Classifications

• • A—HUMAN NECESSITIES
  • A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
  • A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
  • A24B15/00—Chemical features or treatment of tobacco; Tobacco substitutes, e.g. in liquid form
  • A24B15/18—Treatment of tobacco products or tobacco substitutes
  • A24B15/20—Biochemical treatment
• • A—HUMAN NECESSITIES
  • A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
  • A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
  • A24B3/00—Preparing tobacco in the factory
  • A24B3/04—Humidifying or drying tobacco bunches or cut tobacco
• • A—HUMAN NECESSITIES
  • A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
  • A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
  • A24B3/00—Preparing tobacco in the factory
  • A24B3/12—Steaming, curing, or flavouring tobacco
• • A—HUMAN NECESSITIES
  • A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
  • A24B—MANUFACTURE OR PREPARATION OF TOBACCO FOR SMOKING OR CHEWING; TOBACCO; SNUFF
  • A24B9/00—Control of the moisture content of tobacco products, e.g. cigars, cigarettes, pipe tobacco
• • A—HUMAN NECESSITIES
  • A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
  • A24F—SMOKERS' REQUISITES; MATCH BOXES; SIMULATED SMOKING DEVICES
  • A24F40/00—Electrically operated smoking devices; Component parts thereof; Manufacture thereof; Maintenance or testing thereof; Charging means specially adapted therefor
  • A24F40/20—Devices using solid inhalable precursors
• • A—HUMAN NECESSITIES
  • A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
  • A24F—SMOKERS' REQUISITES; MATCH BOXES; SIMULATED SMOKING DEVICES
  • A24F40/00—Electrically operated smoking devices; Component parts thereof; Manufacture thereof; Maintenance or testing thereof; Charging means specially adapted therefor
  • A24F40/40—Constructional details, e.g. connection of cartridges and battery parts
• • A—HUMAN NECESSITIES
  • A24—TOBACCO; CIGARS; CIGARETTES; SIMULATED SMOKING DEVICES; SMOKERS' REQUISITES
  • A24F—SMOKERS' REQUISITES; MATCH BOXES; SIMULATED SMOKING DEVICES
  • A24F40/00—Electrically operated smoking devices; Component parts thereof; Manufacture thereof; Maintenance or testing thereof; Charging means specially adapted therefor
  • A24F40/10—Devices using liquid inhalable precursors

Definitions

• a method for treating tobacco material and treated tobacco material A method for treating tobacco material and treated tobacco material
• the present invention relates to a method of treating tobacco and a treated tobacco by fermentation.
• the fermentation is an anaerobic fermentation.
• Treatments to enhance or add flavours and aromas to the tobacco material at a later stage of tobacco processing often involve the addition of one or more additive(s) to the tobacco and can require additional processing steps and equipment, which can be costly and time-consuming. Furthermore, the addition of additives to the tobacco may be not well perceived by some consumers.
• the invention relates to a method for treating tobacco material, the method comprising fermenting the tobacco material.
• the fermenting step includes: incubating the tobacco material under anaerobic conditions.
• the fermenting step includes: applying a pressure to the tobacco material comprised between 1000 kilograms per square meter and 4000 kilograms per square meter.
• the fermenting step includes: keeping the moisture content of the tobacco material comprised between 25 percent in weight and 40 percent in weight of the total weight of the tobacco material.
• the fermenting step lasts at least one month.
• a fermentation of the tobacco takes place under the claimed conditions. Due to the fermentation, certain chemical compounds present in the tobacco material may change and in turn organoleptic properties of the tobacco material may vary as well. Furthermore, the tobacco material fermented according to the method of the invention may present a lower level of asparagine.
• tobacco material may ferment.
• Tobacco plants may host microorganisms which in turn may include bacteria, mould and actinomyces. Studies have shown that bacteria occupy most of the present microorganisms in the tobacco, while mould and actinomyces are minorities. Yeast is little or cannot be detected at all.
• Fermented tobacco can be made by various suitable techniques known in the art, for example as described in "Research Progress in Tobacco Fermentation” published by Yang Yang et al, Journal of Biosciences and Medicines 2018, 6, 105-114 available on line at: http://www.scirp.org/iournal/ibm; or in US 5372149, or in US 4528993, and others.
• Anaerobic fermentation is defined as the conversion of complex organic compounds into smaller molecules in the absence of oxygen.
