lee

Food Sci. Biotechnol. 22(4): 1039-1046 (2013)

DOI 10.1007/s10068-013-0181-y

Evaluation of Physicochemical Properties and Fermenting Qualities of

Apple Wines Added with Medicinal Herbs

Je-Hyuk Lee, Tae Ho Kang, Byung Hun Um, Eun-Hwa Sohn, Woo-Cheul Han, Seol-Hee Ji, and Ki-Hyo Jang

Received: 11 December 2012 / Revised: 30 January 2013 / Accepted: 30 January 2013 / Published Online: 31 August 2013

KoSFoST and Springer 2013

Abstract To evaluate the possibility for development of

apple wine containing medicinal herbs, we have prepared

Fuji apple (Malus domestica) wines containing pine

(Pinus densiflora Siebold et Zuccarini) needle and

hwanggi (Radix Astragali)/mistletoe (Viscum album).

Normal apple wine was fermented rapidly, but after 40

days fermentation/maturation, the final ethanol content,

pH, total acidity, and contents of sugar/organic acid

showed similar levels in 3 kinds of apple wines. In other

hands, the total phenols content, antocyanins content, and

brightness were higher in apple-pine wine and apple-herb

wine than in normal apple wine. Apple-herb wine had

higher values in total phenol contents, brightness, free

amino acid contents, and quenching activity for ABTS free

radicals than normal apple wine, and had similar sensory

evaluation values with normal apple wine. Taken together,

it is supposed that hwanggi and mistletoe might provide

functional components to normal apple wine and might be

applied to development of functional apple wines.

Keywords: antioxidant, apple wine, hwanggi, mistletoe,

pine needle

Introduction

Recently, increased public interest for health and well-

being life has raised the usage of the natural materials with

health enhancing functions. Some fermented alcoholic

beverages were investigated to possess health enhancing

functionality for relieving hangovers and preventing diseases

(1,2). In most fermented alcoholic beverages, the rice is

used as a major material and leaves/roots of medicinal

herbs, such as ginseng, fruit of corni (Cornus officinalis

Sieb. et Zucc), and skullcap (Scutellaria baicalensis Georgi),

were supplied to improve health enhancing functions of

fermented beverages. Bioactive substances were extracted

from the materials of alcoholic beverages or produced by

microorganisms involved in fermentation (1,2). Free radicals

and reactive oxygen species (ROS) cause the aging,

inflammation, ateriosclerosis, vessel disease, and cancer in

our body. Many studies have been progressed for screening

antioxidant materials from medicinal herbs to scavenge

ROS (3). According to the report analyzing the published

90 reports concerning anti-oxidation of Korean domestic

natural materials from 2003 to 2011, the efficiency of

solvents to extract antioxidant substances showed the

following order; water

1040 Lee et al.

components were sugars, organic acids, minerals, sorbitol,

cellulose, pectic substances, and amino acids (1,4). Acidity

and contents of polyphenolic substances in fruit wines are

related with a taste and act as an important point for

assessment of fruit wine taste. In practical, apples, grapes,

and orange/tangerine have the proper amount of organic

acids and can be utilized for fermentation of fruit wines

without addition of organic acids. Production of great

quality apple wine need the fresh apples, not stored after

harvest, and therefore apple wines should be produced in

harvesting period of apples. Unfortunately the preference

of apple wine is low, and most apples were consumed as a

raw apple in Korea. Only 5% of apple in 28,088 tons of

food-processing apple was utilized for production of apple

wine. Therefore Korean domestic apple wine manufacturing

is lacking in terms of scale (1). In the USA having

advanced technology to produce apple wines, the various

fermenting techniques were developed for apples below

normal for fermenting property, such as mixing several

apple cultivars (5).

