{"id":43638,"date":"2020-10-28T14:30:00","date_gmt":"2020-10-28T14:30:00","guid":{"rendered":"https:\/\/discover.restek.com\/uncategorized\/the-effects-of-lc-particle-choice-on-column-performance-switching-from-3-and-5-micrometer-fully-porous-particles-fpp-to-5-micrometer-superficially-porous-particles-spp\/"},"modified":"2026-01-28T16:01:22","modified_gmt":"2026-01-28T16:01:22","slug":"the-effects-of-lc-particle-choice-on-column-performance-switching-from-3-and-5-micrometer-fully-porous-particles-fpp-to-5-micrometer-superficially-porous-particles-spp","status":"publish","type":"post","link":"https:\/\/discover.restek.com\/zh-hans\/articles-fr\/gnar2109\/the-effects-of-lc-particle-choice-on-column-performance-switching-from-3-and-5-micrometer-fully-porous-particles-fpp-to-5-micrometer-superficially-porous-particles-spp","title":{"rendered":"The Effects of LC Particle Choice on Column Performance: Switching from 3 and 5 \u00b5m Fully Porous Particles (FPP) to 5 \u00b5m Superficially Porous Particles (SPP)"},"content":{"rendered":"\n

Introduction<\/h2>\n\n\n\n

Superficially porous particles (commonly referred to as SPP or “core-shell” particles) have been proven to provide fast and efficient LC separations. These particles feature a solid, impermeable core enveloped by a thin, porous layer of silica that offers significantly higher efficiency and sensitivity than traditional fully porous particles (FPP). In this technical note, we will compare the performance of Raptor 5 \u00b5m SPP LC columns to traditional 3 and 5 \u00b5m FPP LC columns. We will demonstrate how faster, more efficient analyses can be attained with greater sensitivity and reduced system pressure by switching from fully porous particles to superficially porous particles\u2014without changing instrumentation.<\/p>\n\n\n\n

Switching from FPP to SPP: Increased Efficiency, Decreased Pressure<\/h2>\n\n\n\n

One of the primary advantages of SPP is its ability to provide increased column efficiency, often with similar or even reduced backpressure, when compared to conventional FPP. By increasing efficiency while decreasing pressure, the user can achieve faster analysis times without changing instrumentation. As shown in Figure 1, column backpressure decreases by approximately 50% on average across the instrument flow rates tested (0.2 to 2 mL\/min) when switching from a 3 \u00b5m FPP column to a Raptor 5 \u00b5m SPP column. Although the 5 \u00b5m FPP column displays comparable pressure to the Raptor 5 \u00b5m SPP column, it is at the expense of efficiency.<\/p>\n\n\n

Figure 1:<\/strong>\u00a0Switch from a 3 \u00b5m FPP column to a Raptor 5 \u00b5m SPP to cut backpressure in half.<\/div>
\n\n
\"chart,<\/figure>\n\n<\/div><\/div><\/div>\n\n\n

The ratio of theoretical plates (N) at optimal flow divided by pressure can be used as a measure of efficiency for a column. When compared for each column, the Raptor 5 \u00b5m SPP column has double the number of plates per unit pressure as the 5 \u00b5m FPP column, and four times the number of plates per unit pressure as the 3 \u00b5m FPP column (Figure 2).<\/p>\n\n\n\n

Alternatively, the relationship between efficiency and pressure can be illustrated by comparing the theoretical plates of each column while holding pressure relatively constant (Figure 3). Again, the Raptor 5 \u00b5m SPP column shows a dramatic increase in efficiency at equivalent pressure.<\/p>\n\n\n

Figure 2:<\/strong>\u00a0Focusing on plates per unit pressure, switching from 3 and 5 \u00b5m FPP columns to Raptor 5 \u00b5m SPP columns offers clear advantages.<\/div>
\n\n
\"\"<\/figure>\n\n<\/div><\/div><\/div>\n\n
Figure 3:<\/strong>\u00a0With equivalent pressures, Raptor 5 \u00b5m superficially porous particles exhibit significantly more plates than 3 and 5 \u00b5m fully porous particles.<\/div>
\n\n
\"chart,<\/figure>\n\n<\/div><\/div><\/div>\n\n\n

