were investigated for inhibitory activity
against a-amylase and a-glucosidase, as well as angiotensin I-converting
enzyme (ACE), which has been linked to hyperglycemia-associated hyperten-
sion. Porcine pancreatic a-amylase (PPA) was allowed to react with each
phenolic-optimized food extract, and the derivatized enzyme–phytochemical
mixtures obtained were characterized for residual amylase activity. The a-
glucosidase and ACE activities were determined in the presence of each
phenolic-optimized food extract. The amylase activity was inhibited more than
the glucosidase activity in the presence of these phytochemical extracts, and
more so by Asian foods than by American foods. The Asian spice ginger was
found to possess strong ACE inhibitory activity in addition to significant anti-
amylase activity. The a-amylase enzyme inhibition was positively associated
with extract antioxidant activity and negatively with extract protein content.
The significance of food-grade, plant-based amylase inhibitors for modulation
of carbohydrate breakdown and control of glycemic index of foods in the
context of preventing hyperglycemia and diabetes mellitus complications in
the long term and ACE inhibitors for modulation of associated hypertension
is hypothesized and discussed.
3
Corresponding author. TEL: (413) 545-1022; FAX: (413) 545-1262; EMAIL:
kalidasshetty@yahoo.com

Journal of Food Biochemistry 29 (2005) 278–294. All Rights Reserved.

278 © Copyright 2005, Blackwell Publishing
 ANTI-HYPERGLYCEMIC POTENTIAL OF SELECTED FOODS 279

INTRODUCTION

Hyperglycemia is a condition characterized by an abnormal excess of

sugar in the blood. Elevated postprandial blood glucose levels are widely
recognized as one of the earliest disease markers in the prediction of subse-
quent microvascular and macrovascular complications that can progress to full
symptomatic type 2 diabetes (T2DM) (Ratner 2001; Zimmerman 2001;
Fonseca 2003). While the majority of available synthetic antidiabetic drugs
target the dual metabolic defects that characterize T2DM, impaired insulin
secretion and insulin resistance, some of these drugs (e.g., metformin) can
have negative side effects at high doses (Ohmura et al. 1998; Mudaliar and
Henry 2001; Carroll et al. 2003; Fonseca 2003). Thus, a major goal of antidi-
abetic research is the discovery of anti-hyperglycemic agents that are safe and
that lack any negative side effects.
To this end, research has begun to embrace traditional medicines and
foods from various cultures as scientists search for clues to discover new
therapeutic drugs for T2DM and associated hypertension. Traditional
Indian and Chinese medicines have long used plant and herbal extracts as
antidiabetic agents (Chen et al. 2001; Grover et al. 2002). South American
(e.g., Suriname) traditional medicine uses plant extracts to treat hyperten-
sion to great effect (Hasrat et al. 2004). These plants, which include holy
basil (Ocimum sanctum) and oregano (Origanum vulgare), are typically
rich in phenolic compounds. Phenolic compounds are known to interact
with proteins and can inhibit enzymatic activity (Dawra et al. 1988).
Indeed, many medicinal plant and herbal extracts have been found to
inhibit the enzymatic activity of a-glucosidase and a-amylase (Kim et al.
2000; Grover et al. 2002; Vats et al. 2002; McCue and Shetty 2004;
McCue et al. 2004). Dietary a-glucosidase and a-amylase inhibitors that
act in the gut by inhibiting the enzymatic breakdown of starch, soluble car-
bohydrates and their derived and digested products have been identified as
a potentially natural and safe approach for controlling hyperglycemia
through modulation (i.e., decrease) of meal-derived glucose absorption
(Fonseca 2003).
In this study, we investigated the potential of phenolic-optimized aqueous
extracts of selected foods consistent with typical American and Asian diets
for anti-amylase and anti-glucosidase activities (antidiabetic potential). The
food extracts were analyzed with respect to the soluble phenolic and protein
contents and free radical scavenging antioxidant activities of the extracts.
Further, selected extracts identified with high-antidiabetic potential were fur-
ther investigated for antihypertensive potential via inhibitory activity against
angiotensin I-converting enzyme (ACE), which has been linked to hypergly-
cemia-associated hypertension (DiCarli et al. 2003).
280 P. MCCUE ET AL.

