Altria Group, Inc. · NYSE:MO
Altria — Technical Presentation (2019)
19 slides from Altria's technical presentation, 2019
Slide 1: Optimization of DNPH derivatization conditions for carbonyl compounds in e-vapor products
Optimization of 2,4- dinitrophenylhydrazine (DNPH) derivatization conditions for the determination of carbonyl compounds in e-vapor products Lena Jeong, John Miller, Niti Shah Altria Client Services I Lena Jeong | Postdoctoral Fellow l Tobacco Science Research Conference, 2019 I 1
Slide 2: Background
Background ▪ FDA requires reporting of carbonyls in e-vapor products*: ▪ No CRM available for measuring carbonyls in e-vapor products - CRM 74: mainstream cigarette smoke - CRM 86: tobacco and tobacco products ▪ Carbonyl compounds react with 2,4-dinitrophenylhydrazine (DNPH) the presence of an acidi
Slide 3: Current Methods
Current Methods DNPH conc. Acid Diluent CRM 74 11.65 mM 2.05 M phosphoric acid 50/50 ACN/H2O Altria 17.5 mM 1.82 M perchloric acid ACN (published) Challenges with current methods: • High background for formaldehyde in current DNPH • Low and unstable recovery for acrolein J.W. Flora et al., Method fo
Slide 4: 1. Formaldehyde Contamination in DNPH
1. Formaldehyde Contamination in DNPH DNPH ~30% H2O High background signals for Formaldehdye • Problematic for low level quantitation • Often requires recrystallization • Lot-to-lot variation in background levels 0.12 µg/puff • Limited availability (issue for high volume testing) Alternate DNPH type
Slide 5: 1. Formaldehyde Contamination in DNPH
1. Formaldehyde Contamination in DNPH DNPH ~30% H2O DNPH-HCl 0.12 µg/puff < 0.01 µg/puff Switching to HCl salt form dramatically reduced background carbonyl levels Altria Client Services I Lena Jeong | Postdoctoral Fellow l Tobacco Science Research Conference, 2019 I 5
Slide 6: 2. Low and Unstable Acrolein Recovery
2. Low and Unstable Acrolein Recovery CRM 74 Altria published 100 Desired recovery above 80% 80 Acrolein recovery (%) 60 Unstable acrolein-DNPH complex 40 20 Matrix: E-Liq pH 9.36 n=3 0 0 10 20 30 40 50 60 70 80 90 100 Time (min) J.W. Flora et al., Method for the Determination of Carbonyl Compounds
Slide 7: Investigation into Low Acrolein Recovery
Investigation into Low Acrolein Recovery Polyderivatization of Acrolein* Polyderivatization increases under strong acidic conditions *S. Uchiyama, Y. Inaba, N. Kunugita, J. Chromatogr. A, 1217 (2010) 4383-4388. doi: 10.1016/j.chroma.2010.04.056 Altria Client Services I Lena Jeong | Postdoctoral Fell
Slide 8: Method Optimization
Method Optimization ▪ Need to optimize method to: - Reduce formaldehyde background using new DNPH-HCl - Obtain higher and more stable acrolein recovery ▪ Evaluate preparation of DNPH solution: - Acid type/concentration - DNPH concentration - Solvent ratio Altria Client Services I Lena Jeong | Postdo
Slide 9: Derivatization Optimization
Derivatization Optimization Evaluation of pH and DNPH concentration ▪ Concentration of DNPH (9 mM; 4.5 mM; 1.8 mM) in ACN solution prepared with 1.5 % (v/v) of - 1.82 M perchloric acid pH 0.04 - 0.1 M sodium citrate buffer pH 3 - 0.1 M sodium citrate buffer pH 6 Literature reports that acidity of de
Slide 10: Effect of pH
