Asahipak - Hplc.eu

Calibration curves for KW400 series and KW-800 seriesMolecular weight (Protein)10 610 510 410 3KW402.5-4FKW404-4FKW405-4FKW403-4FMolecular weight (Protein)10 610 510 4KW-802.510 3KW-803KW-804Recovery rate of proteinsRecovery (%)ProteinKW402.5-4F KW403-4Fγ- GlobulinBovine serum albuminOvalbuminMyoglobinCytochrome cLysozyme988989909287969697899298Standard analysiscolumns10 7 Elution volume (mL)10 2 1.5 2.0 2.5 3.0 3.5 4.0 4.510 7 4510 25 6 7 8 9 10 11 12 13Elution volume (mL)α-Chymotrypsinogen A9594Comparison of KW402.5-4F and KW-802.5Sample : 10μL1. Blue dextran 2000 0.2mg/mL2. γ-Globulin 0.8mg/mL3. Ovalbumin 0.8mg/mL4. Myogrobin 0.56mg/mL5. Uridine 0.04mg/mLN=17,000(A) KW402.5-4F 1 2 34(B) KW-802.5Column : Shodex KW400-4F series, Shodex PROTEIN KW-800 seriesEluent : 50mM Sodium phosphate buffer + 0.3M NaCl(pH7.0)Flow rate : (KW400) 0.33mL/min(KW-800) 1.0mL/minDetector : (KW400) UV(280nm) (small cell volume)(KW-800) UV(280nm) (conventional type)Column temp. : 25˚CKW400 series are high-performance semi-micro columnswith theoretical plate number which is approximately 1.5times larger and a detection sensitivity (peak height) thatis 3 to 4 times higher than what KW-800 columns have.N=8,2001 2 3 455Milk whey in yogurtSample : Whey, 5μLColumn : Shodex KW402.5-4F, KW403-4FEluent : 50mM Sodium phosphate buffer+ 0.3M NaCl(pH7.0)Flow rate : 0.33mL/minDetector : UV(280nm) (small cell volume)Column temp. : 25˚CLectinsSample : 5μL1. Lectin from Soybean 0.6mg/mL2. Lectin from Arachis hypogaea 1.1mg/mL3. Lectin from Canavalia ensiformis (Con A) 0.9mg/mL4. Lectin from Lens culinaris (LCA) 0.7mg/mL5. Lectin from Triticum vulgaris (WGA) 0.8mg/mL213 4 5Aqueous SEC (GFC) columns : Silica-based05 10 15 min 20 30 4050 min 0 5 10 minColumn : (A) Shodex KW402.5-4F(B) Shodex PROTEIN KW-802.5Eluent : 50mM Sodium phosphate buffer+ 0.3M NaCl(pH7.0)Flow rate : (A) 0.33mL/min, (B) 1.0mL/minDetector : UV(280nm) (small cell volume)Column temp. : 25˚CColumn : Shodex KW402.5-4F + KW403-4FEluent : 50mM Sodium phosphate buffer+ 0.3M NaCl(pH7.0)Flow rate : 0.20mL/minDetector : UV(280nm) (small cell volume)Column temp. : 30˚CColumn : Shodex KW402.5-4FEluent : 50mM Sodium phosphate buffer+ 0.3M NaCl(pH7.0)Flow rate : 0.33mL/minDetector : UV(220nm) (small cell volume)Column temp. : 30˚CComparison of standard protein separation usingvarious GFC columnsSample :A : Thyroglobulin (bovine)B : γ-Globulin (bovine)C : Ovalbumin (chicken)D : Nyoglobin (horse)E : CyanocobalaminSB-803 HQGF-510 HQKW-803BAACBACDB0 5 10 15 20 25 30minDCDEESEC columns for an aqueous system, i.e., SB-803 HQ,GF-510 HQ and KW-803, were compared with eachother in terms of the separation of standard proteins.The result demonstrates that the KW-803, a silica-basedcolumn, is the most suitable for such an application.E35Molecular weight (Protein)10 610 5KW-803GF-510 HQSB-803 HQ10 411 13 15 17 19 21 minColumn : Shodex OHpak SB-803 HQ, Asahipak GF-510 HQ,PROTEIN KW-803Eluent : 0.2M Phosphate buffer(pH6.9)Flow rate : 0.5mL/minDetector : UV(280nm)Column temp. : 30˚CProteins in human blood serum3610 15 20 min 10 15 20 30 minColumn : Shodex PROTEIN KW-803 x 2Eluent : 50mM Phosphate buffer + 0.3M NaCl(pH7.0)Flow rate : 1.0mL/minDetector : UV(280nm)Column temp. : Room temp.12345678Sample : Human serum 0.1% each1. Fibrinogen 50μL2. α 2 -Macroglobulin 50μL3. IgG 50μL4. Transferrin 50μL5. Plasminogen 50μL6. Albumin 100μL7. Antitrypsin 100μL8. Hemoglobin 100μL