MATERIALS AND METHODS

American Foods
The fresh green pepper (GP), string beans (SB), baby spinach (BB),
broccoli sprouts (BS), red pepper (RP), fresh carrot (FC), Romaine lettuce
(RL), red grape (GR), tomato (TM) and basil leaves (BL) were purchased at
a local supermarket as sources of dietary phenolics. Graham cracker (GC)
(Keebler Co., Elmhurst, IL), Chips Ahoy cookies (CA) (Nabisco Inc., East
Hanover, NJ) and Wheat Thins crackers (WT) (Nabisco) were also used in
this study as selected samples of processed starch foods.

Asian Foods
The powdered Asian spices were purchased at a local market. The
fenugreek (FN), mustard (M), ginger (G), cinnamon (CN) and turmeric (TR)
were from SWAD Brand (Raja Foods, Skokie, IL). The fennel powder (FL)
was of the Laxmi, Shamiana and Joy Brand (House of Spices, Flushing, NY).
The cardamom powder (CM) was from Gaban Spice Co., Tokyo, Japan. The
fresh eggplant (EP), coccinia (CX), bittergourd (BG) and small brinjal (BJ)
were purchased at a local market. All are sources of dietary phenolics.

Extract Preparation
For each of the American food products, 10 g of product (5 g for BL)
was homogenized in 50 mL of distilled water (dH2O) for 1 min using a Waring
laboratory blender set on “HIGH.” The homogenate was centrifuged at
10,000 rpm at 4C for 20 min. The supernatant was vacuum filtered through
Whatman filter paper #1 and then used as the crude extract for each product.
Extracts for the Asian food products were similarly prepared, except that 10 g
of each product was homogenized in 100 mL of dH2O. The extracts were
subsequently optimized for phenolic content as described below.

Total Soluble Phenolic Content Assay

The total soluble phenolic content in each extract was determined using
a previously described method (McCue et al. 2000). A phenolic standard
curve was established at 725 nm with gallic acid (25–200 mg/mL) in 95%
ethanol (EtOH).

Treatment of a-Amylase with Food Extracts

The treatment of amylase was performed as previously described, with
some modifications (McCue and Shetty 2004). Fifty milligrams of powdered
 ANTI-HYPERGLYCEMIC POTENTIAL OF SELECTED FOODS 281

porcine pancreatic a-amylase (PPA) (Sigma Aldrich Chemical Co., St. Louis,
MO, USA) was added to 27 mL of dH2O. For each food extract, a volume
equivalent to 400 mg total phenolic content was added to the above solution,
and the mixture was adjusted to pH 6.9. After dilution to 30-mL total volume,
the amylase–food extract mixtures were incubated for 24 h at 4C with stirring.
The control mixture used was 1 mL of dH2O in place of extract.

Characterization of a-Amylase Activity

The a-amylase activity was determined by the method of McCue and
Shetty (2004), using starch as a substrate in a colorimetric reaction using 3,5-
dinitrosalicylic acid. A standard curve was generated for the hydrolyzed prod-
ucts (reducing groups) using D-(+)-maltose monohydrate. The activity was
calculated as units per milligram of protein, where 1 unit was defined as the
amount of enzyme required to liberate 1 mmol of maltose under assay condi-
tions. The protein content was determined using the Bio-Rad protein assay
kit. Data were reported as amylase inhibition (AI) index values, defined herein
as the ratio of the amylase activity of the control (enzyme alone) to that of
the enzyme/clonal extract mixture (Correia et al. 2004). Values greater than 1
indicate AI.

Characterization of a-Glucosidase Inhibition by Food Extracts

The inhibitory activity of the selected American and Asian food extracts
against yeast a-glucosidase was measured according to Kim et al. (2000),
with slight modification. Briefly, 0.5 U of yeast a-glucosidase dissolved in
100 mM phosphate buffer (pH 7), 0.2% bovine serum albumin (BSA) and
0.02% sodium azide (NaN3) was pipetted into an empty well of a flat-bot-
tomed 96-well microplate. To this well was added 50 mg of food extract in a
10-mL volume. The control was dH2O. The mixture was incubated for 5 min
at room temperature (RT). Fifty microliters of 5 mM para-nitrophenyl-a-D-
glucopyranoside substrate solution was added, and the increase in absorbance
at l= 405 nm was determined over a 5-min period. Data were expressed as
percentage (%) inhibition = (DA405, control - DA405, test)/(DA405, control) ¥ 100.