• the term can be also defined as the conditions in which, as a result of both chemical equilibria and biochemical activities, oxygen is not available for redox reactions. Instead, other oxidized compounds may be present which can be used by micro-organisms for specific types of energy metabolism.
• Anaerobic conditions may coexist with aerobic ones: oxygen in gaseous form may be unavailable to microorganisms in micro-environments (such as aggregates of detritus suspended in water) while at the same time it may be present in the macro-environment (water).
• micro-environments such as aggregates of detritus suspended in water
• the main energy extraction pathway may be coming from glycolysis, some amino acids being also used as carbon/nitrogen sources.
• the preferred nitrogenous compounds usually include glutamine, alanine, serine, threonine, aspartate, asparagine, urea, and arginine.
• pressure is applied in order to remove the air from the tobacco material.
• the applied pressure "squeezes out" the air from the tobacco material, so that, after the container is closed, oxygen is not present anymore, or it is present only in minimal quantity, in the container.
• the tobacco material in a container and closing the container, after the air has been removed from the closed container, allow reaching quickly the anaerobic conditions. This way of achieving anaerobic condition is preferable as it is cost effective and easy to implement.
• the tobacco material is placed in a container and air is removed and replaced with water.
• the container in which the tobacco material is placed is for example a barrel.
• the barrel is made of wood, or concrete, or metal or a combination of any of these three materials.
• the anaerobic conditions are kept for the desired fermenting duration.
• the tobacco material has a moisture content comprised between 25 percent in weight and 40 percent in weight of the total weight of the tobacco material (weight by weight percentage). More preferably, the tobacco material has a moisture content comprised between 25 percent in weight and 35 percent in weight of the total weight of the tobacco material (weight by weight percentage). More preferably, the tobacco material has a moisture content comprised between 28 percent in weight and 32 percent in weight of the total weight of the tobacco material (weight by weight percentage). More preferably, the tobacco material has a moisture content of 30 percent in weight of the total weight of the tobacco material (weight by weight percentage).
• the tobacco material is wetted with water. Water is added to the tobacco material. Preferably, the tobacco material is wetted before being introduced in the container where the anaerobic conditions are obtained and kept.
• this moisture content is maintained. Therefore, preferably, during the fermentation, the moisture content of the tobacco material is monitored. For example, if the tobacco material is introduced in a container where the fermentation takes place, the container may be opened, and the moisture of the tobacco material may be measured when the container is opened. Preferably, the container is opened at regular intervals in order to perform the measurement of the tobacco moisture.
• the moisture may be measured by a moisture sensor provided inside the container. In this way, the moisture may be measured also when the tobacco material is in the closed container.
• moisture of the tobacco material is measured at regular intervals.
• the tobacco material is subject to a pressure.
• the pressure the tobacco material is subjected to is preferably comprised between 1000 kilograms per square meter (kg/m 2 ) and 4000 kilograms per square meter (kg/m 2 ).
• the pressure applied to the tobacco material is maintained in the above range during fermentation.
• the pressure may be applied to the tobacco material by any means.
• the pressure may be applied pumping an inert gas in the container.
• the pressure may be applied putting a weight on the tobacco material causing the desired pressure range to be applied to the tobacco material.
• the container may be filled with wet tobacco material and, as a "lid" of the container, a weight is placed in contact to the tobacco material till water seeps out of the container.
• the tobacco material is inserted in a container and a weight is located on top or above the tobacco material to exert the desired pressure.
• a weight is located on top or above the tobacco material to exert the desired pressure.
• the container is closed, leaving the weight inside the container, so that the weight may keep applying pressure to the tobacco material.
• the anaerobic conditions an amount of moisture in the tobacco material comprised between 25 percent in weight and 40 percent in weight of the total weight of the tobacco material, and an applied pressure comprised between 1000 kilograms per square meter and 4000 kilograms per square meter. Therefore, stating that the tobacco material is subject to the fermentation conditions, means that the tobacco material is subjected to anaerobic conditions, an amount of moisture in the tobacco material comprised between 25 percent in weight and 40 percent in weight of the total weight of the tobacco material, and an applied pressure comprised between 1000 kilograms per square meter and 4000 kilograms per square meter.
• the tobacco material is subjected to the fermentation conditions above described for at least one month.
• tobacco material is subjected to the fermentation conditions for at least two months.
• tobacco material is subjected to the fermentation conditions for at least six months.
• tobacco material is subjected to the fermentation conditions for at least 12 months.
• tobacco material is subjected to the fermentation conditions for at least 24 months.
