Glycative Stress Research
Online edition : ISSN 2188-3610
Print edition : ISSN 2188-3602
Received : September 26, 2018
Accepted : October 29, 2018
Published online : December 31, 2018
doi:10.24659/gsr.5.4_163

Original article

Antiglycative effect of Kaempferia parviflora Wall. Ex. Baker (Zingiberaceae) :Prevention of advanced glycation end product formation.
Masayuki Yagi 1), Yukari Tateiwa 1), Kayoko Inoue 2), Yasuo Sato 2), Wakako Takabe 1),
Kaori Ishizaki 1), Yoshikazu Yonei 1)
1) Anti-Aging Medical Research Center and Glycative Stress Research Center, Graduate School of Life and Medical Sciences,
Doshisha University, Kyoto, Japan
2) Rene Co. Ltd., Osaka, Japan

Abstract

Advanced glycation end products (AGEs) are formed due to non-enzymatic reactions between protein and glucose.
The in vivo-formed and -accumulated AGEs are a risk factor of accelerating the aging process or causing onsets of lifestylediseases, which is considered as glycative stress. Some plant materials, such as vegetables and herbs, are recognized to haveanti-glycation effects. It has been reported that Zingiber officinale Roscoe has an inhibitory effect on glycation. Rhizomes of
Kaempferia parviflora Wall. Ex. Baker (KP), one of the plants of the Zingiberaceae family, have an anti-glycation effect, andpolymethoxy flavonoid (PMF) is involved in anti-glycation activities as an active component of KP, as has been reported. This
study aimed to analyze active constituents of KP as an inhibitor on glycation, comparing, regarding inhibitory effectiveness,extracts of KP rhizomes sold or marketed as health foods in Japan and examining the correlation between inhibitory
effectiveness and 5 types of PMF contained in KP.
Ten types of KP materials and 5 types of PMF were investigated as test samples. Dried powder samples were extractedwith hot water at 80 °C and with 70% ethanol. Sample extract solution or sample solution was added to the glycation model ofhuman serum albumin (HSA) and glucose. The formed fluorescent AGEs (excitation wavelength 370 nm/detection wavelength
440 nm) were measured, and inhibition rates of AGEs formation were calculated with the measurement values of AGEs.
The effectiveness of inhibition was assessed with AGEs at an inhibitory concentration of 50% (IC 50). Following this, hot water extracts of the powder sample were examined with fractionation and purification using hydrophobic cartridge columns
and the inhibitory effect on glycation of each effluent fraction were examined. The content of PMF in sample solutions were
measured, using HPLC.
Anti-glycation effects were recognized in 4 types of PMF and all of the 10 types of KP materials. Inhibitory effects on
glycation tended to be stronger in hot-water extracts of dried powder than in the 70% ethanol extract solution and essence
extract solution. Larger amounts of PMF contained in KP were detected in 70% ethanol extracts than in hot-water extracts( p < 0.03). There was no recognized correlation between the IC 50 of the sample solution and content of the PMF. Analyzing
hot-water extracts of dried powder using hydrophobic cartridge columns with fractionation and purification, purified water
not containing PMF, 50% acetonitrile containing PMF, and 75% acetonitrile containing PMF showed an anti-glycation effectwith eluate fraction.
Hydrophilic substances were assumed to have existed, as anti-glycation constitutes of KP, excluding for PMF. Furthermore,
there was a possibility that components, except for PMF, could greatly contribute to the inhibitory effects of KP.

