Gelatin / Albumin

Gelatin

Gelatin contains alpha, beta and gamma chains, and the ratio of their content is said to have a significant effect on the performance of gelatin. Asahipak GS-620 7G-P is especially suitable for analysis of the components of gelatin by the PAGI method (test method of gelatin for photo film in Japan).

Sample : Gelatin

Columns      : Shodex Asahipak GS-620 7G-P (7.5mmID*500mm) x 2
Eluent       : 0.1M KH2PO4 aq./0.1M Na2HPO4 aq.=50/50
Flow rate    : 1.0mL/min
Detector     : UV(230nm)
Column temp. : 50deg-C

Gelatin contains alpha, beta and gamma chains, and its molecular weight distribution has a significant influence on the characteristics of photographic films and on the crystallization of silver halides in an emulsion. Asahipak GS-620 7G-P is especially suitable for analysis of the components of gelatin and the combination of OHpak SB-805 HQ and SB-804 HQ is suitable for molecular weight distribution analysis by the PAGI method (test method of gelatin for photo film in Japan).

Sample : Gelatin (Koepff 16922) 0.2%
100micro-L

Columns      : Shodex Asahipak GS-620 7G-P (7.5mmID*500mm) x 2
               Shodex OHpak SB-805 HQ x 3 + SB-804 HQ (8.0mmID*300mm each)
Eluent       : 0.1M KH2PO4 aq./0.1M Na2HPO4 aq.=50/50
Flow rate    : 1.0mL/min
Detector     : UV(230nm)
Column temp. : 50deg-C

Gelatin contains alpha, beta and gamma chains, and the ratio of their content is said to have a significant effect on the performance of gelatin. The Asahipak GS-620 HQ column is suitable for general analysis of the components of gelatin and Asahipak GS-620 7G-P is especially suitable for analysis of the components of gelatin by the PAGI method(test method of gelatin for photo film in Japan). When using an Asahipak GS-620 7G-P, the column should be pretreated in accordance with “Precautions in Analyzing Gelatin” attached to the operation manual so as to obtain reproducible data.

Sample : Gelatin

 
Column       : Shodex Asahipak GS-620 7G-P (7.5mmID*500mm) x 2 
Eluent       : 0.2M Sodium phosphate(pH7.0) 
Flow rate    : 0.3mL/min 
Detector     : UV(220nm) 
Column temp. : 30deg-C

Courtesy of Dr. Mizusawa and Dr. Ohno, Chiba University

Gelatin contains alpha, beta and gamma chains, and the ratio of their content is said to have a significant effect on the performance of gelatin. The Asahipak GS-620 HQ column is suitable for general analysis of the components of gelatin and Asahipak GS-620 7G-P is especially suitable for analysis of the components of gelatin by the PAGI method(test method of gelatin for photo film in Japan). When using a Asahipak GS-620 7G-P, the column should be pretreated in accordance with “Precautions in Analyzing Gelatin” attached to the operation manual so as to obtain reproducible data.

Sample : Gelatin

Columns      : Shodex Asahipak GS-620 7G-P (7.5mmID*500mm) x 2
Eluent       : 0.2M NaH2PO4(pH6.8)
Flow rate    : 0.57mL/min
Detector     : UV(220nm)
Column temp. : 50deg-C

Courtesy of Dr. Ohno, Chiba University

Albumin

Asahipak GS-520 7G, a multimode column, separates proteins by GFC mode. However, albumins such as bovine serumalbumin and human serum albumin are eluted slower because the albumins have affinity with the molecular structure of poly(vinylalcohol) gel which is packed in GS-520. Therefore, albumins can separate with components of similar molecular weight. Here, albumin (ca, MW 69,000) is separated with tranferrin (ca. MW 80,000).
GS-520 7G is an old type of GS-520 HQ.

