FUNCTIONAL FOOD

JIANHUA ZHOU*, NA FENG *Corresponding author College of Life Science, Shandong Normal University, Jinan 250014, China

Jianhua Zhou

Preparation of Chinese herbal oral liquid for alleviation of drunkenness

KEYWORDS: flowers of Honeysuckle (Flos Lonicera), flowers of Kudzu vine (Flos Pueraiae lobata), honey, alleviation of drunkenness.

Two animal tests on mice were conducted – righting reflex test to observe the effect of herbal extracts on Abstractdrunk rate, drunk latent time and sober up time, and blood alcohol content test using head-space gas chromatography method to measure the effect of herbal extracts on alcohol metabolism rate. By the means of optimizing experimental condition, the best composition of the compound oral liquid in terms of volume ratio of Honeysuckle flower extract : Kudzu vine flower extract : honey solution : citric acid solution is 3 : 7 : 0.2 : 0.01. This compound oral liquid developed has a unique taste with effect of drunkenness alleviation and long term health benefits, and thus can be applied in industrial production.

INTRODUCTION and protect the liver from damages caused by alcohol (17- 18). As such, Kudzu vine fl ower has long been considered as Excessive drinking damages body function and causes health a traditional Chinese medicine to treat alcoholic intoxication. problems such as fatty liver, alcoholic hepatitis, etc. In severe Research has shown that extract of fl owers of Kudzu vine has cases, it may lead to shock or even brain death. Therefore, the effect of protecting gastric mucous, facilitating digestion, research on fi nding an effective oral liquid which prevents improving immunity as well as lowering blood fat and sugar drunkenness has socially applicable value. content (19-21). Honeysuckle is a half-evergreen perennial plant belongs to Honey contains many types of sugar - glucose, fructose, the family of caprifoliaceae (1). One of the main chemical sucrose and maltose. There are also trace amount of vitamin, constituents in the extract of fl owers of Honeysuckle is amino acid, enzyme, yeast, pollen grain, wax, etc. (22). chlorogenic acid which promotes bile secretion (2-3). Honey has constituents which make it antisepsis and anti- Triterpenoid saponin in the extract also shows signifi cant infl ammatory (23). It also has the effect of distoxication and protective effect on liver (4-5). With the effects of promoting promotes tissue regeneration (24). Research has shown that bile secretion and protecting the liver, fl owers of Honeysuckle honey is effective in strengthening immune system, improving has been commonly applied for medical purposes (6). sleep, promoting digestion, protecting the liver and resisting Research has also shown that extract from honeysuckle fatigue. Besides high nutritive value, honey contains a rare fl ower induces immune tolerance thus can be used as fructose which promotes alcohol absorption and shortens the immunosuppressant (7-8). This protects transplanted organs sober up time (25-26). Furthermore, honey also has protective hence can be used to avoid transplant rejection in organ effect on cardiovascular system and immune system (27-28). transplantation. Furthermore, modern pharmacology Based on the pharmacological properties of fl owers of experiments and clinical practices all show that honeysuckle Honeysuckle, fl owers of Kudzu vine and honey in traditional fl ower has antibiosis function towards various bacteria (9-14). Chinese medicine, a compound oral liquid with these three This reduces the need of adding preservatives in products materials as main ingredients to relieve drunkenness was containing honeysuckle fl ower extracts. proposed. Two animal tests were conducted - righting refl ex Kudzu vine is a wild leguminous plant in the genus pueraria. test to observe effect of herbal extracts on mice drunk rate, Clinical trials have confi rmed that fl owers of Kudzu vine can drunk latent time and sober up time, and blood alcohol be used in making tea drinks to alleviate symptoms of been content test using head-space gas chromatography drunk such as vomiting (15). Modern pharmacology study method to measure the effect of herbal extracts on alcohol also shows that isofl avone in fl owers of Kudzu vine can affect metabolism rate of mice. Based on animal test results and metabolic system and endocrine system to relief intoxication the method of optimizing experimental condition, the best symptoms caused by alcohol drinking (16), reduce composition of the compound oral liquid was obtained. The spontaneous alcohol intake of alcohol dependence animal, compound oral liquid developed used honeysuckle fl owers

