TN-2094 -Optimal Separation of PAH Compounds Including

Home , Triphenylene

TN-2094 APPLICATIONS Ramkumar Dhandapani, Ph.D. TN-2094 Optimal Separation of PAH Compounds including Product Manager - Gas Chromatography ™ He has a PhD in Analytical Chemistry and Chrysene & Triphenylene using Zebron ZB-PAH-CT over 15 years experience in chromatographic method development and troubleshooting. GC column Ramkumar loves to write poems, read Ramkumar Dhandapani and Rola Elabaji Shakespeare, and attend Shakespeare plays. Phenomenex, Inc., 411 Madrid Ave., Torrance, CA 90501 USA

Introduction Polycyclic aromatic hydrocarbons (PAHs) are carcinogenic sub- Figure 1. stances that must be monitored in food and environment as per Master Resolution Equation local regulations. There are numerous PAH compounds that have structural similarities including Chrysene and Triphenylene that are hard to resolve on a traditional GC column. Chromatographically, there are 3 components that contributes to improve resolution of peaks. It comprises of Efficiency term, Selectivity term, and Reten- tion term. Among the three terms, Selectivity is most influential in drastic resolution improvement. For optimal resolution of Chrysene and Triphenylene, several combinations of stationary phases were explored. Presented in this study is an optimal stationary phase Ze- Efﬁciency Selectivity Retention bron ZB-PAH-CT that was utilized for separation of EU 15+1 PAHs along with Triphenylene to evaluate overall separation of PAH and separation of critical pairs Chrysene and Triphenylene. N = theoretical plates α = k2/k1 k = (tR-t0)/t0

Figure 2. EU 15+1 PAH and Triphenylene on a Zebron ZB-PAH-CT GC column

Complete Separation of Triphenylene and Chrysene Separation of Benzo[b,j,k]

5 3 4 9 12 7 8 11 2

40 41 20 21 22 29

4 5 12 3

13 1 11 2 9 8 15 16 17 6 7 14 10 App ID 25715

15 20 25 30 35 40 45 50 min

Sample: Column: Zebron ZB-PAH-CT 1. Benzo[c]fluorene 10. Benzo[a]pyrene Dimensions: 40 meter x 0.18 mm x 0.14 µm 2. Cyclopenta[c,d]pyrene 11. Indeno[1,2,3-c,d]pyrene Part No.: 7PD-G044-47 3. Benz[a]anthracene 12. Dibenz[a,h]anthracene Injection: Split 30:1 @ 320 °C, 1 μL 4. Triphenylene 13. Benzo[g,h,i]perylene ™ Recommended Liner: Zebron PLUS Single Taper Z-Liner 5. Chrysene 14. Dibenzo[a,l]pyrene ® Liner Part No.: AG2-4B13-05 (for Shimadzu 2010 GC) 6. 5-MethylChrysene 15. Dibenzo[a,e]pyrene Carrier Gas: Helium @ 78 psi (constant pressure) 7. Benzo[b]fluoranthene 16. Dibenzo[a,i]pyrene Oven Program: 45 °C for 0.8 min to 200 °C @ 45 °C/min to 265 °C @ 3 °C/ min for 8. Benzo[j]fluoranthene 17. Dibenzo[a,h]pyrene 5 min to 270 °C @ 1 °C/min to 320 °C @ 10 °C/min for 15 min 9. Benzo[k]fluoranthene Detector: MSD (Shimadzu® GC-MS-QP2010 Ultra) Mode: SIM SIM Ions: 216, 226, 228, 242, 252, 276, 278, 302 m/z Transfer line Temperature: 300 °C Source Temperature: 300 °C

Having trouble reproducing this method? We would love to help! Visit www.phenomenex.com/LiveChat to get in touch with one of our Technical Specialists Page 1 of 4 1.25 6 7 8 1.00

0.75 TN-2094 0.50 0.25

25 25.1 25.2 25.3 25.4 25.5 25.6 25.7 25.8 25.9 26 26.1 26.2

1.25 6 7 8 1.00 4 2 3 0.75 0.50 Figure 3. 0.25 EU 15+1 PAH on a Zebron ZB-PAH-EU GC column 0

25 25.1 25.2 25.3 25.4 25.5 25.6 25.7 25.8 25.9 26 26.1 26.2

17.8 17.9 18 18.1 18.2 18.3 18.4 18.5 18.6 18.7 18.8 18.9 19 19.1

11 1.25 4 10 2 7 3 6 8 1.00 1.5 0.75

1 0.50

0.25 0.5 0 0

17.8 17.9 18 18.1 18.2 18.3 18.4 18.5 18.629 18.7 25 18.8 25.129.518.9 25.2 19 25.3 19.130 25.4 25.5 30.525.6 25.7 25.831 25.9 26 31.526.1 31.7526.2 32