KEY WORDS: anti-glycation effect, Kaempferia parviflora, polymethoxy flavonoid

Contact Address: Professor Masayuki Yagi, PhD
Anti-Aging Medical Research Center / Glycative Stress Research Center,
Graduate School of Life and Medical Sciences, Doshisha University
1-3, Tatara Miyakodani, Kyotanabe-shi, Kyoto, 610-0394 Japan
Phone/Fax: +81-774-65-6394 Email: myagi@mail.doshisha.ac.jp
Co-authors; Tateiwa Y, bmn2089@mail4.doshisha.ac.jp ; Inoue K, tmkayoko@miracle.ocn.ne.jp ;Sato Y, ceo.gran@gmail.com; Takabe Y, wtakabe@mail.doshisha.ac.jp ;Ishizaki K, ishizaki@antiaging-com.co.jp ; Yonei Y, yyonei@mail.doshisha.ac.jp_ 163 _

Glycative Stress Research 2018; 5 (4): 163-170(c) Society for Glycative Stress ResearchAntiglycation effect of Kaempferia parviflora

Introduction

Non-enzymatic glycation, which is the bonding of
reducing sugar such as glucose to protein, forms advanced
glycation end products (AGEs). The in vivo formation andaccumulation of advanced glycation end products induces
glycative stress, accelerating the aging process or causing
an onset risk of lifestyle diseases 1, 2). The inhibition of
postprandial hyperglycemia, the inhibition of glycation andthe degradation and excretion of AGEs are measures for
reducing glycative stress. It has previously been reported
that aminoguanidine (AG) has inhibitory effects of AGEs
formation and effects of preventing disease progression ofnephropathy, retinopathy and neuropathy 3, 4). However, AG
has not reached a stage for commercialization, as AG wasfound to have adverse effects, such as anemia, hepatopathy
and vitamin B6 deficiency. It has been reported that tea,
herb tea 5), vegetables, herbs 6), fruits 7) and yoghurt 8) arefoods with inhibitory effects on glycation. Furthermore,
Zingiber officinale Roscoe, which has long been utilized asa vegetable and herbal medicine, have inhibitory effects on
glycation, as has been reported 9, 10). Rhizomes of Kaempferia
parviflora Wall. Ex. Baker (KP), one of the plants of theZingiberaceae family, has been utilized recently as a healthfood. KP is characterized by the blackish purple of theroot inside, and its products sold in Japan are called black
galangal, black ginger or black turmeric. KP is distributedin Thailand, Laos, India and Myanmar. The appellation oforigin KP in Thailand is Krachai Dam 11). The extracted
liquid of KP roots in Bangladesh is traditionally utilized
as an alternative medicine for vomiting and diarrhea 12).
A decoction of KP with alcohol is a traditional medical
treatment for asthma, erectile dysfunction, gout, diarrhea,
dysentery and diabetes mellitus 13, 14). It has been reported
that extracts of KP roots function for the activation of lipid
metabolism, brown adipose tissue and lipolysis of white
adipose tissue 15). The characteristic components contained in
KP roots are anthocyanin and polymethoxy flavonoid (PMF).

It has been reported that PMFs, which are contained in KP,
have effects to improve metabolic syndromes, the metabolic
function of muscle 15) and obesity 16), as well as to enhancephysical fitness 17, 18). KP roots have anti-glycation effects, andPMFs are involved in its action component 19).
This study aimed to analyze active constituents of KP
as an inhibitor on glycation, comparing, regarding inhibitory
effectiveness, extracts of KP rhizomes sold as health foods
in Japan and examining correlations between inhibitory
effectiveness and PMF contained in KP.

Materials and methods
1) Reagent

Human serum albumins (HSA), lyophilized powder, ≥96% (agarose gel electrophoresis) was purchased from SigmaAldrich Japan (Meguro-ku, Tokyo, Japan). Aminoguanidinehydrochloride (AG) was purchased from Wako Pure
Chemical Industries (Chu-ku, Osaka, Japan). Reagents of
5, 7 - dimethoxyflavone, 5, 7, 4’ - trimethoxyflavone, 3, 5, 7 trimethoxyflavone, 3, 5, 7, 4’ - tetramethoxyflavone and3, 5, 7, 3’, 4’ - pentamethoxyflavone were purchased from
TOKIWA Phytochemical (Sakura, Chiba, Japan). Other
reagents of first or HPLC grade were purchased from Wako
Pure Chemical Industries or nacalai tesque (Nakagyo-ku,
Kyoto, Japan).