Effect of pH 99mM mMTCI DNPH HCl;pH DNPH; pH0.04 0.04 99mM mMTCI DNPH; DNPH HCl;pH3 pH 3 99mM mMTCI DNPH; DNPH HCl;pH6 pH 6 100 Acrolein recovery (%) 80 60 40 20 Matrix: Water n=3 0 0 10 20 30 40 50 60 70 80 90 100 Time (min) Low pH results in decomposition of acrolein-DNPH complex Altria Client Ser
Slide 11: Effect of DNPH Concentration
Effect of DNPH Concentration 99mM mMTCI DNPH; DNPH pH33 HCl; pH 4.5 4.5mM mM TCI DNPHDNPH; pH33 HCl; pH 1.8 1.8mM mMTCI DNPH; DNPH pH33 HCl; pH 100 Acrolein recovery (%) 80 60 40 20 Matrix: Water n=3 0 0 20 40 60 80 100 Time (min) DNPH concentration is directly related to the derivatization rate Alt
Slide 12: Derivatization Optimization: Effect of Water Content
Derivatization Optimization Effect of Water Content DNPH conc. Acid Diluent CRM 74 11.65 mM 2.05 M phosphoric acid 50/50 ACN/H2O Altria 17.5 mM 1.82 M perchloric acid ACN (published) ▪ How is derivatization rate affected in presence of added water? Varying diluent ratios tested: - 0/100 H2O/ACN - 25
Slide 13: Water Content Comparison
Water Content Comparison 99 mM mM TCI DNPHDNPH; HCl;pH6; pH 6;ACN ACN DNPH 9 mM TCI 9 mM HCl;TCI DNPH; pH6;DNPH; pH 6; 25/75pH6; 25/75 ACN H2O/ACN H2O/ACN 9 mM mM TCI DNPHDNPH; HCl; pH6; pH 6;50/50 50/50H2O/ACN H2O/ACN 100 80 Acrolein recovery (%) 60 40 20 Matrix: Water n=3 0 0 10 20 30 40 50 60 70
Slide 14: Optimized Method vs. CRM
Optimized Method vs. CRM 74 99mM mMTCI DNPH; DNPH CRM HCl;pH 74 pH3;3;ACN ACN CRM 74 100 80 Acrolein recovery (%) 60 40 20 Derivatization too slow. pH shifting with sample addition. n=3 0 0 10 20 30 40 50 60 70 80 90 100 Time (min) Sample matrix: 50/50 PG/GLY with 2.5% nicotine (pH 9.36) Altria Clie
Slide 15: Comparison of Methods: Acrolein Recovery
Comparison of Methods: Acrolein Recovery Optimized Method Method Optimized CRMMethod Optimized 74 Altria published Altria published 100 Needed buffer capacity achieved! 80 Acrolein recovery (%) 60 DNPH conc. Acid (type; %; Diluent (vendor) conc.) 40 CORESTA 11.65 mM 2.9% of 2.05 M 50/50 (No. 74) pho
Slide 16: New Method Performance with Varying Sample pH
New Method Performance with Varying Sample pH Water Basic e-liq Acidic e-liq 100 Acrolein recovery (%) 80 60 40 20 n=3 0 0 20 40 60 80 100 Time (min) Basic e-liquid: 50/50 PG/GLY with 2.5% nicotine (pH 9.36) Acidic e-liquid: 50/50 PG/GLY with benzoic acid (pH 3.72) Altria Client Services I Lena Jeon
Slide 17: Comparison of Methods: Percent Recovery
Comparison of Methods: Percent Recovery Derivatization time: 10 min n=3 Formaldehyde Acetaldehyde Acrolein Crotonaldehyde CRM 74 100% ± 2.8% 78.7% ± 0.91% 74.5% ± 1.9% 100% ± 2.4% Altria published 87.2% ± 1.3% 77.7% ± 2.6% 72.5% ± 2.3% 103 ± 2.0% Optimized method 102% ± 0.84% 80.7% ± 2.9% 85.6% ± 2.
Slide 18: Summary - Learnings
Summary - Learnings ▪ Switching to DNPH HCl form dramatically reduced background levels of formaldehyde ▪ Highly acidic DNPH solution results in polyderivatization of acrolein-DNPH (formation of AD1) ▪ Use of buffer to control the pH improves and stablizes acrolein recovery for over 90 min derivatiz
Slide 19: Conclusions
Conclusions ▪ DNPH-HCl form reduces background levels of formaldehyde and improves quantitation of carbonyls in e-vapor aerosol ▪ The DNPH derivatization method was optimized to give acceptable recovery levels for all aldehydes including acrolein ▪ New conditions allow for better stability of acrole