automat ic analyzer reagents lipids - veredus …...addition concentration 20 mg/dl 500 mg/dl 50...

Automatic Analyzer Reagents

Lipids

02 Cholestest® TG

04 Cholestest® CHO

06 Cholestest® N HDL

08 Cholestest® LDL

Lipids Automatic Analyzer Reagents

2

1. Purpose of use

2. Features

3. Components and Ingredients

4. Measurement principle (Enzyme/Free glycerol eliminated method)

Free glycerol is eliminated in the first reaction, and triglyceride is measured in the second reaction.

For the measurement of Triglyceride(TG) in serum or plasma

TG, the main constituent of body fat, is formed by the esterfication of glycerol and three fatty acids. Measurement of TG is useful in the diagnosis of abnormal lipid metabolism.TG has been attracting attention for its correlation with arterial sclerosis and coronary arteriopathy.

Cholestest® TGTriglyceride kit

1. Minimally affected by cross-contamination.2. Free glycerol is eliminated by the enzymatic method.3. There are no influence of Bilirubin, Hemolysis, Ascorbic acid

Main reaction(by Reagent2)

Neutral fat Glycerol Fatty acid＋LPL

ATP ADPGlycerol-3-phosphoric acidGlycerol ＋＋ GK

O₂O₂

dihydroxyacetone phosphate

dihydroxyacetone phosphate

Glycerol-3-phosphoric acid H₂O₂H₂O₂

H₂O O₂

＋＋ GPOGK

GPO

ESBmTred-purple colorH₂O₂ ＋ POD

4-aminoantipyrine

*LPL：Lipoprotein lipase*GPO：Glycerol-3-phosphoric acid oxidase

*ATP：Adenosine triphosphatase*GK：Glycerol kinase

Pre reaction by Reagent1(Eliminatio of Glycerol)

Glycerol ATP Glycerol-3-phosphoric acid ADP＋

＋

＋

＋

＋H₂O₂ Catalase

⃝ Reagent 1 　 N-ethyl-N-sulfobutyl-m-toluidine sodium, good buffer(pH7.0) 　 Glycerol kinase, Glycerol-3-phosphoric acid oxidase

⃝ Reagent 2　 4-aminoantipyrine, Lipoprotein lipase, 　 Peroxidase, good buffer(pH6.5)

Glycerol-3-phosphoric acid

*ESBmT：N-Ethyl-N-Sulfobutyl-m-Toluidine

3

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Chorestest® TG

5. Data

6. TraceabilityTriglyceride

secondary standard within the company

daily method

JCCRM224-5a

Cholestest N calibrator

standard method within the company


JSCC recommended method

ReCCs

SEKISUI MEDICAL

SEKISUI MEDICAL

each laboratory

daily method

■ Within-run reproducibility

■ Stability

■ LinearitySample1 Sample2 Sample3

n 20 20 20 Mean 103.6 254.6 119.6 S.D. 0.89 1.70 1.19 C.V.（％） 0.86 0.67 0.99 Max. 106 258 121 Min. 102 252 117 Range 4 6 4

0/10 2/10 4/10 6/10 8/10 10/10

3,000

2,700

2,400

2,100

1,800

1,500

1,200

900

600

300

0

150

135

120

105

90

75

60

45

30

15

00 200 400 600 800100 300 500 700 900 1,000

1,000

900

800

700

600

500

400

300

200

100

0

（mg/dL）

y = 0.9946x + 0.3r = 0.9997n = 50X平均 = 175.8Y平均 = 174.6σ = 2.70

（mg/dL）（ 1 ）（ 2 ）

Dilution

（mg/dL）

■ InterferenceF-BIL C-BIL Hb Ascorbic acid Rheumatoid factor Glycerol

addition concentration 20 mg/dL 20 mg/dL 500 mg/dL 50 mg/dL 500 U/mL 5000mg/dL

measurement value

Base Serum

104.0 104.0 105.0 108.0 118.0 101.5

Including interfering substance

103.0 103.5 106.5 105.0 117.0 102.5

（mg/dL）

0 7 14 21 28 35

300.0

250.0

200.0

150.0

100.0

50.0

0.0

On board day

（mg/dL）

materials method operatorCalibration

value assignment

results

4


4. Measurement principle (enzyme method/COD-POD method)

⃝ Reagent 1 　 4-aminoantipyrine, Cholesterol Esterase, Peroxidase

⃝ Reagent 2 　Cholesterol oxydase, N-ethyl-N-sulfobutyl-m-toluidine sodium

*CE：Cholesterol Esterase*COD：Cholesterol oxydase

*POD：Peroxidase*ESBmT：N-Ethyl-N-Sulfobutyl-m-Toluidine

Cholestest® CHOCholesterol kit

Esterified cholesterol Free cholesterol

Free cholesterol ⊿4-Cholestenone

4-aminoantipyrine

H₂O₂

H₂O₂ ESBmT

fatty acid

red-purple color

＋

＋

＋＋

COD

CE

POD

1. Purpose of use

2. Features

For the measurement of total cholesterol (CHO) in serum or plasma.