2) Rhizome of Kaempferia parviflora (KP)

Samples of 10 types of KP rhizomes were KP extracts
sold in Japan: dried matters and essence extract powder(Table 1). Powdery samples were used with no alteration, andslice dried samples were used after being crushed to powder
by a laboratory mixer (Osaka Chemical, Kita-ku, Osaka,
Japan).

Table 1. The rhizome sample of Kaempferia parviflora.
SamplenumberSampleform

Product nameblack galangal(cooled-vacuum drying)v216 -1

Maker
RENE Co., Ltd., Osaka Japan

ManufacturingmethodSolidcontent

dried (35°C, −98 MPa)rhzome powder−

black galangal(hot-air drying)

RENE Co., Ltd., Osaka Japan

dried (100°C, air)rhzome powder−

black ginger powder - MF5JAPAN TBLET Co., Ltd., Kyoto Japan

dried rhzome powder

−

v220

black ginger (Krachaidum)

AKAJIN Co., Ltd.

dried slice rhzome

−

v217

black ginger extract - WSP

ORYZA OIL & FAT CHEMICAL Co., Ltd., Aichi Japan

hydrous ethanol extract

5%

v218

black ginger extract - P

ORYZA OIL & FAT CHEMICAL Co., Ltd., Aichi Japan

hydrous ethanol extract

30%

v221

black ginger extract powder

TOYO SHINYAKU Co., Ltd., Fukuoka JapanSIRTMAX(black turmeric extract)

TOKIWA Phytochemical Co., Ltd., Chiba Japanv223

black ginger extract

MARUZEN Phamaceuticals Co., Ltd., Hiroshima Japan

100%

v224

black ginger extract

Sabinsa Japan Corporation, Tokyo Japan100%

v216 -2
v219

v222

dried

extract_ 16 4 _100%
hydrous ethanol extract

100%Glycative Stress Research

3) Preparation for KP extract

Preparation for dried powder samples was as follows:
extracts were obtained with 2 g of powder and a mixtureof 40 mL of purified water or 70 % ethanol solution. For
hot-water extracts, the mixed solution was incubated for
75 minutes in a water bath where the temperature wasmaintained at 80 °C. For 70% ethanol extracts, the mixed
solution was incubated for 4 hours in a water bath at 50 °C.
Samples were obtained after both extract solutions were
centrifuged at 2,500 rpm (800 × g) for 10 minutes and filtered.
To measure the concentrations of the solid samples, 5 mL of
the sample solutions were put in an aluminum tray and thendried and vaporized at 120 °C for 1 hour. Concentrations of
the solid samples were calculated by weighing the amountof evaporation residues. For the preparation of the extract
powder, by obtaining as precise information as possible
regarding the solid content ratio of the extract powder fromthe manufacturing makers of the samples, the samples ofextract powder were dissolved at the level of 10 mg/mL ofsolid concentration, adding a 70% ethanol solution.

4) Measurement of anti-glycation activity

The measurement of anti-glycation activities employed
the glycation model of HSA and glucose 20). The inhibitory
concentrations of the AGE formation were measured asfollows: Solution A was a mixture of the sample solution,
0.1 mol/L phosphoric acid buffer (pH 7.4), 40 mg/mL HSA,
2.0 mol/L glucose solution and distilled water at a ratio of
1 : 5 : 2 : 1 : 1. Solution B was a mixed solution containing
purified water instead of glucose solution in Solution A.
Solution C was an incubated solution which was arranged
with purified water instead of the sample solution in
Solution A. Solution D was a mixed solution containing
purified water instead of the glucose solution in Solution C,
which was incubated at 60 °C for 40 hours. To measure thevolume of the AGEs of the reaction liquid after incubation, afluorescent microplate reader was employed, measuring theAGE-derived fluorescence (excitation wavelength: 370 nm;
detection wavelength: 440 nm). The inhibitory ratio of theAGEs formation was calculated with the following formula.
The inhibitory concentration of AGEs formation (%) =[1 − (A− B) / (C − D)] × 100
Furthermore, a 50% inhibitory concentration (IC 50) wasused; mg/mL was calculated based on the inhibitory rateof the AGE formation of each KP extract solution at three
concentrations 21). Smaller values of IC 50 indicate a stronger
anti-glycation effect.