• tobacco material is subjected to the fermentation conditions for less than 36 months.
• the application of the fermentation conditions may be continuous for all the claimed time (for example, longer than one month, or longer than two months, or longer than 6 months, or longer than 12 months, or longer than 24 months).
• the fermentation conditions may be applied during a plurality of time intervals forming a sequence of time intervals.
• the time intervals are separated one from the other by "interruptions".
• one or more interruptions of one or more of: presence of anaerobic conditions, amount of moisture comprised between 25 percent in weight and 40 percent in weight of the total weight of the tobacco material, application of a pressure comprised between 1000 kilograms per square meter and 4000 kilograms per square meter may take place.
• the interruption may take place in order check the tobacco material.
• the moisture of the tobacco material may be measured during an interruption.
• the interruption may take place to turn or mix the tobacco material, so that a uniform treated tobacco material may be obtained.
• the fermentation conditions are applied continuously for a total duration equal to T.
• an interruption is present between two consecutive time intervals in which the fermentation conditions are applied.
• the interruption does not last longer than 6 hours.
• the fermentation conditions are applied to the tobacco material for at least a month to see a desired chemical modification of the tobacco material.
• the fermentation condition may be applied till the desired quantity of one or more chemical substances is reached in the tobacco material.
• a chemical substance comprised in the tobacco material may lower or increase due to the fermentation conditions.
• the fermentation conditions are therefore stopped when the chemical substance reaches the desired quantity.
• the fermentation condition can be applied till the desired colour of the tobacco material is obtained.
• the amount of reducing sugars and free amino acids present in the tobacco material has been monitored. Most abundant naturally present sugars in tobacco leaves are glucose, fructose and sucrose. Differences in sugar content may be present among tobacco varieties. For example, Virginia has high level of sugar (generally in a range from 8 percent to 30 percent) while Burley is characterized with low content of sugars (generally in a range of 1 percent to 2 percent). However, regardless of the tobacco type which is used in the tobacco material, a reduction in the content of reducing sugars during the fermentation under the fermentation conditions of the invention has been found.
• tobacco material contains levels of amino acids.
• the amino acids may contribute substantially to the level of certain components in the smoke or aerosol produced by the final product where the fermented tobacco material is contained, and to the sensory properties of the smoke or aerosol.
• Different type of tobaccos may contain different quantities of amino acids.
• the growing location for the tobacco may alter the ratios of the levels of different amino acids, but rather similar profiles for the same tobacco amino acid are generally maintained. Regardless of the tobacco type and origin, during fermentation under the fermentation conditions of the invention, it has been observed that the asparagine content in the tobacco material decreases.
• Asparagine may be converted thermally into acrylamide.
• Acrylamide is considered a potentially harmful substance. It is desired to obtain a decrease in the content of asparagine in the tobacco material because it may concur in a decrease in the acrylamide formation.
• the above changes in the amount of asparagine and in the reducing sugars in the tobacco material may be seen after one month from the moment in which the fermentation conditions are applied to the tobacco material.
• the fermentation conditions are applied and remain applied to the tobacco material for at least one month.
• anaerobic fermentation of the tobacco material takes place when the tobacco material is subjected to the fermentation conditions.
• This anaerobic fermentation alters the reducing sugars and the asparagine amount in the tobacco material. Therefore, with a natural process such as fermentation and without the addition of additives or external microorganisms to the tobacco material, a reduction of certain potentially harmful substances may be achieved. These substances may include acrylamide.
• the fermentation in addition may change the organoleptic properties of the tobacco material. These organoleptic changes may take place because reducing sugars are converted to pyruvate and pyruvic acid, which are precursors of many other flavour compounds. This means that there may be a significant change in the organoleptic properties of the tobacco material after the fermentation of the invention.
• the taste characteristics of the tobacco material may be changed compared to those of the same tobacco material following conventional curing and without the application of the fermentation under fermentation conditions according to the invention.
• the terms "change” or “changed” are used in the context of the flavour or organoleptic properties to mean that there is a modification from one overall taste or sensory character to another, as identified by expert smokers. This may include an improvement.
• the method includes drying the tobacco material to obtain a dried tobacco material having a moisture content comprised between 1 percent and 15 percent in weight of the total weight of the tobacco material.
