Abstract
Cyanobacterial glycogen has gained interest as a valuable biomass feedstock for biofuel production. However, an ideal method for native glycogen quantification has not been developed. Here, we have proposed a simple methodology that enables the quantitative determination of cyanobacterial glycogen concentration with high repeatability using aqueous size-exclusion chromatography with a differential refractive index detector (SEC/RID). Our SEC/RID system also allows size distributions for native glycogen based on hydrodynamic volumes (Vh), which is proportional to the product of the molecular mass (M) and intrinsic viscosity [η], obtained by universal calibration using linear homopolymers of known M with Mark-Houwink-Sakurada parameters. The universal calibration curve achieved a broad linear range (Vh parameter [η]M=2×102-8×108mLg-1) with a high correlation coefficient (R2=0.9942), because the developed system is equipped with an OHpak SB-806M HQ aqueous column containing four types of polyhydroxy methacrylate-based particles with different particle and pore sizes. Based on the SEC/RID system, response of molecular size distribution of glycogen in microalgae to the cultivation condition was first observed. Our established SEC/RID method has several advantages over conventional techniques, including the simultaneous quantitative and size distribution analyses of glycogen, and represents a potentially useful tool to elucidate the relationship between structural properties and the roles of glycogen in metabolism.
| Original language | English |
|---|---|
| Pages (from-to) | 90-97 |
| Number of pages | 8 |
| Journal | Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences |
| Volume | 930 |
| DOIs | |
| Publication status | Published - Jul 1 2013 |
| Externally published | Yes |
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All Science Journal Classification (ASJC) codes
- Biochemistry
- Analytical Chemistry
- Cell Biology
- Clinical Biochemistry
Cite this
Aqueous size-exclusion chromatographic method for the quantification of cyanobacterial native glycogen. / Izumi, Yoshihiro; Aikawa, Shimpei; Matsuda, Fumio; Hasunuma, Tomohisa; Kondo, Akihiko.
In: Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, Vol. 930, 01.07.2013, p. 90-97.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Aqueous size-exclusion chromatographic method for the quantification of cyanobacterial native glycogen
AU - Izumi, Yoshihiro
AU - Aikawa, Shimpei
AU - Matsuda, Fumio
AU - Hasunuma, Tomohisa
AU - Kondo, Akihiko
PY - 2013/7/1
Y1 - 2013/7/1
N2 - Cyanobacterial glycogen has gained interest as a valuable biomass feedstock for biofuel production. However, an ideal method for native glycogen quantification has not been developed. Here, we have proposed a simple methodology that enables the quantitative determination of cyanobacterial glycogen concentration with high repeatability using aqueous size-exclusion chromatography with a differential refractive index detector (SEC/RID). Our SEC/RID system also allows size distributions for native glycogen based on hydrodynamic volumes (Vh), which is proportional to the product of the molecular mass (M) and intrinsic viscosity [η], obtained by universal calibration using linear homopolymers of known M with Mark-Houwink-Sakurada parameters. The universal calibration curve achieved a broad linear range (Vh parameter [η]M=2×102-8×108mLg-1) with a high correlation coefficient (R2=0.9942), because the developed system is equipped with an OHpak SB-806M HQ aqueous column containing four types of polyhydroxy methacrylate-based particles with different particle and pore sizes. Based on the SEC/RID system, response of molecular size distribution of glycogen in microalgae to the cultivation condition was first observed. Our established SEC/RID method has several advantages over conventional techniques, including the simultaneous quantitative and size distribution analyses of glycogen, and represents a potentially useful tool to elucidate the relationship between structural properties and the roles of glycogen in metabolism.
AB - Cyanobacterial glycogen has gained interest as a valuable biomass feedstock for biofuel production. However, an ideal method for native glycogen quantification has not been developed. Here, we have proposed a simple methodology that enables the quantitative determination of cyanobacterial glycogen concentration with high repeatability using aqueous size-exclusion chromatography with a differential refractive index detector (SEC/RID). Our SEC/RID system also allows size distributions for native glycogen based on hydrodynamic volumes (Vh), which is proportional to the product of the molecular mass (M) and intrinsic viscosity [η], obtained by universal calibration using linear homopolymers of known M with Mark-Houwink-Sakurada parameters. The universal calibration curve achieved a broad linear range (Vh parameter [η]M=2×102-8×108mLg-1) with a high correlation coefficient (R2=0.9942), because the developed system is equipped with an OHpak SB-806M HQ aqueous column containing four types of polyhydroxy methacrylate-based particles with different particle and pore sizes. Based on the SEC/RID system, response of molecular size distribution of glycogen in microalgae to the cultivation condition was first observed. Our established SEC/RID method has several advantages over conventional techniques, including the simultaneous quantitative and size distribution analyses of glycogen, and represents a potentially useful tool to elucidate the relationship between structural properties and the roles of glycogen in metabolism.
UR - http://www.scopus.com/inward/record.url?scp=84878543992&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84878543992&partnerID=8YFLogxK
U2 - 10.1016/j.jchromb.2013.04.037
DO - 10.1016/j.jchromb.2013.04.037
M3 - Article
C2 - 23727872
AN - SCOPUS:84878543992
VL - 930
SP - 90
EP - 97
JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
SN - 1570-0232
ER -