5) Measurement of polymethoxylated flavonoid (PMF)

After being diluted two times, PMF was measured using
HPLC as follows: Column: Inerstil ODS- 4, 3 μm, 100 × 4.6
mm I.D. (GL Sciences, Shinjuku-ku Tokyo, Japan), Eluent:
0.1% formic acid/0.1% formic acid - acetonitrile (ACN)(70/30), Flow rate: 1.0 mL/min, Detection wavelength: UV
at 260 nm, Column temperature: 25°C, Sample injection
volume: 10 μL.

6) Fractionation of extracts using hydrophobic
cartridge column

Fractionation and purification of hot-water extracts
employed hydrophobic cartridge column (Oasis HLB, Nihon
Waters, Shinagawa-ku, Tokyo, Japan). Hydrophobic cartridge
columns were conditioned with 3 mL of ACN and 3 mLof distilled water. Following this step, 1 mL of the sample
solution was injected. Subsequently, 3 mL of elution was
examined step by step with ACN concentrations from 0%,10%, 20%, 50% to 75%, and then collected.

Statistical analysis

The inhibitory rates of AGE formation were shownas mean value ± standard deviation. Comparisons between
groups were performed in measurements using the Tukey’s
test. The correlation analysis between the measured values
was performed using the Pearson's correlation coefficient.
Interpretation of correlation was at a range of 0.4 < |r| <=1.0, and weak correlation at a range of − 0.2 < |r| <= 0.4. A
hazard rate of less than 5% was considered to be statisticallysignificant.

Results

Anti-glycation effect of KP

Ten types of KP samples were examined: 4 types ofdried powder of rhizomes and 6 types of essence extract
powder of rhizomes. The IC 50 of each sample as antiglycation activities was shown, classifying each into hotwater extract of dried powder, 70% ethanol extract solution,and extract powder (Fig. 1).
The values of the IC 50 were 0.027 ± 0.010 mg/mL (mean± standard deviation, 4 samples) in the hot-water extracts,
0.029 ± 0.011 mg/mL (4 samples) in the 70% ethanol extracts,
and 0.064 ± 0.053 mg/mL (6 samples) in the powder extracts.
The mean value of the hot-water extracts was the smallest.
However, there was no statistically significant difference inthe measured values of the extracts among the 3 groups.
All of the IC 50 in the hot-water extracts and 70 %ethanol extracts of the 4 types of dried powder samples were
smaller than in AG. Particularly, the IC 50 (0.014 mg/mL) of
the v216 - 1 in the hot-water extraction had the smallest valueof the 10 types of KP materials; the value was 1/3.4 of AG.
The IC 50 values of the 70% ethanol extracts were smaller
than the AG, except for v 244.

Anti-glycation activity of PMF

The inhibition rate of the AGE formation was measured
in 5 types of PMF contained in KP: 5, 7 - dimethoxyflavone,
5, 7, 4’ - trimethoxyflavone, 3, 5, 7 - trimethoxyflavone,
3, 5, 7, 4’ - tetramethoxyf lavone and 3, 5, 7, 3’, 4’ pentamethoxyflavone. As a result, in the 4 types of PMF,
excluding that of 3, 5, 7 - trimethoxyflavone, anti-glycation
activity was recognized (IC 50; 0.028 ± 0.024 mg/mL, 4
samples, Table 2 ). There was no correlation between the IC 50
and the number, or the binding site of the methoxy groups
within the PMF molecule.

_ 165 _

Antiglycation effect of Kaempferia parviflora

The amount of PMF in the KP extract solution

The amount of PMF in the hot -water extracts of dried
powder sample (0.20 ± 0.04 mg/mL, 4 samples) were 3.6
times (v220) - 8.2 (v 216-1) times larger than that in theethanol extracts (1.11 ± 0.54 mg/mL, 4 samples, p < 0.03,
Fig. 2). The amount of PMF in the extract powder solution
was recorded to be 5.7 times larger than the differences,
depending on the types of samples. Composition ratios of
the 5 types of PMF contained in the sample extract solution
were almost the same both in the ethanol extract solution andextract powder solution. However, regarding the hot-water
extract, the ratio of 3, 5, 7 - trimethoxyflavone was one-sixthof others (1.3 ± 2.6 mg/mL, 4 samples).

