TC

2. Test condition (parameters)

CHOLESTEROL REAGENT KIT（ENZYME METHOD） Temperature 37℃ Reagent1 volume 240 µL
◆Product Name Main Wavelength 500~520nm Sample volume 3 µL
CHOLESTEROL REAGENT KIT（ENZYME METHOD） Absorbance range 0~2A Reaction time 300-600s
◆Intended Use
Optical path 1.0 cm Test mode 2 point end assay
This reagent is applied to the invitro quantitive determination of total cholesterol concentration in human serum.
Serum cholesterol determination can be used as indicator to diagnose liver function, biliary function, intestinal absorption, Sub-wavelength 660nm Reagent2 volume 60 µL
coronary artery disease, thyroid function and adrenal disease. Cholesterol content on hyperlipoproteinemia diagnosis and 3. Operating procedure
classification is very important. Normal cholesterol level is affected by stress, age, sex, hormone balance and pregnancy. Sample volume: 3μL Absorbance A1 Temperature: 37℃
Reagent1 volume: 240μL Reagent2 volume: 60μL Main Wavelength:
◆Principle 500nm
Cholesterol ester in the sample, under the existence of lipoproteinesterase , hydrolyzed into cholesterol and free fatty acid.
Total cholesterol oxidized by cholesterol oxidase to generate cholest -4 - ene -3 - ketone and hydrogen peroxide. The Absorbance A2
generated hydrogen peroxide, under the existence of peroxidase, react with hydroxybenzoic acid and 4 - amino-antipyrine to 300s 300-600s
produce H2O and quinone imine pigments. The generated volume of quinone imine pigment is proportional to total cholesterol
volume in the sample, by measuring the generated pigment volume at specific wavelength, total cholesterol concentration can
be calculated. 4. Calibration
Cholesterol ester + H2O LPL Cholesterol + fatty acid It is suggested to use supplementary calibrator: conduct 2 points calibration determination with purified water and the
calibrator. After the determination, the instrument will generate calibration curve automatically. When lot number is changed
Cholesterol + O2 Cholesterol oxidase Cholesteric -4 - en -3 - keto + H2O2 or QC is invalid, calibration shall be conducted again.
H2O2 + 4 - aminoantipyrine + Para hydraxy benzoic acid POD Quinone imine pigments + H2O 5. QC
It is suggested to use QC products produced by Dirui. The laboratory shall establish its own QC area and limit. If QC value is
◆Reagent Composition
out of control, correction measures shall be taken.
Reagent 1 6. Calculating method
Lipoprotein lipase > 300 U / L Sample tube △A
Peroxidase > 750 U / L Sample concentration(mmol/L)= × Calibrator concentration(mmol/L)
p-hydroxybenzoic acid 45 mmol / L Calibration tube △A
Triton X-100 0.3%
It’s suggested to use Calibrator for testing. The results obtained with Calibrator are more reliable than those without
Buffer 50 mmol / L
Calibrator.

Reagent 2 ◆Reference range

4-aminoantipyrine 0.3 mmol / L Optimal range: ＜5.2mmol/L
Cholesterol oxidase > 300 U / L Brim rise: 5.2mmol/L～5.71mmol/L
Buffer 50 mmol / L Rise: ≥5.72mmol/L
Containing non-reactive filler and stabilizer / The reference range applied is the expected value for this method, which is for reference only. It is recommended for all
Each component in different lot kits cannot be exchanged. laboratories to do relevant tests to validate such range or establish their own reference range.
◆Storage conditions and shelf life ◆Explanation of Results
1. The reagent should be kept at temperature of 2℃~8℃ and sealed in dry place without sunshine. The shelf life is 18 months. 1. If cholesterol concentration＞20 mmol/L (or 774 mg/dL), it can be diluted by physiological saline, and multiply result by
2. Under condition of 2℃~8℃, the open vial stability is 30 days. diluting times.
2. Cholesterol determination is only one of the indicators of clinical diagnosis for patients, and clinicians also conduct a
◆Suitable Device
comprehensive diagnosis including body, history diagnosis, as well as other items and diagnostic methods.
All semi-auto and auto-chemistry analyzers are suitable. All kinds of parameters of auto-chemistry analyzers are prepared for
reference. ◆Limit
1. The accuracy of results relies on the control of calibration, testing temperature and time.
◆Sample Requirements
2. When jaundice is ＞68.8μmol/L, hemoglobin is ＞10 g/L, triglyceride is ＞50mmol/L, ascorbic acid is ＞0.57mmol/L,
Serum sample should be fresh and not be hemolyzed.
the test result will be affected.
The serum sample is stable for 3~4 days at 2~8℃ and is stable for several months at -20℃.
◆Procedure ◆Product performance index
1. Reagent preparation: both R1 and R2 are liquid and ready for use, also can be used as working reagent in proportions of 1. Linearity: up to 20 mmol/L.
4:1. 2. Blank absorbance: A≤0.100.
3. Limit of detection: test normal saline 20 times repeatedly, and the minimum test limit is determined as 0.013mmol/L by Time 20 20 10 10 10 13 10 9 12 9
average +2 times SD. Photometric
13~31 13~31 13~31 13~41 11~31 17~41 17~47 9~31 9~31 11~33
4. Precision: test two samples with different concentration on the same test system within 20 work days. point

