CN102980913B - Biological thermochemistry method for safety evaluation and screening of cigarette auxiliary material - Google Patents
Biological thermochemistry method for safety evaluation and screening of cigarette auxiliary material Download PDFInfo
- Publication number
- CN102980913B CN102980913B CN201210452337.7A CN201210452337A CN102980913B CN 102980913 B CN102980913 B CN 102980913B CN 201210452337 A CN201210452337 A CN 201210452337A CN 102980913 B CN102980913 B CN 102980913B
- Authority
- CN
- China
- Prior art keywords
- cigarette
- test
- culture media
- cultivating
- nutrient
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 235000019504 cigarettes Nutrition 0.000 title claims abstract description 90
- 239000000463 material Substances 0.000 title claims abstract description 48
- 238000000034 method Methods 0.000 title claims abstract description 39
- 238000011156 evaluation Methods 0.000 title claims abstract description 12
- 238000012216 screening Methods 0.000 title claims abstract description 12
- 230000000694 effects Effects 0.000 claims abstract description 35
- 230000004060 metabolic process Effects 0.000 claims abstract description 23
- 230000006378 damage Effects 0.000 claims abstract description 12
- 238000004458 analytical method Methods 0.000 claims abstract description 11
- 230000008569 process Effects 0.000 claims abstract description 10
- 230000001603 reducing effect Effects 0.000 claims abstract description 8
- 235000015097 nutrients Nutrition 0.000 claims description 54
- 239000001963 growth medium Substances 0.000 claims description 49
- 239000000243 solution Substances 0.000 claims description 33
- 238000012360 testing method Methods 0.000 claims description 32
- UGFAIRIUMAVXCW-UHFFFAOYSA-N Carbon monoxide Chemical compound [O+]#[C-] UGFAIRIUMAVXCW-UHFFFAOYSA-N 0.000 claims description 17
- 239000003546 flue gas Substances 0.000 claims description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- 239000007788 liquid Substances 0.000 claims description 13
- 239000012530 fluid Substances 0.000 claims description 12
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 10
- 239000002609 medium Substances 0.000 claims description 10
- 241000222178 Candida tropicalis Species 0.000 claims description 9
- 239000000779 smoke Substances 0.000 claims description 9
- 239000001888 Peptone Substances 0.000 claims description 8
- 108010080698 Peptones Proteins 0.000 claims description 8
- 241000223892 Tetrahymena Species 0.000 claims description 8
- 210000002919 epithelial cell Anatomy 0.000 claims description 8
- 210000002950 fibroblast Anatomy 0.000 claims description 8
- 238000001914 filtration Methods 0.000 claims description 8
- 239000008103 glucose Substances 0.000 claims description 8
- 238000004519 manufacturing process Methods 0.000 claims description 8
- 235000019319 peptone Nutrition 0.000 claims description 8
- 101100060558 Schizosaccharomyces pombe (strain 972 / ATCC 24843) coa3 gene Proteins 0.000 claims description 7
- 210000003470 mitochondria Anatomy 0.000 claims description 7
- 150000003839 salts Chemical class 0.000 claims description 7
- 241000588724 Escherichia coli Species 0.000 claims description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 6
- 229940024606 amino acid Drugs 0.000 claims description 6
- 235000001014 amino acid Nutrition 0.000 claims description 6
- 150000001413 amino acids Chemical class 0.000 claims description 6
- 230000000391 smoking effect Effects 0.000 claims description 6
- 230000002503 metabolic effect Effects 0.000 claims description 5
- CWGFSQJQIHRAAE-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol tetrahydrochloride Chemical compound Cl.Cl.Cl.Cl.OCC(N)(CO)CO CWGFSQJQIHRAAE-UHFFFAOYSA-N 0.000 claims description 4
- 244000063299 Bacillus subtilis Species 0.000 claims description 4
- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 4
- 241000894006 Bacteria Species 0.000 claims description 4
- 239000003109 Disodium ethylene diamine tetraacetate Substances 0.000 claims description 4
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 4
- 240000006024 Lactobacillus plantarum Species 0.000 claims description 4
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims description 4
- 241000235070 Saccharomyces Species 0.000 claims description 4
- 241000191967 Staphylococcus aureus Species 0.000 claims description 4
- 239000007640 basal medium Substances 0.000 claims description 4
- 229940041514 candida albicans extract Drugs 0.000 claims description 4
- 239000012153 distilled water Substances 0.000 claims description 4
- 239000003797 essential amino acid Substances 0.000 claims description 4
- 239000000284 extract Substances 0.000 claims description 4
- 230000002401 inhibitory effect Effects 0.000 claims description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 4
- 229940072205 lactobacillus plantarum Drugs 0.000 claims description 4
- 238000003860 storage Methods 0.000 claims description 4
- 239000005720 sucrose Substances 0.000 claims description 4
- 239000000725 suspension Substances 0.000 claims description 4
- 238000002137 ultrasound extraction Methods 0.000 claims description 4
- 239000012138 yeast extract Substances 0.000 claims description 4
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 claims description 2
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 claims description 2
- 229930024421 Adenine Natural products 0.000 claims description 2
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 claims description 2
- 239000004475 Arginine Substances 0.000 claims description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims description 2
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 claims description 2
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 claims description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 2
- 239000007995 HEPES buffer Substances 0.000 claims description 2
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 claims description 2
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims description 2
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 claims description 2
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 claims description 2
- LEVWYRKDKASIDU-IMJSIDKUSA-N L-cystine Chemical compound [O-]C(=O)[C@@H]([NH3+])CSSC[C@H]([NH3+])C([O-])=O LEVWYRKDKASIDU-IMJSIDKUSA-N 0.000 claims description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 claims description 2
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 claims description 2
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 claims description 2
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 claims description 2
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 claims description 2
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 claims description 2
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 claims description 2
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 claims description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims description 2
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 claims description 2
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 claims description 2
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims description 2
- 239000004472 Lysine Substances 0.000 claims description 2
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 claims description 2
- 108010009736 Protein Hydrolysates Proteins 0.000 claims description 2
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 claims description 2
- 229930006000 Sucrose Natural products 0.000 claims description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 2
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 claims description 2
- 239000004473 Threonine Substances 0.000 claims description 2
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 claims description 2
- 239000006035 Tryptophane Substances 0.000 claims description 2
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 claims description 2
- 239000002253 acid Substances 0.000 claims description 2
- 229960000643 adenine Drugs 0.000 claims description 2
- 235000004279 alanine Nutrition 0.000 claims description 2
- GZCGUPFRVQAUEE-SLPGGIOYSA-N aldehydo-D-glucose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O GZCGUPFRVQAUEE-SLPGGIOYSA-N 0.000 claims description 2
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 2
- 235000003704 aspartic acid Nutrition 0.000 claims description 2
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 claims description 2
- 239000011575 calcium Substances 0.000 claims description 2