Antioxidant Activity
The antioxidant activity of each food extract was determined as the
ability of the extract to scavenge 2,2-diphenyl-1-picrylhydrazyl (DPPH) free
radicals. A 0.1 mM DPPH radical solution in 95% ethanol was prepared. One
milliliter of ethanolic DPPH solution was mixed with 1 mL of sample or 95%
ethanol (as control), vortexed well and then incubated for 30 min at RT. The
samples were then centrifuged for 30 s at 13 500 rpm at RT. The absorbance
 282 P. MCCUE ET AL.

Green pepper

String bean

Baby spinach

Broccoli sprouts

Red pepper

Carrot

Romaine lettuce

Red grape

Tomato

Basil leaf

Graham crackers

Chips Ahoy

Wheat Thins

Control

0 0.5 1 1.5 2 2.5 3 3.5

AI index

FIG. 1. INHIBITION OF PORCINE PANCREATIC a-AMYLASE IN VITRO BY EXTRACTS

OF SELECTED AMERICAN FOODS
Values above 1 suggest amylase inhibitory activity. Values below 1 suggest amylase stimulatory
activity. AI, amylase inhibition.

of each sample at l= 517 nm was measured. This antioxidant activity was

given as percentage (%) of DPPH scavenging, calculated as ([control
absorbance - extract absorbance]/[control absorbance] ¥ 100).

Angiotensin I-converting Enzyme Inhibition Assay

An in vitro assay based on the method of Cushman and Cheung (1971)
was used to determine the ACE-inhibitory activity of selected food extracts.
The in vitro ACE-inhibitory activity is quantified by measuring the formation
of hippuric acid (peak absorbance at 228 nm) from hippuryl-histidyl-leucine
(HHL) after reaction with ACE in the presence and absence of an inhibitor.
The decrease in A228nm is proportional to the inhibition resulting from the food
extract. Various concentrations (total phenolic basis) of food extract (adjusted
to pH 8.3) were added to 2 mU of ACE and 100 mL of HHL dissolved in
100 mM borate, 300 mM sodium chloride (NaCl) (pH 8.3) and the mixture
was incubated at 37C for 60 min. The reaction was stopped by the addition
of 150 mL of 0.5 N hydrochloric acid (HCl) and analyzed at 228 nm by high-
 ANTI-HYPERGLYCEMIC POTENTIAL OF SELECTED FOODS 283

Cinnamon

Brinjal

Bittergourd

Coccinia

Eggplant

Turmeric

Ginger

Cardamom

Mustard

Fennel

Fenugreek

Control

0 0.5 1 1.5 2 2.5 3 3.5

AI index

FIG. 2. INHIBITION OF PORCINE PANCREATIC a-AMYLASE IN VITRO BY EXTRACTS

OF SELECTED ASIAN FOODS
Values above 1 suggest amylase inhibitory activity. Values below 1 suggest amylase stimulatory
activity. AI, amylase inhibition.

performance liquid chromatography (HPLC) using a C18 column, a 20 mM

o-phosphoric acid (pH 2.5) mobile phase and a flow rate of 1 mL/min.

RESULTS AND DISCUSSION

Antidiabetic Effect of Selected American and Asian Foods

In this study, we have investigated the potential antidiabetic effect of
selected American and Asian foods optimized for phenolic content as natural
sources of a-amylase and a-glucosidase inhibitors for modulation of post-
prandial hyperglycemia by decreasing meal-derived carbohydrate absorption.
Figure 1 shows the anti-amylase activity of extracts of selected American
foods on an equivalent phenolic basis. Notably, for the majority of vegetables
tested, the AI was relatively high (AI index values between 1.25 and 2.5)
compared to the extracts of processed starch foods such as cookies and snack
284 P. MCCUE ET AL.

