1.Reagent preparation：Reagent is liquid reagent ready to use.

LDL-C 2.Test condition (parameters)

LOW DENSITY LIPOPROTEIN CHOLESTEROL REAGENT KIT（DIRECT METHOD） Temperature 37℃ Reagent 1 300 μL
Main Wavelength 540～570nm Reagent 2 100μL
◆Product Name Sub Wavelength 660nm Sample volume 4 μL
Test mode 2 point end assay Reaction time 300 s
LOW DENSITY LIPOPROTEIN CHOLESTEROL REAGENT KIT（DIRECT METHOD）
Absorbance Range 0~2 A Optical path 1.0cm
◆Intended Use 3.Testing procedure
The reagent is applied to the invitro quantitative determination of Low Density Lipoprotein-Cholesterol (LDL-C) Record absorbency A1 Temperature: 37℃
Sample volume: 4μL
concentration of human serum. Main wavelength: 546nm
Reagent R1volume:300μL
The rise of LDL-C is the dangerous factor of atherosclerosis and especially of coronary heart disease. The determination of Reagent R2volume:100μL Sub wavelength: 660nm
total-cholesterol was commonly used for hyperlipidemia diagnosis which is one of the pathogeny of atherosclerosis.
Compared to total-cholesterol value, LDL-C value is more relevant to deplethoric heart disease. Record absorbance A2

◆Principle 300s 300s

The low-density lipoprotein cholesterol in the sample, under the action of surfactant in the reagent, was selectively catalyze
hydrolyze to cholesterol and free fatty acids. The generated cholesterol oxidized by cholesterol oxidase to generate cholest -4 -
ene - 3 - ketone and hydrogen peroxide, hydrogen peroxide catalyzed by peroxidase react with aniline color of the original
material and 4 - amino-antipyrine to produce H2O and quinone imine pigments. The volume of quinone imine pigments is 4. Calibration
proportional to low-density lipoprotein cholesterol. By measuring the generated pigment volume at specific wavelength, the It is suggested to use supplementary calibrator to conduct 2-point calibration. The calibration curve is formed automatically.
concentration of low-density lipoprotein cholesterol in the sample can be calculated. When lot number is changed or QC is invalid, calibration shall be conducted again.
Cholesterol esterase 5. QC
Low-density lipoprotein cholesterol ester ＋ H2O Cholesterol ＋ Free fatty acid It is suggested to use QC products produced by Dirui. The laboratory shall establish its own QC area and limit. If QC value is
Cholesterol oxidase out of control, correction measures shall be taken.
6. Calculation
Cholesterol + O2 Cholest-4- ene -3- ketone ＋ H2O2
Sample tube △A
Peroxidase
Sample concentration(mg/dL) = ×Calibrator concentration (mg/dL)
H2O2 + 4-Aminoantipyrine + Aniline color of the original material Quinone imine pigment + H2O
Calibration tube△ A
◆Reagent Composition
◆Reference Range
Reagent 1
4-aminoantipyrine 1mmol/L Suitable Range and Hypercholesterolemia Judging Standard
Cholesterol Oxidase 500U/L Serum total-cholesterol LDL-cholesterol
Peroxidase 800U/L Suitable range Below 200.08mg/dL Below 119.97mg/dL
Cholesterol Esterase 800U/L
Critical range 200.08mg/dL ~219mg/dL 119.97mg/dL ~138.93mg/dL
Non-ionic Surfacetant 0.5%
MOPS Buffer 100mmol/L Hypercholesterolemia Above 219.82 mg/dL Above 140.09 mg/dL
Polymeric Compound Appropriate The reference range applied is the expected value for this method, which is for reference only. It is recommended for all
Reagent 2 laboratories to do relevant tests to validate such range or establish their own reference range.
DSBmT 1.2% ◆Explanation of Results
Non-ionic Surfacetant 0.5%
1. If the density of the sample exceeds detection range, please retest after diluted with normal saline, multiplied by the dilution
MOPS Buffer 100mmol/L
multiple results.
Each component in different lot kits can not be exchanged
2. The result of the test on the low density lipoprotein-cholesterol concentration is only one of the indicators for clinicians’
◆Storage Conditions and Shelf Life diagnosis. Clinicians should conduct a comprehensive diagnosis according to the patient’s body, historical diagnosis, as well
When stored at 2~8℃ and protected from light, the reagent is valid for 12 months. The used reagent is valid for 30 days at as other items and diagnostic methods.
2~8℃.
◆Limit
◆Suitable Device
1. The accuracy of results relies on the calibration of instrument, control of testing temperature and time.
Any kind of semi-auto and auto-chemistry analyzers; all kinds of parameters of auto-chemistry analyzers are prepared for
2. If jaundice＞68.8μmol/L, , hemoglobin＞10g/L, triglyceride＞37mmol/L, VC＞1.7mmol/L probably have interference on
reference.
the result. .
◆Sample Requirements ◆Specifications
Serum sample. Hemolysis, fat or polluted sample can not be used. 1. Linearity: up to 450mg/dL
Serum sample should not be hemdytic. Avoid repeated freezing and thrawing sample. The samples are valid for 7 days at 2. Blank absorbance: A≤0.150
2~8℃ and 1 month at -20℃. 3. The minimum test limit: test normal saline 20 times repeatedly, and the minimum test limit is determined as 3.26 mg/dL
◆Procedure average +2 times SD.
4. Precision: test two samples with different concentration on the same test system within 20 work days. Method
2 point 2 point 2 point 2 point 2 point 2 point 2 point 2 point 2 point 2 point
end end end end end end end end end end
Precision of the same lot number n=20 Precision between days n=20
QC serum Time 20 20 10 10 10 13 10 9 12 9
X ( mg/dL) SD CV% X ( mg/dL) SD CV% Photometric
13~31 13~31 13~31 13~41 11~31 17~41 17~47 9~31 9~31 11~33
point
Sample 1 92.7 1.35 1.46 92.3 1.41 1.53 Main
546 546 546 546 546 546 546 546 546 546
wavelength
Sample 2 134.2 2.07 1.54 132.7 1.51 1.14
Sub
660 660 660 660 660 660 660 660 660 660
5. Method comparison: Conduct tests upon 200 samples with reagent y of the company and the on the market reagent x, the wavelength
relevance between our reagent (y) and approved market reagent (x) is: y=0.753x+20.686 r=0.982 Reagent
300 300 300 300 300 300 300 300 300 300
◆Standardized Traceability R1/T1

