DESCRIPTION
BIO-CATALYTIC FILTER
This invention refers to the development of a filter from conventional materials, on which specific biological catalysts (oxidoreductases, methemoglo- bin) have been immobilized. This filter removes harmful substances contained in tobacco smoke, indoors and outdoors polluted air and engine exhaust.
Tobacco smoking addiction is more prevailing worldwide than alcohol and drugs put together1 causing serious health problems and killing more peo- pie. Despite this, the number of people that smoke cigarettes increases every day.
There are numerous substances in the cigarette smoke. Among the harmful ones are2: ammonia, benzopyrene, dimethylnitrosamine, catechol, meth- ylquinolines, methylnaphthalene, nitrosopyrrolidine, quinoline, pyrene, hy- drogen cyanide, pyridine, nicotine, hydrazine-6.2-naphthylamine, hydrogen cyanide, phenols. Also, as harmful substances have been reported acetaldehyde3, acetone4, acrolein5, benzene6, 1,3 butadiene7, 2-butanone8, free radicals9, isoprene10, methanethiol11, nitric oxide12, carbon monoxide 2, toluene13, hydrogen sulphide14 and peroxides9. In the outdoor (atmospheric) and indoor (interior) polluted air there are many harmful gases. In the atmospheric air, the most important are: benzene6, 1,3 butadiene7, nitric oxide1 , carbon dioxide16, lead17, carbon monoxide2, polycyclic aromatic hydrocarbon hydrocarbons18 and ozone19. Among indoor air pollutants, besides nitrogen dioxide and carbon monox- ide, are asbestos , pesticides , radon gas , cigarette smoke, formalde- hyde and several volatile organic toxic compounds.
Finally, the main exhaust gases from combustion of fossil fuels in engines
• * *7 are carbon monoxide , simple (1,3 butadiene ) and cyclic hydrocarbons (benzene2). The Diagram A depicts a cigarette with the conventional and the BC-F I filter. The BC-F I consists of 200 mg of conventional materials (activated carbon particles) on which the enzymes and hemoglobin have been immobilized (adsorbed).
Enzymes Superoxide dismutase (SOD). It was prepared from cells of the yeast Sac- charomyces cerevisiae. The intracellular enzyme was partially purified with organic solvents, lyophilized and stored in the freezer.
Peroxidase (Horse Radish Peroxidase, HRP). The enzyme was a product of the Sigma Chemical Co., USA.
Hemoglobin
It was a commercial product (methemoglobin), provided by the HARTMEX BV Co., Holland. It contained (μmole/gr): Methemoglobin (MetHb) > 36 and Oxyhemoglobin (OxyHb) < 4.5. Cigarettes
The cigarettes KENTUCKY 1R3F (research cigarettes, tobacco health research, University of Kentucky, Code 1R3F, tar 15.0 mg/cigarette, nicotine 1.16 mg/cigarette) and KENTUCKY 1R4F (research cigarettes, tobacco health research, University of Kentucky, Code 1R4F, tar 9.2 mg/cigarette, nicotine 0.8 mg/cigarette), internationally recognized as reference cigarettes, were used. For comparison reasons, the cigarettes with a biological filter (BIOLOGICAL FILTER) BF Full Flavor (SEKAP Co., Athens, Xanthi, Greece, tar 14.0 mg/cigarette, nicotine 1.0 mg cigarette) and BF Lights (SEKAP Co., Athens, Xanthi, Greece, tar 8.0 mg/cigarette, nicotine 0.7 mg/cigarette)..
Enrichment of filter with enzymes and hemoglobin
The enzymes and hemoglobin were immobilized by adsoφtion on conventional materials (activated carbon particles) in the form of a water solution. The latter contained 100 Units of SOD, 100 Units of HRP and 2 mg hemo- globin. Following adsoφtion, the materials were dried under vacuum at 35°C and assembled on the cigarette filter as shown in Diagram A.
Analytical methods
Cigarette smoke contains gaseous and solid compounds. The latter were separated with the aid of the special filter Cambridge, located between the filter and the syringe used for sampling (Diagram B)
Gases. Samples of 35 ml were taken at the 2nd, 4th and 6th puff. The puffs were separated by one minute intervals and each puff lasted 2 sec. This method was preferred over the collection of puffs in a special container (balloon) due to significant losses of measured compounds encountered with the latter device.
Gas Chromatography - Ultraviolet Spectroscopy (GC-UV). A new apparatus24 was employed for the determination of the gaseous substances acetaldehyde, acetone, benzene, 1,3 butadiene, 2-butanone, hydrogen sul- fide, isoprene, methanethiol, nitric oxide and toluene. For quantitative comparison reasons, the analysis was carried out under the same conditions, that is:
Chromatography column : PRP3, 8 cm length, 1.6 mm diameter Carrying gas : Nitrogen, lO ml/min Temperature range : 55-130°C (10 °C/min) 130-160°C (4 °C/min)
Sample volume: 450 μl (room temperature) Detector : UV spectrophotometer (166-330 nm)
Solid substances. They are contained in the tar collected on an appropriate filter (Cambridge) during the first 6 puffs. The benzo[α]pyrene was deter- mined as reported m the literature
The results appear in the Figures la(la/l , la/2, la/3, l a/4), lb(lb/l , l b/2, lb/3, lb/4), lc(lc/l , lc/2, lc/3), 2a(2a/l , 2a/2, 2a/3, 2a/4), 2b(2b/l , 2b/2, 2b/3, 2b/4), 2c(2c/l , 2c/2, 2c/3), 3 and 4. The comparison of constituents in the smoke was made between cigarettes with the conventional and BC-F I filter. Three replicates, randomly designed, were used in measurement. In all measurements, the standard deviation was practically <10%.
In the Figures l a-lc and 2a-2c a comparison was made between KENTUCKY 1R3F and BF Full Flavor and between KENTUCKY 1R4F and BF Lights, respectively. The results concern the retention of the harmful compounds acetaldehyde, acetone, acrolein, benzene, 1 ,3 butadiene, 2- butanone, hydrogen sulfide, isoprene, methanethiol, nitric oxide and toluene by the BC-F I filter during the 2nd, 4th and 6th puffs is concerned. The Figure 3 shows the retention of benzo[α]pyrene by the BC-F I filter during the first six puffs. Finally, in the Figure 4, the average retention of harmful constitu- ents in the smoke of tested cigarettes by BC-F I is depicted.
LITERATURE
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