An HPLC assay for rat liver ferrochelatase activity | Zendy

Li Famei | Zendy; Lim Chang Kee | Zendy; Peters Timothy J. | Zendy

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An HPLC assay for rat liver ferrochelatase activity

Author(s) -

Li Famei,

Lim Chang Kee,

Peters Timothy J.

Publication year - 1987

Publication title -

biomedical chromatography

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.4

H-Index - 65

eISSN - 1099-0801

pISSN - 0269-3879

DOI - 10.1002/bmc.1130020408

Subject(s) - chemistry , ferrochelatase , protoporphyrin ix , chromatography , protoporphyrin , high performance liquid chromatography , zinc , methanol , zinc protoporphyrin , biochemistry , heme , porphyrin , photodynamic therapy , enzyme , organic chemistry

A rapid, reliable, sensitive and reproducible HPLC method was developed for the assay of ferrochelatase activity in rat liver. The assay was carried out aerobically with Zn 2+ and mesoporphyrin or protoporphyrin IX as substrates. Zn‐porphyrins formed were extracted with dimethyl sulphoxide/methanol (30:70, v/v) containing Zn‐deuteroporphyrin as the internal standard for separation and quantification by reversed‐phase chromatography. The K m for mesoporphyrin was 5.9 μM, for protoporphyrin IX 8.8 μM and for zinc 6.0 μM. The specific activities were 33.1 ± 5.0 nmol Zn‐mesoporphyrin or 13.4 ± 2.0 nmol Zn‐protoporphyrin formed per hour per mg of protein for mitochondria and 12.3 ± 2.2 nmol Zn‐mesoporphyrin or 4.6 ± 0.9 nmol Zn‐protoporphyrin per hour per mg of protein for liver homogenate.

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