The Raptor 5 \u00b5m column has nearly twice the plates (i.e., greater efficiency) as the fully porous 3 and 5 \u00b5m particle columns.<\/p>\n\n\n\n

Switching from FPP to SPP: Increased Efficiency, Faster Run Times<\/h2>\n\n\n\n

The invention of SPP has provided analysts with fast separations without the need for expensive Ultra High Performance Liquid Chromatography (UHPLC) instruments, thereby increasing sample throughput without capital investment. To investigate the increased speed and efficiency of superficially porous particles, the Raptor Biphenyl 5 \u00b5m SPP column was compared to columns packed with fully porous 3 and 5 \u00b5m particles by performing an assay using identical instrumentation and isocratic method conditions (Figure 4).<\/p>\n\n\n

Figure 4:<\/strong>\u00a0Cut analysis times almost in half by switching to 5 \u00b5m SPP with lower pressure over 3 \u00b5m FPP and far superior efficiency over 5 \u00b5m FPP.<\/div>
\n
\"Comparison:<\/div>

LC_GN0556<\/p>

Peaks<\/h4>\n
<\/th>Peaks<\/th>Conc.
(mg\/mL)<\/th><\/tr><\/thead>\n
1.<\/td>Uracil<\/a><\/td>0.02<\/td><\/tr>\n
2.<\/td>Benzene<\/a><\/td>3.0<\/td><\/tr>\n
3.<\/td>Naphthalene<\/a><\/td>0.5<\/td><\/tr>\n
4.<\/td>Biphenyl<\/a><\/td>0.06<\/td><\/tr>\n<\/tbody><\/table><\/div>

Conditions<\/h4>
Column<\/th>
Temp.:<\/th>30 \u00b0C<\/td><\/tr>
Standard\/Sample<\/th>LC Reversed Phase Test Mix #1 (cat.# 35005<\/a>)<\/td><\/tr>\n
Diluent:<\/th>Methanol:water (75:25)<\/td><\/tr><\/td><\/tr>
Inj. Vol.:<\/th>2 \u00b5L <\/td><\/tr>
Mobile Phase<\/th><\/td><\/tr>
A:<\/th>Water <\/td><\/tr>
B:<\/th>Acetonitrile <\/td><\/tr>
<\/td>
Time (min)<\/th>Flow (mL\/min)<\/th>%A<\/th>%B<\/th><\/tr><\/thead>
0.00<\/td>0.8<\/td>45<\/td>55<\/td><\/tr>
20<\/td>0.8<\/td>45<\/td>55<\/td><\/tr><\/tbody><\/table><\/td><\/tr><\/table>
Detector<\/th>DAD @ 254 nm<\/td><\/tr>
Instrument<\/th>HPLC<\/td><\/tr>
Notes<\/th>All columns were 150 mm x 4.6 mm ID. Values for efficiency (N) were calculated from the Biphenyl peak.<\/td><\/tr><\/table><\/div><\/div><\/div>
<\/path><\/g><\/path><\/g><\/svg>Download PDF<\/a><\/div><\/div><\/div>\n<\/div><\/div><\/div>\n\n\n

<\/p>\n\n\n\n

The last analyte eluted on the Raptor Biphenyl 5 \u00b5m column in 7.1 minutes resulting in a 45% decrease in analysis time compared to the 3 \u00b5m and 5 \u00b5m fully porous particle columns, which had run times of 13.6 and 12.9 minutes respectively. In addition, a 43% improvement in efficiency was observed for the Raptor Biphenyl 5 \u00b5m SPP column over the 5 \u00b5m FPP column. The 3 \u00b5m FPP column displayed only slightly less efficiency but nearly double the backpressure of the Raptor Biphenyl 5 \u00b5m SPP column.<\/p>\n\n\n\n

<\/div>\n\n\n\n

Switching from FPP to SPP: Increased Sensitivity, Better Performance<\/h2>\n\n\n\n