CHO is produced in the liver and is also dietary-derived. Measurement of CHO is useful for the diagnosis of abnormal lipid metabolism, liver disease, and metabolic disease.

1. Minimally affected by cross-contamination.2. There are no influence of Bilirubin, Hemolysis, Ascorbic acid

Cholesterol can be categorized into ester type and free type. Ester type cholesterol is converted to free type cholesterol by cholesterol esterase. Free type cholesterol produces H₂O₂ during the reaction, and ultimately developing a red-purple color. CHO can be calculated from the change in absorbance.

5

Cholestest® CHO

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NIST SRM911

daily samples

results


daily method

Scientific analyticalmethod

SI unit

NIST

SEKISUI MEDICAL

each laboratory


■ Stability


n 20 20 20Mean 114.1 252.1 174.8S.D. 0.85 2.02 1.47C.V.（％） 0.75 0.80 0.84Max. 116 255 178Min. 113 249 173Range 3 6 5

5. Data

6. TraceabilityCholesterol (CHO)

materials method operator

0/10 2/10 4/10 6/10 8/10 10/10

1,250

1,125

1,000

875

750

625

500

375

250

125

0

300

270

240

210

180

150

120

90

60

30

0

Dilution

（mg/dL）（ 1 ）（ 2 ）

0 40 80 120 160 200 240 280 320 360 400

400

360

320

280

240

200

160

120

80

40

0

（mg/dL）

y = 1.0069x - 1.7r = 0.9988n = 50X平均 = 194.4Y平均 = 194.1σ = 1.50

（mg/dL）

■ InterferenceF-BIL C-BIL Hb Chyle Ascorbic acid Rheumatoid factor

addition concentration 20 mg/dL 20 mg/dL 500 mg/dL 3000 formazin turbidity 50 mg/dL 500 U/mL

measurement value

Base plasma

161.5 160.0 161.5 164.0 167.0 181.0


160.0 161.5 161.0 172.5 165.5 181.0

（mg/dL）

0 7 14 21 28 35

300.0

250.0

200.0

150.0

100.0

50.0

0.0

On board day

（mg/dL）

Calibrationvalue assignment

6

1. Purpose of use

2. Features


4. Measurement principle (direct method)

⃝ Reagent 1 　 N,N-bis(4-sulfobutyl)-m-toluidine2Na(DSBmT)　 Choresterol oxidase(from bacteria), peroxidase, good buffer(pH6.0)

⃝ Reagent 2 　4-aminoantipyrine,Cholesterol Esterase,Surface acting agent, good buffer(pH6.0)

Unique detergent selectively solubilizes HDL to specifically measure HDL.

For the measurement of HDL-C in serum or plasma

The decreases in HDL cholesterol concentration is associated with coronary artery disease, hyperlipidemia, smoking, obesity, diabetes and hepatic diseases; increase in the concentration is associated with alcohol intake or moderate exercise.In addition, factors such as age, gender, and genetic predisposition are known to affect the concentration of HDL cholesterol.

HDL

HDL

LDL VLDL

CODCE

1. Accurate value can be obtained from highly chylous samples2. Accurate value can be obtained from samples with high immune globulin levels3. No burden on the instrument as the reagent does not contain metal ions such as Mg salt 4. There are no influence of Bilirubin, Hemolysis, Ascorbic acid

HDL will be solublization

H₂O₂

ChylomicronSurface acting agent

*CE：Cholesterol Esterase*POD：Peroxidase

*COD：Cholesterol oxydase*ESBmT：N-Ethyl-N-Sulfobutyl-m-Toluidine

⊿4-Chorestenone ＋

4-aminoantipyrineH₂O₂ ESBmT red-purple color＋＋ POD

Cholestest® N HDLHDL-C kit

7

Chorestest® N HDL

（mg/dL）

0 15 30 45 60 75 90 105 120 135 150

150

135

120

105

90

75

60

45

30

15

0

（mg/dL）

0/10 2/10 4/10 6/10 8/10 10/10

200

180

160

140

120

100

80

60

40

20

0

100

90

80

70

60

50

40

30

20

10

0

y = 0.9904x - 0.3r = 0.9994n = 50X平均 = 52.16Y平均 = 51.96σ = 0.50

（ 1 ）（ 2 ）

Dilution

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6. TraceabilityHDL-C

JCCRM224-5a

results


■ Stability


n 20 20 20 Mean 38.72 90.78 53.77 S.D. 0.38 0.54 0.47 C.V.（％） 0.98 0.59 0.87 Max. 39.6 91.9 54.5 Min. 38.1 89.8 53.0 Range 1.5 2.1 1.5

