The metabolism of the plant growth regulator maleic hydrazide (MH) has been studied in sterile cell suspension cultures of tobacco, soybean, maize and wheat under standardized conditions. Maleic hydrazide was converted to its O-b -D-glucoside in yields between 9.0 % (tobacco) 15.0 % (soybean), 5.1 % (maize) and 2.2 % (wheat) respectively. This glucoside was completely cleaved under simulated conditions (pH 1, 37°C, 24 h) of a ruminant stomach. From these results it is concluded that MH-O-b-D-glucoside (MHG) belongs to a small group of acid-labile pesticide conjugates (11). The participation of a glucosyltransferase (GT) (EC 2.4.1-) in this conjugation reaction of MH was demonstrated in vitro for the first time. In addition, up to 18 % of the applied maleic hydrazide became associated with nonextractable residues (NER) in soybean, whereas in tobacco only 0.2 % could be detected in this fraction. The residue from soybean cells was solubilized only to a low degree (about 3 %) under simulated stomach conditions, but up to 20 % by the white rot fungus Phanerochaetechrysosporium.
For the first time, the role(s) of selected amino acids and sugars in the production of volatile compounds in microwave prepared heat treated tobacco suspensions (HTTS) has been unambiguously defined. The role can be expressed in two major reaction pathways:
1) Strecker degradation of amino acids and 2) amino acid/sugar Maillard-type reactions. The mechanism of the Strecker degradation was confirmed by the addition of selected 13C- labelled amino acids to the reaction suspension. The addition of the amino acid resulted in a dramatic increase in the concentration of the specific, appropriate, low molecular weight Strecker aldehyde in the headspace above the reaction suspension. The mechanism of the amino acid/sugar Maillard-type reactions was substantiated by: 1) adding a wide array of amino acids to the suspension;
2) by employing 15N-labelled amino acids; and
3) employing 13C-labelled sugars. Addition of amino acids to the suspension followed by heat treatment resulted in significant increases (2 ×) in the concentration of volatile pyrazines relative to the control. Most amino acids significantly increased the concentration of headspace pyrazines. Reactivity differences, as a function of sugars, was also demonstrated. Analysis of the headspace above suspensions incorporating 15N-labelled amino acids and 13C-labelled sugars revealed the 15N and 13C atoms to have been distributed throughout a wide array of volatile pyrazines. The distribution of the 15N atoms within the pyrazines was unique to the amino acid and the distribution of the 13C atoms was linked to the type of sugar employed. These results have documented for the first time in a heat treated tobacco formulation the mechanisms of amino acids and sugars in the production of volatile compounds which have documented intense sensory characteristics.
Smoking machines are mechanical devices for the standardized smoking of cigarettes, facilitating the determination of nicotine and condensate yields of cigarettes according to strictly controlled parameters such as puff duration, puff volume, number of puffs and butt length. The development of smoking machines dates back some 50 years. The first - a four channel smoking machine - was built by Bradford, Harlan and Hammer in 1936.
The metabolism of the plant growth regulator maleic hydrazide (MH) has been studied in sterile cell suspension cultures of tobacco, soybean, maize and wheat under standardized conditions. Maleic hydrazide was converted to its O-b -D-glucoside in yields between 9.0 % (tobacco) 15.0 % (soybean), 5.1 % (maize) and 2.2 % (wheat) respectively. This glucoside was completely cleaved under simulated conditions (pH 1, 37°C, 24 h) of a ruminant stomach. From these results it is concluded that MH-O-b-D-glucoside (MHG) belongs to a small group of acid-labile pesticide conjugates (11). The participation of a glucosyltransferase (GT) (EC 2.4.1-) in this conjugation reaction of MH was demonstrated in vitro for the first time. In addition, up to 18 % of the applied maleic hydrazide became associated with nonextractable residues (NER) in soybean, whereas in tobacco only 0.2 % could be detected in this fraction. The residue from soybean cells was solubilized only to a low degree (about 3 %) under simulated stomach conditions, but up to 20 % by the white rot fungus Phanerochaetechrysosporium.
For the first time, the role(s) of selected amino acids and sugars in the production of volatile compounds in microwave prepared heat treated tobacco suspensions (HTTS) has been unambiguously defined. The role can be expressed in two major reaction pathways:
1) Strecker degradation of amino acids and 2) amino acid/sugar Maillard-type reactions. The mechanism of the Strecker degradation was confirmed by the addition of selected 13C- labelled amino acids to the reaction suspension. The addition of the amino acid resulted in a dramatic increase in the concentration of the specific, appropriate, low molecular weight Strecker aldehyde in the headspace above the reaction suspension. The mechanism of the amino acid/sugar Maillard-type reactions was substantiated by: 1) adding a wide array of amino acids to the suspension;
2) by employing 15N-labelled amino acids; and
3) employing 13C-labelled sugars. Addition of amino acids to the suspension followed by heat treatment resulted in significant increases (2 ×) in the concentration of volatile pyrazines relative to the control. Most amino acids significantly increased the concentration of headspace pyrazines. Reactivity differences, as a function of sugars, was also demonstrated. Analysis of the headspace above suspensions incorporating 15N-labelled amino acids and 13C-labelled sugars revealed the 15N and 13C atoms to have been distributed throughout a wide array of volatile pyrazines. The distribution of the 15N atoms within the pyrazines was unique to the amino acid and the distribution of the 13C atoms was linked to the type of sugar employed. These results have documented for the first time in a heat treated tobacco formulation the mechanisms of amino acids and sugars in the production of volatile compounds which have documented intense sensory characteristics.
Smoking machines are mechanical devices for the standardized smoking of cigarettes, facilitating the determination of nicotine and condensate yields of cigarettes according to strictly controlled parameters such as puff duration, puff volume, number of puffs and butt length. The development of smoking machines dates back some 50 years. The first - a four channel smoking machine - was built by Bradford, Harlan and Hammer in 1936.