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1. Introduction
-Lipoic acid (thioctic acid) is a potent anti-oxidant that has been widely used in food supplement
preparations. -Lipoic acid has been used to alleviate peripheral pain in severe diabetic patients and its
application in food preparations is getting popular. According to Standards Concerning the Scope of
Pharmaceutical Products by PFSB Notification No. 0331009 dated March 31, 2004, the Ministry of Health,
Labor & Welfare has revised and re-categorized -lipoic acid as an additive to be used in general food
preparations or beverages.
COOH
COOH
S S
HS SH
Oxidized form
Reduced form
At Oryza Oil & Fat Chemical Co., Ltd., innovative process has been developed for the
production and commercialization of -lipoic acid enabling its application in the food industry.
In addition, liquid form -lipoic acid with high water dispersibility has been developed for its
increasing demand in the beverages industry. Meanwhile, studies have been carried out in
Oryza Oil & Fat Chemical Co., Ltd. to evaluate the various beneficial effects of -lipoic acid,
e.g. skin whitening effect, inhibition of adipocytes production and growth promoting effect on
muscle cells.
Glucose
[Energy source]
Glycolysis system
Pyruvate dehydrogenase
-lipoic acid
[Metabolization ]
Acetyl CoA [A starter of energy production, a fuel molecule ]
CO2
H 2O
e- [Electron]
H 2O
Energy
ATP*
* Adenosine triphosphate
Fig. 2. Energy Production from Glucose and Site of Action of -Lipoic Acid
Creatine is important for the production of energy and lipid catabolism in muscles. As skeletal
muscle tissue is the major site for glucose following a meal, -lipoic acid that enhances glucose
uptake by skeletal muscle is potentially useful in weight reduction and long term prevention
against obesity.
1) Saengsirisuwan V., Perez F. R., Sloniger J. A., Maier T., Henriksen E. J. Interaction of exercise training
and -lipoic acid on insulin signaling in skeletal muscle of obese Zucker rats. Am. J. Physiol.
Endocrinol. Metab. 287E529-536 (2004).
2) Burke D. G., Chilibeck P. D., Parise G., Tarnopolsky M. A., Candow D. G. Effect of -lipoic acid
combined with creatine monohydrate on human skeletal muscle creatine and phosphagen concentration.
Int. J. Spot. Nutr. Exerc. Metab. 13, 294-302 (2003).
The effect of -lipoic acid on muscle cell lines, L6 cells, was examined. L6 cells were
cultured with -lipoic acid for 24 hours. Fig. 3 shown that cell production increases in the
presence of -lipoic acid. -lipoic acid promote muscle growth and maintenance of healthy lean
muscle.
107
106
105
104
103
102
101
100
99
98
97
0
3
10
30
Concentrationg/mL
100
Method
L6 cells (5x104 cells/ml) were suspended and cultured in D-MEM medium containing 10%
bovine fetal serum, 100 units/ml of penicillin G and 100g/ml of streptomycin. 100l of the
above suspension was distributed into 96-well microplate. Different concentrations of -lipoic
acid was added and the mixture was incubated for 24 hours. Degree of cell growth was
determined by MTT assay.
Muscle weightg
0.18
0.17
0.16
0.15
0.14
Control
Fig. 4. Effects of Continuous Intake of -Lipoic Acid on the Weight of the Soleus Muscle in Mice (mean
S.E., n=6)
Method
Mice (ddy strain, male, 5 weeks old) were fed with diet (MF: Oriental Yeast Co., Ltd)
containing -lipoic acid (concentration 0.05% & 0.1%) for 24 days. Weight of soleus muscle
was measured.
In another study conducted by Dicter N et. al.3, -lipoic acid shown to cause mitochondrial
uncoupling and inhibition of glycogen synthesis. Glucose metabolism is regulated and weight
maintenance is achieved.