The performance of existing methods can be instantly improved by replacing conventional 3 \u00b5m or 5 \u00b5m FPP columns with the Raptor 5 \u00b5m column. Enhanced method efficiency, peak width, and analysis time can be achieved under gradient conditions, as demonstrated in Figure 5.<\/p>\n\n\n

Figure 5:<\/strong>\u00a0Increase resolution and decrease analysis time by switching from FPP to Raptor SPP columns.<\/div>
\n
\"Comparison:<\/div>

LC_GN0555<\/p>

Peaks<\/h4>\n
<\/th>Peaks<\/th><\/tr><\/thead>\n
1.<\/td>Norfentanyl<\/a><\/td><\/tr>\n
2.<\/td>Acetyl fentanyl<\/a><\/td><\/tr>\n
3.<\/td>Fentanyl<\/a><\/td><\/tr>\n
4.<\/td>Sufentanyl<\/a><\/td><\/tr>\n<\/tbody><\/table><\/div>

Conditions<\/h4>
Column<\/th>
Temp.:<\/th>30 \u00b0C<\/td><\/tr>
Standard\/Sample<\/th><\/td><\/tr>
Diluent:<\/th>Water<\/td><\/tr>
Conc.:<\/th>25 \u00b5g\/mL<\/td><\/tr><\/td><\/tr>
Inj. Vol.:<\/th>1 \u00b5L <\/td><\/tr>
Mobile Phase<\/th><\/td><\/tr>
A:<\/th>Water + 0.1% formic acid <\/td><\/tr>
B:<\/th>Methanol + 0.1% formic acid <\/td><\/tr>
<\/td>
Time (min)<\/th>Flow (mL\/min)<\/th>%A<\/th>%B<\/th><\/tr><\/thead>
0.00<\/td>0.6<\/td>65<\/td>35<\/td><\/tr>
2.00<\/td>0.6<\/td>15<\/td>85<\/td><\/tr>
2.01<\/td>0.6<\/td>65<\/td>35<\/td><\/tr>
4.00<\/td>0.6<\/td>65<\/td>35<\/td><\/tr><\/tbody><\/table><\/td><\/tr><\/table>
Detector<\/th>PDA @ 210 nm<\/td><\/tr>
Instrument<\/th>HPLC<\/td><\/tr>
Notes<\/th>All columns were 50 mm x 2.1 mm ID.<\/td><\/tr><\/table><\/div><\/div><\/div>
<\/path><\/g><\/path><\/g><\/svg>Download PDF<\/a><\/div><\/div><\/div>\n<\/div><\/div><\/div>\n\n\n

<\/p>\n\n\n\n

Average resolution values were 6.0 on the Raptor Biphenyl 5 \u00b5m SPP column and 4.1 on the 5 \u00b5m FPP column. In addition to a shorter run time, average analyte resolution increased 46% when switching to superficially porous particles of the same diameter.<\/p>\n\n\n\n

In another example using gradient conditions and mass spectrometry, the analysis time decreased from 2.3 minutes on the 5 \u00b5m FPP column to 1.5 min on the Raptor Biphenyl SPP 5 \u00b5m column (Figure 6). Meanwhile, average sensitivity for the 5 \u00b5m SPP column increased by 64% over the 3 \u00b5m FPP column and 128% over the 5 \u00b5m FPP column.<\/p>\n\n\n

Figure 6:<\/strong>\u00a0Raptor 5 \u00b5m SPP columns also provide increased sensitivity over 3 or 5 \u00b5m FPP columns.<\/div>
\n
\"Comparison:<\/div>

LC_CF0598<\/p>

Peaks<\/h4>\n
<\/th>Peaks<\/th><\/tr><\/thead>\n
1.<\/td>3,3′,5-Triiodo-L-thyronine (T3)<\/td><\/tr>\n
2.<\/td>3,3′,5-Triiodo-L-thyronine (rT3)<\/td><\/tr>\n
3.<\/td>L-Thyroxine (T4)<\/td><\/tr>\n<\/tbody><\/table><\/div>

