Effect of Aqueous Extract from - American Chemical Society

Chapter 18

Downloaded by UNIV OF SOUTHERN CALIFORNIA on August 7, 2013 | http://pubs.acs.org Publication Date: December 17, 2001 | doi: 10.1021/bk-2002-0803.ch018

Effect of Aqueous Extract from Lepidium meyenii on Mouse Behavior in Forced Swimming Test 1

1

2

Bo Lin Zheng , Kan H e , Zhen Yen Hwang , Yang Lu2, Sui Jun Y a n , Calvin Hyungchan K i m , and Qun Yi Zheng 3

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1

1

Pure World Botanicals, Inc., 375 Huyler Street, South Hackensack, NJ 07606 Shenyang Medical College, Shenyang, People's republic of China Liaoning College of Traditional Chinese Medicine, Shenyang, Liaoning, People's Republic of China 2

3

The plant Lepidium meyenii (Walp.), with the common name of maca, is a less known domesticated plant of Peru. In the present study, we have investigated the activity of energy enhancement of aqueous extracts from roots of maca on the behavior in mice using forced swimming test. The results of dose-response study with the aqueous extract, MacaForce™ AQ-1, in a range of daily dose at 4, 10, 20 and 40 mg/g body weight of mouse indicated that the maximum activity was reached by two weeks during a 21-day study. The energy enhancement was especially evident at the daily dose level greater than 20 mg/g body weight. The swim time in mice with 7-day oral administration with MacaForce™ AQ-1 at daily dose of 20 and 40 mg/g body weight were 13.50 ± 3.18 minutes and 16.72±2.94 minutes, respectively (control group at 10.47±2.38 minutes); with 14-day oral administration of MacaForce™ AQ-1 at the same doses were 17.42±4.22 minutes and 19.33±3.86 minutes, respectively (control group at 11.4512.46 minutes). In addition, the swim time in mice

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© 2002 American Chemical Society

In Quality Management of Nutraceuticals; Ho, C., et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2001.


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with 7-day oral administration of aqueous extracts, MacaForce™ AQ-3 and MacaForce™ AQ-4, at daily dose 40 mg/g body weight were 10.89±2.74 minutes and 14.38±3.09 minutes, respectively (control group at 11.74±1.38 minutes); with 14-day oral administration of MacaForce™ AQ-3 and AQ-4 at the same dose were 14.81±2.11 minutes and 15.34±2.60 minutes, respectively (control group at 10.90±2.04 minutes). The study with aqueous extracts, MacaForce™ A Q 1, AQ-2, AQ-3, and AQ-4, showed that the increase in swim time was directly related to the increase in content of polysaccharides in the aqueous extracts. Increase in serum lactate dehydrogenase (LDH) activity and decrease in the concentration of serum lactic acid were dose related with the content of potential active constituents.

Lepidium meyenii (Walp.), known as maca, is a plant that grows in the Andean Mountains at altitudes of more then 10,000 feet (/). Maca is one of the few plants that learned to flourish in this harsh environment of intense sunlight, violent winds, poor soil and low temperature. Native Peruvians have traditionally utilized maca for both nutritional and medicinal purposes. For centuries, maca roots have been used to enhance the fertility and sexual performance of men and women, as well as to treat women with menopausal symptoms. In 1960's, some studies were conducted to determine the effect of maca in rats, but the design and results were far from being satisfactory (2). A recent in vivo animal study has demonstrated that oral administration of purified lipidic extract of maca enhanced the sexual function in mice and rats (5). The present study used a forced swimming test followed by measurements of lactic acid, lactate dehydrogenase and melonic acids in serum to evaluate the energy enhancement property of aqueous maca extracts, MacaForce™, in mice. Unlike the lipidic extract, the major constituents of aqueous extract of maca include a number of water soluble materials, such as amino acids, trace mineral, sugars, polysaccharides, partial fatty acids and sterols. The results of the study indicated that aqueous extract of Lepidium meyenii (Walp.) significantly enhanced motor tolerance and relieved fatigue of the mice. The present study reveals for the first time an energy enhancement property of Lepidium meyenii.