Green pepper

String beans

Baby spinach

Broccoli sprouts

Red pepper

Carrot

Romaine lettuce

Red grape

Tomato

Basil leaf

Graham crackers

Chips Ahoy

Wheat Thins

0.8 0.85 0.9 0.95 1 1.05 1.1 1.15 1.2 1.25

aGI index

FIG. 3. INHIBITION OF YEAST a-GLUCOSIDASE IN VITRO BY EXTRACTS OF SELECTED

AMERICAN FOODS
Values above 1 suggest a-glucosidase inhibitory activity. Values below 1 suggest a-glucosidase
stimulatory activity. aGI, a-glucosidase inhibition.

crackers (with values below 1, or slightly above). We speculate that the higher
AI index value observed for the extract of WT may be related to the presence
of residual wheat phenolics remaining in the product or to Maillard reaction
products that could have formed during processing (Hahnemann et al. 1989;
Schumacher and Kroh 1994; Schumacher et al. 1996). The GR extract had
the strongest anti-amylase activity of the American foods (AI index value of
2.2 ± 0.4), followed by GP (1.8 ± 0.2), BS (1.8 ± 0.3) and FC (1.7 ± 0.3).
Figure 2 shows the anti-amylase activity of extracts of selected Asian
foods on an equivalent phenolic basis. As observed with nonstarch, plant-
based American food extracts, all of the selected Asian food extracts possessed
anti-amylase activity (with AI index values slightly above 1.0 to well over
2.5). The G extract possessed the strongest anti-amylase activity (AI index
value = 2.7 ± 0.5), followed by CX (2.4 ± 0.6), M (2.0 ± 0.4) and CN
(1.8 ± 0.4).
Although the selected American and Asian food extracts showed moder-
ate to strong anti-amylase activities, the same extracts did not perform as well
 ANTI-HYPERGLYCEMIC POTENTIAL OF SELECTED FOODS 285

Cinnamon

Brinjal

Bittergourd

Coccinia

Eggplant

Turmeric

Ginger

Cardamom

Mustard

Fennel

Fenugreek

0.8 0.85 0.9 0.95 1 1.05 1.1 1.15 1.2 1.25

aGI index

FIG. 4. INHIBITION OF YEAST a-GLUCOSIDASE IN VITRO BY EXTRACTS OF SELECTED

ASIAN FOODS
Values above 1 suggest a-glucosidase inhibitory activity. Values below 1 suggest a-glucosidase
stimulatory activity. aGI, a-glucosidase inhibition.

against a-glucosidase. Figure 3 shows the anti-glucosidase activity of the

American food extracts on an equivalent phenolic basis. Only the extracts of
WT, GR, BS and SB displayed anti-glucosidase activity, albeit slight (with
a-glucosidase inhibition (aGI) index values = 1.1 ± 0.01, 1.04 ± 0.003,
1.04 ± 0.01 and 1.04 ± 0.01, respectively). The majority of these extracts also
displayed significant anti-amylase activity (Fig. 1). Figure 4 shows the anti-
glucosidase activity of the Asian food extracts on an equivalent phenolic basis.
Similar to the American food extracts, the majority of the Asian food extracts
did not possess significant a-glucosidase inhibitory activity. The only excep-
tion was CN extract, which possessed the strongest a-glucosidase inhibitory
activity of the extracts tested (aGI index value = 1.19 ± 0.03), followed by FN
(1.09 ± 0.01), FL (1.04 ± 0.01) and G (1.02 ± 0.02).

Antihypertensive Effect of Selected American and Asian Foods

Four extracts were selected from the phenolic-optimized American and
Asian food extracts that possessed the most significant a-amylase and a-
286 P. MCCUE ET AL.

100

90

80

70
% ACE inhibition

60

50

40

30

20

10

0
1.45 7.25 14.5
Ginger extract phenolic content (mg)

FIG. 5. ANGIOTENSIN I-CONVERTING ENZYME (ACE) INHIBITION BY GINGER EXTRACT

glucosidase inhibitory activities and were further tested for potential to inhibit
the activity of ACE. The four extracts tested for anti-ACE activity were GR,
G, CN and FN. Only G extract possessed ACE inhibitory activity (Fig. 5). The
anti-ACE activity of the G extract was very strong, with 47 ± 3.0% inhibition
occurring after the addition of a volume of extract equivalent to only 7.25 mg
of the total phenolic content.

