Automat ic Analyzer Reagents Lipids - Veredus …...addition concentration 20 mg/dL 500 mg/dL 50...
Transcript of 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
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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
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
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800
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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
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3. Components and Ingredients
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.
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Cholestest® CHO
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Cholestest N calibrator
NIST SRM911
daily samples
results
standard method within the company
daily method
Scientific analyticalmethod
SI unit
NIST
SEKISUI MEDICAL
each laboratory
■ Within-run reproducibility
■ Stability
■ LinearitySample1 Sample2 Sample3
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
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320
280
240
200
160
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80
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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
Including interfering substance
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
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1. Purpose of use
2. Features
3. Components and Ingredients
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
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Chorestest® N HDL
(mg/dL)
0 15 30 45 60 75 90 105 120 135 150
150
135
120
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(mg/dL)
0/10 2/10 4/10 6/10 8/10 10/10
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90
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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
■ Within-run reproducibility
■ Stability
■ LinearitySample1 Sample2 Sample3
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)
■ 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
49.0 49.3 49.7 49.8 50.6 55.8
Including interfering substance
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)
materials method operator
secondary standard within the company
daily samples
Cholestest N calibrator
ReCCs
SEKISUI MEDICAL
SEKISUI MEDICAL
each laboratory
standard method within the company
standard method within the company
ultracentrifugal separationheparin-Mn method
daily method
Calibrationvalue assignment
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3. Components and Ingredients
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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JCCRM224-5a
Beta Quantification
ReCCs
SEKISUI MEDICAL
SEKISUI MEDICAL
each laboratoryeach laboratory
5. Data■ Within-run reproducibility
■ Stability
■ LinearitySample1 Sample2 Sample3
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
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60
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(mg/dL)
y = 1.0082x - 1.3r = 0.9989n = 50X平均 = 113.7Y平均 = 113.4σ = 1.20
700
630
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420
350
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0
200
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160
140
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0
( 1 ) ( 2 )
Dilution
■ 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
95.5 96.0 95.0 96.5 97.0 105.0
Including interfering substance
93.5 93.5 96.0 95.5 96.0 105.5
(mg/dL)
6. Traceability
materials method operator
LDL-C
Cholestest N calibrator
secondary standard within the company
daily samples
standard method within the company
standard method within the company
daily method
Cholestest® LDL
Calibrationvalue assignment
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