Conditions<\/h4>
Column<\/th>
Temp.:<\/th>40 \u00b0C<\/td><\/tr>
Standard\/Sample<\/th><\/td><\/tr>
Diluent:<\/th>Water:methanol (35:65)<\/td><\/tr>
Conc.:<\/th>1 ng\/mL<\/td><\/tr><\/td><\/tr>
Inj. Vol.:<\/th>2 \u00b5L <\/td><\/tr>
Mobile Phase<\/th><\/td><\/tr>
A:<\/th>0.1% Formic acid in water <\/td><\/tr>
B:<\/th>0.1% Formic acid in methanol <\/td><\/tr>
<\/td>
Time (min)<\/th>Flow (mL\/min)<\/th>%A<\/th>%B<\/th><\/tr><\/thead>
0.00<\/td>0.4<\/td>35<\/td>65<\/td><\/tr>
2.00<\/td>0.4<\/td>20<\/td>80<\/td><\/tr>
2.01<\/td>0.4<\/td>35<\/td>65<\/td><\/tr>
3.50<\/td>0.4<\/td>35<\/td>65<\/td><\/tr><\/tbody><\/table><\/td><\/tr><\/table>
Detector<\/th>MS\/MS<\/td><\/tr>
Ion Mode:<\/th>ESI+ <\/td><\/tr>
Instrument<\/th>HPLC<\/td><\/tr>
Notes<\/th>All columns were 100 mm x 2.1 mm ID.<\/td><\/tr><\/table><\/div><\/div><\/div>
<\/path><\/g><\/path><\/g><\/svg>Download PDF<\/a><\/div><\/div><\/div>\n<\/div><\/div><\/div>\n\n\n
<\/div>\n\n\n\n

Conclusion<\/h2>\n\n\n\n

Raptor 5 \u00b5m LC columns with superficially porous particles (SPP) display lower backpressure and increased efficiency when compared to columns packed with 3 and 5 \u00b5m fully porous particles (FPP) of similar dimension. When used in the development of new assays, Raptor 5 \u00b5m SPP LC columns offer fast run times and excellent method performance without upgrading instrumentation. When substituted into existing methodologies, switching from conventional 3 \u00b5m and 5 \u00b5m FPP columns to Raptor 5 \u00b5m SPP LC columns has the potential to dramatically decrease analysis times while improving efficiency and sensitivity. Certain assays may require some degree of method development to achieve optimal results, but Raptor 5 \u00b5m LC columns are compatible with most assays and offer an excellent way to increase performance without extra cost or labor.<\/p>\n\n\n\n

<\/div>\n","protected":false},"excerpt":{"rendered":"

In this technical note, we will compare the performance of Raptor 5 \u00b5m superficially porous particle (SPP) LC columns to traditional 3 and 5 \u00b5m fully porous particle (FPP) LC columns. We will demonstrate how faster, more efficient analyses can be attained with greater sensitivity and reduced system pressure by switching from FPP to SPP\u2014without altering method conditions.<\/p>\n","protected":false},"author":53,"featured_media":0,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"_kad_blocks_custom_css":"","_kad_blocks_head_custom_js":"","_kad_blocks_body_custom_js":"","_kad_blocks_footer_custom_js":"","_kadence_starter_templates_imported_post":false,"_kad_post_transparent":"","_kad_post_title":"","_kad_post_layout":"","_kad_post_sidebar_id":"","_kad_post_content_style":"","_kad_post_vertical_padding":"","_kad_post_feature":"","_kad_post_feature_position":"","_kad_post_header":false,"_kad_post_footer":false,"footnotes":""},"categories":[462],"tags":[],"industries-application":[],"post-badge":[],"resource-type":[],"product-library":[2463,2445],"resource-technique":[2318,2320,2362],"ppma_author":[606,760,608],"class_list":["post-43638","post","type-post","status-publish","format-standard","hentry","category-articles-fr","product-library-colonnes-lc","product-library-produits-pour-la-chromatographie-en-phase-liquide","resource-technique-chromatographie-en-phase-liquide","resource-technique-spectrometrie-de-masse-ms","resource-technique-ms-ms-fr"],"acf":[],"taxonomy_info":{"category":[{"value":462,"label":"Articles"}],"product-library":[{"value":2463,"label":"Colonnes 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