Roles of Phenolic Content, Antioxidant Activity and Protein

Concentration
To understand better the mechanism(s) of action of the selected phenolic-
optimized food extracts against amylase, glucosidase and ACE, the total
soluble phenolic content, antioxidant activity and protein concentration data
were measured for all of the extracts. Figures 6 and 7 show the total soluble
phenolic contents of the selected American and Asian food extracts, respec-
tively, determined as gallic acid equivalents (GAE) by reaction with Folin-
Ciocalteu reagent. Most apparent at first glance is that the American food
extracts possessed much lower phenolic contents than the Asian food extracts.
The GAE values for the American food extracts ranged from 27.6 ± 0.5 mg/
mL (TM) to 164.7 ± 1.8 mg/mL (BS), whereas the GAE values for the Asian
 ANTI-HYPERGLYCEMIC POTENTIAL OF SELECTED FOODS 287

180

160

140
Average gram of GAE/mL extract

120

100

80

60

40

20

0
WT CA GC BL TM GR RL FC RP BS BB SB GP
American foods

FIG. 6. TOTAL PHENOLIC CONTENT OF EXTRACTS OF SELECTED AMERICAN FOODS

GAE, gallic acid equivalents; WT, Wheat Thins crackers; CA, Chips Ahoy cookies; GC, Graham
cracker; BL, basil leaves; TM, tomato; GR, red grape; RL, Romaine lettuce; FC, fresh carrot; RP, red
pepper; BS, broccoli sprouts; BB, baby spinach; SB, string beans; GP, green pepper.

food extracts ranged from 17.9 ± 0.4 mg/mL (CX) to 916.6 ± 13.8 mg/mL
(CN). The data suggest that high phenolic content does not always confer a
high anti-amylase or anti-glucosidase activity of a food extract. The snack
cracker and cookie extracts had moderate phenolic contents (Fig. 6), yet low
enzyme inhibition index values (Figs. 1 and 3). Similarly, the FC extract had
relatively low phenolic content (Fig. 6), and yet moderately high anti-amylase
activity (Fig. 1). It is more likely that the higher enzyme inhibition indices
observed for certain food extracts (such as CN or G) may be representative
of certain key phenolic compounds that may not be present in other extracts.
Figures 8 and 9 show the antioxidant activities of the selected American
and Asian food extracts, respectively, determined as percentage (%) DPPH
free radical-scavenging activity. The antioxidant activity was high in most
extracts and ranged from 0.0 ± 2.6% (GC) to 90.5 ± 0.3% (TM) for the Amer-
ican foods (Fig. 8), and from 14.5 ± 1.4% (BJ) to 84.9 ± 0.3% (M) for Asian
foods (Fig. 9). The data in Figs. 8 and 9 suggest that higher extract antioxidant
activity is associated with higher extract anti-amylase activity, as extracts that
288 P. MCCUE ET AL.

1000

900

800
Average gram of GAE/mL extract

700

600

500

400

300

200

100

0
FN FL M CM G TR EP CX BG BJ CN
Asian foods

FIG. 7. TOTAL PHENOLIC CONTENT OF EXTRACTS OF SELECTED ASIAN FOODS

GAE, gallic acid equivalents; FN, fenugreek; FL, fennel; M, mustard; CM, cardamom; G, ginger; TR,
turmeric; EP, eggplant; CX, coccinia; BG, bittergourd; BJ, small brinjal; CN, cinnamon.

had low antioxidant activity (i.e., RL, GC and BJ) also possessed low anti-
amylase activity (Figs. 1 and 2). Similarly, the extracts with higher antioxidant
activity (i.e., G, M and FC) also possessed higher anti-amylase activity. For
the American food extracts, the antioxidant activity was moderately correlated
to AI index value (coefficient = 0.53). For the Asian food extracts, the antiox-
idant activity also correlated to the AI index value (coefficient = 0.35; 0.48
when excluding FL and FN data). This finding is in accordance with our
previous observation of a correlation between antioxidant activity and anti-
amylase activity in clonal herbal extracts as well as the synthetic antioxidants
butylated hydroxytoluene (BHT) and Trolox (McCue et al. 2004).
Figures 10 and 11 show the protein concentrations of the selected Amer-
ican and Asian food extracts, respectively, determined as BSA equivalents. In
the American food extracts (Fig. 10), the protein concentration was high in
extracts that possessed low anti-amylase activity (0.292 ± 0.002 mg/mL
[CA]-0.580 ± 0.003 mg/mL [RL]), and low in extracts that had high anti-
amylase activity (0.038 ± 0.001 mg/mL [RP]-0.315 ± 0.007 mg/mL [BB]). In
Asian food extracts (Fig. 11), the trend was similar, except for the M extract
which had both a high protein content and a high anti-amylase activity. The
 ANTI-HYPERGLYCEMIC POTENTIAL OF SELECTED FOODS 289