Pine (Pinus densiflora Siebold et Zuccarini) needles

have been used for the treatment of cardiovascular diseases

and liver diseases in Korean traditional medicines, and was

reported to have antioxidant functions by improvement of

lipid metabolism and of regulating reactive oxygen in

animal study. Hwanggi (Radix Astragali) is a dried root

part of a perennial herb, Astragalus membranaceus Bunge,

belonged in Leguminosae. In Korean traditional medicines,

hwanggi has warming property in body, sweet taste, and

notoxicity, and has been used for energy supplement to a

body. In addition, it is reported that hwanggi has a diuretic

function, immune improvement, a strengthening of kidney

function, and antioxidant activity, and that its bioactive

substances are formononectin, triterpenoide glycosides,

and saponins such as astragaloside (6,7). Perennial plant

mistletoe (Viscum album) is parasitic on orinamu [Alnus

japonica (Thunb.) Steud.] and oak (Quercus acutissima),

and has been used for treatment of cancers by its anti-

carcinogenic components, such as lectin, viscototix, and

alkaloid. Additionally mistletoe has abundant polyphenols

and shows high antioxidant activity (8,9).

Recently the fermented rice wines containing fruits

produced abundantly in Korea, such as grape and apple,

have been receiving consumers interests more and more.

Generally grape wines have high antioxidant activity caused

from polyphenol compounds, however despite of apple

wines great taste, the research data for physiological

functions, such as antioxidant activity, are insufficient for

apple wines.

In this study, the pine needles, hwanggi, and mistletoe

with antioxidant activity were added to apple juice and

were fermented using addition of yeasts. The aim of this

study is to investigate the physicochemical characteristics

and antioxidant activity of fermented apple wine with

natural health enhancing herbs.

Materials and Methods

Materials Apple (Fuji, Malus domestica), hwanggi

(Radix Astragali, Astragalus membranaceus Bunge), and

mistletoe (Viscum album) were harvested in Jecheon

(Chungbuk, Korea) in 2006. Hwanggi was used as dried

root part of A. membranaceus Bunge. Pine (Pinus densiflora

Siebold et Zuccarini) needles were harvested in Samchock

(Kangwon, Korea) 2006, and were simply washed and

dried. Yeast (Saccharomyces cerevisiae) was from Red Star

Premier Cuvee (LeSaffre, France). Sucrose and K2S2O5were purchased from Cheiljedang (Seoul, Korea) and

Sigma-Aldrich (St. Louis, MO, USA), respectively. And

pectinase (Pectinex 100 L, 5,000 FDU/mL at 55oC) was

from Novozyme (Bagsvaerd, Denmark).

Fermentation of apple wines As shown in Table 1, 3

different compositions of apple fermentation materials (15

L) were added to the plastic fermentation jar (26-L) with

an airlock apparatus. And the initial sugar content was set

to 24oBx by adding sucrose. Fermentation mixtures were

sterilized by addition of sulfite (K2S2O5) (2 g) for 16 h at

room temperature. After sterilization, 3 kinds of apple

wines were fermented by 5 g of Saccharomyces cerevisiae

and 5 mL of pectinase at 23-24oC for 14 days. During

fermentation period, the fermentation mixture was mixed in

totality twice a day for the 1st week, and once a day for a

next week. After 14 day-fermentation of apple wines, the

non-soluble materials in apple wines were removed by

filtration, and were aged at 19-20oC for 26 days. Samples

(50 mL) of apple wines were took, stored at 20oC, and

analyzed after thawing.

Measurement of pH, sugar content, and total acidity

The pH of apple wines was measured with a pH meter

(model 725p; Istek Co., Seoul, Korea). Total sugar content

of sample was determined by a hand-held refractometer

(model N-1; Atago, Tokyo, Japan) using 50 L sample.

Table 1. Proportion of apple-herb wines

Component Apple wineApple-pine

wineApple-herb

wine

Apple juice (mL)

Pine leave (g)

Mistletoe (g)

Hwanggi (g)

K2S2O5 (g)

Pectinase (mL)

Dry yeast (g)

15,000

-

-

-

2

5

5

15,000

60

-

-

2

5

5

15,000

-

60

60

2

5

5

Physicochemical Characteristics of Apple-Herb Wines 1041

For determination of total titratable acidity (TTA), samples

(10 mL) were filtered with 0.45-mm syringe filter. And

then samples were titrated with 1-2 drops of phenolphthalein

to pH 8.3 with 0.1 N NaOH. TTA was calculated by the

following equation using consumed amounts (mL) of 0.1 N

NaOH at the end-point (pink color).

