Spectrophotometric determinations of serum phospholipids with preconcentrations of molybdophosphate-Malachite Green aggregates using a membrane filter

• Published in: BUNSEKI KAGAKU Vol. 38; no. 11; pp. 627 - 631
• Main Authors: MATSUBARA, Chiyo, TAKAMURA, Kiyoko, YONEHARA, Yoshiyuki, YOSHIHARA, Kazuhiro, KOBAYASHI, Narumi
• Format: Journal Article
• Language: Japanese
• Published: The Japan Society for Analytical Chemistry 05.11.1989
• Subjects: enzymatic hydrolysis of phospholipids; molybdophosphate-Malachite Green aggregate; preconcentration using a membrane filter; spectrophotometric determinations of serum phospholipids
• ISSN: 0525-1931
• DOI: 10.2116/bunsekikagaku.38.11_627

High sensitive spectrophotometric determinations of serum phospholipids were conducted. The levels of serum phospholipids reflect liver functions, so thier determinations are clinically important. Phospholipids are hydrolyzed by phospholipase C and alkakine phosphatase with the release of phosphate. The resulting phosphate associated with Mo-MG reagent (mixed solution of ammonium molybdate and Malachite Green) to form molybdophosphate-Malachite Green aggregates. The aggregates were selectively collected on a nitro cellulose membrane filter (pore size:3.0 μm) and dissolved in methylcellosolve along with the membrane filter. Absorbance (λ: 627 nm) was proportional to the amount of phospholipids. Molar absorptivity of the aggregates in methylcellosolve (ε=2.8 × 105 M-1 cm-1) was large enough to minimize the serum sample for determination. Ascorbic acid, bilirubin, Na+, K+, Ca2+ and Cl- in serum did not affect the results obtained by this method.