100

90

80

70
% DPPH scavenging

60

50

40

30

20

10

0
WT CA GC BL TM GR RL FC RP BS BB SB GP

American foods

FIG. 8. ANTIOXIDANT ACTIVITY (AS DPPH FREE RADICAL SCAVENGING) OF EXTRACTS

OF SELECTED AMERICAN FOODS
DPPH, 2,2-diphenyl-1-picrylhydrazyl; WT, Wheat Thins crackers; CA, Chips Ahoy cookies; GC,
Graham cracker; BL, basil leaves; TM, tomato; GR, red grape; RL, Romaine lettuce; FC, fresh carrot;
RP, red pepper; BS, broccoli sprouts; BB, baby spinach; SB, string beans; GP, green pepper.

protein content ranged from 0.005 ± 0.001 mg/mL (CX) to 1.427 ± 0.067 mg/
mL (FN).
The onset of T2DM and its associated long-term complications, such as
hypertension, has been linked to persistent postprandial hyperglycemia
(Haffner 1998; DiCarli et al. 2003; Fonseca 2003). A number of synthetic
drugs is available to treat T2DM and mainly work to affect insulin resistance
or defective insulin secretion (Ohmura et al. 1998; Mudaliar and Henry 2001;
Fonseca 2003). However, the use of many of these drugs has been associated
with negative side effects, leading researchers to seek safe and/or natural
sources for new therapeutics (Dawra et al. 1988; McCarty 2000). Currently,
much focus has been on the discovery of dietary sources of mild a-amylase
and/or a-glucosidase inhibitors to delay the intestinal absorption of digested
carbohydrates (Scheen 2003; McCue and Shetty 2004).
Here, we report the ability of phenolic-optimized aqueous extracts of
selected foods common to American and Asian diets to inhibit a-amylase and
a-glucosidase. GR, GP, BS and FC extracts had the strongest anti-amylase
activities of the American foods (Fig. 1), whereas G, CX, M and CN extracts
had the strongest anti-amylase activities of the Asian foods (Fig. 2). Compared
290 P. MCCUE ET AL.

100

90

80

70
% DPPH scavenging

60

50

40

30

20

10

0
FN FL M CM G TR EP CX BG BJ CN

Asian foods

FIG. 9. ANTIOXIDANT ACTIVITY (AS DPPH FREE RADICAL SCAVENGING) OF EXTRACTS

OF SELECTED ASIAN FOODS
DPPH, 2,2-diphenyl-1-picrylhydrazyl; FN, fenugreek; FL, fennel; M, mustard; CM, cardamom; G,
ginger; TR, turmeric; EP, eggplant; CX, coccinia; BG, bittergourd; BJ, small brinjal; CN, cinnamon.

to the a-amylase inhibition, the a-glucosidase inhibition by the American and

Asian food extracts was not as significant (Figs. 3 and 4). For the American
foods, WT, GR, BS and GP extracts had minor a-glucosidase inhibitory
activity (Fig. 3). Similarly, the extracts of the Asian foods CN, FN, FL and G
also had minor a-glucosidase inhibitory activity (Fig. 4). Interestingly, the G
extract was also found to possess significant anti-ACE activity (Fig. 5), which
suggests that G may also have a strong potential as an antihypertensive agent.
The antioxidant activity (Figs. 8 and 9), but not necessarily the total phenolic
content (Figs. 6 and 7), was associated with amylase inhibition. The protein
content seemed to be inversely associated with the amylase inhibition. We
speculate that protein–phenolic and/or phenolic–phenolic synergies may be
involved in the food extract enzyme-inhibition mechanism.