- 229910052791 calcium Inorganic materials 0.000 claims description 2
- 239000006143 cell culture medium Substances 0.000 claims description 2
- 239000007859 condensation product Substances 0.000 claims description 2
- 229960003067 cystine Drugs 0.000 claims description 2
- 238000013461 design Methods 0.000 claims description 2
- 235000020776 essential amino acid Nutrition 0.000 claims description 2
- 230000003328 fibroblastic effect Effects 0.000 claims description 2
- 239000011724 folic acid Substances 0.000 claims description 2
- 235000013922 glutamic acid Nutrition 0.000 claims description 2
- 239000004220 glutamic acid Substances 0.000 claims description 2
- 235000013905 glycine and its sodium salt Nutrition 0.000 claims description 2
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims description 2
- 229960000310 isoleucine Drugs 0.000 claims description 2
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 claims description 2
- 239000004310 lactic acid Substances 0.000 claims description 2
- 235000014655 lactic acid Nutrition 0.000 claims description 2
- 229930182817 methionine Natural products 0.000 claims description 2
- 239000011570 nicotinamide Substances 0.000 claims description 2
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 claims description 2
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 2
- 229920000053 polysorbate 80 Polymers 0.000 claims description 2
- 235000013930 proline Nutrition 0.000 claims description 2
- ZUFQODAHGAHPFQ-UHFFFAOYSA-N pyridoxine hydrochloride Chemical compound Cl.CC1=NC=C(CO)C(CO)=C1O ZUFQODAHGAHPFQ-UHFFFAOYSA-N 0.000 claims description 2
- 239000011764 pyridoxine hydrochloride Substances 0.000 claims description 2
- 235000004400 serine Nutrition 0.000 claims description 2
- ZDQYSKICYIVCPN-UHFFFAOYSA-L sodium succinate (anhydrous) Chemical compound [Na+].[Na+].[O-]C(=O)CCC([O-])=O ZDQYSKICYIVCPN-UHFFFAOYSA-L 0.000 claims description 2
- KDYFGRWQOYBRFD-UHFFFAOYSA-N succinic acid Chemical compound OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 claims description 2
- 239000001384 succinic acid Substances 0.000 claims description 2
- 229940095064 tartrate Drugs 0.000 claims description 2
- 239000011747 thiamine hydrochloride Substances 0.000 claims description 2
- 235000015193 tomato juice Nutrition 0.000 claims description 2
- 229960004799 tryptophan Drugs 0.000 claims description 2
- 235000002374 tyrosine Nutrition 0.000 claims description 2
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 claims description 2
- 239000004474 valine Substances 0.000 claims description 2
- 238000002474 experimental method Methods 0.000 abstract description 13
- 230000008859 change Effects 0.000 abstract description 2
- 238000007792 addition Methods 0.000 abstract 1
- 239000000126 substance Substances 0.000 description 17
- 241000208125 Nicotiana Species 0.000 description 9
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 9
- 210000004027 cell Anatomy 0.000 description 9
- 238000005516 engineering process Methods 0.000 description 9
- 238000007707 calorimetry Methods 0.000 description 7
- 244000005700 microbiome Species 0.000 description 7
- 238000012545 processing Methods 0.000 description 7
- 238000011160 research Methods 0.000 description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 6
- 230000007269 microbial metabolism Effects 0.000 description 6
- 238000012544 monitoring process Methods 0.000 description 6
- 230000009467 reduction Effects 0.000 description 6
- 241000193388 Bacillus thuringiensis Species 0.000 description 5
- 229940097012 bacillus thuringiensis Drugs 0.000 description 5
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 description 4
- 239000000470 constituent Substances 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 239000002028 Biomass Substances 0.000 description 3
- -1 carbon nano tube compound Chemical class 0.000 description 3
- 239000002041 carbon nanotube Substances 0.000 description 3
- 229910021393 carbon nanotube Inorganic materials 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000002574 poison Substances 0.000 description 3
- 231100000614 poison Toxicity 0.000 description 3
- 231100000820 toxicity test Toxicity 0.000 description 3
- XMWRBQBLMFGWIX-UHFFFAOYSA-N C60 fullerene Chemical compound C12=C3C(C4=C56)=C7C8=C5C5=C9C%10=C6C6=C4C1=C1C4=C6C6=C%10C%10=C9C9=C%11C5=C8C5=C8C7=C3C3=C7C2=C1C1=C2C4=C6C4=C%10C6=C9C9=C%11C5=C5C8=C3C3=C7C1=C1C2=C4C6=C2C9=C5C3=C12 XMWRBQBLMFGWIX-UHFFFAOYSA-N 0.000 description 2
- 230000000274 adsorptive effect Effects 0.000 description 2
- 239000003708 ampul Substances 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 238000002485 combustion reaction Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 229910003472 fullerene Inorganic materials 0.000 description 2
- 238000007654 immersion Methods 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 210000002311 liver mitochondria Anatomy 0.000 description 2
- 230000001473 noxious effect Effects 0.000 description 2
- 210000003463 organelle Anatomy 0.000 description 2
- 230000002572 peristaltic effect Effects 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 231100000027 toxicology Toxicity 0.000 description 2
- JIAARYAFYJHUJI-UHFFFAOYSA-L zinc dichloride Chemical compound [Cl-].[Cl-].[Zn+2] JIAARYAFYJHUJI-UHFFFAOYSA-L 0.000 description 2
- 241000203069 Archaea Species 0.000 description 1
- 231100000750 In vitro toxicology Toxicity 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- 239000004743 Polypropylene Substances 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 230000002745 absorbent Effects 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- WNROFYMDJYEPJX-UHFFFAOYSA-K aluminium hydroxide Chemical compound [OH-].[OH-].[OH-].[Al+3] WNROFYMDJYEPJX-UHFFFAOYSA-K 0.000 description 1
- 229910021502 aluminium hydroxide Inorganic materials 0.000 description 1
- 229960003121 arginine Drugs 0.000 description 1
- 239000003610 charcoal Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical class CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000003063 flame retardant Substances 0.000 description 1
- 229960002885 histidine Drugs 0.000 description 1
- 239000013067 intermediate product Substances 0.000 description 1
- 230000009878 intermolecular interaction Effects 0.000 description 1
- 229960003136 leucine Drugs 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 229960003646 lysine Drugs 0.000 description 1
- VTHJTEIRLNZDEV-UHFFFAOYSA-L magnesium dihydroxide Chemical compound [OH-].[OH-].[Mg+2] VTHJTEIRLNZDEV-UHFFFAOYSA-L 0.000 description 1
- 239000000347 magnesium hydroxide Substances 0.000 description 1
- 229910001862 magnesium hydroxide Inorganic materials 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 229960004452 methionine Drugs 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000002438 mitochondrial effect Effects 0.000 description 1
- 230000006677 mitochondrial metabolism Effects 0.000 description 1
- 239000002048 multi walled nanotube Substances 0.000 description 1
- 229940066779 peptones Drugs 0.000 description 1
- 229960005190 phenylalanine Drugs 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 238000011020 pilot scale process Methods 0.000 description 1
- 229920001155 polypropylene Polymers 0.000 description 1
- 239000002109 single walled nanotube Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 229960002898 threonine Drugs 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 229960004295 valine Drugs 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 239000011592 zinc chloride Substances 0.000 description 1
- 235000005074 zinc chloride Nutrition 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a biological thermochemistry method for safety evaluation and screening of cigarette auxiliary material. According to the method, an external macro micro-thermal experiment is adopted, the whole metabolic process of a microexamination object after disposed is monitored in real time through an isothermal thermal activity detector, and heat change in the gathering module collecting process is utilized to calculate relative physicochemical parameters. The method is easy in operation, extremely low in cost, short in experiment period, high in efficiency and reliable in result accuracy. The whole metabolic process can be observed in real time, a new method is created for evaluating cigarette harm reducing effects and screening additions and the method provides an important reference basis for further analysis evaluations in the following steps. The method not only promotes efficiency but also greatly reduces the cost.