Correlation between the volume of PMF andthe anti-glycation activity of KP extract solution

There was no correlation between the total amount of
PMF contained in KP (except for 3, 5, 7 - trimethoxyflavone),
0.2

and IC 50 of KP (Fig. 3 ). Although hot-water extracts
contained less amount of PMF content in comparison withother sample solution, they showed smaller IC 50.

Anti-glycation activities of KP extracts in fractionatedeluate with hydrophobic cartridge column

A sample of v216 -1, which had the smallest IC 50 in
hot-water extraction for dried powder, was examined withfractionation and purification of the hydrophobic cartridge
column, and then the anti-glycation activity of the eluate
was measured. As a result, on water fractions, 50% ACN
fractions and 75% ACN fractions, the inhibition ratios of theAGEs formation were more than 50% (Fig. 4 ). Measurementof the amount of PMF in each eluted fraction indicated that
PMF existed in 50% ACN and 75% ACN fractions. The
amount of PMF contained in 50% ACN fraction was almost
the same as that of 75% ACN. However, the inhibition
concentration of the AGEs formation at 75% ACN fraction
was 1.8 times stronger than that of 50% ACN ( p < 0.01).

hot - water extract

70% ethanol extract

IC 50 (mg/mL)

0.15

0.1

0.05

0
v216-1v216-2

v219

v220

AG

Sample number

0.2

IC 50 (mg/mL)

0.15

0.1

0.05

0
v217

v218

v221

v222

v223

v224

AG

Sample number

Fig. 1. Anti-glycation activity of Kaempferia parviflora.

Sample number; reference of Table 1. IC 50 calculated HSA-glucose glycation model test result. IC 50 ; 50 %inhibitory concentration of AGE formation; AGE, advanced glycation end product; HSA, human serum albumin.

_ 16 6 _Glycative Stress Research

Table 2. Anti-glycation activity of polymethoxyflavone involvement Kaempferia parviflora rhzome.

polymethoxyflavonoid

IC 50 (mg/mL)

5, 7- dimethoxyflavone

0.061

5, 7, 4’-trimethoxyflavone

0.009

3, 5, 7 -tridimethoxyflavone> 0.1

3, 5, 7, 4’-tetramethoxyflavone

0.012

3, 5, 7, 3’, 4’-pentamethoxyflavone

0.029

IC 50 ; 50% inhibitory concentration of AGE formation; AGE, advanced glycation end product.

1.80

5, 7- dimethoxyflavone
5, 7, 4 - trimethoxyflavone

1.60

3, 5, 7 - trimethoxyflavone
3, 5, 7, 4 - tetramethoxyflavone

Concentration (mg/mL)

1.40

3, 5, 7, 3 , 4 - pentamethoxyflavone

1.20
1.00
0.80
0.60
0.40
0.20
0.00
v216-1 v216-2 v219
hot - water extract

v220 v216-1 v216-2 v219

v220

v217

v218

70% ethanol extract

Fig. 2. Polymethoxyflavonoid concentration of Kaempferia parviflora rhzome extract.

v221

v222

v223

v224

extracted sample

Sample number; reference of Table 1. Polymethoxyflavonoid measured by HPLC method. Columun; Inerstil ODS- 4, 3 μm, 100 x 4.6 mm
I.D., eluent; 0.1% formic acid / 0.1% formic acid -acetonitrile (70/30), flow rate; 1.0 mL/min, detection; UV at 260 nm, temperature; 25°C,
Sample number
injection volume; 10 μL.

_ 167 _

Antiglycation effect of Kaempferia parviflora

0.18
0.16
0.14

IC 50 (mg/mL)

0.12
0.10
0.08
0.06
0.04
0.02
0.00
0

0.5

1

1.5

2

Polymethoxyf lavonoid concentration (mg/mL)

Fig. 3. Correlation of IC50 to concentration of polymethoxyflavonoids.