3) Dicter N., Madar Z., Tirosh O. -lipoic acid inhibits glycogen synthesis in rat soleus muscle via its
oxidative activity and the uncoupling of mitochondria. J. Nutr. 132, 3001-3006 (2002).
Control
1g/mL
3g/mL
10g/mL
Method
3T3-L1 adipocytes (5x104 cells/ml) were incubated in D-MEM medium (high glucose)
containing 10% bovine fetal serum for 2 days. The medium was then replaced by another
medium containing insulin (1g/ml), dexamethasone (0.25M), isobutylmethylxanthine (0.5
mM) and different concentrations of -lipoic acid. The new medium was further incubated for a
total of 7 days. -lipoic acid and insulin (1 g/ml) was replaced every 2 days.
In adipocytes, an enzyme exists that converts glucose that is taken by insulin to triglyceride.
The enzyme, glycerol 3-phosphate dehydrogenase (GPDH), is involved in this process to store
GPDH
Activity(Unit/mL)
GPDHUnit/mL
excessive glucose-derived energy in fat cells. We studied the activity of-lipoic acid on crude
GPDH prepared from 3T3-L1 adipocytes and discovered that it has inhibitory activity (Fig. 6).
Namely, --lipoic acid prevents fat accumulation converted from excessive sugar.
0.06
0.05
0.04
0.03
0.02
0.01
0
Control
100
300
1000
g/mL
Concentration(g/L)
The lysate prepared from differentiated 3T3-L1 cells was used as the enzyme source. GPDH
activity was measured by commercioally available kit (Primary Cell), Japan.
Control
Control
5
4
3
2
1
0
0
6
8
Day
10
12
14
Fig. 7 Change in Mice Weight Fed -Lipoic Acid Continuously with or without Exercise (n=5)
6
Method
Mice (ddY, male, 5 weeks old) were fed the diet (MF, Oriental Yeast) that includes -lipoic acid (0.1%) for
13 days. Exercise was loaded with a treadmill (MK-770M, Muromachi Kikai) for ten minutes (5 rpm/min)
once a day.
Before Taking
Wightkg
65.512.2
65.512.1
4/7
Body Fat%
19.15.8
18.65.9
6/7
22.24.0
22.24.0
3/7
BMIkg/m
After Taking
Impedance
49066
48055
5/7
Fat Content%
12.72.6
12.53.2
6/7
0.918.3
0.918.2
4/7
Waist Sizecm
77.210.9
77.612.1
2/7
Hip Sizecm
95.58.9
92.17.3
7/7
Waist / Hips
0.810.06
0.840.08
2/7
13.34.0
14.64.3
Blood Sugarmg/dL
87.3100.7
69.318.0
Cholesterolmg/dL
199.428.1
HDL- Cholesterolmg/dL
54.616.3
Triglyceridemg/dL
Phosphatidemg/dL
Creatininemg/dL
1/7
p<0.01
204.036.1
6/7
2/7
54.1144.8
1/7
155.0127.5
198.3206.3
1/7
219.933.6
228.750.4
4/7
0.820.11
0.870.131
p<0.05
6/7
Total Proteing/dL
7.110.34
7.160.29
4/7
Values are shown with the average of 7 subjects (one subject stopped the test because of epigastric distress)
with standard deviation.
Table 2: Obesity Indexes and Blood Parameters before and after Taking - Lipoic Acid (200 mg)
Parameters
Before Taking
After Taking
Wightkg
71.015.3
70.414.0
6 / 10
Body Fat%
22.36.6
21.96.4
6 / 10
24.45.4
24.34.9
4 / 10
BMIkg/m
Impedance
48977
48168
5 / 10
Fat Content%
14.65.8
14.35.2
6 / 7
4.413.4
4.412.4
4 / 10
Waist Sizecm
82.111.4
82.29.5
3 / 10
Hip Sizecm
96.08.11
95.48.8
7 / 10
Waist / Hips
0.850.05
0.860.03
1 / 10
16.77.3
16.37.6
6 / 10
Blood Sugarmg/dL
97.028.4
94.029.9
5 / 10
Cholesterolmg/dL
200.226.6
203.126.5
4 / 10
HDL- Cholesterolmg/dL
58.422.7
57.320.3
3 / 10
Triglyceridemg/dL
164.4117.7
118.165.5
4 / 10
Phosphatidemg/dL
238.341.4
230.633.5
5 / 10
Creatininemg/dL
0.860.16p<0.05
0.840.15
8 / 10
Total Proteing/dL
7.210.2
7.320.1
Value are shown with the average of 10 subjects with standard deviation.