Sample :
1. IgM
2. IgG
3. Transferrin
4. Mercaptoalbumin
5. Nonmercaptoalbumin
6. Creatinine
7. Uric acid

Columns      : Shodex Asahipak GS-520 7G (7.5mmID*500mm) x 4
Eluent       : 0.03M Sodium phosphate buffer(pH7.0) + 0.15M Na2SO4
Flow rate    : 1.0mL/min
Detector     : UV(250nm)
Column temp. : 30deg-C

Asahipak GS-520 7G, a multimode column, separates proteins by GFC mode. However, serum albumins such as bovine and human albumins are eluted slowly because the albumins have affinity with the molecular structure of poly(vinyl alcohol) gel which is packed in GS-520 7G. Therefore, albumins can separate with components of similar molecular weight. Here, albumin is separated with tranferrin, IgG, IgM and IgA. With PROTEIN KW-800 series, albumins and transferrin cannot be separated because their molecular weights are too close to be separated by GFC mode.
GS-520 7G is an old type of GS-520 HQ.

Sample :
Albumin
Transferrin
IgG
IgA
IgM
Mercaptoalbumin
Nonmercaptoalbumin

Columns      : Shodex Asahipak  GS-520 7G (7.5mmID*500mm) x 4  
Eluent       : 0.1M Sodium phosphate buffer(pH7.0) + 0.3M NaCl
Flow rate    : 1.0mL/min
Detector     : UV(250nm)
Column temp. : 30deg-C

Mercaptpalbumin and nonmercaptoalbumin were analyzed using Asahipak ES-502N (a column for weak anion exchange chromatography). The ratio of human mercaptoalbumin (HMA) to human nonmercaptoalbumin (HNA) in serum is thought to have some relation with aging. The correlation between the ratio and various types of illness is being studied.

Sample :
Globulin
Uric acid
HMA, Human Mercaptoalbumin
HNA, Human Nonmercaptoalbumin

Column       : Shodex Asahipak ES-502N 7C (7.5mmID*100mm)
Eluent       : 50mM N-methylpiperazine-HCl buffer(pH4.8) + 400mM Na2SO4 + 0.3% C2H5OH
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : 35deg-C

The ratio of human mercaptoalbumin(HMA) to human nonmercaptoalbumin(HNA) in serum is thought to have some relation with aging. The correlation between the ratio and various types of illness is being studied. Ion exchange mode has been used most commonly for the analysis of albumin, however, as isocratic elution widens the peaks, gradient elution must be applied in ion exchange mode. In contrast, one of the characteristics of Asahipak GS-520 ( a multimode column ) is its capacity of enabling isocratic elution of HMA and HNA with isocratic elution using simple eluent such as phosphate buffer. This is the original Asahipak GS-520 column separation method which allows proteins other than albumin to elute first in the GFC mode and only albumin to elute later.
GS-520 7E is an old type of GS-520 HQ.

Sample : Human serum
HMA (Human Mercaptoalbumin)
HNA (Human Nonmercaptoalbumin)

fHMA = (HMA)/(HMA + HNA)

Columns      : Shodex Asahipak GS-520 7E (7.5mmID*250mm) x 8
Eluent       : 0.03M Sodium phosphate buffer(pH6.87) + 0.15M Na2SO4
Flow rate    : 0.8mL/min
Detector     : UV(280nm)
Column temp. : 28deg-C

Coutesy of Dr. Sogami, Fujita Health University, School of health Science

Albumins in human serum were analyzed using Asahipak GS-520 7G (a multimode column).
GS-520 7G is an old type of GS-520 HQ.

Sample : Human serum
Globulin
Uric acid
HMA (Human Mercaptoalbumin)
HNA (Human Nonmercaptoalbumin)

Column       : Shodex Asahipak GS-520 7G (7.5mmID*500mm)
Eluent       : 30mM Sodium phosphate buffer(pH6.9) + 300mM Sodium citrate
Flow rate    : 1.0mL/min
Detector     : UV(280nm)
Column temp. : 30deg-C

Serum in multiple myeloma patients was analyzed using Asahipak GS-520 7G ( a multimode column ).
GS-520 7G is an old type of GS-520 HQ.

Sample : Human serum
AlbuminIgA Myeloma, IgD Myeloma
IgE Myeloma, IgG Myeloma, IgMTransferrin

Columns      : Shodex Asahipak GS-520 7G (7.5mmID*500mm) x 4
Eluent       : 0.1M Sodium phosphate buffer(pH7.0) + 0.3M NaCl
Flow rate    : 1.0mL/min
Detector     : UV(254nm)
Column temp. : 37deg-C

Courtesy of Dr. Kozuru,, Kyushu Cancer Center

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