The amount of polymethoxyflavonoids measured by HPLC method. IC50 calculated HSA-glucose glycation model test result. IC50;
50% inhibitory concentration of AGE formation; AGE, advanced glycation end product.

120

Inhibition ratio (%)

100

80

60

40

20

0
water

10% ACN

20% ACN

Eluted fruction

50% ACN

75% ACN

beforefructionFig. 4. Inhibition ratio of AGE formation to eluted fraction by the hydrophobic cartridge column.

Sample; hot-water extract of v216-1 (reference of Table 1). Hydrophobic cartridge column: Oasis HLB, Inhibition ratio of AGEs
tested HSA-glucose glycation model, **; p < 0.01 by Tukey’ s test, ACN; acetonitrile; AGEs, advanced glycation end products;
HSA, human serum albumin.

_ 16 8 _Glycative Stress Research

Discussion

PMF and Anti-glycation activity of KP

This study aimed to analyze active constituents of KP
as an inhibitor on glycation, comparing, regarding inhibitory
effectiveness, 10 types of extracts of KP rhizomes (4 types ofdried powder and 6 types of extract powder), and examining
the correlation between inhibitory effects on glycation and 5
types of PMF contained in KP.
The inhibitory effects on glycation tended to be stronger
in hot-water extracts of dried powder than in 70% ethanol
extract solutions and essence extract solutions of dried
powder. The amount of PMF, which was contained in KP,
was larger in 70% ethanol extracts than in hot-water extracts(p < 0.03). There was no recognized correlation between
the IC 50 of the KP sample solutions and content quantityof PMF. Moisture ethanol extract of KP has anti-glycation
activity, as previously reported, and PMF is reported to be
its active component 19). This study confirmed that 4 typesof PMF, excluding that of 3, 5, 7 - trimethoxyflavone, had
anti-glycation activities as well as involvement of PMF
in the anti-glycation effect of KP. Furthermore, this study
examined fractions which did not contain PMF (water
elution), other than fractions which contained PMF (50 %ACN and 75% ACN), regarded eluted fractions of hot-water
extracts by the hydrophobic cartridge column, and clarified
that anti-glycation activities were detected in the fractions
without PMF (water elution). Therefore, it is assumed thatcomponents other than PMF would greatly contribute to antiglycation activities of KP. Other than for PMF, there were
two possibilities, at most, for components which have antiglycation effects; one possibility was a hydrophilic substancewhich was maintained in the hydrophobic cartridge
column, and the other was a substance which existed
in the eluted fraction of 75% ACN in the hydrophobic
cartridge column. The total amount of PMF (except for
3, 5,7 - tridimethoxyflavone), which contained in solution
eluted with fractionation using the hydrophobic cartridge
column, was 0.266 mg/mL in 50 % ACN and 0.245 mg/
mL in 75% ACN. However, the inhibition concentration of
the AGEs formation was 52.2% in 50 % ACN and 93.6%
in 75% ACN. That is, anti-glycation activities of eluted
fractions did not depend on PMF concentrations. The reason
that the inhibition ratio of 75% ACN was 1.8 times larger
than that of 50% ACN may have been due to the possibility
that the substance which should promote the glycation wasremoved through fractionation and purification. Thus, further
examinations are required for verification on purificationof eluates. It has been previously reported that KP rhizomes
contain 11 types of flavonoid 22) and phenolic glycosides 23).
However, it has not been identified that these constitutes
would fulfil the role of anti-glycation activity.

