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Zenix SEC 3µm phases are made of uniform, hydrophilic, and neutral nanometer thick proprietary surface coating chemically bonded on high purity and mechanically stabilized silica. Zenix SEC 3µm unique packing combined with large pore volume delivers highest and unrivaled separation efficiency and resolution. The well-controlled surface chemistry results in excellent lot-to-lot reproducibility. Our unique bonding chemistry, coupled with the maximized bonding density, allows Zenix SEC to provide high stability and negligible non-specific interactions. The available pore sizes of Zenix packings are 100, 150 and 300Å. Typical applications for Zenix SEC columns include separation and analysis of biological molecules and water soluble polymers in aqueous buffers.
- Particle size of 3 µm
- Selection of pore size: 100, 150 and 300 Å
- Highest separation efficiency and resolution
- High capacity
- High stability over low and high concentration salt
- Lot-to-lot reproducibility
- High protein recovery with intact biological activity
- Negligible non-specific interactions
- Ideal for separation and analysis of biological molecules: proteins, nucleic acids, oligonucleotides, peptides and virus
- Ideal for separation and analysis of natural polymers, e.g. polysaccharides, synthetic polymers, and nanomaterials, e.g. nanoparticles
Zenix SEC Technical Specifications
| Phase | Zenix SEC-100 | Zenix SEC-150 | Zenix SEC-300 |
| Material | Neutral, hydrophilic film bonded silica |
| Particle size | 3 µm |
| Pore size | ~ 100 Å | ~ 150 Å | ~ 300 Å |
| Protein MW range (native) | 100 - 100,000 | 500 - 150,000 | 5,000 - 1,250,000 |
| pH stability | 2 - 8.5 (pH 8.5-9.5 can be tolerated temporarily.) |
| Backpressure (psi for a 7.8×300 mm) | ~ 1,500 | ~ 1,500 | ~ 1,500 |
| Maximum backpressure (psi) | ~ 4,500 | ~ 4,500 | ~ 3,500 |
| Salt concentration range | 20 mM - 2.0 M |
| Maximum temperature (°C) | ~ 80 |
| Mobile phase compatibility | Aqueous and organic |
| Conditions | |
| Column: | Zenix SEC-300, 7.8×300mm |
| Mobile phase: | 150 mM PBS, pH 7.0 |
| Flow rate: | 1.0 mL/min |
| Temperature: | Ambient (~23 °C) |
| Detector: | UV 214 nm |
| Injection: | 5 µL |
| Sample: | 1. Thyroglobulin (1.0 mg/mL), 670 kD
2. BSA dimer, 132 kD
3. BSA (1.0 mg/mL), 66 kD
4. Ribonuclease A (1.0 mg/mL), 13.7 kD
5. Uracil (0.1 mg/mL), 120 D |
High Separation Efficiency
The advantages of developing small particle size are higher efficiency and higher resolution. When particle size is decreased to 3 µm from 5 µm, the column efficiency is almost doubled. As shown in Table 1, the plate numbers of BSA dimmer, BSA, ribonuclease A increased from 2720 to 4600, 6590 to 13090, 11160 to 22000 when the particle size decreased from 5 µm to 3 µm. Fig. 2 and Fig. 3 further show that high efficiency has been achieved by 3 µm Zenix columns with various proteins. The efficiency of p-aminobenzoic acid reached to the plate number of 40,000 for 30 cm long Zenix column.
Figure 1. Separation of protein mixture A by Zenix SEC-300 and SRT SEC-300 columns.
| Conditions | |
| Column: | Zenix SEC-300 and SRT SEC-300 |
| Mobile phase: | 150 mM PBS, pH 7.0 |
| Flow rate: | 1.0 mL/min for 7.8×300mm, 0.35 mL/min for 4.6×300mm |
| Temperature: | Ambient (~23 °C) |
| Detector: | UV 214 nm |
| Injection: | 10 µL for 7.8×300mm, 3 µL for 4.6×300mm |
| Sample: | 1. Thyroglobulin (1.0 mg/mL), 670 kD
2. BSA dimer, 132 kD
3. BSA (1.0 mg/mL), 66 kD
4. Ribonuclease A (1.0 mg/mL), 13.7 kD
5. Uracil (0.1 mg/mL), 120 D |
Table 1. Efficiency of Zenix SEC-300 and SRT SEC-300 columns.