Wight
Impedance
30
80
Fat
Content
Body
Fat
90
550
5 / 10
BMI
BMI
28
25
26
20
25
24
kg
20
10
450
15
50
40
After
Before
After
18
16
Before
After
22
20
10
350
Before
kg/m2
60
15
kg
70
Before
After
Before
After
Waist
Size
115
95
110
20
90
10
cm
cm
80
-30
Before
85
60
80
Before
110
17
0.85
After
7
0.7
Before
270
HDL-Cholesterol
HDL-
90
700
250
80
600
230
70
500
210
60
190
50
50
170
40
40
150
30
60
Before
Before
After
Creatinine
After
After
Before
Triglyceride
Phosphatide
330
310
290
400
270
250
300
200
230
210
100
190
170
Before
After
Before
Total
Protein
7.6
1.1
7.4
g/dL
1.0
0.9
0.8
7.2
7
6.8
0.7
6.6
6.4
0.6
Before
After
Before
After
Fig. 8 Changes in Obesity Indexes and Blood parameters before and after Taking -Lipoic Acid (100 mg)
After
350
Before
After
7.8
1.2
mg/dL
After
mg/dL
70
0.75
Before
mg/dL
mg/dL
mg/dL
80
13
11
100
90
15
0.8
Cholesterol
Blood
Sugar
120
19
90
65
After
21
95
70
-20
23
0.9
100
75
-10
Thickness
of Abdominal Fat
/
0.95
105
85
Waist / Hip
100
cm
30
Hip Size
mg/dL
Degree of Obese
After
3. Cosmeceutical Effects
(1) Skin-Whitening Effect
Effect on melanin formation (in vitro)
The effect of -lipoic acid on B16 melanoma cells was examined. As illustrated in Fig. 9,
-lipoic acid demonstrated a dose-dependent suppression effect on melanin formation. -lipoic
acid is a potentially useful skin whitening agent.
120
100
80
60
40
20
0
0
25
50
Concentration g/mL)
100
Fig. 9. Effects of -Lipoic Acid on Melanocyte (B16) Growth (mean S.E., n=6)
Method
B16 melanoma cells (5x104 cells/ml) were suspended in MEM medium (containing 10% fetal bovine serum,
100 units/ml penicillin and 100g/ml streptomycin) containing 2mM theophylline, and 500l of the suspension
was placed into a 24-well plate. Different concentration of -lipoic acid (55l) was added and the mixtures
were incubated for 3 days. After incubation, PBS (300 l) was added and cells were crushed by ultrasonication.
Crushed cell mixture was recovered on a 96-well plate and absorbance was determined at
wavelength 415 nm (reference wavelength 700 nm).
UV ray exposure
UV ray exposure
UV ray exposure
End of Administration
-2
UV ray exposure
Start of Administration
pigmentation, proving that it performs skin-lightening activity with oral intake both in vitro and
in vivo.
10
12
Change
the color value(L*
L*in
Light
0
Day after first UV ray exposure(Day)
-2
-4
-6
-8
Dark
-10
-12
0 mg/kg
1 mg/kg
25 mg/kg
50 mg/kg
-14
Industries Co., Ltd.) before UV ray radiation (day 0) and 8th and 10th days after starting the radiation.