Precision of the same lot number n=20 Precision between days n=20 Main
QC serum 505 505 505 505 505 505 505 505 505 505
wavelength
X ( mmol/L) SD CV% X ( mmol/L) SD CV%
Sub wavelength 660 660 660 660 660 660 660 660 660 660
Sample 1 4.19 0.07 1.57% 4.37 0.10 2.34% Reagent R1/T1 240 240 240 240 240 240 240 240 240 240
Sample 2 7.31 0.12 1.65% 7.67 0.18 2.39% R2/T2 60 60 0 60 0 60 60 60 60 0
5. Method comparison: after 200 samples are clinical tested and compared with approved market reagent (x), the relevance R3/T3 —— —— 60 —— 60 —— —— —— —— 60
between our reagent (y) and approved market reagent (x) is: y=0.959x-0.1272，r=0.986. R4/T4 —— —— 0 —— 0 —— —— —— —— 0
◆Standardization Traceability Normal volume
3 3 3 3 3 3 3 3 3 3
The constant value of calibrator can be traced to international reference material SRM 909b. of serum sample

Absorbance limit 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3
◆Matters Need Attention
1. Cautions for Operation Reaction type
Positive Positive Positive Positive Positive Positive Positive Positive Positive Positive
reaction reaction reaction reaction reaction reaction reaction reaction reaction reaction
1.1 The product is only for in vitro diagnosis.
1.2 Do not add additive and avoid sunshine during the test. Prozone check
-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
limit limit limit limit limit limit limit limit limit limit
1.3 Volume of reagent and sample can be changed proportionally in accordance with the requirements of instrument.
1.4 The cover shall be tightened up to prevent CO2. Calibration 2-point 2-point 2-point 2-point 2-point 2-point 2-point 2-point 2-point 2-point
1.5 Reagent cannot be used if it is substituted by water and blank absorbance value at 500 nm is more than 0.100A. method linearity linearity linearity linearity linearity linearity linearity linearity linearity linearity

1.6 Unit Conversion: mmol/L×38.7= mg/dL Deflection check 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3
1.7 Four R1 and one R2 can be used as single reagent after being mixed. The mixed reagent can be stable for 7 days if kept Discreteness
0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1
away from sunshine at 2~8℃. check
2. Cautions for safety Sensitivity check 0.03 0.03 0.03 0.03 0.03 0.03 0.03 0.03 0.03 0.03
2.1 Consider the product as dangerous materials that may cause HIV, HBV, HCV and other infections. To avoid the risk, use Blank horizontal
disposable single-use gloves. 0-0.1 0-0.1 0-0.1 0-0.1 0-0.1 0-0.1 0-0.1 0-0.1 0-0.1 0-0.1
check
2.2 Avoid contact with skin, clothes, and eyes. Once in contact with skin or clothes, rinse the contact part with plenty of water,
Linearity range 0~20 0~20 0~20 0~20 0~20 0~20 0~20 0~20 0~20 0~20
and go to see a doctor.
2.3 The samples and waste liquid have potential infectious risk, and the user should manage them according to the laboratory
safety operation rule, local laws and regulations.
◆Reference
1. Young DS. Et al. Clin Chem 1975; 21.
2. NCCLS . Interference Testing in Clinical Chemistry;Approved Guideline, 2005.
◆Date of Approval and Revision: 12/2014

◆Packaging Specification
No. Product Name Package Type

232012502008 R1: 4×50mL R2: 1×50mL Standard: 2mL DIRUI CS-400/600/800/1200/1300/1600/6400 Package

232012502002 R1: 4×80mL R2: 4×20mL Standard: 2mL DIRUI CS-240/300 Package

232012502012 R1: 2×60mL R2: 2×15mL Standard: 2mL DIRUI CS-T Package

232012502013 R1: 4×40mL R2: 4×10mL Standard: 2mL DIRUI CS-T Package

232012502019 R1: 4×150mL R2: 1×150mL Standard: 2mL 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 TC TC TC TC TC TC TC TC TC TC

Unit mmol/L mmol/L mmol/L mmol/L mmol/L mmol/L mmol/L mmol/L mmol/L mmol/L

2 point 2 point 2 point 2 point 2 point 2 point 2 point 2 point 2 point 2 point
Method
end end end end end end end end end end