The constant value of calibrator can be traced to international reference SRM1951b. R2/T2 100 100 0 100 0 100 100 100 100 0

R3/T3 —— —— 100 —— 100 —— —— —— —— 100

◆Matters Need Attention
R4/T4 —— —— 0 —— 0 —— —— —— —— 0
1. Cautions for operation
Normal
1.1 The product is only for in vitro diagnosis.
volume of
1.2 Do not add additive during the test. serum
4 4 4 4 4 4 4 4 4 4

1.3 Volume of reagent and sample can be changed proportionally in accordance with the requirements of instrument. sample
1.4 Avoid storing the reagent at 0℃ or below. Absorbance
3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3
limit
1.5 Units conversion: mmol/L × 38.7 = mg/dL
Reaction Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive
2. Cautions for safety type reaction reaction reaction reaction reaction reaction reaction reaction reaction reaction
2.1 Consider the product as dangerous materials that may cause HIV, HBV, HCV and other infections. To avoid the risk, use Prozone -3.3 lower -3.3 lower -3.3 lower -3.3 lower -3.3 lower -3.3 lower -3.3 lower -3.3 lower -3.3 lower -3.3 lower
disposable single-use gloves. check limit limit limit limit limit limit limit limit limit limit
2.2 Avoid contact with skin, clothes, and eyes. Once in contact with skin or clothes, rinse the contact part with plenty of water, Calibration 2-point 2-point 2-point 2-point 2-point 2-point 2-point 2-point 2-point 2-point
method linearity linearity linearity linearity linearity linearity linearity linearity linearity linearity
and go to see a doctor.
Deflection
2.3 The samples and waste liquid have potential infectious risk, and the user should manage them according to the laboratory check
3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3
safety operation rule, local laws and regulations. Discreteness
0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1
check
◆Reference
Sensitivity
0.03 0.03 0.03 0.03 0.03 0.03 0.03 0.03 0.03 0.03
1. Young DS. Effects of Drugs on Clinical Laboratory Tests. AACC Press, 1990. check
Blank
2. Guo Zhanjun, etc.，The Study Progress of Detection Method of Low Density Lipoprotein Cholesterol, Clinical
horizontal -0.01~0.15 -0.01~0.15 -0.01~0.15 -0.01~0.15 -0.01~0.15 -0.01~0.15 -0.01~0.15 -0.01~0.15 -0.01~0.15 -0.01~0.15
Biochemistry and Ecsomatics Fascicle of Foreign Medical, 21/6，p289～290（2000）. check
Linearity
3. Liu Zhaoyin,etc.，The Study Progress of Analysis Method of Serum LDL-C Automatic，Clinical Biochemistry and 3~450 3~450 3~450 3~450 3~450 3~450 3~450 3~450 3~450 3~450
range
Ecsomatics Fascicle of Foreign Medical，20/2，51～52（1999）.
4. Miki,etc.，Measurement of LDL-Cholesterol，U.S. Pat.5,814,472.
5. NCCLS.Interference Testing in Clinical Chemistry;Approved Guideline,2005.
◆Date of Approval and Revision: 02/2014
◆Packaging Specification
No. Specifications Type

232012802007 R1:3×50mL R2:2×25mL Standard：0.8mL Dirui CS-400/600/800/1200/1300/1600/6400 package

232012802001 R1:4×60mL R2:4×20mL Standard：0.8mL Dirui CS-240/300 package

232012802012 R1:2×60mL R2:2×20mL Standard：0.8mL Dirui CS-T series package

232012802013 R1:4×30mL R2:4×10mL Standard：0.8mL Dirui CS-T series package

232012802015 R1:3×150mL R2:1×150mL Standard：0.8mL Dirui CS-1600/6400 package

◆P.S.：CS Series Auto-Chemistry analyzer parameters

Model CS-240 CS-300 CS-400 CS-600 CS-1200 T240 T300 CS-6400 CS-1600 CS-1300

Item LDL-C LDL-C LDL-C LDL-C LDL-C LDL-C LDL-C LDL-C LDL-C LDL-C

Unit mg/dL mg/dL mg/dL mg/dL mg/dL mg/dL mg/dL mg/dL mg/dL mg/dL