11
Control
1 mg/kg
25 mg/kg
50 mg/kg
Fig. 11 Radiation Area on the 8th Day after Starting the Radiation
12
Cell proliferation%
120
110
100
90
80
Control
1
5
Concentrationg/mL
25
Fig. 12. Effects of -Lipoic Acid on NB1RGB Fibroblast Growth (mean S.E., n=6)
Method
NB1RGB cells (2x105 cells/ml) were suspended in -MEM medium (containing 10% bovine fetal serum,
100 units/ml penicillin and 100g/ml streptomycin), and 100l of the suspension was placed into a 96-well
plate. Different concentration of -lipoic acid (10 l) was added and incubated for 2 days. Degree of cell
growth was determined using MTT assay.
13
Control
Corneum
Epidermis
Dispersed region
of granule cells
Dermis
Granules
-lipoic acid
(1 g/mL)
Corneum
Epidermis
Dispersed region of
granule cells
Dermis
Method
Reconstructed human skin model (TESTSKINTM: Toyobo Co., Ltd) was used. -lipoic acid was injected
into the dermal layer of skin and incubated for 6 days. The medium was replaced every 3 days. Cross section of
tissue specimens were prepared after treatment in 10% formalin. Changes in specimens was observed under
microscopic enlargement.
14
4. Antioxidative Effects
The antioxidative effect of -lipoic acid has been renown for years. Studies were carried out
50
40
30
20
10
0
75
100
250
500
1000
DPPH Radical
Scavenging Activity(%)
SOD-like Activities
to confirm its anti-oxidative effect. As illustrated in Fig. 14 and Fig. 15, -lipoic acid showed a
dose-dependent antioxidative effect. -lipoic acid is a potentially useful antioxidant for the
prevention of degenerative diseases.
Final concentrationppm)
35.0
30.0
25.0
20.0
15.0
10.0
5.0
0.0
-5.0
10
100
1000
Final concentrationppm
5000
Modified product
conventional product
15
Relative content
% of the initial value
120
100
80
60
80
100
120
60
40
20
0
0
1
Time (hr)
(2) pH Stability
Relative content
% of the initial value obtained
without pH adjustment
The effect of pH on -lipoic acid was examined at room temperature in dark for 1 day and 1
week. -lipoic acid is highly remained stable at wide pH range, pH 3-10 (as shown in Fig. 18).
100.0
80.0
After 1 day
After 1 week
60.0
40.0
20.0
0.0
3
10
pH
16
7. Nutritional Information
Results
0.2 g/100 g
0.0 g/100 g
99.8 g/100 g
0.0 g/100 g
0.0 g/100 g
898 kcal/100 g
0.0 g/100>
1 mg/100 g
Method
Moisture
Karl Fischer Reagent
*1
Protein
Kieldahl method
Fat
Direct extraction method
Ash
Direct ashing method
*2
Carbohydrate
Energy*3
Dietary fiber
Prosky method
Sodium
Atomic absorption spectrophotometory
*1 N=6.25
*2 100 (moisture + protein + fat + ash)
*3 Factors for calculating the energy valueprotein, 4; fat, 9; carbohydrate, 4; dietary fiber, 2
Tested by:SRL, Inc.
Date of issue of the test result reportSeptember 2, 2004
Research result issue numberNo. 200408200016
8. Safety Profile
(1) Acute Toxicity (LD50)
In the single-dose toxicity test in ddY mice, the LD50 values of -lipoic acid were 405 mg/kg and 277
mg/kg in male and female mice, respectively. These values correspond to ingestion of 16.6 g and 24.3 g, of
-lipoic acid, in adult humans weighing 60 kg.
17
18
Results
Detection Limits
Assay Method
Cyclohexane
Not Detected
5 ppm
Ethyl acetate
Not Detected
Toluene
Not Detected
5 ppm
5 ppm
GC-MS
GC-MS
Acetone
Not Detected
5 ppm
GC-MS
Hexane
Not Detected
5 ppm
GC-MS
Dichloromethane
Not Detected
1 ppm
GC-MS
19
GC-MS
S
n
C O OH
C O OR 3
Polymer A
A : R = R = H or
Polymer
B
B : R = R = H or
3
R = HH oor
r E tEtO H
Fig. 18. Structure of -lipoic acid Polymers A and B.