58 Agro FOOD Industry Hi Tech - vol 25(2) - March/April 2014 extract, Kudzu vine flowers extract and honey solution as main was put under water bath at 75°C and a centrifugal evaporator ingredients with no adding of preservative. This oral liquid was applied until there is no scent of alcohol, after which, the has significant effect in alleviation of drunkenness. Long term contents were sealed to standby. intake of the liquid has health benefits in terms of improving liver function and immunity. The collected extracts of flowers of Honeysuckle and flowers of Kudzu vine, together with honey solution and citric acid solution were made into oral liquid by mixing under the best ratio ORAL LIQUID PREPARATION METHOD deducted according to experimental results. Deployed oral liquid would then go through filling, sealing, sterilization and be packed Materials used into final product after cooling. Flowers of honeysuckle Flowers of Kudzu vine Black Locust Honey (produced by Shanghai Guan Sheng RESULTS AND DISCUSSIONS Garden bee products co. LTD) Citric acid (food grade) Animal test 1 56 ° spirit Er Guo Tou liquor (a strong liquor produced by Righting reflex test (29-30) was used to observe the drunk and Beijing red star co. LTD.) sober states of mice. After been washed with alcohol, the 240 Kunming mice, all male, aged 2 months, weight experimental mice were put on the experimental desk with 28-32 g (from Center for New Drug Evaluation, Shandong backside facing down. If a mouse kept such posture and did University, China.) not flips back for over 30 s, righting reflex was deemed to be lost and the mouse was deemed to be drunk. Otherwise the mouse Process flow chart would be deemed as not drunk. Drunk rate was taken as the percentage of mice been drunk in a group. Trailing condition was recognized when a mouse, after been washed with alcohol, was unable to walk stably with its tail dragging on the ground. The latent time for drunkenness was defined as the time taken for righting reflex to disappear and sober up time was the time taken for the tested mouse to return to normal state after righting reflex disappears. 40 healthy 2-month-old male Kunming mice were divided into four groups – control group, experimental group 1, experimental group 2 and experimental group 3 – with ten mice per group. Before the test, the experimental mice were fed for 3 days and starved for 12 hours in the laboratory. The mice of control group were fed with normal saline and mice in experimental groups were fed with compound herbal extracts of volume ratio of honeysuckle flower extract: Kudzu vine flower extract at 5:5, 3:7 and 1:9 for group 1, group 2 and group 3 respectively. Each mouse was washed with 0.25mL/10g weight of the specified liquid according to their groups. 30 minutes later each mouse was washed with 0.15mL/10g weight of 56 ° spirit Er Guo Tou liquor. Observations were taken once every minute and data on trailing time, latent time and sober up time were taken. SPSS11.0 software was used to analysis the data. Results were recorded in Table 1 and Table 2 below. Key technical points High quality flowers of Honeysuckle were selected, washed then dried in oven at 60°C for 3 h. The dry flowers were then ground and passed through a 40 mesh sieve. 12 times the volume of deionized water was added to the sieved sample and the mixture was leached for 1hr under constant temperature at 75°C. The mixture was then filtered three times and the filtrate was collected. The filtrate was water bathed at 80℃ and compressed to 1/2 its original volume using a centrifugal evaporator, after which, the contents were sealed to standby. High quality flowers of Kudzu vine were selected, washed then dried in oven at 60°C for 3 h. The dried flowers were then ground and passed through a 40 mesh sieve. 14 times the volume of 40% ethanol solution was added to the sieved sample and the mixture was leached for 1.5hr. The mixture was then filtered and both the filtrate and residue were collected. 12 times 40% ethanol solution was added to the residue and the resulted solution was left to leach for another 1hr. This solution was filtered Table 1. Data on mice drunk rate and filtrate was collected. The total amount of collected filtrate

Agro FOOD Industry Hi Tech - vol 25(2) - March/April 2014 59 Table 2. Data on mice trailing time, latent time and sober up time Table 3. Record of blood ethanol content (uL/mL) Note: Compare with control group, * indicates moderate Note: Within same time expand, compare result of compound significance ( P<0.05), ** indicates large significance (

From Table 3, for control group, significant increase in blood ethanol content was shown 30 minutes after mice were washed with liquor. The value reached a maximum point at around 120 minutes then started to drop. Compare with control group, honeysuckle flower extract group and Kudzu vine flower extract group showed significantly higher blood ethanol content around 30-60 minute. However, after 90 minutes blood ethanol content of honeysuckle flower extract group and Kudzu vine flower extract group were both lower than that of control group. This indicates that extracted liquid of honeysuckle flower and Kudzu vine flower both has significant effect on speeding up Table 4. Factors level in orthogonal test alcohol decomposing rate in mice.

60 Agro FOOD Industry Hi Tech - vol 25(2) - March/April 2014 of drunkenness alleviation. Since animal tests were conducted by first feed the mice with specified liquid then lavage them with alcohol, experiment results deduced not only indicates disintoxication effect of the tested liquid but also its effect on preventing drunkenness. Therefore, the compound oral liquid investigated has socially applicable value.

