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 ( <P 0.01). herbal extracts group with control group, ** indicates significant Died mouse were not taken into account when calculating difference(P<0.01); compare with honeysuckle flowers extract the drunk rate. group, ##indicates significant difference(P<0.01); compare with Kudzu vine flower extract group,DD indicates significant difference(P<0.01). Died mouse were not taken into account when calculating the drunk rate. From Table 1 and Table 2, it can be observed that experimental group 2 showed a significant drop in drunk rate compared with control group, reaching the level of large For compound herbal extract group, blood ethanol content significance.