CONCLUSIONS AND IMPLICATIONS

While reports in the literature are building evidence on medicinal

plant and herbal extracts as potential T2DM therapeutic agents, informa-
 ANTI-HYPERGLYCEMIC POTENTIAL OF SELECTED FOODS 291

0.7

0.6

0.5
BSA equivalents (mg/mL)

0.4

0.3

0.2

0.1

0
WT CA GC BL TM GR RL FC RP BS BB SB GP
American foods

FIG. 10. PROTEIN CONCENTRATION OF EXTRACTS OF SELECTED

AMERICAN FOODS
BSA, bovine serum albumin; WT, Wheat Thins crackers; CA, Chips Ahoy cookies; GC, Graham
cracker; BL, basil leaves; TM, tomato; GR, red grape; RL, Romaine lettuce; FC, fresh carrot; RP,
red pepper; BS, broccoli sprouts; BB, baby spinach; SB, string beans; GP, green pepper.

tion concerning common everyday foods is lacking. A healthy lifestyle,

including a proper diet and exercise, is known to be an important compo-
nent in achieving glycemic control. Our results showed that common veg-
etables and spices contained significant antidiabetic activity in vitro, as
well as anti-ACE activity in one case, and suggest that diet modification
to include these types of foods along with balancing carbohydrate intake
throughout the day may represent a promising strategy to help control
postprandial hyperglycemia through modulation of carbohydrate absorp-
tion. Dietary a-amylase and a-glucosidase inhibitors from common foods
are potentially safer, and therefore, may be a preferred alternative for the
inhibition of carbohydrate breakdown and control of glycemic index of
food products. Future research is aimed at investigating the potential
antidiabetic and antihypertensive activities of other foods and food-grade
herbal extracts and at elucidating putative phenolic component synergies
and mechanisms.
292 P. MCCUE ET AL.

1.6

1.4

1.2
BSA equivalents (mg/mL)

0.8

0.6

0.4

0.2

0
FN FL M CM G TR EP CX BG BJ CN
Asian foods

FIG. 11. PROTEIN CONCENTRATION OF EXTRACTS OF SELECTED ASIAN FOODS

BSA, bovine serum albumin; FN, fenugreek; FL, fennel; M, mustard; CM, cardamom; G, ginger; TR,
turmeric; EP, eggplant; CX, coccinia; BG, bittergourd; BJ, small brinjal; CN, cinnamon.

REFERENCES

CARROLL, M.F., GUTIERREZ, A., CASTRO, M., TSEWANG, D. and

SCHADE, D.S. 2003. Targeting postprandial hyperglycemia: A compar-
ative study of insulinotropic agents in type 2 diabetes. J. Clin. Endo-
crinol. Metab. 88(11), 5248–5254.
CHEN, H., FENG, R., GUO, Y., SUN, L. and JIANG, J. 2001. Hypoglycemic
effects of aqueous extract of Rhizoma polygonati odorati in mice and
rats. J. Ethnopharmacol. 74, 225–229.
CORREIA, R.T.P., MCCUE, P., VATTEM, D.A., MAGALHÃESA, M.M.A.,
MACÊDOA, G.R. and SHETTY, K. 2004. Amylase and Helicobacter
pylori inhibition by phenolic extracts of pineapple wastes bioprocessed
by Rhizopus oligosporus. J. Food Biochem. 28, 419–434.
CUSHMAN, D.W. and CHEUNG, H.S. 1971. Spectrophotometric assay and
properties of the angiotensin-converting enzyme of rabbit lung. Biochem.
Pharmacol. 20(7), 1637–1648.
 ANTI-HYPERGLYCEMIC POTENTIAL OF SELECTED FOODS 293