REFERENCES AND NOTES

1. Beijing Institute of Botany, Chinese Academy of Sciences, Higher plant field guide (Book IV), Science Publishing House, 296, 758 (1975). 2. Wang Benxiang, Modern Pharmacology [M], Tianjin Publishing House, 204-208 (1997). 3. Lin Qishou, Herbal chemical composition [M], Science and Technology Publishing House, 145-158 (1977). 4. Zhou Chunhui, Su Ye, Li Fengzhi, Modern Food Science and Technology, 26(4), 351-353 (2010). 5. Zhang Huimin, Shi Junying, Food and Drug, 8(12), 11-14 (2006). 6. Hu Chengmu, Jiang Hiu, Liu Hongfeng, et al, Anhui Medical Table 5. Results for L (33)orthogonal test 9 and Pharmaceutical Journal, 12(4), 295-297 (2008). 7. Hou Huina, Zeng Yaoying, Huang Xiuyan, et al, Immunological Journal, 24(2), 178 (2008). From Table 5, it can be seen that ranking of influence of 8. Zhuang Li, Zhang Chao, Ali Muse, Liaoning Traditional Chinese the factors A B C on sensory quality was A > C > B thus Medicine, 40(2), 378-380 (2013). 9. Chaovanalikit Arusa, Thompson Maxine M., Wrolstad Ronald best recipe was A2B3C3. Under verifying experiment, oral E., Journal of Agricultural and Food Chemistry, 52(4), 848-852 liquid with recipe A2B3C3 received a score of 92. Thus the best recipe of honeysuckle flower and Kudzu vine flower (2004). compound oral liquid in terms of volume ratio of honeysuckle 10. Wang Qiang, Chen Donghui, Deng Wenlong, Pharmacology and Clinics, 23(3), 40-45 (2007). flowers extract : Kudzu vine flowers extract : honey solution : 11. Wang Huifang, Dong Yushan, La Wanying, Chinese Journal of acetic acid solution is 3: 7: 0.2 : 0.01. Coal Industry Medicine, 13(5), 820-822 (2010). 12. Kwon C. S., Son K. H., Kim J. S. Food Science and Biotechnology, 13(6), 768-772 (2004). 13. Liu Dan, Peng Dingwei, Qi Gang, Acta Academiae Medicinae CPAPF, 14(1), 78-80 (2005). 14. Zhang Huimin, Shi Junying, Food and Drug, 8(12), 11-14 (2006) 15. State Administration of Traditional Chinese Materia Medica Editorial Board, Chinese herbal medicine (4), Shanghai Science and Technology Publishing House, 619 (1999). 16. Zhen Pan, Zhang Yanjie, Zhang Ying, Journal of Hebei North University, 24(5), 6~18 (2008). 17. Yeo O. H., Myung J. H., et al, J Pharmacol Sci, 93, 331-336 (2003). 18. Hae U. L., Eun A. B., et al, J Pharmacol Sci, 97, 541-544 (2005). 19. Kim C., Shin S., Ha H., et al, Archives of Pharmacal Research, 26(3), 210-213 (2003). 20. Choi, Shin M. H., Park K. Y., et al., Journal of Medicinal Food, 7(1), 31-37 (2004). 21. Ouyang Changhan, Zhang Ruixue, Ye Tao, Food Research And Development, 33(11), 57 (2012). 22. Xue Wentong, Song Ruixia, Chen Xiangning, Food Science and Technology, (5). 22-23 (2004). CONCLUSIONS 23. Wang Weiwu, Journal of Bee, (11), 31 (2001). 24. Mckay Becky, Journal of Agricultural & Food Information, The best recipe for honeysuckle flowers and Kudzu 12(2), 130-140 (2011). vine flowers compound oral liquid for alleviation of 25. Onyesom I., European Journal of Scientific Research, 13(3), drunkenness has volume ratio of honeysuckle flower 325-331 (2006). extract : Kudzu vine flower extract : honey solution : 26. Wei Guoxue, Huang Jikun, Yang Jun, Journal of Integrative acetic acid solution at 3 : 7 : 0.2 : 0.01. This compound Agriculture, 11(4), 684-693 (2012). oral liquid can effectively elongate drunk latent time, 27. Xiong Penghui, Chinese Journal of Ethnomedicine and shorten sober up time and improve alcohol tolerance, Ethnopharmacy, (19), 49 (2011). 28. Wang Xinliang, Wu Jiliang, Yang Jing, Chinese Traditional thus can effectively reduce mice drunk rate and has Patent Medicine, 31(7), 1120-1122 (2009). significant alleviation effect on drunkenness. Animal 29. Jia Xin, Household Technology, (2), 20 (2002). tests also prove that there is synergy between extracts 30. Yang Muxiang, Yu Wentao, Hu Jinkua, Health Digest Medical of honeysuckle flowers and Kudzu vine flowers in terms Monthly, 4(3), 37-39 (2007).

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