A more accurate estimation of free-base nicotine in moist snuff
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1 A more accurate estimation of free-base nicotine in moist snuff Mikael Staaf Fredrik Hodin Jörgen Lundin October,
2 Objective The term free-base nicotine (FBN) refers to the unprotonated form of nicotine which is readily available for uptake through the mucous membranes. The concept of FBN is commonly used in the research literature and mandated by some regulators. There is limited evidence for the relevance of FBN estimated with current approach to human physiology. The presentation will show another approach for estimating FBN that may be more relevant compared to the current method October,
3 Estimations of FBN using the Henderson-Hasselbalch equation ph = pka + log([b]/[bh + ]) (1) [B]/[BH + ] = 10 (ph-pka) (2) %FBN = ([B]/[BH + ] 100 / ([B]/[BH + ] + 1) (3) Estimations of FBN commonly use: ph-value of an aqueous extract Logarithmic constant (pk a ) for nicotine at 25 C and infinitely dilute solution ( 8.02) October,
4 What is questionable with this approach? Temperature saliva temperatures are generally around 35 C pk a value salt are added to moist snuff products Sample preparation change of product integrity October,
5 Temperature and ionic strength The pk a of nicotine decreases with increasing temperature 1 The ph is affected by temperature 2 The pk a increases with increasing ionic strength 3 1 González et al., 1980; Fowler, Ciolino at al., González et al., 1980; Lauterbach et al., October,
6 Alternative approach ph and conductivity are measured in snuff liquid at 35 C Ionic strength is calculated from the conductivity pk a is corrected for Ionic strength using titration of a model liquid October,
7 Ionic strength (M) ph, conductivity and ionic strength of moist snuff liquid 1) Liquid preparation using a garlic press 2) Measurement of ph and conductivity at 35 C 3) Calculation of ionic strength using standard curve for conductivity vs ionic strength of NaCl solutions October, R² = Conductivity (ms/cm)
8 Determination of corrected pk a Titration of model liquids Titration of 0.05 M Nicotine in NaCl solutions of different ionic strengths: 0 M, 0.5 M, 1.0 M, 1.5 M and 2 M Titrand solution were adjusted to ph 11 using NaOH Titrant of HCl with the corresponding ionic strength Titrations were performed at 35 C October,
9 V H C l Determination of corrected pk a Estimation of pk a2 using two methods 1) Displacement of the HHE to fit the titration curve. 2) Curve fitting of the titration data using a 1 5 Nicotine in 1 M NaCl solution sigmoidal equation October, p H LogEC Std. Error R Curve fitting with sigmoidal equation
10 pka Determination of corrected pk a Standard curve for pk a vs ionic strength y = x x R² = Ionic strength (M) October, Our data González González 1 Our data I (M) pka pka * * * 8.34 * Extrapolated values 1 González et al., 1980
11 Results Current method Alternative method Product name ph FBN (%) ph (35C) I (M) pka FBN (%) General Classic Portion % % General Classic Portion Extra Strong % % Ettan Orignal Portion % % Grov Portion % % Göteborgs Rapé White Large % % LD Original Portion % % Skruf Stark White % % Granit Portion % % Copenhagen Natural Pouches % % Skoal Wintergreen Pouches % % Grizzly Wintergreen Pouches % % Timber Wolf Pouches Natural % % October,
12 Results Current method Alternative method Product name ph FBN (%) ph (35C) I (M) pka FBN (%) General Classic Portion % % General Classic Portion Extra Strong % % Ettan Orignal Portion % % Grov Portion % % Göteborgs Rapé White Large % % LD Original Portion % % Skruf Stark White % % Granit Portion % % Copenhagen Natural Pouches % % Skoal Wintergreen Pouches % % Grizzly Wintergreen Pouches % % Timber Wolf Pouches Natural % % October,
13 Results Current method Alternative method Product name ph FBN (%) ph (35C) I (M) pka FBN (%) General Classic Portion % % General Classic Portion Extra Strong % % Ettan Orignal Portion % % Grov Portion % % Göteborgs Rapé White Large % % LD Original Portion % % Skruf Stark White % % Granit Portion % % Copenhagen Natural Pouches % % Skoal Wintergreen Pouches % % Grizzly Wintergreen Pouches % % Timber Wolf Pouches Natural % % October,
14 Discussion The alternative method for estimating FBN should be more relevant compared to the current approach. To fully assess the potential for nicotine uptake the FBN estimate require complementary addition of other parameters: Change of ph and ionic strength during use Product specific factors (e.g., pouch size and shape) Clinical studies are required to confirm if the alternative method provides better correlation to real nicotine uptake compared to the current method October, Contact:
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