| Peak | Protein | Zenix 300 (4.6×300) | Zenix 300 (7.8×300) | SRT 300 (7.8×300) |
| 1 | Thyroglobulin | 2180 | 1730 | 1120 |
| 2 | BSA Dimer | 4390 | 4600 | 2720 |
| 3 | BSA | 10280 | 13090 | 6590 |
| 4 | Ribonuclease A | 16490 | 22000 | 11160 |
| 5 | Uracil | 33640 | 38500 | 27860 |
Figure 2. Separation of protein mixture B by Zenix SEC-150 and 300 columns with 7.8 mm ID.
| Conditions | |
| Column: | 7.8×300 mm, 3 µm |
| Mobile phase: | 150 mM PBS, pH 7.0 |
| Flow rate: | 1.0 mL/min |
| Temperature: | Ambient (~23 °C) |
| Detector: | UV 214 nm |
| Injection: | 10 µL |
| Sample: | 1. Thyroglobulin, 670 kD
2. γ-Globulin, 158 kD
3. Ovalbumin, 44 kD
4. Ribonuclease A, 13.7 kD
5. p-Aminobenzoic acid, 137 D |
Table 2. Efficiency of 7.8×300 mm Zenix SEC-150 and 300 columns
| Peak | Protein | Zenix 150 | Zenix 300 |
| 1 | Thyroglobulin | 12420 | 1295 |
| 2 | γ-Globulin | 2860 | 3650 |
| 3 | Ovalbumin | 6620 | 11760 |
| 4 | Ribonuclease A | 16450 | 21690 |
| 5 | p-Aminobenzoic acid | 40550 | 39400 |
Figure 3. Separation of protein mixture B by Zenix SEC-150 and 300 columns with 4.6 mm ID.
| Conditions | |
| Column: | 4.6×300 mm, 3 µm |
| Mobile phase: | 150 mM PBS, pH 7.0 |
| Flow rate: | 0.35 mL/min |
| Temperature: | Ambient (~23 °C) |
| Detector: | UV 214 nm |
| Injection: | 5 µL |
| Sample: | 1. Thyroglobulin, 670 kD
2. γ-Globulin, 158 kD
3. Ovalbumin, 44 kD
4. Ribonuclease A, 13.7 kD
5. p-Aminobenzoic acid, 137 D |
Table 3. Efficiency of 4.6×300 mm Zenix SEC-150 and 300 columns.
| Peak | Protein | Zenix 150 | Zenix 300 |
| 1 | Thyroglobulin | 6020 | 2410 |
| 2 | γ-Globulin | 2560 | 3000 |
| 3 | Ovalbumin | 6030 | 10260 |
| 4 | Ribonuclease A | 13350 | 17020 |
| 5 | p-Aminobenzoic acid | 35500 | 33480 |
High Stability
The proprietary stationary phases of Zenix SEC packings utilize densely bonded chemistry on the silica surface, which greatly hinders the diffusion of the molecules that would attack the bond of silica-stationary phase layer, thus enabling high stability over a wide range of pH from 2 to 8.5.
Mobile Phase Compatibility
Zenix SEC phases are compatible with most aqueous buffers, such as ammonium acetate, phosphate, trizma and so on. Zenix SEC phases can tolerate high concentration of salts, such as 2.0 M. Furthermore, Zenix SEC columns are stable in both organic solvents, such as methanol, ethanol, THF, DMF, DMSO, and so on; as well as the mixture of water and organic solvents.
High Loading Capacity
Loading capacity is critical for size exclusion separation and purification. Figure 4 shows high loading capacity for BSA as one example (>500 µg for an analytical column).
