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If the cholesterol concentration of samples is higher than 2 mmol/L (800 mg/L), then protocol CHO2D should be used (same protocol with pre-set 1:10 sample predilution). 

Test definition	CHO2B (751)	CHO2D (752)
Measuring range      mmol/L      mg/L	Standard 0.01 – 2.07 3.87 – 800	High 0.1 – 20.7(1)  38.7 – 8,000(1)
Control level	B 1, 2, 3	Custom
Sample predilution	No	1:10
Measuring mode	Absorbance	Absorbance
Abs. calc. mode	Endpoint	Endpoint
Reaction mode	R-S	R-S
Reaction direction	Increase	Increase
Wavelength A/B	512/659 nm	512/659 nm
Calc. first/last	17/69	17/69
Unit	mmol/L, mg/L	mmol/L, mg/L

1. Up to 207 mmol/L (80 g/L) with automatic dilution.

Pipetting parameters	CHO2B (751)	CHO2D (752)
Water Diluent (Sample Predilution)(1)	No	20 μL sample + 180 μL water
R	47 μL + 55 μL water	47 μL + 55 μL water
Sample (S)	20 μL + 20 μL water	20 μL + 20 μL water
Total Volume	142 μL	142 μL

1. Sample predilution is performed in a prior step, after which the reaction mix is prepared in the same way as the unmodified sample. 

Calibration
Calibrator	Calibrator B Bio (CAL B, 07-1105-5)
Calibration mode	Linear regression
Calibration interval	Each new lot, and if recalibration is required due to QC results.

• Control B Level 1 Bio (CONB1, 07-1106-3)
• Control B Level 2 Bio (CONB2, 07-1107-1)
• Control B Level 3 Bio (CONB3, 07-1109-8)

• Custom control: For quality control of CHO2D, use custom controls with cholesterol concentrations in the high range of 0.15 to 7.5 g/L. Ensure that the cholesterol in the control material stays in solution during the automatic 1:10 dilution step with water.
  The cholesterol in the Controls B Level 1-3 Bio is stabilized but begins to precipitate if diluted with water. Therefore, these controls are not suitable for control of the CHO2D protocol because the automatic dilution step may cause too low recovery of cholesterol.

Use the recommended control material. Other suitable control material can also be used.
Control intervals and acceptance limits should be adapted to each lab\u00adoratory's individual requirements. If values do not fall within the defined limits, corrective measures and recalibration are required.Conversion factors for cholesterol concentration:

• 1 mmol/L = 386.6 mg/L
• 1 g/L = 2.59 mmol/L

This method has been standardized by isotope dilution mass spectrometry (ID-MS) and according to Abell-Kendall.Representative performance data on Cedex Bio HT Analyzers are shown. Results obtained in individual laboratories may differ. Precision was determined in samples of three concentration levels. Coefficients of variation (CV) were calculated for in-run precision (n=21) and inter-run precision (on 10 days).

	Level 1	Level 2	Level 3
Mean	0.051 mmol/L (20 mg/L)	0.12 mmol/L (46 mg/L)	1.39 mmol/L (537 mg/L)
CV in-run [%]	1.4	1.3	0.2
CV inter-run [%]	1.9	1.4	1.2

", "Country": "XG", "Code": "Protocols", "Name": "Protocols" }, { "Language": "en", "Value": "

Vial	Content	Composition
CHO2B	Reagent cassette, 400 tests	Reagent: PIPES, 225 mmol/L, pH 6.8; choline esterase (Pseudomonas spec.), ≥1.5 kU/L; cholesterol oxidase (E. coli), ≥0.45 kU/L; horseradish peroxidase, ≥0.75 kU/L; Mg2+, 10 mmol/L; 4-aminoantipyrine, ≥0.45 mmol/L; phenol, ≥12.6 mmol/L; sodium cholate, 0.6 mmol/L; non-reactive preservative; stabilizer.

", "Country": "XG", "Code": "Content", "Name": "Content" }, { "Language": "en", "Value": "Unit of measure is \"piece\".", "Country": "XG", "Code": "CB - Order Information", "Name": "CB - Order Information" }, { "Language": "en", "Value": "

• Cedex Bio HT Analyzer*, including accessories and disposables
• Standard laboratory equipment

• Calibrator B Bio*
• Control B Level 1 Bio*
• Control B Level 2 Bio*
• Control B Level 3 Bio*

", "Country": "XG", "Code": "Additional Equipment and Reagent Required", "Name": "Additional Equipment and Reagent Required" }, { "Language": "en", "Value": "Cholesterol is a major component of all biological membranes. In cell cultures, cholesterol can be produced by the cells, nevertheless, a certain level of cholesterol in the culture medium is advantageous for cell growth and viability. With a controlled cholesterol concentration, cells grow faster and to higher density. For control of culture media and for monitoring in a culture process, the cholesterol concentration can be determined on a Cedex Bio or Cedex Bio HT Analyzer with high reliability and convenience.", "Country": "XG", "Code": "Applications", "Name": "Applications" }, { "Language": "en", "Value": "Method: Enzymatic photometric assay using cholesterol oxidase.
Recovery: ±10%", "Country": "XG", "Code": "Specification", "Name": "Specification" }, { "Language": "en", "Value": "

• Handle all samples as if potentially infectious, using safe laboratory procedures. As the sensitivity and titer of potential pathogens in the sample material can vary, the operator must optimize pathogen inactivation and follow the appropriate measures according to local safety regulations.
• Do not eat, drink, or smoke in the laboratory work area.
• Do not pipette by mouth.
• Wear protective disposable gloves, laboratory coats, and eye protection, when handling samples and kit reagents.
• Wash hands thoroughly after handling samples and reagents.

• Discard unused reagents and waste in accordance with country, federal, state, and local regulations.
• Safety Data Sheets (SDS) are available online at documentation.roche.com, or upon request from the local Roche office.

", "Country": "XG", "Code": "Safety Information", "Name": "Safety Information" } ] } } ] }