• the drying step is preferably performed after the fermentation under fermentation conditions has been terminated. After the total fermentation period T has elapsed, the treated tobacco material is preferably removed from the container where it was placed, and the pressure applied to the tobacco is decreased. The treated tobacco material is then dried to a water content of between 1 percent and 15 percent in weight of the total weight of the tobacco material, more preferably between 5 percent and 10 percent. The drying is performed so that the treated tobacco material may be easily processed is subsequent steps.
• the method comprises the step of: curing the tobacco material before fermenting.
• the tobacco material processed according to the method of the invention may comprise post-curing tobacco.
• post-curing tobacco refers to tobacco that has been cured.
• the curing of the tobacco is preferably realized according to standard procedures and may depend on the type of tobacco which is included in the tobacco material.
• the tobacco material may include tobacco of different types and having had different curing.
• the tobacco of different types may be blended and then treated according to the invention.
• the tobacco material treated according to the method of the invention may comprise tobacco that has been re-graded, green-leaf blended, conditioned, de-stemmed or threshed (or not in the case of whole leaf), dried or packed.
• the method includes keeping the temperature of the tobacco material comprised between 25 degrees Celsius and 35 degrees Celsius.
• the temperature of the tobacco material is kept between 25 degrees Celsius and 35 degrees Celsius while the tobacco material is subjected to the fermentation conditions.
• the temperature of the tobacco material is automatically kept within this range by the fermentation process. No additional devices are needed to cool or heat the tobacco material.
• the method comprises the step of turning the tobacco material.
• the turning of the tobacco material may provide an improved homogenization.
• Turning the tobacco material may means turning the tobacco material upside down.
• Turning the tobacco material may means overturning the tobacco material.
• the interruption of the fermentation conditions caused by the turning may be used also to measure certain parameters of the tobacco material, for example the moisture content.
• the fermentation conditions may be not applied any more. All the three fermentation conditions may not be applied during the turning, or only some of them. The fermentation process is thus "interrupted”. After turning, preferably the fermentation conditions are re-applied to the tobacco material.
• the method comprises: securing the tobacco material within a moisture retaining material.
• This step of securing the tobacco material preferably takes place before the tobacco material is subjected to the fermentation conditions. It is desirable for the moisture-retaining material to be resistant to degradation during the tobacco treatment process (the fermentation).
• the moisture-retaining material may comprise a flexible material. This flexible material may be wrapped around the tobacco material.
• the moisture retaining material preferably comprises plastic material. Alternatively, or in addition, the moisture-retaining material may comprise a rigid material.
• the container in which the tobacco material is introduced may function as a moisture retaining material. In this case, the material of the container may include for example metal, wood, plastic, or concrete.
• the method comprises: wetting the tobacco material in water before fermenting, so that a moisture content of the tobacco material comprised between 25 percent in weight and 40 percent in weight of the total weight of the tobacco material is achieved.
• the moisture of the tobacco material is generally low. Therefore, preferably water is added to the tobacco material to reach a moisture level between 25 percent and 35 percent in weight. More preferably, water is added also during the fermentation process in order to keep the moisture of the tobacco material comprised between 25 percent in weight and 40 percent in weight of the total weight of the tobacco material for at least one month, more preferably for at least 2 months, preferably for at least 6 months, preferably for at least 12 months, preferably for at least 24 months.
• the invention relates to a tobacco material treated according to the method of the previous aspect, wherein the treated tobacco material comprises an amount of asparagine at least 50 percent, more preferably 60 percent, even more preferably 80 percent lower than the amount of asparagine contained in the same tobacco material before the treatment according to the previous aspect.
• the amount of asparagine is at least 50 percent, more preferably 60 percent, even more preferably 80 percent lower than the amount of asparagine contained in the same tobacco material before the treatment.
• the treated tobacco according to the method of the invention may change its chemical composition with respect to the untreated tobacco.
• Treated tobacco material in the present context means tobacco material that underwent the treatment as described in the previous process, that is, tobacco material that has been subjected for at least one month to the fermentation conditions.
• Untreated tobacco material in the present context means tobacco material that did not undergo the treatment as described in the previous process, that is, tobacco material that has not been subjected to the fermentation conditions. Untreated tobacco material is for example the tobacco material which is inserted in the container before the treatment of the invention starts. The treated tobacco material is compared with the same tobacco material which did not undergo the treatment according to the invention (the untreated tobacco material). The decrease in asparagine may be associated with an increase in aspartate. This suggests that fermenting bacteria produce specific asparaginase(s) to assimilate C and N from amino acid resources. This reaction may produce ammonia.