Description
Technical field
The present invention relates to Harm reduction techniques effect and security fields, refer to particularly a kind of biothermochemistry method for cigarette auxiliary material safety evaluatio and screening thereof.
Background technology
Nearly decades, the concerned degree of food-safety problem was the state that reaches unprecedented, although the most situations of tobacco are not directly eaten, but a lot of countries still divide foodstuff or accurate foodstuff into tobacco, more and more concerned as " Special food " its security, trace routine and method are also more and more stricter.The international tobacco scientific research of CORESTA(Cooperation Centre) set up relevant smoke of tobacco in vitro toxicity test job group, its main task is in vitro toxicity test philosophy the correct according to international endorsement, to adapt to character and the unique property of smoke of tobacco, method of testing settles the standard simultaneously.In vitro toxicity test is the direct mensuration of harm not, but can guarantee that test products can not increase potential harm completely, and toxicology data can be to a certain degree for cigarette product safety evaluatio provides theoretical foundation and information.Yet, security and Evaluation of Functional for novel cigarette with auxiliary material, even conventional detection often also needs to expend very large manpower and materials, and testing cost is high, the time cycle is long, efficiency is low etc., problem has exposed out.Along with cigarette with auxiliary material increase fast, require stricter, industry be badly in need of to want some new high speeds accurately method assist, cigarette is carried out to preliminary screening with auxiliary material, reduce the scope, reduce costs, raise the efficiency.
All chemistry, physics and life process are all accompanied by thermal effect.These thermal effect are measured accurately, and done reasonably to analyze, this just becomes thermochemical marrow.Along with the development of Science and Technology, the updating of instrument and equipment, the particularly development of modern micro-thermal technology, thermochemistry is penetrated into other field more and more, thereby has formed many emerging cross disciplines.Wherein most characteristic biothermochemistry is familiar with by people gradually and is paid attention to.
The thermal effect of the chemical reaction in biosome is generally all very little, and carries out under Isothermal Condition.Modern isothermal trace thermal technology is exactly the method growing up on this basis.From the basic research content of life science, can relate to biological grow, Physiology and biochemistry, matter and energy metabolic process.
Calorimetry (Calorimetry) is the important research method of biothermochemistry, and it has many distinctive features for the thermal effect research of living things system change procedure.The principle of work of constant temperature calorimeter can be sketched as putting into the sample of measuring cell, because of the certain thermal effect of producing of a certain process, thereby the temperature of himself is changed.Measuring cell has a heat abstractor that maintains temperature constant state around.As there is certain temperature difference between sample and heat radiator, heat just flows to heat radiator from sample, and the speed of warm extent and heat flow is proportional.Highly sensitive thermoelectric element is distributed in reaction vessel around, can measure the temperature difference between sample and environment and be converted into certain voltage, output after amplifying.If the reaction terminating of sample, it just remains on identical temperature with environment so, no longer includes thermal effect output, and the voltage that thermoelectric element produces is zero.In calorimetry process, do not need to add any reagent, so just can not introduce living things system normal activity and the influential factor of metabolism, but also can on purpose add a certain material, to study the impact of this material on biosome activity.In addition, calorimetry can be used for the system of any complexity, as chaos, high dispersive system, to the solvent property of studied system, spectral quality and electrochemical properties without any restriction, as long as there is thermal effect to apply, therefore, can be used for directly in vitro tissue and the suspending liquid of research.Amount of samples is few, and after calorimetric experiment, to research system, without any destruction, can also carry out generate subsequent fractional analysis.Although calorimetry lacks specificity, itself has specificity research object, so can obtain some significant results by this non-specific method.Calorimetry can be for the various special process in life system, as between the growth metabolism process in each level in living organism, enzymatic reaction, biomacromolecule and biomacromolecule and little intermolecular interaction etc.Certainly, calorimetry also has its weak point, can not follow the trail of the growth and decline process of a certain specific indivedual intermediate products and the individual instances in meta system, the summation that can only show various process thermal effect, but researcher can be in conjunction with other means, as various electron microscope techniques, spectrum and wave spectrum etc., resolve, learn from other's strong points to offset one's weaknesses.Biomass thermal technology and theory are gradually improved, and have now basically formed the cross disciplines such as biothermochemistry and biological chemistry thermodynamics.And biomass thermal technology has been penetrated into the every field of life science, at aspects such as biological chemistry, clinical medicine, environmental science, Agro-ecology, Food Sciences, obtain widespread use.