Treatment process to utilize KP rhizomes

For utilization of KP as health foods, diversified
treatment processes are performed. The test samples used inthis study were dried products and essence extract powders
of KP rhizomes which are sold or marketed. The KP
rhizomes were for drying as slices (v220) or powder (v216 -1,
v216 -2, v219). For methods for drying KP, low-temperature
vacuum dehydration (v216 -1) and high-temperature drying(v216 -1, v216 -2 and v219) were performed. In addition,hydrous ethanol extract method was employed to produce

essence powder (v217, v218, v221, v222, v223 and v224).
Powder samples that were manufactured in high-temperaturedrying (v216-2 and v219) appeared black in color. Contrarily,
powder samples with low-temperature dehydration (v2161) appeared in a color of dark purple. It was assumed that
thermal denaturation of anthocyanin contained in KP would
affect the black color of the sample powder dried at a hightemperature (v216-2 and v219) depending on the drying
condition. Regarding the hot-water extract, the IC 50 of the
low-temperature vacuum dehydration (v216 -1) was lower
than the high-temperature drying (v216 -2). Contrarily,
regarding the 70% ethanol extract, the high-temperaturedrying (v216-2) was lower than the low-temperature vacuum
dehydration (v216 -1). It was assumed that the hydrophiliccomponents of anti-glycation activities contained in the KP
could be easily affected by heat.
KP contains PMF as a characteristic component. It has
been reported that the KP have beneficial effects, suchas improvement of metabolic syndrome, improvement ofmuscular metabolism dysfunction 15), obesity amelioration 16)
and enhancement of physical strength 17, 18). As PMF ishydrophobic, ethanol is utilized to obtain PMF efficiently
from KP. This study also indicated that larger amounts of
PMF extract were obtained through 75% ethanol dried
powder extraction and essence extraction rather than throughhot-water extraction.
In the case where the content quantity of PMF was
focused as the usage of KP, there was a possibility that 70%
ethanol extraction of low-temperature dehydration would be
advantageous. Contrarily, focusing on anti-glycation activity,
there was a possibility that the hot-water extraction of the
low-temperature dehydration powder would be advantageous.
In the case where two activities must be covered, direct
ingestion of dried powder was recognized to be a usage
which has the least influence of extraction. It was suggested
that proper treatment process would be required to suit its
purpose.

Conclusion

This study compared 10 types of rhizomes of KP(Kaempferia parviflora Wall. Ex. Baker) and 5 types of
PMF contained in KP regarding anti-glycation effects, andexamined correlations between quantity of PMF and antiglycation activities. There was no recognized correlation
between content quantity of PMF and inhibitory effects of
KP extract solutions. The examinations for anti-glycation
effects employing the fractionation and purification of KP
extracts suggested that active components, except for that of
PMF, would exist as a hydrophilic substance. A possibility
was indicated that some components greatly contributed tothe anti-glycation activity of KP excluding PMF.

Statement of conflict of interest

This research received support from Rene Co. Ltd.
(Chuo-ku, Osaka, Japan).

_ 169 _

Antiglycation effect of Kaempferia parviflora

Acknowledgments

This study was presented at “the 9th Meeting of theSociety for Glycative Stress Research” on September 5th,
2015, at Kyoto, Japan. This study was supported by theJapanese Council for Science, Technology and Innovation,
SIP (Project ID 14533567 ), “ Technologies for creatingnext-generation agriculture, forestry and fisheries ” (funding
agency: Bio-oriented Technology Research Advancement
Institution, NARO).