• the treated tobacco material comprises an amount of glutamine at least 50 percent, more preferably 60 percent, even more preferably 80 percent lower than the amount of glutamine contained in the same tobacco material before the treatment according to the method of the previous aspect.
• the amount of glutamine is at least 50 percent, more preferably 60 percent, even more preferably 80 percent lower than the amount of glutamine contained in the same tobacco material before the treatment.
• the treated tobacco material according to the method of the invention may change its chemical composition with respect to the untreated tobacco material.
• the treated tobacco material is compared with the same tobacco material which did not undergo the treatment according to the invention.
• the decrease in glutamine may be associated with an increase in glutamate. This suggests that fermenting bacteria produce specific glutaminase(s) to assimilate C and N from amino acid resources. This reaction may produce ammonia.
• the treated tobacco material comprises an amount of total reducing sugars at least 50 percent, more preferably 60 percent, even more preferably 85 percent lower than the amount of total reducing sugars contained in the same tobacco material before the treatment according to the method of the previous aspect.
• the amount of reducing sugars is at least 50 percent, more preferably 60 percent, even more preferably 85 percent lower than the amount of reducing sugars contained in the same tobacco material before the treatment.
• Reducing sugars are the sum of the following: glucose, fructose, sucrose, maltose. The majority of the reducing sugars in the treated tobacco material may be converted.
• Reducing sugar resources such as glucose and fructose present in the starting tobacco material may be used as a source of energy by the anaerobic bacteria.
• the glycolysis pathway transforms glucose (or fructose) into pyruvate.
• the altered levels of these compounds may contribute to the desirable taste and aroma of the treated tobacco material.
• the treated tobacco material is at least 100 times more acidic than the untreated tobacco material.
• the pH of the treated tobacco material and the pH of the untreated tobacco material may differ of at least 2 pH units. In different tobacco material, the pH may remain substantially unchanged.
• the treated tobacco comprises lactic acid.
• lactic acid is known to be a related catabolic product.
• the lactic acid may have a "smoothing effect" regarding nicotine harshness.
• the lactic acid may be responsible in the lowering of the pH of the treated tobacco material.
• the invention relates to a tobacco material comprising: less than 3 percent of total reducing sugars in total dry weight basis. More preferably, the tobacco material comprises less than 2 percent of total reducing sugars in total dry weight basis. Even more preferably, the tobacco material comprises less than 1 percent of total reducing sugars in total dry weight basis. Preferably, the tobacco material comprises less than 300 milligrams per kilogram of asparagine in total dry weight basis.
• the tobacco material is fermented tobacco material.
• the tobacco material is preferably the tobacco material treated according to the method of the invention.
• the fermentation is obtained without the addition of any microorganisms besides those already included in the tobacco material before fermentation.
• the tobacco material comprises less than 70 milligrams per kilogram of glutamine in total dry weight basis.
• the tobacco material comprises more than 10000 milligrams per kilogram in total dry weight basis of total free amino acids.
• the tobacco material includes hand - stripped leaves from which ribs have been removed.
• the tobacco material is cured.
• the curing is performed before fermentation.
• the tobacco material comprises Kasturi tobacco.
• the invention relates to an aerosol generating article comprising a tobacco material according to the previous aspect.
• tobacco material refers to any part of a tobacco plant or a mixture of different tobacco plants and includes without limitation tobacco leaf scraps, tobacco green leaf scraps, tobacco stems, tobacco dust created during tobacco processing, and tobacco leaf prime lamina strip and a combination thereof.
• the tobacco material can have the form of processed tobacco parts or pieces, cured and aged tobacco in essentially natural lamina or stem form, a tobacco extract or a mixture of the foregoing, for example, a mixture that combines extracted tobacco pulp with granulated cured and aged natural tobacco lamina.
• the tobacco material can be in solid form, in liquid form, in semi-solid form, or the like.
• the term "tobacco material” includes any part and any related by-product, such as for example the leaves or stems, of any member of the genus Nicotiana.
• the tobacco material for use in the present invention is preferably from the species Nicotiana tabacum. Any type, style or variety of tobacco may be treated. Examples of tobacco which may be used include but are not limited to Virginia, Burley, and Oriental tobaccos, and blends of any of these types.
• the tobacco material comprises Kasturi tobacco.
• the tobacco material to be treated may comprise or consist of post-curing tobacco.
• post-curing tobacco refers to tobacco that has been cured but has not undergone any further treatment process to alter the taste or aroma of the tobacco material.
