DETERMINATION OF PHOSPHATIDYLCHOLINE IN A FLOW-INJECTION SYSTEM USING IMMOBILIZED ENZYME REACTORS

被引：33

作者：

MASOOM, M ^{[1
]}

ROBERTI, R ^{[1
]}

BINAGLIA, L ^{[1
]}

机构：

[1] UNIV PERUGIA, SCH MED, DEPT BIOCHEM, I-06100 PERUGIA, ITALY

来源：

ANALYTICAL BIOCHEMISTRY | 1990年 / 187卷 / 02期

关键词：

D O I：

10.1016/0003-2697(90)90450-N

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Two alternative procedures are described for the quantitative determination of phosphatidylcholine in a flow-injection system utilizing immobilized enzymes. Phospholipase C from Bacillus cereus and phospholipase D from cabbage were covalently bound to the surface of controlled-pore glass beads and the enzyme-derivatized beads were packed in small columns. In the first procedure, the phospholipase C column was connected with a second column containing coimmobilized alkaline phosphatase and choline oxidase. In the alternative procedure, the column packed with immobilized phospholipase D was connected with a column packed with immobilized choline oxidase. The hydrogen peroxide produced through the action of choline oxidase in both flow-injection systems was detected amperometrically. Both procedures are suitable for an accurate and rapid quantitation of phosphatidylcholine. The sensitivity of the method based on phospholipase C and alkaline phosphatase is higher than that using phospholipase D. Quantitation of phosphatidylcholine at the nanomole level can be easily obtained using the first method. © 1990.

引用

页码：240 / 245

页数：6

共 11 条

[1]

BLATON V, 1983, CLIN CHEM, V29, P806

[2]

FOLCH J, 1957, J BIOL CHEM, V226, P497

[3]

GURANTZ D, 1981, J LIPID RES, V22, P373

[4] COMPLETE SEPARATION OF PHOSPHOLIPIDS FROM HUMAN-HEART COMBINING 2 HPLC METHODS [J].

JUANEDA, P ;

ROCQUELIN, G .

LIPIDS, 1986, 21 (03) :239-240

[5]

LOWRY OH, 1951, J BIOL CHEM, V193, P265

[6] SOME CHARACTERISTICS OF PHOSPHOLIPASE-C FROM BACILLUS-CEREUS [J].

OTNAESS, AB ;

LITTLE, C ;

SLETTEN, K ;

WALLIN, R ;

JOHNSEN, S ;

FLENGSRUD, R ;

PRYDZ, H .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 1977, 79 (02) :459-468

[7] POROUS GLASS AS A SOLID SUPPORT FOR IMMOBILISATION OR AFFINITY CHROMATOGRAPHY OF ENZYMES [J].

ROBINSON, PJ ;

DUNNILL, P ;

LILLY, MD .

BIOCHIMICA ET BIOPHYSICA ACTA, 1971, 242 (03) :659-+

[8]

SCHIEFER S, 1985, METHOD ENZYMAT AN, V8, P87

[9]

SCHRIEWER H, 1983, J CLIN CHEM CLIN BIO, V21, P611

[10]

TAKAYAMA M, 1977, CLIN CHIM ACTA, V79, P93, DOI 10.1016/0009-8981(77)90465-X

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