Total titratable acidity% (TTA%)=[(mL of 0.1 N NaOH)

(N NaOH)0.067 (malic acid coefficient)100]/mL sample

Analysis of ethanol content For determination of ethanol

content, the samples of apple wines were passed through

0.45-m filter, and were injected to a GC (6890; Agilent

Technologies Inc., Santa Clara, CA, USA) equipped with

flame ionization detector. HP-Innowax column (0.25 m,

30 m0.25 mm, Agilent Technologies Inc.) was used for

the analysis of ethanol. Temperature of column oven was

programmed to be hold at 35oC for 5 min, to be increased

to 150oC at a rate of 5oC/min, to be increased to 250oC at

a rate of 20oC/min, and to be hold at 250oC for 2 min.

Injection volume was 10 L and split ratio was 10:1.

Temperatures of injector and detector were 225 and 260oC,

respectively.

Analysis of wine color The color of apple wines was

determined by a UV-Vis spectrophotometer (UV-1650 PC;

Shimadzu, Tokyo, Japan) using 10-mm quartz cuvette, and

the distilled water was used as the blank. Total phenol

content, hydroxycinnamate, the brownness, the anthocyanin

content, and chromaticity/brightness were expressed as

A280, A320, A420, A420+A520, and A420/A520, respectively.

Analysis of organic acids To determine the composition

of organic acids, apple wines were centrifuged at 10,000g

for 10 min, and colorants and proteins were removed from

apple wine samples by Sep-pak C18 cartridge. And then

samples were analyzed by HPLC equipped with C-610

column (30 cm7.8 mm; Supelco Co., Bellafonte, PA, USA).

Organic acids in samples were isolated with a mobile

phase, 0.1% phosphoric acid at a flow rate of 0.5 mL/min,

and were detected on 210 nm by UV detector.

Analysis of amino acids For determination of amino

acids in apple wines, samples were centrifuged at 10,000g

for 10 min. And supernatant was passed through 0.45-m

filter, and 20 L of sample was injected to amino acid

analyzer (L-8800; Hitachi, Tokyo, Japan).

On-line analysis of antioxidant activity Antioxidant

property of apple wines containing medicinal herbs was

detected by scavenging ABTS radical. ABTS reagent

(2 mM) containing 3.5 mM of potassium persulfate was

diluted to 8-fold in a brown bottle and was induced to

produce ABTS radicals at room temperature in a dark

place for 16 h for the analysis of antioxidant activity.

Samples of apple wines were passed through 0.45-m

filter, and 20 L of sample was injected to HPLC equipped

with an YMC Hydrosphere C18 column (4.6250 mm,

5L; Wilmington, NC, USA). Mobile phase was programmed

to flow as following; water for 5 min, a gradient flow to

acetonitrile for 35 min, and acetonitrile for 10 min at a rate

of 0.4 mL/min. And isolated phenolic compounds were

detected on 320 nm by diode array detector. Sample passed

through the first detector was mixed with an ABTS reagent

supplied at a flow rate of 0.2 mL/min from the second

pump. Intensity of scavenging ABTS radical by isolated

fermentation substances was determined by absorbance on

734 nm. For calculation of antioxidant capacity, the

standard curve was plotted using Trolox, as an antioxidant

reference in 0-400 M, and ABTS scavenging capacity

was expressed as Trolox equivalent.

Sensory evaluation of apple wines Fermented- and

aged-apple wines were stored in a refrigerator for 20 h. The

sensory evaluation, using the 9-point hedonic scales (from

1=dislike extremely to 9=like extremely), was conducted

by 29 students panel of university for color, taste, and

flavor of apple wines. Data of sensory evaluation were

analyzed by one-way analysis of variance (ANOVA) (10).