• the post curing tobacco may have been blended with other styles, varieties or types of tobacco.
• the tobacco material to be treated may comprise or consist of tobacco that has been re-graded, green-leaf blended, conditioned, de-stemmed or threshed (or not in the case of whole leaf), dried or packed.
• the tobacco material comprises lamina tobacco material.
• the tobacco may comprise between about 70% and 100% lamina material.
• the lamina may be in whole leaf form.
• the tobacco material comprises cured whole leaf tobacco.
• the tobacco material substantially comprises cured whole leaf tobacco.
• the tobacco material consists essentially of cured whole leaf tobacco.
• the tobacco material comprises stem tobacco material.
• the tobacco may comprise up to a 30 percent of stem material.
• the process of "curing" green tobacco depends on the type of tobacco harvested. For example, Virginia flue (bright) tobacco is typically flue-cured, whereas Burley and certain dark strains are usually air-cured.
• the flue-curing of tobacco typically takes place over a period of five to seven days compared to one to two months for air-curing. Many major chemical and biochemical changes begin during the curing process and continue through the early phases of leaf drying.
• the conversion of the tobacco from its yellow to brown colour generally results in formation and substantial accumulation of nitrosamines, and an increased microbial content.
• Virginia tobacco is generally 'flue-cured.' The tobacco leaves are hung in curing barns, where heated air is generated to dry the leaves. As the leaves lose moisture, they develop their distinct aroma, texture, and colour. The farmer must carefully guide this process, which takes up to a week, during which time the temperature of the heated air must be constantly monitored and gradually increased. Too much or too little heat at any stage of the process will have a negative impact on the quality of the tobacco.
• Burley and oriental tobaccos are cured differently. Burley is 'air-cured' in barns where the heat and humidity come from natural ventilation. The curing process takes up to two months. Oriental tobacco is 'sun-cured' by hanging the leaves outdoors in the sun for about two weeks.
• Example Exl A method for treating tobacco material, the method comprising: o fermenting the tobacco material to obtain treated tobacco material, including:
• Example Ex 2 The method according to Exl, including the step of: o drying the tobacco material to obtain a dried tobacco material having a moisture content comprised between 5 percent and 10 percent in weight of the total weight of the tobacco material.
• Example Ex 3 The method according to Ex 1 or Ex 2, comprising the step of: o curing the tobacco material before fermenting.
• Example Ex 4 The method according to one or more of the preceding Ex 1 - Ex 3, wherein the fermenting lasts less than three years.
• Example Ex 5 The method according to one or more of the preceding Ex 1 - Ex 4, comprising the step of o turning the tobacco material.
• Example Ex 6 The method according to one or more of the preceding Ex 1 - Ex 5, including: o adding water to the tobacco material to maintain the moisture content of the tobacco material comprised between 25 percent in weight and 40 percent in weight of the total weight of the tobacco material.
• Example Ex 7 The method according to one or more of the preceding Ex 1 - Ex 6, including keeping the temperature of the tobacco material comprised between 25 degrees Celsius and 35 degrees Celsius.
• Example Ex 8 The method according to one or more of the preceding Ex 1 - Ex 7, comprising: o securing the tobacco material within a moisture retaining material.
• Example Ex 9 The method according to one or more of the preceding Ex 1 - Ex 8, comprising: o wetting the tobacco material in water before fermenting, so that a moisture content of the tobacco material comprised between 25 percent in weight and 40 percent in weight of the total weight of the tobacco material is achieved.
• Example ExlO Method according to one or more of the preceding Ex 1 - Ex 9, wherein the amount of asparagine in the treated tobacco material becomes at least 50 percent lower than the amount of asparagine contained in the same tobacco material before treatment according to Exl - Ex9.
• Example Exll Method according to one or more of the preceding Ex 1 - Ex 10, wherein the amount of asparagine in the treated tobacco material becomes at least 50 percent lower than the amount of asparagine contained in the same tobacco material before treatment according to Exl - ExlO.
• Example Ex 15 Treated tobacco material according to Ex 13 or Ex 14, wherein the treated tobacco material comprises an amount of total reducing sugars at least 85 percent lower than the amount of total reducing sugars contained in the same tobacco material before the treatment according to Ex 1 - Ex 12.
• Example Ex 16 Treated tobacco according to any of the preceding Ex 13 - Ex 15, wherein the treated tobacco material is at least 100 times more acidic than the same tobacco material before the treatment according to Ex 1 - Ex 12.