5. Data（mg/dL）



measurement value

Base plasma

49.0 49.3 49.7 49.8 50.6 55.8


50.8 50.0 50.5 49.0 50.2 55.0

（mg/dL）

0 7 14 21 28 35

125.00

100.00

75.00

50.00

25.00

0.00

On board day

（mg/dL）



daily samples


ReCCs

SEKISUI MEDICAL

SEKISUI MEDICAL

each laboratory



ultracentrifugal separationheparin-Mn method

daily method


8


4. Measurement principle (direct method)

⃝ Reagent 1 　4-aminoantipyrine,Choresterol oxidase, Cholesterol Esterase 　Peroxidase, Surface acting agent1, good buffer(pH6.3) ⃝ Reagent 2 　N,N-bis(4-sulfobutyl)-m-toluidine2Na(DSBmT) 　Surface acting agent2, good buffer(pH6.3)

The use of 2 types of unique detergents enables highly specific measurement of LDL-C.

For the measurement of LDL-C in serum or plasma

Traditionally, measurement of total cholesterol had been widely used for the diagnosis of hyperlipidemia, one of the causes of arteriosclerotic disease. However, in 1986, Health, Labor, and Welfare Ministry reported that ischemic heart disease has a stronger correlation with LDL cholesterol level.

HDL

LDL

VLDL

COD

COD

CE

POD

CE

1. Correct value can be obtained from highly chylous samples (up to TG 1500mg/dL)2. Good correlation with βquantification3. There are no influence of Bilirubin, Hemolysis, Ascorbic acid

Micellar cholesterol

Micellar cholesterol

Micellar cholesterol H₂O₂

Micellar cholesterol H₂O₂

H₂O₂

ChylomicronSurface acting agent1

DSBmT red-purple color＋

H₂O₂ no color4-aminoantipyrine

4-aminoantipyrine

＋

＋ POD

*CE：Cholesterol Esterase*COD：Cholesterol oxydase

*POD：Peroxidase*DSBｍT：N,N-bis(4-sulfobutyl)-m-Toluidine

LDL LDLSurface acting agent1

Surface acting agent2

Cholestest® LDLLDL-C kit

1. Purpose of use

2. Features

Reaction on Reagent1：Surface acting agent1 get choresterol except LDL erased. Reaction on Reagent2(Only LDL will be to color)

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9

JCCRM224-5a

Beta Quantification

ReCCs

SEKISUI MEDICAL

SEKISUI MEDICAL

each laboratoryeach laboratory

5. Data■ Within-run reproducibility

■ Stability


n 20 20 20 Mean 65.1 115.5 106.2 S.D. 0.69 1.00 0.49 C.V.（％） 1.06 0.87 0.46 Max. 66 117 107 Min. 64 114 105 Range 2 3 2

（mg/dL）

0 7 14 21 28 35

150.00

125.00

100.00

75.00

50.00

25.00

0.00

On board day

（mg/dL）

（mg/dL）

0/10 2/10 4/10 6/10 8/10 10/10 0 30 60 90 120 150 180 210 240 270 300

300

270

240

210

180

150

120

90

60

30

0

（mg/dL）

y = 1.0082x - 1.3r = 0.9989n = 50X平均 = 113.7Y平均 = 113.4σ = 1.20

700

630

560

490

420

350

280

210

140

70

0

200

180

160

140

120

100

80

60

40

20

0

（ 1 ）（ 2 ）

Dilution



measurement value

Base plasma

95.5 96.0 95.0 96.5 97.0 105.0


93.5 93.5 96.0 95.5 96.0 105.5

（mg/dL）

6. Traceability


LDL-C



daily samples



daily method

Cholestest® LDL


For more information contact:

International Business Department Diagnostics Division

13-5, Nihonbashi 3-chome, Chuo-ku, Tokyo 103-0027 JAPANTel: +81-3-3272-0828, Fax: +81-3-3272-0907E-mail: [email protected]: www.sekisuimedical.jp/english

automat ic analyzer reagents lipids - veredus …...addition concentration 20 mg/dl 500 mg/dl 50...

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