PhospholipidsCholine oxidase-DAOS method

• Phospholipid specific enzymatic assay

• Measures Sphingomyelin, Lecithin, and Lysolecithin

• Linear up to 1000 mg/dL

Phospholipids play an important function in the composition of cell membranes and in the

emulsification and absorption of fat in the body. Serum phospholipids are formed in the

liver, where they become bound to apolipoproteins, which makes them soluble in plasma.

The Wako Phospholipids C assay is an enzymatic method utilizing choline oxidase in a

reaction which can be measured photometrically. This assay has been shown not to be

significantly influenced by coexisting substances such as ascorbic acid and bilirubin.

Catalog No. Product Name Pkg Size Storage

997-01801 Phospholipids C

8 X 50 mL 2-10°C

Color Reagent 8 X for 50 mL 2-10°C

Standard Solution 2 X 10 mL 2-10°C

Buffer Solution

For Research Use Only. Not for use in diagnostic procedures.

Performance characteristics

Principle

When a sample is added to the Color Reagent, phospholipids (lecithin, lysolecithin, sphingomyelin) in the sample are hydrolyzed by phospholipase D to produce choline, which in turn is oxidized by choline oxidase to betaine and hydrogen peroxide. The hydrogen peroxide produced causes N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline (DAOS) and 4-aminoantipyrine to undergo a quantitative oxidative condensation catalyzed by peroxidase (POD), producing a blue pigment. The amount of phospholipids in the sample is determined by measuring the absor-bance of the blue color.

Correlation

For additional information, please contact customer service at:Email: wakodx-customerservice@fuji�lm.comPhone: 877-714-1924

© 2018 FUJIFILM Wako Diagnostics U.S.A. Corporation

WLS-1956-3

Method

Specimen collection and preparation• Use serum as a specimen

• Hemolysis, ascorbic acid and bilirubin do not havesignificant influence on the assay

• Lecithin, lysolecithin and sphingomyelin of phospholipids are measured, but phosphatidyl ethanolamine in the sample is not measured.

0 100 200 300 400 500 6000

100

200

300

400

500

600

Pho

spho

lipid

s C

-Tes

t W

ako,

mg/

dL

Phospholipids B-Test Wako, mg/dL

y = 1.008x - 3.8r = 0.994

PrecisionWithin-run precision

Run # Sample # Replicates Mean (U/mL) SD CV (%)

Run 1 Serum A 10 163.2 0.63 0.39

Run 1 Serum B 10 191.9 0.74 0.39

InstrumentsVarious automated analyzer applications are available.

PhospholipidsPhospholipase D

+ H2OLecithinLysolecithinSphingomyelin

Choline oxidase

POD

HOCH2CH2N(CH3)3 +OH- +

Choline

HOCH2CH2N(CH3)3 +OH- + 202

OOC-CH2N+(CH3)3 + 2H2O2

H3C C C NH2

2H2O2 + H3C N N O

Phosphatidic acidN-Acyl-sphingosylphosphateLysophosphatidic acid

H3C C C N

N

H3C N C O

N

N+

H3CO

H3CO

OCH3 OH

C2H5

OCH3

C2H5

CH2CHCH2SO3Na

OH

4-Aminoantipyrine DOAS

Blue pigment

+

CH2CHCH2SO3NA OH- + 3H2O

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