phospholipids - cedarlane · phospholipids play an important function in the composition of cell...

ww

w.w

ako

dia

gn

ost

ics.

com

Wako Diagnostics

• 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

Buffer Solution 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

Choline oxidase-DAOS method

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

© Copyright 2017 Wako Life Sciences, Inc. Wako Life Sciences, Inc. 1025 Terra Bella Ave. Mountain View, CA 94043

E: [email protected] T: 650.210.9153 www.wakodiagnostics.com

Correlation

0 100 200 300 400 500 6000

100

200

300

400

500

600

Phos

phol

ipid

s C-

Test

Wak

o, m

g/dL

Phospholipids B-Test Wako, mg/dL

y = 1.008x - 3.8r = 0.994

Method

WLS-1956-2

Performance characteristics

PrincipleWhen 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 absorbance of the blue color.

Specimen collection and preparation• Use serum as a specimen

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

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

�

��

��

��

��

��

��

��

��

��

��

��

��

��

� ��

��

��

��

��

��

��

��

��

��

��

��

� ��

��

��

��

��

��

+

+POD

C2H5

N+

OCH3

H3CO

H3CO

OCH3

N

N

CN

CC

OH3C

H3C

Blue pigment

C2H5

OH

N

NH2

2H2O2 +

H3C

H3C O

C C

OH

N C

N CH2CHCH2SO3Na

DAOS

CH2CHCH2SO3NaOH- 3H2O

4-Aminoantipyrine

Phospholipids +LecithinLysolecithinSphingomyelin

H2O

2O2

HOCH2CH2N(CH3)3+OH-

Phospholipase D

+

+

+

Choline oxidase2H2O2

Choline

HOCH2CH2N(CH3)3+OH-

-OOC-CH2N+(CH3)3

Phosphatidic acidN-Acyl-sphingosylphosphateLysophosphatidic acid

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

��

� ��

��

��

��

��

��

� ��

��

��

��

��

��

��

� � ��

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.

For more product information, please contact customer service at 1-877-714-1924.

phospholipids - cedarlane · phospholipids play an important function in the composition of cell...

Documents