Altria Group, Inc. · NYSE:MO

Altria — Technical Presentation (October 2019)

21 slides from Altria's technical presentation, Oct 9, 2019

  1. Slide 1: Optimized Method for Determination of Selected Phenolic Compounds in Cigarette and Cigar Smoke by UPLC-FLD

    Optimized Method for Determination of Selected Phenolic Compounds in Cigarette and Cigar Smoke by UPLC-FLD Xiaohong Cathy Jin, Thomas J. Hurst and Karl A. Wagner CORESTA SSPT CONFERENCE OCT. 6th-10th 2019 Xiaohong Cathy Jin l Regulatory Affairs l Altria Client Services l CORESTA Oct 6th-10th l Final

  2. Slide 2: Introduction

    Introduction  Selected phenolic compounds are included in the FDA “Established List of HPHCs in Tobacco Products and Tobacco Smoke1”.  Analytical methods for phenols in tobacco and cigarette smoke: - Wu, J. et al. reported an HPLC-FLD method in 20122 - CORESTA recommended method 78 (CRM 78), 34 mi

  3. Slide 3: Objective

    Objective  To develop a rapid method suitable for analysis of phenols in both cigarette and cigar smoke using Ultra-High Pressure Liquid Chromatography (UPLC) with Fluorescence detector (FLD) Xiaohong Cathy Jin l Regulatory Affairs l Altria Client Services l CORESTA Oct 6th-10th l Final l 3

  4. Slide 4: Selected phenolic compounds

    Selected phenolic compounds OH OH OH OH OH Catechol* Phenol* m-Cresol* OH OH Hydroquinone OH OH * Included in FDA’s list of HPHCs OH o-Cresol* p-Cresol* Resorcinol 4 Xiaohong Cathy Jin l Regulatory Affairs l Altria Client Services l CORESTA Oct 6th-10th l Final l

  5. Slide 5: Experimental – Smoke collections

    Experimental – Smoke collections  Cigarette or cigar smoke was collected on a Cambridge filter pad (CFP) per smoking regime Puff Puff Puff Samples Smoking regime volume frequency duration (mL) (sec) (sec) Cigarette ISO (ISO 3308) 35 60 2 Intense (ISO 20778) 55 30 2 Cigar CORESTA1 202 40 1.5 1. CORE

  6. Slide 6: Experimental – Sample preparations

    Experimental – Sample preparations • Collect smoke sample on a CFP 1 • Transfer pad to an amber vial • Add 40 mL of extraction solution (1% acetic acid) • Shake for 30 min at 350 RPM 2 • Filter extract through 0.2 µm PVDF syringe filter • Inject 2 µL into UPLC-FLD 3 Xiaohong Cathy Jin l Regulatory A

  7. Slide 7: Column selection

    Column selection -Chromatograms of standard 2 phenol hydroqu catech 100 Acquity UPLC®HSS m-cresol p-cresol PFP*, 1.8 µm, o-cresol 2.1 x 100 mm resor % min 1.00 2.00 3.00 4.00 5.00 6.00 7.00 100 phenol hydroqu Kinetex® Core-shell catech m-cresol F5, 1.7µm, o-cresol p-cresol 2.1 x 100 mm resor % min 1

  8. Slide 8: Analytical method – Parameters

    Analytical method -Parameters  Acquity I-Class UPLC system: binary pump, temperature controlled autosampler, fluorescence detector  Kinetex® Core-shell column F5, 1.7µm, 100 x 2.1mm  Autosampler temperature: 5 oC  Injection volume: 2 µL  Mobile phase, gradient elution (flow rate 0.35 mL/min) -

  9. Slide 9: Analytical method – Instrument settings

    Analytical method -Instrument settings Mobile phase gradients 100 95 78 composition (%) Mobile phase 80 70 60 A (%) B (%) 40 30 20 min 0 5 0 2 4 6 8 10 12 Detector settings Excitation Emission Time (min) Analytes wavelength (nm) wavelength (nm) Hydroquinone, resorcinol, 0.0 - 2.2 280 310 catechol 2.

  10. Slide 10: Representative chromatograms of smoke samples

    Representative chromatograms of smoke samples catech hydroqu 100 1R6F ISO phenol o-cresol p-cresol m-cresol resor % -0 min 1.00 2.00 3.00 4.00 5.00 6.00 7.00 catech hydroqu 100 phenol Cigar p-cresol resor m-cresol o-cresol % min -0 1.00 2.00 3.00 4.00 5.00 6.00 7.00 Xiaohong Cathy Jin l Regulatory A

  11. Slide 11: Method validation – Calibration

    Method validation -Calibration Standards Conc. in smoke* Elements Analytes (µg/mL) (µg per cig. or cigar) 0.4 - 160 (cig. ISO) Hydroquinone, 0.05 - 20 0.7 - 267 (cig. Intense) phenol, catechol 2 - 800 (cigar) Calibration ranges Resorcinol, 0.08 - 32 (cig. ISO) p-cresol, m- 0.01 - 4 0.13 - 53 (cig In

  12. Slide 12: Method validation – Precision

    Method validation -Precision1 TPM 3 days Phenols intra- Phenols inter- Smoking Samples average day precision day precision regime (mg/cig) (%RSD) (%RSD) ISO 9.6 3.1 - 9.8 2.0 - 8.1 3R4F Intense 48.5 4.2 - 10.2 2.8 - 13.9 ISO 10.4 2.4 - 6.5 4.1 - 6.8 1R6F Intense 48.0 2.3 - 10.4 3.3 - 8.9 CORESTA mon

  13. Slide 13: Method validation – Phenols yields in validation samples

    Method validation -Phenols yields in validation samples (n=18 for cig. and n=12 for cigar) 250 3R4F ISO 3R4F Intense Phenols conc. (µg/cig. or cigar) 200 1R6F ISO 150 1R6F Intense CM7 ISO CM7 Intense 100 Cigar 50 0 Hydroqu. Resor. Catech. Phenol p-Cresol m-Cresol o-Cresol Xiaohong Cathy Jin l Regula

  14. Slide 14: Method validation – Recovery

    Method validation -Recovery (2 fortification levels at 0.5x & 2x of native analytes conc., n=3) Fortification level 1R6F ISO CM 7 ISO Cigar Low level fortification 74% - 96% 86% - 95% 75% - 83% ~0.5X of native conc. High level fortification 75% - 96% 88% - 97% 77% - 97% ~2X of native conc. Xiaohong

  15. Slide 15: Compared to CRM – Chromatograms of 3R4F ISO sample

    Compared to CRM 78 -Chromatograms of 3R4F ISO sample hydroqu catech 100 UPLC phenol p-cresol m-cresol resor o-cresol % min 0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 catech 100 hydroqu phenol CRM 78 p-cresol m-cresol resor o-cresol % 0.0 5.0 10.0 15.0 20.0 min  UPLC method is 3 times faster than CRM 78. Xiaoh

  16. Slide 16: Compared to CRM78 – Calibration ranges

    Compared to CRM78 -Calibration ranges unit UPLC CRM 78 Hydroquinone, µg/mL 0.05 - 20 0.2 - 8 catechol, phenol Resorcinol µg/mL 0.01 - 4 0.04 - 1.6 p-cresol, m- µg/mL 0.01 - 4 0.06 - 2.4 cresol, o-cresol  UPLC method has wider dynamic calibration ranges than CRM 78. Xiaohong Cathy Jin l Regulatory A

  17. Slide 17: Compared to CRM78 – Phenols yields in 3R4F and CM smoke

    Compared to CRM78 -Phenols yields in 3R4F and CM 7 smoke 3R4F intense 50 3R4F ISO 120 100 Conc. (µg/cig.) 40 UPLC Conc. (µg/cig.) UPLC CRM 78 80 30 CRM 78 60 20 40 10 20 0 0 Hydroqu. Resor. Catech. Phenol p-Cresol m-Cresol o-Cresol Hydroqu. Resor. Catech. Phenol p-Cresol m-Cresol o-Cresol CM7 ISO CM

  18. Slide 18: Phenols yields in market cigarettes ISO smoke

    Phenols yields in market cigarettes ISO smoke -Commercially marketed American blended cigarettes (n=4) 120 25 A B C D E F 100 20 Phenols conc. (µg/cig.) 80 TPM (mg/cig.) 15 60 10 40 5 20 0 0 Hydroqu. Resor. Catech. Phenol p-Cresol m-Cresol o-Cresol TPM Xiaohong Cathy Jin l Regulatory Affairs l Altri

  19. Slide 19: Phenols yields in machine-made cigar smoke

    Phenols yields in machine-made cigar smoke -Machine made cigars vs. 3R4F 350 3R4F ISO 3R4F Intense G H I J K L Phenols conc. (µg/cig. or cigar) 300 250 200 150 100 GH I J K L 50 0 Hydroqu. Resor. Catech. Phenol p-Cresol m-Cresol o-Cresol  Phenols yields and variability higher in cigar smoke (%RSD:2

  20. Slide 20: Conclusions

    Conclusions  The UPLC-FLD method met the acceptable validation requirements.  The UPLC method is 3 times faster and has wider dynamic calibration ranges compared to CRM 78.  Phenols yields per stick in cigar smoke were higher and more variable than in cigarette smoke. The UPLC-FLD method was foun

  21. Slide 21: Questions?

    Thank You Questions? For copies of this presentation visit the Altria’s Science Website at www.altria.com/alcs-science Altria has a new Altria Science Twitter account Follow us @AltriaScience Xiaohong Cathy Jin l Regulatory Affairs l Altria Client Services l CORESTA Oct 6th-10th l Final l 21

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