Figure 4. BSA loading test on a Zenix SEC-150 column.
| Conditions | |
| Column: | Zenix SEC-150 (3 µm, 7.8×300 mm) |
| Mobile phase: | 150 mM PBS, pH 7.0 |
| Flow rate: | 1.0 mL/min |
| Temperature: | Ambient (~23 °C) |
| Detector: | UV 214 nm |
| Injection: | 10 µL |
Figure 5. BSA loading test on a Zenix SEC-300 column.
| Conditions | |
| Column: | Zenix SEC-300 (3 µm, 7.8×300 mm) |
| Mobile phase: | 150 mM PBS, pH 7.0 |
| Flow rate: | 1.0 mL/min |
| Temperature: | Ambient (~23 °C) |
| Detector: | UV 214 nm |
| Injection: | 10 µL |
Lot-to-Lot Reproducibility
The controlled surface chemistry used to synthesize Zenix SEC phases makes the surface coating highly reproducible, leading to consistent column manufacturing. Separation variation from batch to batch is controlled to be within 5% for retention time.
High Protein Recovery
Zenix SEC phases are hydrophilic and neutral. Proteins and other biological molecules have negligible nonspecific interactions with Zenix stationary phases. The protein adsorption to the silica surface is suppressed, leading to high recovery of intact proteins, maintaining the protein activity after separation. More than 95% recovery is achieved for BSA and lysozyme, the representatives for acidic and basic proteins, respectively.
Pore size vs. MW exclusion limit
| Phases (3 µm) | Pore Size | Protein MW Exclusion Limit |
| Zenix SEC-100 | 100 Å | 100,000 |
| Zenix SEC-150 | 150 Å | 150,000 |
| Zenix SEC-300 | 300 Å | 1,250,000 |
Column Dimension Availability
Available Zenix SEC column dimensions are 0.75, 1.0, 2.1, 3.0, 4.6, 7.8, 10, 21.2 and 30 mm I.D., and 20, 30, 50, 100, 150, 250, 300 and 600 mm length. Sepax also offers custom-made columns. Both stainless steel and PEEK tubes are available.
Zenix phases have wide applications for separation, identification and purification of proteins, protein variants, peptide fragments, phosphorylated, sialylated, pegylated, and other derivatized proteins. They are well suited for studies such as molecular weight estimation, purification and analysis of biological molecules.
Separation and Analysis
- Proteins
- Monoclonal antibodies
- Cell lysates
- Nucleic acids
- Nucleotides
- Peptides
- Water soluble polymers
- Nanoparticles
- Nanotubes
Separation of protein and peptide mixture
Separation of E. coli Lysate
Separation of Biorad standard protein sample
Sepax Zenix SEC-100 | Part Number | Particle Size | Pore Size | ID×Length | Price |
|---|
| | 213100-4605 * | 3 µm | 100Å | 4.6×50mm | $495.00 | | 213100-4615 | 3 µm | 100Å | 4.6×150mm | $1,230.00 | | 213100-4625 | 3 µm | 100Å | 4.6×250mm | $1,310.00 | | 213100-4630 | 3 µm | 100Å | 4.6×300mm | $1,390.00 | | 213100-7805 * | 3 µm | 100Å | 7.8×50mm | $520.00 | | 213100-7815 | 3 µm | 100Å | 7.8×150mm | $1,150.00 | | 213100-7825 | 3 µm | 100Å | 7.8×250mm | $1,325.00 | | 213100-7830 | 3 µm | 100Å | 7.8×300mm | $1,590.00 |