• Example Ex 17 Tobacco material comprising: o less than 3 percent of total reducing sugars in total dry weight basis; o less than 300 milligrams per kilogram of asparagine in total dry weight basis.
• Example Ex 18 Tobacco material according to Exl7, comprising: o less than 1 percent of total reducing sugars in total dry weight basis.
• Example Ex 19 Tobacco material according to Ex 17 or Ex 18, comprising: o less than 70 milligrams per kilo of glutamine in total dry weight basis.
• Example Ex 20 Tobacco material according to one or more of Ex 17 - Ex 19, wherein the fermented tobacco material comprises: o more than 10000 milligrams per kilogram in total dry weight basis of total free amino acids.
• Example Ex 21 Tobacco according to any of the preceding Ex 17 - Ex 20, comprising lactic acid.
• Example Ex 22 Tobacco material according to one or more of Ex 17 - Ex 21, wherein the tobacco material includes hand -stripped leaves from which ribs have been removed.
• Example Ex 23 Tobacco material according to one or more of Ex 17 - Ex 22, wherein the tobacco material is cured.
• Example Ex 25 Aerosol generating article comprising the tobacco material according to any one of Ex 17 - Ex 24.
• FIG 1 and FIG. 2 are histograms representing the quantity of lactic acid in the tobacco material of Example 1 and Example 2, respectively, measured before (0T) and after 6 months (3T) of fermentation according to the invention;
• FIG. 3 and FIG. 4 are histograms representing the quantity of Total alkaloid (TA) levels (percent in total dry weight basis DW) in the tobacco material of Example 1 and Example 2, respectively, measured before (0T) and during fermentation according to the invention;
• TA Total alkaloid
• FIG. 5 and FIG. 6 are histograms representing the quantity of glutamine and glutamic acid in the tobacco material (in total dry weight basis, DW) of Example 1 and Example 2, respectively, measured before (0T) and during fermentation according to the invention;
• FIG. 7 and FIG. 8 are histograms representing the quantity of asparagine and aspartic acid in the tobacco material (in total dry weight basis, DW) of Example 1 and Example 2, respectively, measured before (0T) and during fermentation according to the invention;
• FIG. 9 and FIG. 10 are histograms representing the quantity of total alkaloids and reducing sugars in the tobacco material (in total dry weight basis, DW) of Example 3, respectively, measured before (VG-BF), during fermentation and after fermentation (VG-AF) according to the invention.
• a first and a second tobacco material of the same tobacco type but having a different processing before fermentation have been prepared.
• the tobacco material is Kasturi tobacco.
• the tobacco material was conditioned to obtain a moisture of circa 30 percent. Samples of this tobacco material conditioned but not fermented yet are called 0T ("starting material").
• the conditioned tobacco material is then introduced in three barrels, in each barrel circa 100 kilograms of tobacco material is present. Before the introduction, the tobacco material is wrapped in a material maintaining the acquired moisture. Pressure is applied to each barrel. The pressure is comprised between between 1000 kilograms per square meter and 4000 kilograms per square meter.
• sample called IT After 1 month (sample called IT), 2.5 months (sample called 2T), 6 months (sample called 3T) and 8.5 months (sample called 4T), the barrels were opened, and the samples collected at least in triplicate in each barrel before tobacco turning and readjustment of the moisture content to approximatively 30 percent ⁇ 5 percent.
• the tobacco material was conditioned to obtain a moisture content of circa 30 percent. Samples of this tobacco material conditioned but not fermented yet are called 0T ("starting material").
• Pressure is applied to each barrel.
• the pressure is comprised between between 1000 kilograms over square meter and 4000 kilograms over square meter.
• sample called IT After 1 month (sample called IT), 2.5 months (sample called 2T), 6 months (sample called 3T) and 8.5 months (sample called 4T), the barrels were opened, and the samples collected at least in triplicate in each barrel before tobacco turning and readjustment of the moisture content to approximatively 30 percent ⁇ 5 percent.
• the given value represents an average of several values obtained for each sample of the same type.
• the pH of the samples of tobacco material became acidic reaching 3.2, after fermentation conditions have been applied for 2.5 months (as found in sample 2T). This reflects the process of anaerobic fermentation involving sugars degradation, which usually produces organic acids like (acetic and/or) lactic acids.
• the starting pH of the tobacco material is generally comprised between 5 pH and 6 pH.