20
ALPHA LIPOIC
ACID-P, P80
Applications
Examples
ALPHA LIPOIC
ACID-PC, PC80
ALPHA LIPOIC The product is water-soluble powder for cosmetics. It is Body care (body lotion, body cream, soap
etc.)
ACID-WSPC80 suitable for toners and conditioning lotions.
Makeup (lipstick, foundation, etc.)
ALPHA LIPOIC The product is liquid of -lipoic acid for cosmetics. It is
Makeup (lipstick, etc.)
suitable for cosmetics.
ACID-LC1
12. Packaging
ALPHA LIPOIC ACID-P, P80, WSP8 (Powder, Food Grade)
ALPHA LIPOIC ACID-PC, PC80, WSPC8 (Powder, Cosmetic Grade)
5kg
21
13. Storage
Store in cool, dry place. Avoid humidity. In particular, ALPHA LIPOIC ACID-L1,and ALPHA LIPOIC
ACID-LC1 is stored under 5.
Cosmetic
ALPHA LIPOIC ACID-PC
INCIName
Thioctic acid
-WSPC8
-LC1
Cyclodextrin
Polyglyceryl-10
Thioctic acid
Myristate
Propylene Glycol
Caprylate
Glycerin
Thioctic acid
Alcohol
22
PRODUCT STANDARD
PRODUCT NAME
6.Ignition Residue
Max. 0.1 %
7.Purity Test
(1)6,8-Epitrithiooctanoic acid
(2)Polymer
(3)Heavy Metals
(4)Arsenic
Max. 0.1 %
Max. 2.0 %
Max. 10 ppm
Max. 1 ppm
1.Appearance
2.Certification Test
(HPLC)
(Precipitation Method)
(The Japanese Standards for Food Additives)
(Standard Methods of Analysis in Food
Safety Regulation)
Max. 1 10 cfu/g
(Analysis for Hygienic Chemists)
Max. 1 10 cfu/g
(Analysis for Hygienic Chemists)
Negative
(Analysis for Hygienic Chemists)
Max. 0.1 %
Not detected
Not detected
Not detected
Not detected
Not detected
Not detected
Not detected
(GC)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
Ingredient
Content
Thioctic Acid
100 %
23
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
PRODUCT STANDARD
PRODUCT NAME
4.Loss on Drying
Max. 0.5 %
5.Ignition Residue
Max. 0.1 %
1.Appearance
2.Certification Test
6.Purity Test
(1)6,8-Epitrithiooctanoic acid Max. 0.1 %
(2)Heavy Metals
Max. 10 ppm
(3)Arsenic
Max. 1 ppm
7.Standard Plate Counts
8.Moulds and Yeasts
9.Coliforms
10.Residual Solvents
(1)Ethanol
(2)Other Solvents
Acetone
Cyclohexane
Dichloromethane
Ethyl acetate
Hexane
MTBE
Toluene
11.Composition
(HPLC)
(The Japanese Standards for Food Additives)
(Standard Methods of Analysis in Food
Safety Regulation)
Max. 1 10 cfu/g
(Analysis for Hygienic Chemists)
Max. 1 10 cfu/g
(Analysis for Hygienic Chemists)
Negative
(Analysis for Hygienic Chemists)
Max. 0.1 %
Not detected
Not detected
Not detected
Not detected
Not detected
Not detected
Not detected
(GC)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
Ingredient
Content
Thioctic Acid
Vegetable Oils & Fats
100%
24
80%
20%
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
PRODUCT STANDARD
PRODUCT NAME
4.Loss on Drying
Max. 9.0 %
5.Ignition Residue
Max. 0.1 0%
6.Purity Test
(1)Heavy Metals
(2)Arsenic
Max. 10 ppm
Max. 1 ppm
1.Appearance
2.Certification Test
Max. 1 10 cfu/g
(Analysis for Hygienic Chemists)
Max. 1 10 cfu/g
(Analysis for Hygienic Chemists)
Negative
(Analysis for Hygienic Chemists)
Max. 0.1 %
Not detected
Not detected
Not detected
Not detected
Not detected
Not detected