DAWRA, R.K., MAKKAR, H.P. and SINGH, B. 1988. Protein-binding

capacity of microquantities of tannins. Anal. Biochem. 170(1), 50–53.
DICARLI, M.F., JANISSE, J., GRUNBERGER, G. and AGER, J. 2003. Role
of chronic hyperglycemia in the pathogenesis of coronary microvascular
dysfunction in diabetes. J. Am. Coll. Cardiol. 41(8), 1387–1393.
FONSECA, V. 2003. Clinical significance of targeting postprandial and fast-
ing hyperglycemia in managing type 2 diabetes mellitus. Curr. Med. Res.
Opin. 19(7), 635–641.
GROVER, J.K., YADAV, S. and VATS, V. 2002. Medicinal plants of India
with anti-diabetic potential. J. Ethnopharm. 81, 81–100.
HAFFNER, S.M. 1998. The importance of hyperglycemia in the nonfasting
state to the development of cardiovascular disease. Endocr. Rev. 19(5),
583–592.
HAHNEMANN, B., LEGRUM, W., KOSS, G. and NETTER, K.J. 1989.
Interactions of heterocyclic Maillard products with the hepatic microso-
mal monooxygenase system. Xenobiotica 19(11), 1319–1326.
HASRAT, J.A., PIETERS, L. and VLIETINCK, A.J. 2004. Medicinal plants
of Suriname: Hypotensive effect of Gossypium barbadense. J. Pharm.
Pharmacol. 56, 381–387.
KIM, J.S., KWON, C.S. and SON, K.H. 2000 Inhibition of alpha glucosidase
and amylase by luteolin, a flavonoid. Biosci. Biotechnol. Biochem.
64(11), 2458–2461.
MCCARTY, M.F. 2000. Toward a wholly nutritional therapy for type 2
diabetes. Med. Hypotheses 54(3), 483–487.
MCCUE, P. and SHETTY, K. 2004. Inhibitory effects of rosmarinic acid
extracts on porcine pancreatic amylase in vitro. Asia Pac. J. Clin. Nutr.
13(1), 101–106.
MCCUE, P., VATTEM, D. and SHETTY, K. 2004. Inhibitory effect of clonal
oregano extracts against porcine pancreatic amylase in vitro. Asia Pac.
J. Clin. Nutr. 13, 401–408.
MCCUE, P., ZHENG, Z., PINKHAM, J.L. and SHETTY, K. 2000. A model
for enhanced pea seedling vigor following low pH and salicylic acid
treatments. Process Biochem. 35, 603–613.
MUDALIAR, S. and HENRY, R.R. 2001. New oral therapies for type 2
diabetes mellitus: The glitazones or insulin sensitizers. Annu. Rev. Med.
52, 239–257.
OHMURA, C., TANAKA, Y., MITSUHASHI, N., ATSUM, Y., MATSUOKA,
K., ONUMA, T. and KAWAMORI, R. 1998. Efficacy of low-dose
metformin in Japanese patients with type 2 diabetes mellitus. Curr. Ther.
Res. 59(12), 889–895.
RATNER, R.E. 2001. Controlling postprandial hyperglycemia. Am. J. Car-
diol. 88(Suppl.), 26H–31H.
294 P. MCCUE ET AL.

SCHEEN, A.J. 2003. Is there a role for a-glucosidase inhibitors in the pre-
vention of type 2 diabetes mellitus? Drugs 63(10), 933–951.
SCHUMACHER, D. and KROH, L.W. 1994. The decomposition of Maillard
reaction products by amylolytic enzymes. 1. Reversible inhibition of
alpha- and glucoamylase and alpha-glucosidase by oligosaccharide Ami-
dori compounds. Z. Lebensm. Unters. For. 199(4), 270–274.
SCHUMACHER, D., HIRSCH, D., CAMMERER, B. and KROH, L.W. 1996.
Degradation of Maillard reaction products by amylolytic enzymes. 3.
Inhibition of glucoamylase, alpha-amylase and alpha-glucosidase by heat
treated alpha-glucans and melanoidins. Z. Lebensm. Unters. For. 203(4),
391–394.
VATS, V., GROVER, J.K. and RATHI, S.S. 2002. Evaluation of anti-hyper-
glycemic and hypoglycemic effect of Trigonella foenum-graecum Linn,
Ocimum sanctum Linn, and Pterocarpus marsupium Linn in normal and
alloxanized diabetic rats. J. Ethnopharmacol. 79, 95–100.
ZIMMERMAN, B.R. 2001. Postprandial hyperglycemia: Implications for
practice. Am. J. Cardiol. 88(Suppl.), 32H–36H.