That microcalorimetric method (Microcalorimetry) has is increasingly automated, real-time online, traceization (mg testing sample just can complete experiment), highly sensitive (being low to moderate the heat of nW), favorable reproducibility, and can not destroy the feature of sample, applying to postgraduate's objects system has many advantages.Microcalorimetric method can directly be measured microorganism or cell, even the susceptibility of organelle to various foreign matters; Can quantitative examination allogene and microorganism, cell or organelle between effect, obtain the information of the aspects such as influence degree, drug effect, bacteriostasis rate and toxicity; In conjunction with other experimental techniques, can also the impact mechanism of Study of Exogenous thing on experimental subjects.This safety evaluation to allogene, instructs synthetic, the screening of medicine or adjuvant and application all to have stronger theory significance.Utilize these characteristics and advantages of biomass thermal technology, can bear the responsibility preferably cigarette is carried out to the just important task of screening with auxiliary material.Before cigarette carries out a series of loaded down with trivial details Evaluation of Biocompatibilities with auxiliary material, utilize microcalorimetric method, can on cell (representative pattern cell or microorganism) eubolism, have the auxiliary material of appreciable impact to screen the overwhelming majority fast and accurately.Can significantly reduce later experiments cost, manpower and time like this, play a multiplier effect.
Summary of the invention
The object of the invention is to have for present auxiliary material in vitro toxicology detection method the defect that cost is high, the cycle long, efficiency is lower, a kind of biothermochemistry method for cigarette auxiliary material safety evaluatio and screening thereof is provided.For the screening of new cigarette admixture and safety evaluatio.Before implementing existing standard toxicology method of determination and evaluation, cigarette is classified by influence degree with auxiliary material, the material that has remarkable toxicity or destructive material and do not reach effect is directly excluded, avoid the waste on man power and material.
For solving the problems of the technologies described above, the invention provides a kind of safety evaluatio of cigarette auxiliary material and the biothermochemistry method of screening, comprise the following steps:
(1) Escherichia coli: LB nutrient culture media is for cultivating and test, and LB culture medium prescription is 5g dusty yeast, 10 g peptones, 5gNaCl is dissolved in 1L redistilled water, regulates pH to 7.4, at 120 ℃, 1.034 * 105Pa autoclaving, processes 20 min, stand-by;
(2) staphylococcus aureus: LB nutrient culture media is for cultivating and test;
(3) saccharomyces albicans: LB nutrient culture media is for cultivating and test;
(4) Candida tropicalis: LB nutrient culture media is for cultivating and test;
(5) S. cervisiae: YPDA nutrient culture media is for cultivating and test, YPDA culture medium prescription is that 20g peptone, 10g yeast extract, 2g glucose dissolve with appropriate redistilled water, 0.2% adenine solution that adds again 15ml, constant volume is to 1L, 120 ℃ of autoclaving 15min, room temperature storage, stand-by;
(6) bacillus subtilis: LB nutrient culture media is for cultivating and test;
(7) thuringiensis: LB nutrient culture media is for cultivating and test;
(8) Lactobacillus plantarum: lactic acid bacteria nutrient culture media is for its cultivation and test; Culture medium prescription is by 7.5g dusty yeast, 7.5g peptone, 10g glucose, 2gKH
2pO
4, 0.5ml Tween 80 is dissolved in 900ml redistilled water, then adds 100ml tomato juice, mixes rear low tempertaure storage, stand-by;
(9) have a liking for salt Archimycetes: having a liking for salt Archimycetes nutrient culture media for cultivating and test, having a liking for salt Archimycetes culture medium prescription is: 250gNaCl, 2gK
2sO
4, 30gMgCl
26H
2o, 2g yeast extract, 2.5g lactoalbumin hydrolysate, is dissolved in 1L redistilled water, stand-by;
(10) tetrahymena: tetrahymena nutrient culture media is for cultivating and test, tetrahymena culture medium prescription is that 15g peptone, 5g dusty yeast, 1g glucose are dissolved in 1L distilled water, pH value is adjusted to 7.0 ~ 7.5, stand-by after autoclaving;
(11) fibroblast: fibroblast nutrient culture media is for cultivating and test, and fibroblast culture medium prescription is MEM basal medium, adds 10% hyclone and 0 ~ 0.4gL
-1nonessential amino acid; MEM nutrient culture media basis is 0.20gL
-1anhydrous CaCl
2, 0.0977gL
-1anhydrous MgSO
4, 0.40gL
-1kCl, 6.8gL
-1naCl, 0.122gL
-1naH
2pO
4, 0 ~ 0.001gL
-1choline tartrate, 0 ~ 0.001gL
-1folic acid, 0 ~ 0.002gL
-1inositol, 0 ~ 0.001gL
-1niacinamide, 0 ~ 0.001gL
-1d-VB5 calcium, 0 ~ 0.001gL
-1pyridoxine hydrochloride, 0 ~ 0.0001gL
-1lactochrome, 0 ~ 0.001gL
-1thiamine hydrochloride, 1gL
-1glucose, 0 ~ 0.001gL
-1succinic acid, 0 ~ 0.001gL
-1sodium succinate, 0.011gL
-1fragrant red, 0 ~ 0.0001gL
-1kanamycins and 0.1 ~ 0.3 gL
-1essential amino acid;
(12) epithelial cell: epithelial cell nutrient culture media is for cultivating and test, and epithelial cell culture medium prescription is MEM basal medium, adds the nonessential amino acid of 10% hyclone and 0 ~ 1%;
(13) mitochondria: mitochondria extract recipe is 0.22molL
-1sweet mellow wine, 0.07molL
-1m sucrose, 0.02molL
-1hydroxyethyl piperazine second thiosulfonic acid HEPES, 0.002molL
-1tri(Hydroxymethyl) Amino Methane Hydrochloride Tris-HCl, 0.001molL
-1disodium EDTA EDTA.2Na, pH is 7.0 ~ 7.4; Mitochondria test fluid formula is 0.22molL
-1sweet mellow wine, 0.07molL
-1sucrose, 0.01molL
-1tri(Hydroxymethyl) Amino Methane Hydrochloride Tris-HCl, 0.001molL
-1disodium EDTA EDTA.2Na, pH is 7.0 ~ 7.4;
2) cigarette smoke condensation product is collected pre-service: by standard smoking machine method of operating; By after flue gas trapping, take water, massfraction as 5 ~ 50% ethanolic solutions or massfraction be 10 ~ 50%N, dinethylformamide solution is extract, vibrates or ultrasonic extraction 10 ~ 60min, stand-by after filtering;
3) the model study object bacterium liquid of the every 10 ~ 500ul in step 1) is added in the fluid nutrient medium that 5ml is corresponding;
4) by step 2) strong solution or suspension be divided into 0 μ l, 5 μ l, 10 μ l, 20 μ l, 50 μ l, 100 μ l, 200 μ l concentration gradients, and the strong solution of variable concentrations gradient or suspension are added in step 3) in fluid nutrient medium, after vibration evenly,
5) the uniform fluid nutrient medium that vibrates in step 4) is positioned in calorimeter TAM air or TAM3, design temperature is 20 ~ 40 ℃ of constant temperature,
6) after temperature reaches setting value and stablizes, nutrient solution is imported to the sensing chamber of micro-calorimeter, start image data;
7) according to image data map, matching and calculating, obtain relevant growth heat production curve and thermokinetic parameters, its parameter comprises the growth rate constant k of metabolic process, maximum heat production power P
m, total thermal discharge Q, generation time t
g, time t that maximum heat power is corresponding
m, appearance time t
pwith inhibiting rate I, 503nhibiting concentration IC
50;
8) relevant growth heat production curve and the thermokinetic parameters of more relevant growth heat production curve and thermokinetic parameters, analysis adds impact and the difference thereof of different materials on model study object eubolism, obtain the entry evaluation of relevant cigarette auxiliary material security, can numerous auxiliary material be screened; Analysis has or not the cigarette that adds new material, and whether the affect otherness of its flue gas on model study object metabolic index can evaluating cigarette auxiliary material has the effect of significant reducing tar and reducing harm.