Reference1) Ichihasi M, Yagi M, Nomoto K, et al, Glycation stress andphoto-aging in skin. Ant-Aging Med. 2011; 8: 23-29.
2) Yagi M, Yonei Y. Glycative stress and anti-aging: 1. Whatis glycative stress? Glycative Stress Res. 2016; 3: 152-155.
3) Brownlee M, Vlassara H, Kooney A, et al. Aminoguanidine
prevents diabetes-induced arterial wall protein crosslinking. Science. 1986; 232: 1629-1632.
4) Bolton WK, Cattran DC, Williams ME, et al. Randomized
trial of an inhibitor of formation of advanced glycation
end products in diabetic nephropathy. Am J Nephrol.
2004; 24: 32-40.
5) Hori M, Yagi Y, Nomoto K, et al. Inhibition of advanced
glycation end product formation by herbal teas and its
relation to anti-skin aging. Anti-Aging Med. 2012; 9: 135148.
6) Ishioka Y, Yagi M, Ogura M, et al. Antiglycation effect ofvarious vegetables: Inhibition of advanced glycation end
product formation in glucose and human serum albumin
reaction system. Glycative Stress Res. 2015; 2: 22-34.
7) Parengkuan L, Yagi M, Matsushima M, et al. Antiglycation activity of various fruits. Anti-Aging Med. 2013;
10: 70-76.
8) Tanaka Y, Yagi M, Takabe W, et al. Anti-glycative effectof yogurt: Prevention of advanced glycation end productformation. Glycative Stress Res. 2017; 4: 25-31.
9) Saraswat M, Reddy PY, Muthenna P, et al. Preventionof non-enzymic glycation of proteins by dietary agents:
prospects for alleviating diabetic complications. Br J Nutr.
2009; 101: 1714-1721.
10) Saraswat M, Suryanarayana P, Reddy PY, et al.
Antiglycating potential of Zingiber officinalis and delayof diabetic cataract in rats. Mol Vis. 2010; 16: 1525-1537.
11) Chayan P, Supachai K. Notes on the genus Kaempferia L.
(Zingiberaceae) in Thailand. Journal of Thai Traditional& Alternative Medicine. 2008; 6: 27-51.
12) Mohammed Y, Wahab MA, Yousuf Md, et al. Some tribal
medicinal plants of Chittagong Hill Tracts, Bangladesh. J
Plant Taxon. 2007; 14: 117-128.
13) Tewtrakul S, Subhadhirasakul S, Kummee S. Anti-allergic
activity of compounds from Kaempferia parviflora. J
Ethnopharmacol. 2008; 116: 191-193.
14) Temkitthawon P, Hinds TR, Beavo JA, et al. Kaempferia
parviflora, a plant used in traditional medicine to enhancesexual performance contains large amounts of low affinity
PDE5 inhibitors. J Ethnopharmacol. 2011; 137: 1437-1441.

15) Toda K, Takeda S, Hitoe S, et al. Enhancement ofenergy production by black ginger extract containing
polymethoxy flavonoids in myocytes through improving
glucose, lactic acid and lipid metabolism. J Nat Med.
2016; 70: 163-172.
16) Shimada T, Horikawa T, Ikeya Y, et al. Preventive effect of
Kaempferia parviflora ethyl acetate extract and its major
components polymethoxyflavonoid on metabolic diseases.
Fitoterapia. 2011; 82: 1272-1278.
17) Promthep K, Eungpinichpong W, Sripanidkulchai B, et al.
Effect of Kaempferia parviflora extract on physical fitnessof soccer players: A randomized double-blind placebocontrolled trial. Med Sci Monit Basic Res. 2015; 21: 100108.
18) Toda K, Hitoe S, Takeda S, et al. Black ginger extract
increases physical fitness performance and muscular
endurance by improving inflammation and energy
metabolism. Heliyon. 2016; 2: e00115.
19) Nakata A, Koike Y, Matsui H, et al. Potent SIRT1
enzyme-stimulating and anti-glycation activity ofpolymethoxyflavonoids from Kaempferia parviflora. Nat
Prod Commun. 2014; 9: 1291-1294.
20) Hori M, Yagi M, Nomoto K, et al. Experimental models
for advanced glycation end product formation using
albumin, collagen, elastin, keratin and proteoglycan. AntiAging Med. 2012; 9: 125-134.
21) Yagi M, Yonei Y. Glycative stress and anti-aging. 4. The
evaluation of glycative Stress: Evaluation for anti-glycative
effect. Glycative Stress Res. 2017; 4: 087-092.
22) Sutthanut K, Sripanidkulchai B, Yenjai C, et al.
Simultaneous identification and quantitation of 11
flavonoid constituents in Kaempferia parviflora by gas
chromatography. J Chromatogr A. 2007; 1143: 227-233.
23) Azuma T, Tanaka Y, Kikuzaki H. Phenolic glycosides
from Kaempferia parviflora. Phytochemistry. 2008; 69:2743-2748.

_ 170 _