Results and Discussion

Composition of apple wines supplemented with medicinal

herbs Three kinds of apple wines were fermented and

aged with supplement of pine needle, hwanggi, and

mistletoe for 40 days, as shown in Table 1. Generally for

fermentation of fruit wines, the K2S2O5 was used for

prevention of oxidation for fruit juice, stability of red

colorant, and the sterilization of fruit juice (1). K2S2O5shows the suppression of contaminated bacteria in a

concentration of 100-200, and 133 ppm of K2S2O5 was

used in this study.

Total sugar contents of apple wines supplemented with

medicinal herbs Generally total sugar content (oBx)

during fermentation of fruit wines is an indicator for

activity of yeasts to use sugars in fermented mixture.

Consumption of sugar in apple-pine wine and apple-herb

wine appeared lately as compared to normal apple wine

(Table 2). In all kinds of apple wines of this study, the

consumption rate of sugars was fast in early phase of

fermentation, but became to be reduced as the fermentation

proceeded. Finally during 8 day-fermentation, most sugar

was consumed and the remained total sugar content was

approximately 7.2-7.8oBx.

1042 Lee et al.

pH, TTA, and organic acid contents of apple wines

supplemented with medicinal herbs before maturation

Organic acids produced during fermentation of fruit wines

decrease the pH of wines. However the pH change of apple

wines in this study was very slight in fermentation and

aging process (Table 2). This is anticipated to be caused

from low pH in early fermentation-stage of apple wines.

The initial TTA was detected to be approximately 0.34-

0.38% in 3 kinds of apple wines, but TTA was increased

during fermentation of apple wines up to approximately

0.46-0.54% when fermentation of apple wines was

finished. Chung et al. (11) reported that TTA value in the

early stage of apple wine-fermentation was 0.18% and

after 2 weeks fermentation, was increased to 0.24%, which

are consistent to result of this study. In addition, the pH of

organic acid is due to its dissociation in wines. Figure 1

shows the increase of succinic acid and lactic acid, the

decrease of malic acid during fermentation and aging. Each

organic acid in apple wines has a different degree of

dissociation. Therefore it is supposed that even though

TTAs increase in all 3 apple wines, their pH were not

changed for this reason.

Major organic acids in 3 kinds of apple wines were

malic acid, succinic acid, and lactic acid, and a small

amount of acetic acid were detected (Fig. 1). Total organic

acids was approximately 1.0-1.2% in early stage of

fermentation, but was decreased to 0.40-0.55% during

fermentation and maturation. Levels of organic acids in

apple wines of this study showed the following order;

malic acid>succinic/lactic acid. Kim et al. (1) reported that

apple Fuji have approximately 0.3-0.5% of TTA and the

stored apples have lower TTA values than fresh apples.

Whang et al. (12) reported that organic acid compositions

of apple juice had the following order; malic acid>citric

acid and could be variable depending on cultivars, grade of

maturity, and analysis method. Additionally Choi et al.

(13) have determined that organic acids of fermented apple

wine contained approximately 1.4-2.1% of succinic acid

and lactic acid, 0.8-1.0% of malic acid, and 0.03-0.08% of

acetic acid, which were relatively higher values as compared

with results of this study. It is supposed that the reason is

to be able to produce these organic acids newly from

carbon sources or other organic acids through brewing.

Most carbohydrates in fermentation mixture converse to

ethanol, and only approximately 5% of carbohydrates

converses to glycerol, acetic acid, lactic acid, and succinic

Table 2. Changes in sugar contents, pH, and total titratableacidity (TTA) during apple wine fermentation for 14 days

Time (day) Apple wineApple-pine

wineApple-herb

wine

Sugar (oBx)

0

4

8

10

14

24.0

15.2

09.0

08.0

08.0

24.0

16.0

10.2

10.0

08.6

24.0

20.0

13.0

11.0

08.8

pH

0

4

8

10

14

04.3

03.9

04.0

04.1

04.1

04.2

04.0

04.0

04.1

04.1

04.1

04.0

04.0

04.0

04.1

TTA (%)

