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HPLC Column
Applications
Material
Pictures below show size uniformity and surface smoothness of the packing particles, characteristics that enable more uniform packing with less channeling effect, resulting in lower back pressure and higher column efficiency.
Trace Amount Metal Contents Test
A useful chromatographic test of trace amount of metal contents in the column is to compare the peak symmetry of one pair of positional isomers, 4,4’-dipyridyl and 2,2’-dipyridyl, and a neutral chelating reagent, 1,2-dihydroxylnaphthalene. 4,4’-dipyridyl, which cannot form chelating complex with metal, is used as a reference. 2,2’-dipyridyl and 1,2-dihydroxyl- naphthalene, which are chelating reagents, are sensitive to trace amount metal in silica. When a C18 column based on type A silica or other so-called type B silica with higher metal content is used, the peaks of 2,2’-dipyridyl and 1,2-dihydroxyl- naphthalene would tail or even totally disappear.
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Column: | C18, 4.6×150mm, 5µm |
| Mobile Phase: | 20 mM phosphate(pH 7.6) / methanol=55/45 |
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| Flow rate: | 1.0mL/min | |
| Detector: | 215nm | |
| Temperature: | 25°C | |
| Injection Volume: | 1µL | |
| Samples: | （1） 4,4’-Dipyridyl （2）2,2’-Dipyridyl （3） 1,2 Dihydroxylnaphthalene |
C18—Universal HPLC Analytical Column
C18 is the most commonly used column in the market. It can substitute Waters Symmetry C18, Agilent Zorbax XDB C18, Phenomenex Luna C18, Supelcosil LC-18-DB, YMC ODS-AM, Alltima C18, GL, Inertsil ODS-2 etc. XB-C18 has high theoretical plates and peak capacity, so it’s suitable for analysis of complex samples.
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Structural Formula |  |
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| pH Range | 1.5-10.0 | ||||||
| Particle Size | 3µm, 5µm, 10µm | ||||||
| Surface Area(m2/g) | 320(120Å), 90(300Å) | ||||||
| Carbon Loading(%) | 17(120Å), 8(300Å) | ||||||
| USP List | L11 | ||||||
| Endcapped | Yes | ||||||
Separation of Basic Compounds Antidepressant at pH 7.0
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Column: | C18, 4.6×150mm, 5µm | |||||
| Mobile Phase: | 20mM phosphate(pH 7.0) / methanol=20/80 |
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| Flow rate: | 1.0mL/min | ||||||
| Detector: | 215nm | ||||||
| Temperature: | 25°C | ||||||
| Samples: |
（1)Uracil （2） Ropranolol |
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Comparison of Peak Shape between Batch to Batch
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Column: | C18, 4.6×250mm, 5µm | |||||
| Mobile Phase: | Methanol/water=75/25 | ||||||
| Flow rate: | 1.0mL/min | ||||||
| Detector: | 254nm | ||||||
| Temperature: | 25°C | ||||||
| Samples: | （1）Uracil （2） Phenol（3） 4-chloronitrobenzen （4） Methylbenzene | ||||||
Capacity Factor(K’) of Batch to Batch Reproducibility
Excellent Stability at Low pH and High pH
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Argatroban
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Column: | C18, 4.6×250mm, 3µm |
Time (min) |
Mobile Phase A(%) | Mobile Phase B(%) | |||||
| Mobile Phase: |
A: 10mmol/L ammonium acetate |
0 | 60 | 40 | ||||||
| Flow rate: | 0.6mL/min | 20 | 60 | 40 | ||||||
| Detector: | 259nm | 35 | 50 | 50 | ||||||
| Temperature: | 50°C | 50 | 20 | 80 | ||||||
| Injection Volume: | 10µL | 60 | 20 | 80 | ||||||
| Reference Sample: |
S-argatroban, R-argatroban |
60.1 | 60 | 40 | ||||||
| 80 | 60 | 40 | ||||||||
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Paracetamol Injection USP 36
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Column: | C18, 4.6×250mm, 5µm |
Time (min) |
Mobile Phase A(%) | Mobile Phase B(%) | |||||
| Mobile Phase: |
A:methanol/water/ |
0 | 82 | 18 | ||||||
| Flow rate: | 0.9mL/min | 8 | 82 | 18 | ||||||
| Detector: | 254nm, 275nm, 317nm | 53 | 0 | 100 | ||||||
| Temperature: | 40°C | 58 | 0 | 100 | ||||||
| Injection Volume: | 20µL | 59 | 82 | 18 | ||||||
| Reference Sample: | L-hydroxyproline, glycine, alanine, L-proline |
73 | 82 | 18 | ||||||
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Progesterone(EP 5.0)
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Column: | C18,4.6×150mm, 5µm |
Time (min) |
Mobile Phase A(%) | Mobile Phase B(%) | |||||
| Mobile Phase: | A: water B: acetonitrile |
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| Flow rate: | 0.9mL/min | 0-20 | 50 | 50 | ||||||
| Detector: | 254nm, 275nm, 317nm | 20-27 | 50-20 | 50-80 | ||||||
| Temperature: | 40°C | 27-45 | 20 | 50 | ||||||
| Injection Volume: | 20µL | 45-50 | 20 | 50 | ||||||
| Reference Sample: | Lhydroxyproline ,glycine,alanine, L-proline | |||||||||
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Aflatoxin
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Column: | C18, 4.6×250mm, 5µm | |||||
| Mobile Phase: | Water/methanol/acetonitrile=46/40/14 | ||||||
| Flow rate: | 1.0mL/min | ||||||
| Detector: | Excitation wavelength: 360nm | ||||||
| Temperature: | Emission wavelength: 450nm Gain:17 | ||||||
| Injection Volume: | 30°C | ||||||
| Reference Sample: | Post -column photo chemical derivation(254 nm) | ||||||