Not detected
(GC)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
Ingredient
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
Content
Thioctic Acid
8%
92%
Cyclodextrin
100%
25
PRODUCT STANDARD
PRODUCT NAME
Min. 10.0 %
(HPLC)
3.Purity Test
(1)Heavy Metals
Max. 10 ppm
Max. 1 ppm
(2)Arsenic
Max. 1 10 cfu/g
Max. 1 10 cfu/g
6.Coliforms
Negative
7.Residual Solvents
Solvents except Ethanol
Not detected
8.Composition
Ingredients
Contents
Thioctic Acid
Glycerin Ester of Fatty Acid
Propylene Glycol Ester of Fatty Acid
Glycerin
Ethanol
Total
10%
50%
23%
15%
2%
100%
26
acid
PRODUCT STANDARD
PRODUCT NAME
2.Certification Test
Max. 0.1 %
Max. 2.0 %
Max. 10 ppm
Max. 1 ppm
(HPLC)
(Precipitation Method)
(The Second Method)
(The Third Method, Apparatus B)
Max. 1 10 cfu/g
Max. 1 10 cfu/g
Negative
11.Residual Solvents
(1)Ethanol
(2)Other Solvents
Acetone
Cyclohexane
Dichloromethane
Ethyl acetate
Hexane
MTBE
Toluene
12.Composition
Max. 0.1 %
Not detected
Not detected
Not detected
Not detected
Not detected
Not detected
Not detected
(GC)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
Ingredient
Content
Thioctic Acid
100%
27
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
PRODUCT STANDARD
PRODUCT NAME
2.Certification Test
Max. 0.1 %
Max. 10 ppm
Max. 1 ppm
Max. 1 10 cfu/g
Max. 1 10 cfu/g
Negative
(HPLC)
(The Second Method)
(The Third Method, Apparatus B)
(Analysis for Hygienic Chemists)
(Analysis for Hygienic Chemists)
(Analysis for Hygienic Chemists)
Max. 0.1 %
(GC)
Not detected
Not detected
Not detected
Not detected
Not detected
Not detected
Not detected
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
Ingredient
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
Content
Thioctic Acid
80%
20%
100%
28
PRODUCT STANDARD
PRODUCT NAME
2. Certification Test
Max. 10 ppm
Max. 1 ppm
Max. 1 102 cfu/g
Negative
Negative
Max. 0.1 %
Not detected
Not detected
Not detected
Not detected
Not detected
Not detected
Not detected
(GC)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
(Max. 1 ppm)
Ingredient
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
(GC/MS)
Content
Thioctic Acid
8%
Cyclodextrin
92%
100%
29
PRODUCT STANDARD
PRODUCT NAME
Min.10.0%
(HPLC)
3.Purity Test
(1)Heavy Metals
(2)Arsenic
Max. 10 ppm
Max. 1 ppm
10.Coliforms
Negative
11.Residual Solvents
Solvents except Ethanol
Not detected
12.Composition
Ingredients
Contents
Polyglyceryl-10 Myristate
Propylene Glycol Caprylate
Glycerin
Thioctic Acid
Alcohol
Total
50 %
23 %
15 %
10 %
2%
100 %
30
acid,
ORYZA OIL & FAT CHEMICAL CO., LTD. striving for the development of the new functional
food materials to promote health and general well-being.
From product planning to OEM - For any additional information or assistance, please contact
*The unapproved copy of this catalogue and appropriation are forbidden except for
the exception on the Copyright Act.
*The contents of this catalogue may be changed without prior notice.
Established Date : October 13, 2004
Revised Date : April 4, 2006