As preferred version, the nonessential amino acid in described fibroblastic culture medium prescription is several arbitrarily in glycocoll, alanine, serine, aspartic acid, glutamic acid, proline, tyrosine and cystine; In MEM nutrient culture media, essential amino acid is several arbitrarily in lysine, tryptophane, valine, leucine, isoleucine, threonine, methionine, phenylalanine, arginine and histidine.
Accompanying drawing explanation
Fig. 1 is Escherichia coli Growth metabolic heat spectrogram under 37 ℃ of conditions,
In figure, 1. blank, 2. cigarette, 3. reference cigarette.
Embodiment
In order to explain better the present invention, below in conjunction with specific embodiment, further illustrate main contents of the present invention, but content of the present invention is not only confined to following examples.
Embodiment 1
For a kind of Novel tobacco, with titania, mix the Preliminary Identification that fullerene harm reduction material carries out biological effect, be numbered CB02.This novel fullerene-based material has extraordinary clean-up effect to the objectionable constituent in flue gas.Take Escherichia coli, staphylococcus aureus, saccharomyces albicans is that model study is biological, utilizes the hot detector of TAM3 trace to monitor in real time their metabolic process.We have gathered the main flume that adds this material cigarette afterwards, simultaneously with not adding the flue gas of the cigarette of the same race of this material to contrast.The cambridge filter that has trapped main flume is put into respectively to the wide-mouth conical flask of numbering, added quantitative 10ml distilled water immersion the 30min that vibrates.After filtration, from each conical flask, all accurately take out 100 μ l filtrate reference numerals and join in the 5ml nutrient culture media of inoculating microorganism.3 bottles of nutrient culture media of every kind of microorganism are respectively that the main flume collection liquid that has added CB02 cigarette to produce, the main flume of reference cigarette are collected liquid and blank nutrient culture media.By flow pumps, nutrient culture media is pumped into the sample chamber of micro-calorimeter, 37 ℃ of constant temperature, we have obtained microbial growth metabolic chart (Escherichia coli Growth metabolic chart is as shown in the figure).By to the analysis of data and processing, drawn the series of physical chemical parameters of Escherichia coli, staphylococcus aureus and saccharomyces albicans metabolic process, list in table 1.K is growth rate; P
mfor exothermic maximum power; Q is total thermal discharge of whole metabolic process; t
git is the generation time.By data, found out, add the cigarette mainstream flue gas collection liquid of CB02 substantially not form appreciable impact to three kinds of microorganism eubolisms, its impact will be far smaller than reference cigarette.And the main flume of reference cigarette collection liquid slows down growth of microorganism speed k, exothermic maximum power P
mobviously reduce, total thermal discharge Q declines, generation time t
gextend.Result shows, adds CB02 material can reduce harmful degree of cigarette mainstream flue gas in cigarette.
The related physical chemical parameters of three kinds of microbial metabolisms of table 1
Embodiment 2
For four kinds of Novel tobaccos, with the filter tip of functional carbon nano tube compound material, carry out filter effect analysis.First two mostly is wall carbon nano tube, and diameter is respectively 5 nm, 10 nm; Latter two is Single Walled Carbon Nanotube, and diameter is respectively 2nm, 1nm.These four kinds of filter tips have suction-operated and selectivity effect in various degree to flue gas in theory, are numbered 1#, 2#, 3# and 4# respectively to them.Utilize micro-thermal technology to compare fast and accurately the filtration capacity of these four kinds of filter tips, evaluate their filtration capacity.The poor filter tip of successful is screened and got rid of, without carrying out the assessments such as pilot scale or industrial applications, saved a large amount of funds and time.Experiment utilizes the pipe tobacco of the same trade mark, and using the cigarette of this trade mark as reference cigarette.Take S. cervisiae, Lactobacillus plantarum, bacillus subtilis, halophilic archaea and tetrahymena as model study biology, utilize the hot detector of TAM air isothermal trace to monitor in real time pattern postgraduate thing metabolic process under sample cigarette and reference cigarette mainstream flue gas function influence.By the analysis and arrangement to the data that collect, obtain each related physical chemical index of microbial metabolism.Smoking machine aspirates each sample cigarette and reference cigarette, and traps its main flume with the ethanolic solution of 10ml20%, after 30min vibration, filters and seal up for safekeeping stand-by.Various modes is studied to biology and proceed to respectively in 5ml fluid nutrient medium, then add the main flume trapping solution of processing in nutrient solution, proceed to ampoule bottle.After vibration evenly, ampoule bottle is put into the sample chamber of TAM air Isothermal Hot activity tester, constant temperature 30 ℃ (S. cervisiae, Lactobacillus plantarum, bacillus subtilis and tetrahymenas) or 37 ℃ (having a liking for salt Archimycetes) carries out Real-Time Monitoring.The data that collect, through processing, can obtain related physical chemical parameters (table 2).K is growth rate; P
mfor exothermic maximum power; Q is total thermal discharge of whole metabolic process.Blank is the model study biological metabolism parameter that does not add any flue gas trapping liquid.Analysis data are known, and 1# trapping solution exists less inhibiting effect to the metabolism of model study biology, and parameters all shows than a little less than the inhibition of reference cigarette, illustrates that the effect of filtering noxious composition has surpassed the original-pack filter tip of reference cigarette; 2# trapping solution shows than the higher inhibiting effect of reference cigarette trapping solution, illustrate that the effect of filtering noxious composition is not as good as the original-pack filter tip of reference cigarette, so this novel filter tip is inadvisable; It is minimum that the parameters of 3# trapping solution shows its inhibition ability, and far below reference cigarette, filter effect is best; The parameters of 4# trapping solution is extremely close with reference cigarette trapping solution, and concrete effect also needs further to analyze.To sum up, can think that carbon nano-tube diameter is better at the filter effect of 2 ~ 5nm, wherein with the multi-walled carbon nano-tubes best results of 5nm caliber.
