['II.', 'Experimental:°', 'The', 'appropriate', 'amino', 'acid:', 'was', 'added!', 'to', 'S1', '(BuST', 'or', 'BrS1', 'were', 'obtained', 'as', 'described', 'previously®', ')', 'in', 'the', 'proportions', 'described', 'in', 'Table', 'III..', 'The', 'solutions:of', 'the', 'AA', 'and', 'S1', 'were', 'diluted!', 'with', '20mL', 'of', 'water', 'and!applied', 'to', 'BrBW', 'using', 'standard', 'methods.‘', 'All', 'of', 'the', 'model', 'fillers', 'were', 'allowed', 'to', 'equilibrate', 'in', 'a', 'conditioned', 'laboratory', '(24°', 'C,', '60%', 'relative:', 'humidity:', 'for', '24', 'hours)', 'and', 'made', 'into:cigarettes', "(1.0'-", '1|.1g', 'each)', 'using', 'the', 'Supermatic®', 'cigarette', 'machine.', 'Random', 'smokings', 'of', 'the', 'model', 'fillers', 'were', 'conducted', 'in', 'wiplicate', "on'a", '30-port', 'Borgwaldt', 'smoking', 'machine.', 'Three', 'cigarettes', 'were', 'smoked', 'per', 'replicate', 'producing', 'replicate', 'IT', 'CSCs', 'for', 'each', 'model', 'filler.', 'Each', 'IT', 'CSC', 'sample', 'was', 'made:into', 'a', '10mg/mL', 'DMSO', 'stock', 'solution', 'and', 'submitted', 'for', 'Salmonella', 'testing..', 'The', 'samples', 'were', 'stored', 'at', '-80°', 'C', 'until', 'tested..', 'Samples', 'of', 'each', 'model', 'filler', 'were', 'submitted', 'to', 'ARD', 'personnel', 'for', 'analysis', '(Table', 'IV', 'and', 'V).', '_', 'TIL.', 'Results.and', 'Discussion:', 'At', 'the', 'levels', 'added,', 'all', 'of', 'the', 'AA', 'were', 'soluble', 'in', 'the', 'S1', 'andicigarette', 'preparation.', 'proceeded', 'routinely..', 'This', 'was', 'fortunate', 'given.', 'the', 'low', 'water', 'solubility', 'of', 'most', 'of', 'the', 'AA', 'chosen:', 'The', 'burley', 'spiked', 'fillers.analyzed', 'for', 'total', 'nitrogen', 'near', 'the', 'target', 'value', 'of', '4%', '(Table', 'IV).', 'This', 'represented', 'an', 'AA', 'load!', 'of', "from'5", 'to.9%', 'of', 'the', 'total', 'weight', 'of', 'filler.', 'As:observed', 'in', 'Table', 'IV,', 'there', 'was', 'very', 'little', 'effect', 'of', 'added', 'AA', 'onitotal', 'ash', 'or', 'OV', 'as', 'compared', 'to', 'the', 'BuS', '1', 'control.', 'As', 'expected,', 'HWS', 'were', 'increased', 'in', 'comparison.', 'to', 'the', 'BuS1', 'control.', 'CSC', 'yields', 'from', 'these', 'cigarettes', 'varied', 'from', '36', 'to.46mg/cigarette', '(Table', 'VI).', 'As', 'further', 'illustrated', 'in', 'Table', 'VI,', 'biologicaliactivity', 'of', 'the', 'spiked', 'BuS1', 'cigarettes', 'were', 'fairly', 'consistent', 'with', 'values', 'for', 'an', 'all', 'burley', 'blend', 'or', 'for', 'cigarettes', 'containing', 'burley', "solubles.'", 'Proline', "spiked'cigarettes", 'generated!', 'CSC', 'with', 'the', 'most', 'activity', 'while', 'glutamic', 'acid', 'spiked', 'cigarettes', 'generated', 'CSC', 'with', 'the', 'least', 'activity.', 'or', 'In', 'the', 'case', 'of', 'BrS1', 'spiked', 'with', 'AA,', 'a', 'total', 'nitrogen', 'value', 'of', 'about', '3%', 'in', 'filler', 'was', 'obtained', '(Table', 'V).', 'This', 'gave', 'an', 'AA', 'percent', 'load', 'comparable', 'to', 'the', 'burley', 'S11', 'spiked', 'fillers', '(Table', 'III).', 'As', 'observed', 'in:', 'Table', 'V,', 'there', 'was', 'very', 'little', 'affect', 'of', 'added', 'AA', 'on', 'total', 'ash', 'and', 'a', 'slight', 'effect', 'on', 'OV', 'as', 'compared!', 'to', 'the', 'BrS1', 'control.', 'As', 'expected,', 'HWS', 'were', 'increased!', 'in', 'comparison:', 'to', 'the', 'BrS1', 'control.', 'CSC', 'delivery', 'of', 'these', 'BrS1', 'spiked', 'cigarettes', '(Table', 'VII)', 'was', 'generally', 'greater', 'than', 'those-of', 'the', 'burley', 'spiked', 'cigarettes', '(Table', 'VI),', 'however,', 'S/M', 'activity:', 'levels.', 'were,', 'with', 'the', 'exception', 'of', 'the', 'proline.', 'spikedicigarettes,', 'lower.', 'From', 'this', 'limited', 'study,', 'it', 'appears', 'that', 'CSCs', 'derived', 'from:', '"neutral"', 'amino', 'acid', 'spiked!', 'cigarettes.', 'were', 'relatively', 'more', 'active', 'in', 'the', 'S/Miassay.', 'It', 'should', 'be', 'noted', 'that', 'aspargine', 'is', 'easily', 'converted!', 'to', 'aspartic', 'acid', 'under', 'acidic', 'conditions.’', 'The', 'approximate', 'pH', 'of', 'BuS1', 'is', '6.', 'It', 'has', 'been', 'shown', 'that', 'the', 'addition', 'of', 'sugars', 'to', 'BuCEL', 'lowers', 'the', 'S/M', 'activity', 'of', 'the', 'resulting', 'CSC’', 'and', "the:observed'decrease", 'in', 'S/M', 'activity', 'of', 'the:CSC', 'obtained!', 'from', 'BrS1', 'spiked', 'cigarettes', 'is', 'most', 'likely', 'derived!', 'from', 'the', 'interaction', 'between', 'the', 'added', 'AA', 'and', 'the', 'naturally', 'occurring', 'sugars', 'in', 'BrS1.', '2s', 'BES6TEL', 'SOS’']