* Guard column. Sepax Zenix SEC-150 | Part Number | Particle Size | Pore Size | ID×Length | Price |
|---|
| | 213150-4605 * | 3 µm | 150Å | 4.6×50mm | $495.00 | | 213150-4615 | 3 µm | 150Å | 4.6×150mm | $1,230.00 | | 213150-4625 | 3 µm | 150Å | 4.6×250mm | $1,310.00 | | 213150-4630 | 3 µm | 150Å | 4.6×300mm | $1,390.00 | | 213150-7805 * | 3 µm | 150Å | 7.8×50mm | $520.00 | | 213150-7815 | 3 µm | 150Å | 7.8×150mm | $1,150.00 | | 213150-7825 | 3 µm | 150Å | 7.8×250mm | $1,325.00 | | 213150-7830 | 3 µm | 150Å | 7.8×300mm | $1,590.00 |
* Guard column. Sepax Zenix SEC-300 | Part Number | Particle Size | Pore Size | ID×Length | Price |
|---|
| | 213300-4605 * | 3 µm | 300Å | 4.6×50mm | $495.00 | | 213300-4615 | 3 µm | 300Å | 4.6×150mm | $1,230.00 | | 213300-4625 | 3 µm | 300Å | 4.6×250mm | $1,310.00 | | 213300-4630 | 3 µm | 300Å | 4.6×300mm | $1,390.00 | | 213300-7805 * | 3 µm | 300Å | 7.8×50mm | $520.00 | | 213300-7815 | 3 µm | 300Å | 7.8×150mm | $1,150.00 | | 213300-7825 | 3 µm | 300Å | 7.8×250mm | $1,325.00 | | 213300-7830 | 3 µm | 300Å | 7.8×300mm | $1,590.00 |
* Guard column. Sepax Zenix SEC-100 | Part Number | Particle Size | Pore Size | ID×Length | Price |
|---|
| | 213100-10005 * | 3 µm | 100Å | 10.0×50mm | $850.00 | | 213100-10010 | 3 µm | 100Å | 10.0×100mm | $1,685.00 | | 213100-10015 | 3 µm | 100Å | 10.0×150mm | $1,970.00 | | 213100-10025 | 3 µm | 100Å | 10.0×250mm | $2,560.00 | | 213100-10030 | 3 µm | 100Å | 10.0×300mm | $2,845.00 | | 213100-21205 * | 3 µm | 100Å | 21.2×50mm | $1,880.00 | | 213100-21225 | 3 µm | 100Å | 21.2×250mm | $6,500.00 | | 213100-21230 | 3 µm | 100Å | 21.2×300mm | $7,870.00 |
* Guard column. Sepax Zenix SEC-150 | Part Number | Particle Size | Pore Size | ID×Length | Price |
|---|
| | 213150-10005 * | 3 µm | 150Å | 10.0×50mm | $850.00 | | 213150-10010 | 3 µm | 150Å | 10.0×100mm | $1,685.00 | | 213150-10015 | 3 µm | 150Å | 10.0×150mm | $1,970.00 | | 213150-10025 | 3 µm | 150Å | 10.0×250mm | $2,560.00 | | 213150-10030 | 3 µm | 150Å | 10.0×300mm | $2,845.00 | | 213150-21205 * | 3 µm | 150Å | 21.2×50mm | $1,880.00 | | 213150-21225 | 3 µm | 150Å | 21.2×250mm | $6,500.00 | | 213150-21230 | 3 µm | 150Å | 21.2×300mm | $7,870.00 |
* Guard column. Sepax Zenix SEC-300 | Part Number | Particle Size | Pore Size | ID×Length | Price |
|---|
| | 213300-10005 * | 3 µm | 300Å | 10.0×50mm | $850.00 | | 213300-10010 | 3 µm | 300Å | 10.0×100mm | $1,685.00 | | 213300-10015 | 3 µm | 300Å | 10.0×150mm | $1,970.00 | | 213300-10025 | 3 µm | 300Å | 10.0×250mm | $2,560.00 | | 213300-10030 | 3 µm | 300Å | 10.0×300mm | $2,845.00 | | 213300-21205 * | 3 µm | 300Å | 21.2×50mm | $1,880.00 | | 213300-21225 | 3 µm | 300Å | 21.2×250mm | $6,500.00 | | 213300-21230 | 3 µm | 300Å | 21.2×300mm | $7,870.00 |
* Guard column. |
Zenix SEC-100 Zenix SEC-150 Zenix SEC-300
Zenix SEC-100 Zenix SEC-150 Zenix SEC-300
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Analytical LC Columns