The related physical chemical parameters of table 2 microbial metabolism
Embodiment 3
For two kinds of cigarette papers, improve combustion efficiency, slow down burning rate, to them, be numbered 1# and 2# respectively.These two kinds of cigarette paper have added respectively 2 kinds of different fire retardants---and 1# has added aluminium hydroxide (1%), and 2# has added magnesium hydroxide (1%).These two kinds of fire retardants all can make cigarette paper slow down burning rate, make combustion process comparatively abundant, can reduce the release of objectionable impurities, and preparation be easier to, with low cost.Before carrying out heavy industrialization test, we first utilize microcalorimetric method to carry out entry evaluation to harm reduction effect.The bacillus thuringiensis of take is biological as model study, utilizes the micro-hot detector of TAM 3 to monitor the impact of two kinds of smoke components after cigarette paper burning on bacillus thuringiensis.The experiment of trace heat obtains each related physical chemical index of microbial metabolism, and the micro-thermal result of these two kinds of cigarette paper and conventional cigarette paper is compared, and can understand and whether have reducing poison function.Smoking machine aspirates each sample cigarette and reference cigarette, and traps its main flume with the DMF solution of 10ml40%, after ultrasonic extraction 30min, filters and seal up for safekeeping stand-by.Bacillus thuringiensis is proceeded in 5ml fluid nutrient medium, then to the flue gas trapping liquid that adds cigarette paper burning in nutrient solution.After vibration evenly, utilize peristaltic pump, nutrient solution is pumped into the sample chamber of TAM3 Isothermal Hot activity tester, 37 ℃ of constant temperature carry out Real-Time Monitoring.By the processing to data, can obtain related physical chemical parameters (table 3).Data demonstration, 1# and 2# cigarette paper are on not significantly impact of the metabolism of bacillus thuringiensis, and influence degree is much smaller compared with regular-size cigarette paper; 1# cigarette paper effect is more outstanding.Experiment showed, when cigarette paper reduces burning rate, also can reduce the harmfulness of cigarette.
The related physical chemical parameters of table 3 bacillus thuringiensis metabolic process
Embodiment 4
For a kind of Novel tobacco, with harm reduction nano titanium oxide/ZnS quantum dots compound substance, carry out the Preliminary Identification of biological effect, be numbered NTQ03.This new material has extraordinary clean-up effect to the objectionable constituent in flue gas.The Candida tropicalis of take is biological as model study, utilizes the hot detector of TAM3 trace to monitor in real time its metabolic process.We have gathered the main flume that adds this material cigarette afterwards, simultaneously with not adding the flue gas of the cigarette of the same race of this material to contrast.The cambridge filter that has trapped main flume is put into respectively to the wide-mouth conical flask of numbering, added quantitative 10ml50% DMF solution (DMF solution) to soak and ultrasonic extraction 30min.After filtration, from each conical flask, all accurately take out 100 μ l filtrate reference numerals and join in the 5ml nutrient culture media of inoculating Candida tropicalis.3 bottles of nutrient culture media are respectively that the main flume that has added NTQ03 cigarette to produce is collected the main flume collection liquid of liquid, reference cigarette and the blank nutrient solution of inoculating.By flow pumps, nutrient culture media is pumped into the sample chamber of micro-calorimeter, 37 ℃ of constant temperature, we have obtained the growth metabolism curve of Candida tropicalis.By to the analysis of data and processing, drawn the related physical chemical parameters of Candida tropicalis metabolic process, list in table 4.K is growth rate; P
mfor exothermic maximum power; Q is total thermal discharge of whole metabolic process; t
pit is appearance time.By data, found out, add the cigarette mainstream flue gas collection liquid of NTQ03 substantially not form appreciable impact to Candida tropicalis eubolism, its impact will be far smaller than reference cigarette.And the main flume of reference cigarette collection liquid slows down Candida tropicalis growth rate k, exothermic maximum power P
mobviously reduce, total thermal discharge Q declines, generation time t
pextend.Result shows, adds NTQ03 material can reduce harmful degree of cigarette mainstream flue gas in cigarette.
The related physical chemical parameters of table 4 Candida tropicalis metabolic process
Embodiment 5
For two kinds of absorbent charcoal materials, increase the adsorptive power of filter tip to objectionable constituent.Be that common zinc chloride mixes an activated charcoal, another kind the has been finishing activated charcoal of titania, is numbered NC2 and NN1 to them respectively.Before carrying out heavy industrialization test, we first utilize microcalorimetric method to carry out entry evaluation to harm reduction effect.Take that to become fibroblast and the epithelial cell of mouse be model study object, utilize smoke components after the hot detector monitoring of TAM3 trace cigarette burning on l cell and epithelial impact.The experiment of trace heat obtains each related physical chemical index of microbial metabolism, by having added the micro-thermal result of this bi-material cigarette and conventional cigarette to compare, can understand and whether have reducing poison function.Smoking machine aspirates each sample cigarette and reference cigarette, and with 10ml distilled water immersion, adsorbs the filter disc of flue gas, after 40min vibration, filters and seal up for safekeeping stand-by.Cell is proceeded in 1ml fluid nutrient medium, then add cigarette smoke trapping solution 20 μ l in nutrient solution.After vibration evenly, put into the sample chamber of TAM3 Isothermal Hot activity tester, 30 ℃ of constant temperature carry out Real-Time Monitoring.By the processing to data, can obtain related physical chemical parameters (table 5).Data show, add the cigarette of NC2 and NN1 on not significantly impact of the metabolism of two kinds of cells, and influence degree is much smaller compared with reference cigarette; NN1 material result is more outstanding.Experiment showed, that new cigarette material can reduce the harmfulness of cigarette.