0

4

8

10

14

00.34

00.35

00.38

00.42

00.46

00.38

00.44

00.44

00.44

00.50

00.36

00.36

00.43

00.47

00.54

Fig. 1. Changes of organic acids concentration (%, w/v) inapple-herb wines during 40 day-fermentation/aging. A, applewine; B, apple-pine wine; C, apple-herb wine. *Total: sum ofmalic acid, lactic acid, succinic acid, and acetic acid


acid, and others (1). In this study, malic acid content of

apple wines was consistently reduced during fermentation

and maturation. It is supposed that this phenomenon was

caused from the similar reason to previous reports, in

which malic acid is conversed to lactic acid or ethanol

through malolactic or maloalcoholic reaction (14,15).

Additionally, differences of organic acid contents in this

study with Choi et al. (13) are anticipated to be due to

diversity of metabolism pathway.

Composition of apple wines supplemented with

medicinal herbs after maturation Apple wines, which

were fermented for 14 days and aged for 26 days, had pH

3.85, 0.56%, 8.0oBx, and 15.6% in pH, TTA, total sugar

content, and ethanol contents, respectively (Table 3). Because

the initial sugar content before fermentation was adjusted

to 24.0oBx, the delayed fermentation of apple-pine wine

and apple-herb wine are anticipated to be caused from

some substances to suppress the growth of yeasts, not from

differences between sugar contents of 3 kinds of apple

wines. Essential oil of pine needles has approximately 7-

12% of terpene and 1.3% of phenolic compounds such as

benzoic acid, caffeic acid, cinnamic acid, ellagic acid, and

ferulic acid to exert antibacterial activity (16). Extract of

pine needles was investigated to exert antibacterial function

against some food-borne pathogens by some reports (16-

20). In addition, Bae et al. (20) have reported that petroleum

ether extract (0.5%) of mistletoe suppressed the growth of

Bacillus cereus and Shigella dysenteriae for 24 and 12 h,

respectively. For the reasons mentioned above, it is anticipated

that apple-pine wine will possess the enhancing activities

of pine needle reported previously. Practically physiological

function of apple-pine wine remains as a matter to be studied

further. Mistletoe contains acetylcholine and flavonoids as

well as a glycoprotein lectin with antimicrobial activity to

suppress the mycelium growth of fungi and the production

of N-acetylglucosamine and galactose (21).

When the rice containing 0.5-20% of hwanggi was

fermented by a starter (Aspergillus oryzae, Aspergillus

awamori, and S. cerevisiae), the pH in the early stage of

fermentation was increased slightly. After termination of

fermentation, there was no difference in pH, total acidity,

reducing sugar, and ethanol content, but a value (redness)

and b value (yellowness) in Hunter color value were increased

(22). Therefore it was anticipated that supplement of

hwanggi might affect the sensory values such as color of

wines, not the fermentation rate (22). In this study, apple

wines containing pine needles and medical herbs (hwanggi

and mistletoe) showed a similar final pH, total acidity,

reducing sugar content, and ethanol content to normal

apple wine, but had a color difference to normal apple

wine, which was consistent to Choi et al. (22) (Table 3).

Though apple-herb wines and normal apple-wine had a

similar value in brownness (A420) and color intensity

(A420+A520) for the chromaticity, total phenolic content

(A280), and brightness (A420/A520) of apple-pine wine and

apple-herb wine had higher values than normal apple wine.

Hunter color difference meter expresses color differences

as a value from 80 (green color) to 100 (red color) and b

value from 70 (blue color) to 70 (yellow color). This

result was not consistent to Choi et al. (22), in which

supplement of hwanggi to rice wine increased the a and b

value. Reason for increase of brightness is supposed to be

caused from difference of material-color between rice and

apple.

