| The health hazards caused by smoking cigarettes have been established, and the development of less harmful cigarette has been focused on by the researcher of tobacco industrial. Due to the foundation of application of Chinese herb as medicine for thousands of years and the rich resources of herb in China, the application of Chinese herbal medicine and its active substances in making less harmful Chinese cigarette has been researched as the special way to develop Chinese cigarette industry. The objective of this paper was to choose herb with the effect on reducing hydrogen cyanide in the cigarette smoke (CS), and coptis alkaloids were standout. The coptis alkaloids were added in the cigarette filter to make coptis alkaloids filter, and the effects of coptis alkaloids filter on the harmful components in the CS were studied. The effect and mechanism of coptis alkaloids filter on reducing the inhibition of CS on oral peroxidase (OPO) activity were researched. The coptis alkaloids filter cigarettes were compared with cellulose acetate filter cigarette, and the effects of CS from different filter cigarette on oxidative stress were also studied. The methods and results were shown as follows:1. The selection and extraction of plant alkaloidsThe extraction of plant alkaloids were used ethanol, and the extractions were determined by Dragendorff reagent, Mayer reagent and Wagner reagent. The alkaloids of Coptis chinensis Franch, Amur Corktree Bark, Fritillariae Cirrhosae Bulbus, Lotus Leaf, Radices stephaniae tetrandrae, Tender Catchweed Bedstraw Herb, Rhizoma Corydalis, Radix Stemonae, Rauwolfia and Cephalotaxus sinensis were chosen to added in the filter with30mg/cig. The results showed that smoke from cigarettes with filter containing coptis alkaloids had less hydrogen cyanide by18.3%, which was compared with the control CS. Coptis alkaloids were chosen as active component to reduce hydrogen cyanide among the10kinds of herb. The Coptis alkaloids were extracted and isolated by resin column and determined by HPLC. The total alkaloids and monomer of Coptis alkaloids were obtained with purity of over95%.2. The effect of coptis alkaloids multi-filter on the harmful compounds in the CSCompared to different carrier of Coptis alkaloids, activated carbon fiber (ACF) was superior as Coptis alkaloids'carrier to make modified filter. The Coptis alkaloids multi-filter was composed by cellulose acetate part and15mg ACF part containing1.5mg Coptis alkaloids. The cigarettes were smoked by machine under the ISO standard. A series of smoking runs was undertaken. The smoke was collected and groups of constituents were measured in the smoke. The content of hydrogen cyanide (HCN) and benzo[a]pyrene (B[a]P) in the Coptis alkaloids multi-filter group were over20%less than the control group respectively. And the free radicals in the Coptis alkaloids multi-filter group were over50%less than the control one. Little effect on smoking quality was perceived.3. The mechanism of the reduction of harmful components in the CS by addition of coptis alkaloids in the cigarette filterThe multi filter containing Coptis alkaloids was used to scavenge cyanide ion in the solution of potassium cyanide. The reductions of cyanide ion were observed by58.7%with multi filter containing coptis alkaloids,5.3%with cellulose acetate filter and17.4%with ACF. The interactions of Coptis alkaloids with B[a]P were investigated by fluorescence quenching. The correlation coefficients of Coptis alkaloids and B[a]P were0.991-0.987, and the binding constants were(2.0-9.0) x103. The reduction trend for B[α]P by coptis alkaloids was Palmatine> Berberine> Coptisine. Spectrophotometry was used to examine the scavenging effect of Coptis alkaloids on DPPH free radicals, nitric oxide free radical and superoxide anion radical. In all the models tested, Coptis alkaloids showed their ability to scavenge the free radicals, and the ability to scavenge were Palmatine> Total Coptis alkaloids> Berberine> Coptisine.4. Reducing the inhibitory effect of cigarette smoke on the activity of oral peroxidase by addition of Coptis alkaloids in cigarette filterThe effect of Coptis alkaloids which were added in the cigarette filter on the activity of oral peroxidase (OPO) was studied by exposure of saliva to the CS. The mechanism of inhibition of OPO by CS was investigated by adding aldehydes, nicotine, cyanide, which were present in CS, to the saliva. The antioxidants, cyanate chelator and dialysis were also applied to saliva in vitro in the presence and absence of CS. The results showed cyanide was likely to be the species in CS responsible for the loss of salivary OPO activity. The reduction in HCN might be related with Coptis alkaloids in the cigarette filter which reduce the inhibition on OPO activity by CS over25%.5. The inhibitory effect of Coptis alkaloids in the cigarette filter on oxidative stress of cell caused by the CSBEAS-2B cells, a model of normal human bronchial epithelial cells, were investigated on their oxidative stress (OS) caused by CS. MTT assay results showed that LD50values of cellulose acetate filter group, ACF multi-filter group and Coptis alkaloids multi-filter group were5.21×10-36.32X10-3,7.26×10-3cig/mL, respectively. Compared to the control CS group, the level of intracellular reactive oxygen species (ROS) and lipid peroxidation (MDA) caused by CS from Coptis alkaloids multi-filter group were much lower. The total antioxidant capacity, catalase activity (CAT) and glutathione reductase (GR) of Coptis alkaloids multi-filter group were induced, but the total activity of SOD was reduced. Compared with the control CS group, CS from cigarette with filter containing Coptis alkaloids could reduce the OS damage by CS.
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