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Materials and Methods


Plant Material, Extraction and Purification The dried maca roots were supplied from Peru in 1998. A voucher specimen is deposited in the Herbrio de Museo de Historia Natural ' J . Prado' Un. H . S., Lima, Peru. The extraction was carried out using ethanol and water as solvent via Pure World's proprietary extraction and purification process (4). The resulting aqueous extracts, MacaForce™ AQ-1, AQ-2, AQ-3, and AQ-4, were formulated with excipient like maltodextrin, or with other herbs such as ginseng or rhodiola for comparison. These extracts were dried to form a powdered extract

Phytochemical Analysis MacaForce™ AQ-1 contains 0.004% ^sitosterol (cholestrane was used as internal standard), 0.045% fatty acids, 4.56% amino acids (0.17% alanine, 0.10% arginine, 0.03% aspartic acid, 0.27% glutamic acid, 0.05% glycine, 0.16% isoleucine, 0.14% leucine, 3.25% proline, 0.03% serine, 0.07% threonine, 0.02% tyrosine, 0.27% valine), 22.0% polysaccharide (hydrolysable carbohydrate: 1.3% glucose, 4.7% fructose and 16.0% sucrose) and 0.03% macaene and macamide. MacaForce™ AQ-2 contains 0.18% benzyl-isothiocyanate, 0.019% sterols (0.006% campesteol, 0.003% stigmasterol, 0.010% ^-sitosterol), 1.11% fatty acids (0.28% capric acid, 0.2% lauric acid, 0.19% palmitic acid, 0.02% stearic acid, 0.06% oleic acid, 0.24% linoleic acid, 0.12% linolenic acid), 5.97% amino acids (0.145% alanine, 0.374% arginine, 0.139% aspartic acid, 0.252% glutamic acid, 0.060% glycine, 0.030% histidine, 0.039% isoleucine, 0.038% leucine, 0.031% lysine, 0.013% methionine, 4.630% proline, 0.028% serine, 0.052% threonine, 0.019% tyrosine, 0.115% valine), 21.0% polysaccharide (hydrolyzable carbohydrate: 1.20% glucose, 4.45% fructose and 15.3% sucrose), and 0.27% macaene and macamide MacaForce™ AQ-3 contains 0.001% β-sitosterol, 0.011% fatty acids, 1.137% amino acids (0.041% alanine, 0.024% arginine, 0.008% aspartic acid, 0.068% glutamic acid, 0.013% glycine, 0.040% isoleucine, 0.035% leucine, 0.811% proline, 0.008% serine, 0.018% threonine, 0.005% tyrosine, 0.066% valine), 2.853% polysaccharide (hydrolysable carbohydrate,0.157% glucose, 0.583% fructose and 2.113% sucrose), 0.006% macaene and macamide, 0.14% ginsenoside from Siberian ginseng, and 0.07% salidroside from rhodiola.



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MacaForce™ AQ-4 contains 0.073% benzyl-isothiocyanate, 0.0039% sterol (0.0012% campesterol, 0.0006% stigmasterol, and 0.0021% ^-sitosterol), 0.234% fatty acids (0.059% capric acid, 0.042% lauric acid, 0.040% palmitic acid, 0.004% stearic acid, 0.013% oleic acid, 0.051% linoleic acid, and 0.025% linolenic acid), 2.218% amino acid (0.056% alanine, 0.143% arginine, 0.053% aspartic acid, 0.097% glutamic acid, 0.023% glycine, 0.012% histidine, 0.015% isoleucine, 0.015% leucine, 0.012% lysine, 0.005% methionine, 1.776% proline, 0.011% serine), 9.465% polysaccharide (hydrolyzable carbohydrates, 0.544% glucose, 2.012% fructose and 6.909% sucrose), and 0.006% macaene and macamide

Animals Male and female Shenyang mice (Grade II) weighing 25 ± 1 g were used in the study. Age of mice at the start of the experiment was 8-10 weeks. A l l mice had access to standard diet and water ad libitum. Each test article was added into 10% ethanol/water solution, and was heated to 30°C to make a suspension. The test article and a blank 10% ethanol/water solution of equal volume at daily dose specified by the experimental design were orally administered to experimental and control groups, respectively, by gavage twice per day. Each group of 15 mice consisted of about 50% each of male and female. Periodic analysis of water was performed, and there was no presence of known contaminants in the diet or water.