11 10 4 11 1.5 2 12 3 10 1

0.5 1.5

0 1 4 29 29.5 30 30.5 31 31.5 31.75 32 9 13 2 5 7 14 3 17.8 17.9 18 18.1 618.28 18.3 18.4 18.5 18.6 18.7 18.8 18.9 19 19.1 1.0 15 16 11 10

1.5 0.5 1 Relative Abundance App ID 25708 0.5

0 0 29 29.5 30 30.5 31 31.5 31.75 32

15 20 25 30 35 40 45 50 min

Column: Zebron ZB-PAH-EU Sample: Dimensions: 30 meter x 0.25 mm x 0.20 µm 1. Benzo[c]fluorene Part No.: 7HG-G043-10 2. Benz[a]anthracene Injection: Split 5:1 @ 330 °C, 1 μL 3. Cyclopenta[c,d]pyrene Recommended Liner: Zebron PLUS Single Taper Z-Liner™ 4. Chrysene Liner Part No.: AG2-4B13-05 (for Shimadzu® 2010 GC) 5. 5-MethylChrysene Carrier Gas: Helium @ 24 psi (constant pressure) 6. Benzo[b]fluoranthene Oven Program: 45 °C for 0.8 min to 200 °C @ 45 °C/min to 7. Benzo[k]fluoranthene 226 °C @ 3 °C/min for 0 min to 320 °C @ 8. Benzo[j]fluoranthene 10 °C/min for 20 min 9. Benzo[a]pyrene Detector: MSD, 50-500 m/z 10. Indeno[1,2,3-cd]pyrene Transfer line Temperature: 300 °C 11. Dibenz[a,h]anthracene Source Temperature: 300 °C 12. Benzo[g,h,i]perylene 13. Dibenzo[a,l]pyrene 14. Dibenzo[a,e]pyrene 15. Dibenzo[a,i]pyrene 16. Dibenzo[a,h]pyrene

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Figure 4. Comparison of Analysis of EU 15+1 PAHs and Triphenylene

3 5 4

2 Great Resolution of Chrysene and Triphenylene

Zebron ZB-PAH-CT 30 meter x 0.25 mm x 0.20 µm

18 19 TIC 12 2.52,5

11 13 2.02,0 Better Sensitivity for Late Eluters 3 5 4 1.51,5

9 10 15 16 2 14 1 8 17 1,01.0 7 6

0,50.5 27 % Faster Analysis App ID 25753

10 15 20 25 30 35 40 45 50 min

GC-MS conditions for both applications: Column: As Indicated Sample: Dimensions: As Indicated 1. Benzo[c]fluorene Part No.: 7HG-G044-10 (Zebron ZB-PAH-CT) 2. Cyclopenta[c,d]pyrene Injection: Split 15:1 @ 320 °C, 1 μL 3. Benz[a]anthracene ™ Recommended Liner: Zebron PLUS Single Taper Z-Liner 4. Triphenylene Liner Part No.: AG2-4B13-05 5. Chrysene Carrier Gas: Helium @ 23.7 psi (constant pressure) 6. 5-MethylChrysene Oven Program: 45 °C for 0.8 min to 200 °C @ 45 °C/min to 7. Benzo[b]fluoranthene vs. 266 °C @ 3 °C/ min for 0 min to 320 °C @ 10 °C/ 8. Benzo[j]fluoranthene min to 320 °C for 20 min 9. Benzo[k]fluoranthene ® Detector: MSD (Shimadzu GC-MS-QP2010 Ultra) 10. Benzo[a]pyrene Mode: SIM 11. Indeno[1,2,3-c,d]pyrene SIM Ions: 216, 226, 228, 242, 252, 276, 278, 302 m/z 12. Dibenz[a,h]anthracene Transfer line Temperature: 300 °C 13. Benzo[g,h,i]perylene Source Temperature: 300 °C 14. Dibenzo[a,l]pyrene 15. Dibenzo[a,e]pyrene 16. Dibenzo[a,i]pyrene 17. Dibenzo[a,h]pyrene

4 5 3

2 Popular Brand A 30 meter x 0.25 mm x 0.15 μm

TIC 2.5 22 23 24 25

2.0

11,12 1.5 9 10 8 2, 4, 5 7 3 1.0 13 1 6 0.5

14 15 16 17 App ID 25717

10 15 20 25 30 35 40 45 50 min

Comparative separations may not be representative of all applications.

