Separation of high-class alkanols and high-class eicosanoic acids in sugar cane wax and activity of reducing blood cholesterol

Fangxin LIU; Yunuo ZHANG; Xiaoyu LI; Juan XIE; Min WANG; Xianjun LIU

ABSTRACT

BACKGROUND:

As a by-product in sugar industry, sugar cane wax has been widely used in non-medical field. Some researches indicate that sugar cane wax plays a great role in reducing blood cholesterol; however, the therapeutical effect and clinical application should be studied further.

OBJECTIVE:

To separate the high-class eicosanoic acid and the high-class alkanols, which are suitable for medical application, and further to observe the effect of them on reducing blood cholesterol of model rets with hyperlipemia.

DESIGN:

Randomized control animal study.

SETTING:

Pharmacological Institute, Chongqing Kangerwei Pharmaceutical Co., Ltd.MATERIALS. The experiment was carried out in the Pharmacological Institute of Chongqing K.E.W Pharmaceutical Go.,Ltd. From April 2005 to January 2006. A total of 65 adult female Wistar rats, aged at 3-6 months, weighing 180-220 g, of SPF grade, were provided by Experimental Animal Center of Chongqing Institute of Traditional Chinese Medicine. Raw sugar cane wax was provided by Beijing Jiade Hongsheng Biochemical Technology Co., Ltd. High-class alkanols C26,C28,C30, C32 and high-class eicosanoic acid C28, C30, C32, C34 were provided by Sigma Company (standard materials of gas phase chromatography), and other reagents were national analytical pure.

METHODS:

① Sugar cane wax was extracted from raw sugar cane wax with ethanol and other organic solution and separated from the mixture of high-class eicosanoic acid and the mixture of high-class alkanols with saponification and calcification. Main components were analyzed with gas phase chromatography. The main components of high-class alkanols were C26, C28, C30 and C32 and the main components of high-class eicosanoic acid were C28, C30, C32 and C34, ② Based on references, rats were fed in 3 days and randomly divided into blank group (n =10) and experimental group (n =55).And then, all rats were cut off their tails to collect blood and the triacylglyoerol (TG), total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C) were measured with automatic biochemistry analyzer. Rats were fed with common granule feeds in blank group or with high-lipid feeds (containing 0.1 mass fraction of oiliness, 0.1 mass fraction of yolk powder, 0.01 mass fraction of cholesterol, 0.002 mass fraction of pig's gall salt, 0.788 mass fraction of common feeds) in experimental group. All rats ate and drank freely. Seven days later, blood was collected again from tail tip to measure the contents of TG, TC and HDL-C. Based on level of serum TG, rats in the experimental group were randomly divided into 5subgroups (n =11) negative control group, low-dosage high-class alkanols group, high-dosage high-class alkanols group,Iowdosage high-class eicosanoic acid and high-dosage high-class eicosanoic acid group. Rats in low-dosage and high-dosage high-class alkanols groups were perfused with 5 and 50 mg/(kg·d) high-class alkanols; meanwhile, rats in low-dosage and high-dosage high-class eicosanoic acid groups were perfused with 20 and 200 mg/ (kg·d) high-class eicosanoic acid. Rats in negative control group and blank group were perfused with the same volume of 0.3% carboxymethylcellulose sodium and distilled water, respectively, once a day for successive 30 days. At 16 hours after the last administration, rats were anesthetized to collect blood from heart to measure contents of TG, TC and HDL-C in serum.MAIN OUTCOME

MEASURES:

① Percentage of main component in separated mixtures of high-class eicosanoic acid high-class alkanols; ② levels of serum cholesterol, HDL and TG.

RESULTS:

A total of 65 experimental rats were involved in the final analysis. ① Gas phase chromatography suggested that the content of C28 high-class alkanols was the most (73.6%), and other three kinds of high-class alkanols were counted for 5.3% (C26), 6.2% (C30) and 5.1% (C32), respectively. The total quantity was 90.2%. In the mixture of high-class eicosanoic acid, content of C28 high-class eicosanoic acid was the most (46.6%) and the other three kinds of high-class eicosanoic acid were counted for 16.7% (C30), 6.8% (C32) and 9.3% (C34), respectively. The total quantity was 79.3%. ②Levels of serum TC were (1.46±0.27), (1.66±0.33), (1.44±0.25) and (2.16±0.52) mmol/L in high-dosage and Iow-dosage high-class alkanols groups and high-dosage and Iow-dosage high-class eicosanoic acid groups, respectively, which were lower than those in negative control group [(2.52±0.83) mmol/L, P＜0.01]. Levels of HDL-C were (0.73±0.09), (0.71±0.07), (0.79±0.10) and (0.70±0.08) mmol/L in the four treatment groups, respectively, which were higher than those in negative control group [(0.58±0.13) mmol/L, P＜0.05-0.01].

CONCLUSION:

The high-class alkanols and the high-class eicosanoic acids separated from sugar cane wax made in China significantly have the activity of reducing blood cholesterol; however, the effect on decreasing TG is not obvious.