Major organic acid in apple wines was known to be malic

acid (1). Consistently to former reports, the abundant malic

acid was determined in all of 3 kinds of apple wines in this

study (Fig. 1). Composition of organic acids in fruit wines

is caused from wine materials and organic acids produced

by yeasts during fermentation. Organic acids influence the

sour taste of fruit wines. In the same total acidity, malic

acid, citric acid, and lactic acid showed the sour tastes of

fruit wines in order, and in the same pH circumstance, the

order of malic acid, lactic acid, and citric acid showed the

sour tastes in order (14,15). Therefore it is supposed that

the content of malic acid is a major factor to affect the sour

tastes of apple wines. In this study, the supplemented

amount of pine needles, hwanggi, and mistletoe was 60 g

in 15 L of fermentation mixture, which was only less than

1% of total materials, and therefore did not influence total

organic acids of apple wines.

In fruit wines, the composition of amino acids was

involved in tastes and flavor of wines. Free amino acids

were detected in following order and had a similar

compositions between apple wine and apple-herb wine;

tyrosine>lysine>asparagine>alanine in apple wine and

tyrosine>glutamic acid>asparagine>lysine>alanine in apple-

herb wine (Table 4). In other hand, the total amino acid

content of apple-pine wine was lower than those of normal

Table 3. Physicochemical characteristics of 3 kinds of applewine after 40 day-fermentation/aging

ComponentApplewine

Apple-pine wine

Apple-herb wine

Ethanol (%)

pH

Total titratable acidity (%)

Sugar (oBx)

15.600

3.85

0.56

8.00

15.900

3.84

0.56

8.20

15.900

3.87

0.55

8.20

A280

A420

A520

A320

Color intensity (A420+A520)

Shade (A420/A520)

7.70

0.34

0.11

0.62

0.45

3.12

11.000

0.33

0.09

0.63

0.42

3.88

11.800

0.37

0.08

0.80

0.45

4.68

1044 Lee et al.

apple wine and apple-herb wine. Apple Fuji, used in this

study, has approximately 57 and 200 mg/100 g apple of

free amino acid (FAA) and structural amino acid (SAA),

respectively (23). During fermentation and aging, protease

converse SAA to FAA, and fermentation microorganisms

metabolite FAA to other biomolecules. Therefore, a low SAA

content of apple-pine wine is supposed to be caused from

changes of protease activity by supplement of pine needle,

as compared to apple wine. Pine needles have 23-26 mg/

100 g of FAA contents and 3.3-4.7, 3.5-8.1, 2.7-3.2, and

1.4-5.1 mg/100 g of glutamic acid, threonine, aspartic acid,

and proline, respectively (24). Additionally pine needles

contain approximately 3-4% of total sugar and show the

following sugar contents; glucose > fructose > sucrose.

Mineral content of pine needles has 514-535, 294-329, and

85-89 mg/100 g of K, Ca, and Mg, respectively. Major

amino acids in 3 kinds of apple wines were tyrosine,

glutamic acid, alanine, glycine, serine, threonine, leucine,

phenylalanine, and valine. Tyrosine and glutamic acid have

a little bitter and a little sour or taste like beef broth,

respectively. In addition, alanine, glycine, serine, and

threonine have a sweet taste, and leucine, phenylalanine,

and valine have a bitter taste (25,26). Three kinds of apple

wines had very low level of total FAA content, as compared

with grape wine (27). Therefore for effective improvement

of commercial apple wines, it is supposed to need some

additional processes such as a mixing with other fruit

wines or a supplying of amino acids.

Analysis for antioxidant substances of apple-herb wines

using on-line antioxidant detection system Three kinds

of apple wines were analyzed using on-line antioxidant

analysis HPLC system, equipped with 2 pumps/detectors

and 1 column (Fig. 2A). Sample of apple wines supplied

from the first pump was separated by analyzing column,

and phenolic compounds in sample were detected by the

first detector. And then the separated sample was mixed

with ABTS free radical from the second pump, and the

scavenging degree of ABTS radical was detected with the

second detector on 734 nm. As shown in Fig. 2B, it is

supposed that 3 kinds of apple wines have some substances

scavenging ABTS radical and contain some antioxidant

substances. Bioactive compounds in apples are tannin acid,

flavonoids, catechin, chlorogenic acid, vitamins, and folic

acid, which exert several activities for antioxidation,

Table 4. Compositions of free amino acids (mg/100 g of wine)in 3 kinds of apple wine after 40 day-fermentation/aging