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Results and Discussion Australia The Netherlands t: Separation of PAH isomers are quite challenging as there are t: +61 (0)2-9428-6444 +31 (0)30-2418700 [email protected] [email protected] many PAH components that have structural similarities. Specifi- cally, Chrysene and Triphenylene are one such hard to separate Austria New Zealand t: +43 (0)1-319-1301 t: +64 (0)9-4780951 critical pairs. In the present study, unique selectivity of ZB-PAH- [email protected] [email protected] CT was utilized to resolve EU 15+1 and Triphenylene. As shown in Figure 2, the 40 meter x 0.18 mm x 0.14 µm ZB-PAH was able Belgium Norway t: +32 (0)2 503 4015 (French) t: +47 810 02 005 to completely resolve Chrysene from Triphenylene. In addition, t: +32 (0)2 511 8666 (Dutch) [email protected] ZB-PAH-CT also provided a peak order switch for certain PAH [email protected] analytes compared to ZB-PAH-EU, including Benz[a]anthracene, Poland Canada t: +48 22 104 21 72 Cyclopenta[c,d]pyrene (Peak 2 and 3 in Figure 2 and 3); Ben- t: +1 (800) 543-3681 [email protected] zo[b,j,k]fluoranthene (Peak 7, 8, 9 inFigure 2 and peak 6, 7, 8 in [email protected] Figure 3) due to the unique selectivity. The selectivity change and Portugal China t: +351 221 450 488 enhanced resolution of Chrysene and Triphenylene helps avoid t: +86 400-606-8099 [email protected] false positives in the analysis. In addition to resolving Chrysene [email protected] Singapore and Triphenylene completely, ZB-PAH-CT in a 40 meter dimension Denmark t: +65 800-852-3944 provided the highest resolution of 2.6 for Benzo[b,k]fluoranthene t: +45 4824 8048 [email protected] [email protected] Spain Further, EU 15+1 and Triphenylene mix was evaluated on a 30 Finland t: +34 91-413-8613 meter ZB-PAH-CT and 30 meter Popular Brand A GC column un- t: +358 (0)9 4789 0063 esp[email protected] der identical chromatographic conditions. As shown in Figure 4, [email protected] Sweden ZB-PAH-CT shows better resolution of Chrysene and Triphenylene, France t: +46 (0)8 611 6950 better sensitivity for heavy PAH, and 27% faster run time because t: +33 (0)1 30 09 21 10 [email protected] of the unique stationary phase selectivity. [email protected] Switzerland Germany t: +41 (0)61 692 20 20 t: +49 (0)6021-58830-0 [email protected] Conclusion [email protected] Zebron ZB-PAH-CT provides complete resolution of Chrysene Taiwan India t: +886 (0) 0801-49-1246 and Triphenylene in addition to other EU 15+1 PAHs with a 40 t: +91 (0)40-3012 2400 [email protected] meter column dimension. In addition to providing great resolu- [email protected] tion of Chrysene and Triphenylene, ZB-PAH-CT provides high United Kingdom Ireland t: +44 (0)1625-501367 resolution for Benzo[b,k]fluoranthene, gives better sensitivity for t: +353 (0)1 247 5405 [email protected] heavy PAH, and provides 27% faster run time than common PAH [email protected] GC columns. USA Italy t: +1 (310) 212-0555 t: +39 051 6327511 [email protected] [email protected] All other countries/regions Luxembourg Corporate Office USA t: +31 (0)30-2418700 t: +1 (310) 212-0555 Ordering Information [email protected] [email protected]

™ Mexico Zebron ZB-PAH-CT GC Column t: 01-800-844-5226 Length ID df [email protected] Temp. Limits (°C) Part No. (meter) (mm) (µm) 20 0.18 0.14 40 to 320/340 7FD-G044-47 30 0.25 0.20 40 to 320/340 7HG-G044-10 40 0.18 0.14 40 to 320/340 7PD-G044-47

www.phenomenex.com ™ Phenomenex products are available worldwide. For the distributor in your country/region, Zebron ZB-PAH-EU GC Column contact Phenomenex USA, International Department at [email protected] Length ID df Temp. Limits (°C) Part No. (meter) (mm) (µm) 10 0.10 0.08 40 to 340/360 7CB-G043-59 20 0.18 0.14 40 to 340/360 7FD-G043-47 30 0.25 0.20 40 to 340/360 7HG-G043-10 60 0.25 0.20 40 to 340/360 7KG-G043-10

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