The related physical chemical parameters of table 5 fibroblast and epithelial cell metabolic process
Embodiment 6
For Novel polypropylene material filter membrane, to the adsorptive power of objectionable constituent and filter effect, be numbered PM-3.These two kinds of filter membranes have specific adsorption ability to some molecule, and that can reduce objectionable impurities enters human body by filter tip, and preparation be easier to, with low cost.Before carrying out heavy industrialization test, we first utilize microcalorimetric method to carry out entry evaluation to harm reduction effect.Take that to become rat liver mitochondria be model study object, utilize smoke components after the burning of the hot detector monitoring of TAM air trace cigarette paper on the mitochondrial impact of rat liver.The experiment of trace heat obtains each related physical chemical index of microbial metabolism, by having added the cigarette of filter membrane material and the micro-thermal result of conventional cigarette to compare, can understand and whether have reducing poison function.Smoking machine aspirates each sample cigarette and reference cigarette, and traps its main flume with the ethanolic solution of 10ml5%, after 20min vibration, filters and seal up for safekeeping stand-by.Mitochondria is proceeded in 5ml fluid nutrient medium, then add cigarette smoke trapping solution 50 μ l in nutrient solution.After vibration evenly, utilize peristaltic pump nutrient solution to be pumped into the sample chamber of TAM air Isothermal Hot activity tester, 30 ℃ of constant temperature carry out Real-Time Monitoring.By the processing to data, can obtain related physical chemical parameters (table 6).Data show, add the cigarette of PM-3 on not significantly impact of mitochondrial metabolism, and influence degree is much smaller compared with reference cigarette.Experiment showed, that new cigarette filter membrane material can reduce the harmfulness of cigarette.
The related physical chemical parameters of table 6 rat liver mitochondria exothermic process
。
Claims (1)
1. a biothermochemistry method for the safety evaluatio of cigarette auxiliary material and screening, comprises the following steps:
(1) Escherichia coli: LB nutrient culture media is for cultivating and test, and LB culture medium prescription is 5g dusty yeast, 10g peptone, 5gNaCl is dissolved in 1L redistilled water, regulates pH to 7.4, at 120 ℃, 1.034 * 105Pa autoclaving, processes 20min, stand-by;
(2) staphylococcus aureus: LB nutrient culture media is for cultivating and test;
(3) saccharomyces albicans: LB nutrient culture media is for cultivating and test;
(4) Candida tropicalis: LB nutrient culture media is for cultivating and test;
(5) S. cervisiae: YPDA nutrient culture media is for cultivating and test, YPDA culture medium prescription is that 20g peptone, 10g yeast extract, 2g glucose dissolve with appropriate redistilled water, 0.2% adenine solution that adds again 15ml, constant volume is to 1L, 120 ℃ of autoclaving 15min, room temperature storage, stand-by;
(6) bacillus subtilis: LB nutrient culture media is for cultivating and test;
(7) thuringiensis: LB nutrient culture media is for cultivating and test;
(8) Lactobacillus plantarum: lactic acid bacteria nutrient culture media is for its cultivation and test, and culture medium prescription is by 7.5g dusty yeast, 7.5g peptone, 10g glucose, 2gKH
2pO
4, 0.5ml Tween 80 is dissolved in 900ml redistilled water, then adds 100ml tomato juice, mixes rear low tempertaure storage, stand-by;
(9) have a liking for salt Archimycetes: having a liking for salt Archimycetes nutrient culture media for cultivating and test, having a liking for salt Archimycetes culture medium prescription is: 250gNaCl, 2gK
2sO
4, 30gMgCl
26H
2o, 2g yeast extract, 2.5g lactoalbumin hydrolysate, is dissolved in 1L redistilled water, stand-by;
(10) tetrahymena: tetrahymena nutrient culture media is for cultivating and test, tetrahymena culture medium prescription is that 15g peptone, 5g dusty yeast, 1g glucose are dissolved in 1L distilled water, pH value is adjusted to 7.0～7.5, stand-by after autoclaving;
(11) fibroblast: fibroblast nutrient culture media is for cultivating and test, and fibroblast culture medium prescription is MEM basal medium, adds 10% hyclone and 0～0.4gL
-1nonessential amino acid; MEM nutrient culture media basis is 0.20gL
-1anhydrous CaCl
2, 0.0977gL
-1anhydrous MgSO
4, 0.40gL
-1kCl, 6.8gL
-1naCl, 0.122gL
-1naH
2pO
4, 0～0.001gL
-1choline tartrate, 0～0.001gL
-1folic acid, 0～0.002gL
-1inositol, 0～0.001gL
-1niacinamide, 0～0.001gL
-1d-VB5 calcium, 0～0.001gL
-1pyridoxine hydrochloride, 0～0.0001gL
-1lactochrome, 0～0.001gL
-1thiamine hydrochloride, 1gL
-1glucose, 0～0.001gL
-1succinic acid, 0～0.001gL
-1sodium succinate, 0.011gL
-1fragrant red, 0～0.0001gL
-1kanamycins and 0.1～0.3gL
-1essential amino acid; Wherein, the nonessential amino acid in fibroblastic culture medium prescription is several arbitrarily in glycocoll, alanine, serine, aspartic acid, glutamic acid, proline, tyrosine and cystine; In MEM nutrient culture media, essential amino acid is several arbitrarily in lysine, tryptophane, valine, leucine, isoleucine, threonine, methionine, phenylalanine, arginine and histidine;
(12) epithelial cell: epithelial cell nutrient culture media is for cultivating and test, and epithelial cell culture medium prescription is MEM basal medium, adds the nonessential amino acid of 10% hyclone and 0～1%;
(13) mitochondria: mitochondria extract recipe is 0.22molL
-1sweet mellow wine, 0.07molL
-1m sucrose, 0.02molL
-1hydroxyethyl piperazine second thiosulfonic acid HEPES, 0.002molL
-1tri(Hydroxymethyl) Amino Methane Hydrochloride Tris-HCl, 0.001molL
-1disodium EDTA EDTA.2Na, pH is 7.0～7.4; Mitochondria test fluid formula is 0.22molL
-1sweet mellow wine, 0.07molL
-1sucrose, 0.01molL
-1tri(Hydroxymethyl) Amino Methane Hydrochloride Tris-HCl, 0.001molL
-1disodium EDTA EDTA.2Na, pH is 7.0～7.4;
2) cigarette smoke condensation product is collected pre-service: by standard smoking machine method of operating; By after flue gas trapping, take water, massfraction as 5～50% ethanolic solutions or massfraction be 10～50%N, dinethylformamide solution is extract, vibrates or ultrasonic extraction 10～60min, stand-by after filtering;
3) by step 1) in the model study object bacterium liquid of every 10～500ul add in the fluid nutrient medium that 5ml is corresponding;