• Alkaloids were not or only slightly degraded during the fermentation.
• the total alkaloids (TA) content in percent in total dry weight basis (indicated as % DW in the figures) is shown in Figure 3 (HS leaves) and Figure 4 (CC leaves).
• the content of total alkaloids remained quite stable during the fermentation. After 8.5 month (4T), only 4 percent were degraded in HS and 9 percent in CC leaves. Although statistically relevant, such small variation may just result from sampling. Some limited alkaloid hydrolase activities may not be excluded.
• Sample 4T of HS leaves and sample 3T of CC leaves have a p-value ⁇ 0.01 and Samples IT and 4T of CC leaves have a p-value ⁇ 0.001.
• TSNA tobacco specific nitrosamines
• NNN N'-nitrosonornicotine
• NNK NNK-derived nitrosamine ketone
• NAT N'-nitrosoanatabine
• Figures 5 and 6 show the level of glutamine (white histograms) and glutamic acid (black histograms) in HS leaves and CC leaves, respectively. It is clear from the figures that during fermentation glutamine decreases and glutamic acid increases.
• Figures 7 and 8 show the level of asparagine (striped histograms) and aspartic acid (black histograms) in HS leaves and CC leaves, respectively. It is clear from the figures that during fermentation asparagine decreases and aspartic acid increases.
• Citrate and malate like reducing sugars, are also metabolized during the tobacco heavy fermentation: it is shown from chemical analysis of the samples that more than 60% of the glucose and fructose, citrate and malate present in the starting tobacco material (samples 0T), hand-stripped and chopped leaves are catabolized after 6 months of heavy fermentation (samples 3T). Another observation that can be coupled to the consumption of such organic molecules is the increase of pyruvate (13-14 times) in both HS and CC fermented tobacco material.
• Pyruvate is the substrate of several reactions that may occur under anaerobic conditions: (1) the production of D-lactate, mostly to regenerate NAD+ for the glycolytic reaction; (2) the production of acetate, diacetyl and 2,3 butanediol that may contribute to the delivery of aromatic compounds and flavours in heavy fermented tobacco. Pyruvate may lead to the generation of aromatic compounds, like 2,3-butanediol or lactate as a product of lactic acid bacteria.
• tryptophan degradation the pathway has been described by Ummadi and Weimer (2001, J. Dairy Sci. 84:1773-1782) for cheese bacteria and adapted accordingly.
• more than 78% of the tryptophan present in the starting tobacco material is catabolized after 6 months of fermentation (samples 3T) in both HS and CC leaves.
• Tobacco leaf material has been fully sun-cured for about 10 days.
• the sun cured leaves have been treated as standard for Virginia tobacco.
• the conditioned tobacco material is then introduced in two barrels, in each barrel circa 100 kilograms of tobacco material is present. Before the introduction, the tobacco material is wrapped in a material maintaining the acquired moisture.
• Pressure is applied to each barrel.
• the pressure is comprised between between 1000 kilograms per square meter and 4000 kilograms per square meter.
• the tobacco material has been turned and the moisture content of the tobacco material has been readjusted to approximatively 30 percent ⁇ 5 percent.
• the moisture content of the tobacco material has been readjusted to approximatively 30 percent ⁇ 5 percent.
• no major changes of temperature were observed during the fermentation process moving linearly from 30 degrees Celsius at the beginning of the fermentation (VG-T1) to 26 degrees Celsius at the end of the fermentation (VG-AF).
• the temperature has been measured inside the barrels using captors.
• the pH of the tobacco material did not change significantly during the fermentation run (T1 to AF) staying at 5.1 ⁇ 0.3.
• the fermentation has been stopped after 8 months.
• VG-AF the color of the tobacco material at the end of the fermentation process
• VG-BF the starting material
• VG-T4 the Virginia tobacco material did not show the same "darkness as the Kasturi tobacco after the same amount of fermentation, indicating possibly that 4 months are not sufficient to get full fermentation of Virginia tobacco material.
• reducing sugars were used as substrate by fermenting bacteria. Therefore about 60% of the reducing sugars (RS) were oxidized during the 8 month of fermentation, moving from 18.3 percent (VG-BF) to 7.4 (VG-AF) percent in dry weight (DW). It is possible that a longer period of fermentation would lead to a higher percentage of RS degradation.
• the starting material (VG-BF) had a content of 185 micrograms per gram (pg/g) of glutamine on a dry weight basis.

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