Amino acid Apple wineApple-pine

wineApple-herb

wine

Tyrosine

Aspartic acid

Threonine

Serine

Asparagine

Glutamic acid

Glycine

Alanine

Valine

Cystine

Methionine

Isoleucine

Leucine

Phenylalanine

Lysine

Histidine

Arginine

Proline

6.85

0.10

0.14

0.15

1.34

0.85

0.82

1.31

0.09

0.26

0.25

0.12

0.14

0.35

1.52

0.13

0.41

0.17

2.09

0.03

0.03

0.13

0.19

2.23

0.12

2.62

0.09

0.17

0.23

0.11

0.14

0.17

0.87

0.01

0.23

0.15

7.36

0.08

0.04

0.08

3.09

3.48

0.43

2.18

0.22

0.09

0.21

0.04

0.16

0.37

2.49

0.08

0.89

1.66

Total 15.000 9.61 22.870

Fig. 2. Schematic diagram of on-line HPLC-ABTS detectionsystem (A) and its chromatogram for apple-herb wines (B). (a),apple wine; (b), apple-pine wine; (c), apple-herb wine. Whitearrows indicate the novel polyphenolic ingredients, which weredetected on 320 nm and not found in normal apple wine; Blackarrows indicate the novel ABTS radical scavenging activities,which were not found in normal apple wine.


improvement of appetite, strengthening blood, stabilization

of plasma, prevention of hypertension, and reduction of

blood cholesterol (28). According to a report concerning

phenolic compounds and vitamins in apple, apples have

33-114 mg/100 g of hydroxycinnic acids containing chlorogenic

acid, 7-151 mg/100 g of flavanols such as catechins and

epigallocatechin, 94-94 mg/100 g of proanthocyanidins,

0.1-11 mg/100 g of flavonols such as quercetin, 0.78-2.45

mg/100 g of anthocyanidins, 4 mg/100 g of vitamin C, and

less than 0.1 mg/100 g of carotenoids such as -carotene,

lutein, and zeaxanthine (28).

Pine needles have antioxidant 30 substances including

-pinene (21.5%), limonene (13.1%), -pinene (9.3%),

and terpinolene (8.4%) were detected in essential oil (29).

Additionally it was reported that hwanggi and mistletoe

have some antioxidant substances. Extract of 2-6 years

aged-hwanggi had 44.7-46.7 and 5.3-6.0 mg/g in total

polyphenolic contents and total flavonoids contents,

respectively, and 30-36% of scavenging activity for ABTS

radical (30). Therefore, it is supposed that the high A280value, which indicates total phenolic contents, is related

with total polyphenolic contents of hwanggi (Table 3). In

addition, the amount of mistletoe extract was correlated

with its antioxidant activity. Extract of mistletoe by

supercritical extraction using CO2 at 200 bar/50oC had

approximately 17.6% of total polyphenolic contents.

Although extract of mistletoe had a low electron donating

activity, it showed a high SOD-like activity and high total

polyphenolic contents (8). The peaks in Fig. 2B are

anticipated to be noble compounds isolated from pine

needles, hwanggi, and mistletoe. Therefore the further

studies are necessary to identify single compounds and its

biofunctions.