4) by step 2) strong solution or suspension be divided into 0 μ l, 5 μ l, 10 μ l, 20 μ l, 50 μ l, 100 μ l, 200 μ l concentration gradients, and the strong solution of variable concentrations gradient or suspension are added to step 3) in fluid nutrient medium, after vibration evenly
5) by step 4) in vibration uniform fluid nutrient medium be positioned in calorimeter TAM air or TAM3, design temperature is 20～40 ℃ of constant temperature,
6) after temperature reaches setting value and stablizes, nutrient solution is imported to the sensing chamber of micro-calorimeter, start image data;
7) according to image data map, matching and calculating, obtain relevant growth heat production curve and thermokinetic parameters, its parameter comprises the growth rate constant k of metabolic process, maximum heat production power P
m, total thermal discharge Q, generation time t
g, time t that maximum heat power is corresponding
m, appearance time t
pwith inhibiting rate I, 503nhibiting concentration IC
50;
8) relevant growth heat production curve and the thermokinetic parameters of more relevant growth heat production curve and thermokinetic parameters, analysis adds impact and the difference thereof of different materials on model study object eubolism, obtain the entry evaluation of relevant cigarette auxiliary material security, can numerous auxiliary material be screened; Analysis has or not the cigarette that adds new material, and whether the affect otherness of its flue gas on model study object metabolic index can evaluating cigarette auxiliary material has the effect of significant reducing tar and reducing harm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210452337.7A CN102980913B (en) | 2012-11-13 | 2012-11-13 | Biological thermochemistry method for safety evaluation and screening of cigarette auxiliary material |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210452337.7A CN102980913B (en) | 2012-11-13 | 2012-11-13 | Biological thermochemistry method for safety evaluation and screening of cigarette auxiliary material |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102980913A CN102980913A (en) | 2013-03-20 |
| CN102980913B true CN102980913B (en) | 2014-11-05 |
Family
ID=47855132
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210452337.7A Active CN102980913B (en) | 2012-11-13 | 2012-11-13 | Biological thermochemistry method for safety evaluation and screening of cigarette auxiliary material |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102980913B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104215486B (en) * | 2014-09-12 | 2017-02-01 | 云南中烟工业有限责任公司 | Cigarette paper combustion product capturing method for biological detection |
| CN104962604B (en) * | 2015-07-31 | 2018-01-30 | 云南中烟工业有限责任公司 | Detection method for cigarette smoke condensates reverse mutation |
| CN114487010B (en) * | 2022-02-15 | 2024-03-01 | 云南中烟工业有限责任公司 | Method for evaluating heating cigarette formula based on substance heating energy change |
-
2012
- 2012-11-13 CN CN201210452337.7A patent/CN102980913B/en active Active
Non-Patent Citations (5)
| Title |
|---|
| 《卷烟烟气浸出物对微生物生长的影响》;苏罗毅等;《昆明学院学报》;20101231;第32卷(第6期);41-42页 * |
| 《微量热技术在生命科学研究中的应用》;李月生等;《化工时刊》;20071231;第21卷(第7期);64-67页 * |
| 叶丙静.《AlCl3对大肠杆菌生长代谢的影响》.《微生物代谢活性的微量热研究》.2009,36-38页. * |
| 李月生等.《微量热技术在生命科学研究中的应用》.《化工时刊》.2007,第21卷(第7期), * |
| 苏罗毅等.《卷烟烟气浸出物对微生物生长的影响》.《昆明学院学报》.2010,第32卷(第6期), * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102980913A (en) | 2013-03-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Schnürer et al. | Microbial biomass and activity in an agricultural soil with different organic matter contents | |
| CN105044171B (en) | A kind of preparation method and application of nanometer of platinum dopant/enzyme modification carbon paste electrode | |
| Li et al. | A nanoscale Zr-based fluorescent metal-organic framework for selective and sensitive detection of hydrogen sulfide | |
| CN105693703B (en) | A kind of novel Ratiometric fluorescent probe for the imaging of intracellular lysosomal pH | |
| CN102980913B (en) | Biological thermochemistry method for safety evaluation and screening of cigarette auxiliary material | |
| Langer et al. | Rapid quantification of bioaerosols containing L. pneumophila by Coriolis® μ air sampler and chemiluminescence antibody microarrays | |
| CN108169303A (en) | Aptamer electrochemical sensor based on metal-organic framework material as signal probe | |
| Chen et al. | A structure-dependent ratiometric fluorescence sensor based on metal-organic framework for detection of 2, 6-pyridinedicarboxylic acid | |
| Kavya et al. | Palladium nanoparticles decorated Ni-MOF nanocomposite as an electrochemical platform for the selective detection of dopamine | |
| CN106117241B (en) | The fluorescence probe of lysosomal pH in a kind of detection cancer cell | |
| CN105001862A (en) | Preparation of carbon quantum dots by use of aloe as carbon source and detection method of lemon yellow | |
| CN105628753B (en) | A kind of vitamin B2Bioelectrochemistry detection method | |
| CN107271590A (en) | A kind of method of headspace gas chromatography quick detection anti-bacteria paper biocidal property | |
| CN102980912A (en) | Bio-thermo-chemical method used in cigarette flavor and fragrance security screening | |
| CN105445248A (en) | Method for rapid determination of cyanobacteria cell apoptosis rate | |
| CN105548114B (en) | A method of based on saccharomycete on-line analysis Atmospheric particulates toxicity | |
| CN109651249A (en) | A kind of fluorescence probe detecting endocytoplasmic reticulum cysteine and its synthesis and application | |
| CN104819966A (en) | Method for applying calixarene fluorescent probes to fluorescent imaging of Zn<2+> and F<-> | |
| He et al. | A rapid method for determining Mycobacterium tuberculosis based on a bulk acoustic wave impedance biosensor | |
| CN104165909B (en) | Biological electrochemical detection method of fumaric acid | |
| CN101349667A (en) | Electrochemical detection device of physiological active substance and special-purpose electrochemical sensor preparation method thereof | |
| CN105348214A (en) | Benzoxazinone thiophenol fluorescent probe and preparation method thereof | |
| CN101671718A (en) | Detection method of toxicity of nanomaterials | |
| Lu et al. | Airborne microbial aerosol detection by combining single particle mass spectrometry and a fluorescent aerosol particle sizer | |
| Azimi et al. | A novel method for extraction of galegine by molecularly imprinted polymer (MIP) technique reinforced with graphene oxide and its evaluation using polarography |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |