Figure 1. Typical structure of polysorbate 80.

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Figure 1. Typical structure of polysorbate 80. From: Analysis of Polysorbate 80 and its Related Compounds by RP-HPLC with ELSD and MS Detection J Chromatogr Sci. 2012;50(7):598-607. doi:10.1093/chromsci/bms035 J Chromatogr Sci | © The Author [2012]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com

Figure 2. Polysorbate 80 synthetic routes and possible products. From: Analysis of Polysorbate 80 and its Related Compounds by RP-HPLC with ELSD and MS Detection J Chromatogr Sci. 2012;50(7):598-607. doi:10.1093/chromsci/bms035 J Chromatogr Sci | © The Author [2012]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com

Figure 3. Chromatogram obtained from polysorbate 80 sample (Well, batch number 20080412): mobile phase: 50% CH<sub>3</sub>CN–water (mobile phase A)–THF (mobile phase B); gradient elution (0–80% B in 40 min) (A); mobile phase: 50% CH<sub>3</sub>CN–water (mobile phase A)–CH<sub>3</sub>CN (mobile phase B) gradient elution (0–100% B, in 40 min) (B). From: Analysis of Polysorbate 80 and its Related Compounds by RP-HPLC with ELSD and MS Detection J Chromatogr Sci. 2012;50(7):598-607. doi:10.1093/chromsci/bms035 J Chromatogr Sci | © The Author [2012]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com

Figure 4. LC–ELSD chromatograms of polysorbate samples of standard grade, batch number 20070408 from Well (A); standard grade, batch number 20091018 from Well (B); standard grade, batch number 20100516 from Well (C); high purity grade, batch number 20080412 from Well (D); high purity grade, batch number 20080418 from Well (E); high purity grade, batch number 20101019 (F); high purity grade, batch number 807367D from NOF (G); standard grade, batch number 0000285529 from CRODA (H); ACN (I). From: Analysis of Polysorbate 80 and its Related Compounds by RP-HPLC with ELSD and MS Detection J Chromatogr Sci. 2012;50(7):598-607. doi:10.1093/chromsci/bms035 J Chromatogr Sci | © The Author [2012]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com

Figure 5. TIC traces obtained from polysorbate 80 sample (Well, batch number 20080412). From: Analysis of Polysorbate 80 and its Related Compounds by RP-HPLC with ELSD and MS Detection J Chromatogr Sci. 2012;50(7):598-607. doi:10.1093/chromsci/bms035 J Chromatogr Sci | © The Author [2012]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com

Figure 6. ESI-MS spectra of representative peaks in polysorbate 80: Peak A (A); peak B (B); Peak C (C); Peak D (D); Peak E (E); Peak F (F); Peak G (G). From: Analysis of Polysorbate 80 and its Related Compounds by RP-HPLC with ELSD and MS Detection J Chromatogr Sci. 2012;50(7):598-607. doi:10.1093/chromsci/bms035 J Chromatogr Sci | © The Author [2012]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com

Figure 6. ESI-MS spectra of representative peaks in polysorbate 80: Peak A (A); peak B (B); Peak C (C); Peak D (D); Peak E (E); Peak F (F); Peak G (G). From: Analysis of Polysorbate 80 and its Related Compounds by RP-HPLC with ELSD and MS Detection J Chromatogr Sci. 2012;50(7):598-607. doi:10.1093/chromsci/bms035 J Chromatogr Sci | © The Author [2012]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com

Figure 6. ESI-MS spectra of representative peaks in polysorbate 80: Peak A (A); peak B (B); Peak C (C); Peak D (D); Peak E (E); Peak F (F); Peak G (G). From: Analysis of Polysorbate 80 and its Related Compounds by RP-HPLC with ELSD and MS Detection J Chromatogr Sci. 2012;50(7):598-607. doi:10.1093/chromsci/bms035 J Chromatogr Sci | © The Author [2012]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com

Figure 6. ESI-MS spectra of representative peaks in polysorbate 80: Peak A (A); peak B (B); Peak C (C); Peak D (D); Peak E (E); Peak F (F); Peak G (G). From: Analysis of Polysorbate 80 and its Related Compounds by RP-HPLC with ELSD and MS Detection J Chromatogr Sci. 2012;50(7):598-607. doi:10.1093/chromsci/bms035 J Chromatogr Sci | © The Author [2012]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com