Sensory characteristics of apple wines supplemented

with medicinal herbs The mean sensory scores for

colors, tastes, and flavors of 3 kinds of apple wines by 29

student panels are given in Table 5. Supplement of pine

needles or hwanggi/mistletoe to apple wine showed similar

sensory scores for colors to normal apple wine, and for

tastes of wines showed a little bit less preference with no

significant differences. However, flavors of apple-herb

wine was preferred more than that of apple-pine wine with

a significant difference (p

1046 Lee et al.

coloratum extracts on food-borne pathogens. J. East Asian Soc.Diet. Life 15: 182-187 (2005)

21. Jang CS, Oh MJ, Roh KS. Purification and biochemicalcharacterization of lectin from Viscum album. Korean J. Biotechnol.Bioeng. 14: 578-584 (1999)

22. Choi JH, Park JH, Kim SR, Lee CH, Park SY, Kim TJ, Jeong ST,Choi HS, Yeo SH. Quality characteristics of fermented alcoholicbeverage with Astragali Radix added. J. East Asian Soc. Diet. Life22: 41-51 (2012)

23. National Academy of Agricultural Science. Tables of FoodFunctional Composition-Amino Acid. Rural DevelopmentAdministration, Suwon, Gyeonggi, Korea. pp. 70-71 (2011)

24. Chung AJ, Hwang GH, Yoo MJ, Rhee SJ. Chemical composition ofthe pine sprouts and pine needles for the production of pine sprouttea. Korean J. Dietary Culture 11: 635-641 (1996)

25. Cho Y, Rhee HS. A study on flavorous taste components in kimchi:On free amino acids. Korean J. Food Sci. Technol. 11: 26-31 (1979)

26. National Tax Service Technology Institute. Productive method for

ordered and delicious liquor using neutral spirits. Korean Patent 10-0373985 (2003)

27. Hernndez-Orte P, Cacho JF, Ferreira V. Relationship betweenvarietal amino acid profile of grapes and wine aromaticcomposition. Experiments with model solutions and chemometricstudy. J. Agr. Food Chem. 50: 2891-2899 (2002)

28. National Academy of Agricultural Science. Tables of FoodFunctional Composition. 1st ed. Rural Development Administration,Suwon, Gyeonggi, Korea. pp. 116-425 (2009)

29. Kim YJ, Cho BJ, Ko MS, Jung JM, Kim HR, Song HS, Lee JY,Sim SS, Kim CJ. Anti-oxidant and anti-aging activities of essentialoils of Pinus densiflora needles and twigs. J. Pharm. Soc. Korea 54:215-225 (2010)

30. Lee KH, Park MH, Park YH, Lee SH, Kim KH, Kim YG, Ahn YS,Kim HY. Antioxidant activity and nitric oxide production of ethanolextracts from Astragali membranaceus Bunge and A. membranaceusBunge var mongholicus Hisiao. J. Korean Soc. Food Sci. Nutr. 40:1793-1796 (2011)

/ColorImageDict > /JPEG2000ColorACSImageDict > /JPEG2000ColorImageDict > /AntiAliasGrayImages false /DownsampleGrayImages true /GrayImageDownsampleType /Bicubic /GrayImageResolution 150 /GrayImageDepth -1 /GrayImageDownsampleThreshold 1.50000 /EncodeGrayImages true /GrayImageFilter /DCTEncode /AutoFilterGrayImages true /GrayImageAutoFilterStrategy /JPEG /GrayACSImageDict > /GrayImageDict > /JPEG2000GrayACSImageDict > /JPEG2000GrayImageDict > /AntiAliasMonoImages false /DownsampleMonoImages true /MonoImageDownsampleType /Bicubic /MonoImageResolution 600 /MonoImageDepth -1 /MonoImageDownsampleThreshold 1.50000 /EncodeMonoImages true /MonoImageFilter /CCITTFaxEncode /MonoImageDict > /AllowPSXObjects false /PDFX1aCheck false /PDFX3Check false /PDFXCompliantPDFOnly false /PDFXNoTrimBoxError true /PDFXTrimBoxToMediaBoxOffset [ 0.00000 0.00000 0.00000 0.00000 ] /PDFXSetBleedBoxToMediaBox true /PDFXBleedBoxToTrimBoxOffset [ 0.00000 0.00000 0.00000 0.00000 ] /PDFXOutputIntentProfile (None) /PDFXOutputCondition () /PDFXRegistryName (http://www.color.org?) /PDFXTrapped /False

/SyntheticBoldness 1.000000 /Description >>> setdistillerparams> setpagedevice

lee

Documents