Forced Swim Tests In the first study, fifteen groups of fifteen mice in each group were used. Five groups each from the fifteen groups were selected randomly for 7-day diet test, 14-day diet test, and 21-day diet test. Each five groups consisted of one control, and four groups each fed with MacaForce™ AQ-1. MacaForce™ AQ-1, an aqueous extract of Lepidium meyenii, was administrated to the four groups by gavage at dose of 4, 10, 20 and 40 mg/g body weight. A 4-g lead weight was attached to tail of each mouse and was placed inside a polypropylene jar (30 χ 15 χ 40 cm) containing 30 cm water maintained at 23 ± 2°C. The total swim time in minutes was recorded from the time the mouse was placed in water to the time of death (Table I). Additional experiments using other aqueous extracts of maca, MacaForce™ AQ-3 and MacaForce™ AQ-4, were conducted to determine their property of energy enhancement by the forced swim test. In this study,



262 eight groups of 15 mice in each group were used. Four groups were selected randomly for 7-day diet test, and the remaining for 14-day diet test. Each four groups consisted of one control group and one group each fed with maltodextrin, MacaForce™ AQ-3, and MacaForce™ AQ-4. A l l test articles were administered twice daily to mice by gavage at dose of 40 mg/g body weight. On Day 7 or Day 14, at 2 hours after the second oral administration, a 4-g lead weight was attached to tail of each mouse and was placed inside a polypropylene jar (30 χ 15 χ 40 cm) containing 30 cm water maintained at 23 ± 2°C. The total swim time in minutes was recorded from the time the mouse was placed in water to the time of death (Table II).

Table I. Effect of L. meyenii Lipidic Extract MacaForce™ AQ-1 and on Swim Time of Mice in Forced Swim Test Day

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Group

Control MacaForce™ AQ-1

14

4 10 20 40


21

Dose (per each mouse, mg/g body wt.)

4 10 20 40


4 10 20 40

Animal Number (n)

Duration of Swim Time in Minutes (mean ±SD)

15

10.4712.38

-

15 15 15 15

10.78 ± 2 . 6 8 11.1413.02 13.5013.18 16.7212.94

>0.05 >0.05 0.05 0.05 O.01

40

15

15.3412.60

O.01

*vs control.



264 In the second study, each of MacaForce™ AQ-2 at daily dose of 40 mg/g body weight was orally administered to a group of randomly selected 15 mice by gavage twice per day for 21 days. Another group of 15 mice received a blank 10% ethanol/water solution by similar manner. On Day 21, two hours after the oral administration, a 4-g lead weight was attached to each mouse and was placed into a vertical glass cylinder (40 χ 40 cm, diameter χ height) containing 30 cm water maintained at 23 ± 2°C. The time at which each mouse was submerged in water for more than 3 seconds denoted the end of the swim time. Total swim time of each mouse was measured (Table III). In this study, the methodology for swim test was different compared to that of the first study in order to collect the fresh blood samples from live mice for the determination of lactic acid, L D H and M D A in blood. Table III. Effect of L. meyenii lipidic Extract on Swim Time of Mice in Forced Swim Test* Day

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Group


Dose (per each mouse, mg/g body wt.) 40

Animal Number (n)

Duration of Swim Time in Seconds (mean ±SD)

15 15

110.07 ± 2 . 5 8 124.07 ± 3.30

P**

Effect of Aqueous Extract from - American Chemical Society

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