Reagent for measuring low-density lipoproteincholesterol and preparation method

A low-density lipoprotein and cholesterol technology, which is applied in the field of reagents and preparations for the determination of serum components, can solve the problems of adverse effects of automatic biochemical analyzers, determination of LDL cholesterol, and direct detection, so as to reduce the detection workload and reduce detection time. , the effect of high accuracy

Active Publication Date: 2005-11-16
浙江伊利康生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the above-mentioned known methods cannot be directly detected on an automatic biochemical analyzer, in recent years, LDL-C reagents that are directly detected on an automatic biochemical analyzer have also appeared one after another, as disclosed in Japanese Patent No. 6-242110 A method for specifically measuring cholesterol in the target lipoprotein. This method is to aggregate lipoproteins other than the target lipoprotein to control the reaction with the enzyme, thereby measuring the target lipoprotein (LDL). This method is very practical in realizing automation Yes, but LDL can only be fractionated from lipoproteins other than HDL, but cannot be quantified and fractionated from a mixture of VLDL (very low-density lipoprotein) and CM (chylomicrons), so it is still necessary to use Determination of LDL cholesterol by separate means
At present, polyanions and divalent metal salt complexes are used to form flocculent complexes with non-measuring lipoproteins to measure lipoproteins. However, after the detection of these divalent metal salt complexes, the automatic biochemical analyzer It is easy to react with other alkaline reagent waste liquid in the drainage pipe to form sediment, block the pipe, and cause adverse effects on the automatic biochemical analyzer

Method used

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  • Reagent for measuring low-density lipoproteincholesterol and preparation method
  • Reagent for measuring low-density lipoproteincholesterol and preparation method
  • Reagent for measuring low-density lipoproteincholesterol and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1: Determination of LDL cholesterol using a reagent prepared from digitonin compound and enzyme-prepared high-affinity enzyme compound and surfactant-nonylphenol polyoxyethylene ether sodium ammonium sulfate

[0062] Reagent I (R1)

[0063] Buffer MES (PH: 7.0) 100mmol / L

[0064] Stabilizer MgCL.6H 2 O 2mmol / L

[0065] Chromogen 4-AAP 0.25mmol / L

[0066] Preservative NaN3 5mmol / L

[0067] Stabilizer BSA 0.5g / L

[0068] Catalyst sodium cholate 20mmol / L

[0069] High affinity cholesterol esterase compound 1500U / L

[0070] High affinity cholesterol oxidase compound 3000U / L

[0071] Anti-interference substance ascorbate oxidase 1000U / L

[0072] Surfactant Polyoxyethylene lauryl ether 13g / L (when reagent I is mixed with reagent II, the content is 10g / L)

[0073] Reagent II (R2)

[0074] Buffer MES (PH: 7.0) 100mmol / L

[0075] Stabilizer BSA 0.5g / L

[0076] Preservative NaN3 5mmol / L

[0077] Stabilizer MgCL.6H2O 2mmol / L

[0078] Catalyst horseradish pero...

Embodiment 2

[0083] Example 2: Determination of LDL cholesterol using a reagent prepared from aescin and enzyme-prepared high-affinity enzyme compound and surfactant-alkyl polyoxyethylene ether hydroxypropyl sulfonate

[0084] Reagent I (R1)

[0085] Buffer MOPS (PH: 6.7) 50mmol / L

[0086] Stabilizer EDTA.2Na 2mmol / L

[0087] Chromogen 4-AAP 0.20mmol / L

[0088] Preservative NaN3 5mmol / L

[0089] Stabilizer BSA 0.5g / L

[0090] Catalyst sodium cholate 15mmol / L

[0091] High affinity cholesterol esterase compound 350U / L

[0092] High affinity cholesterol oxidase compound 3500U / L

[0093] Anti-interference substance ascorbate oxidase 1000U / L

[0094]Surfactant Alkyl polyoxyethylene ether hydroxypropyl sulfonate sodium 30g / L (when reagent I is mixed with reagent II, the content is

[0095] 22.5g / L)

[0096] Reagent II (R2)

[0097] Buffer MOPS buffer (PH: 6.7) 50mmol / L

[0098] Stabilizer BSA 0.5g / L

[0099] Preservative NaN3 5mmol / L

[0100...

Embodiment 3

[0106] Example 3: Determination of LDL cholesterol using a reagent prepared from lipoprotein dehydrated protein, a high-affinity enzyme compound prepared by an enzyme, and a surfactant-polyoxyethylene higher alcohol ether.

[0107] Reagent I (R1)

[0108] Buffer PIPES (PH: 6.7) 200mmol / L

[0109] Stabilizer MgCL.6H2O 2mmol / L

[0110] Chromogen 4-AAP 0.25mmol / L

[0111] Preservative NaN3 5mmol / L

[0112] Stabilizer BSA 0.5g / L

[0113] Catalyst sodium cholate 20mmol / L

[0114] High affinity cholesterol esterase compound 2600U / L

[0115] High affinity cholesterol oxidase compound 260U / L

[0116] Anti-interference substance ascorbate oxidase 1000U / L

[0117] Surfactant Polyoxyethylene higher alcohol ether 66g / L (when reagent I is mixed with reagent II, the content is 50g / L)

[0118] Reagent II (R2)

[0119] Buffer PIPES (PH: 6.7) 200mmol / L

[0120] Stabilizer BSA 0.5g / L

[0121] Preservative NaN3 5mmol / L

[0122] Stabilizer MgCL.6H2O 2mmol / L

[0123] Catalyst horserad...

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Abstract

A reagent for detecting low - density lipoprotein cholesterol is composed of buffer solution, stabilizer, chromogen, high affinity enzyme compound, surface active agent being added in reagent as the ratio of 10 - 50 g / L, catalyst, antiseptic and anti - interference substance. Its preparing method includes activating compound, dissolving enzyme, mixing, reacting and proportioning.

Description

technical field [0001] The invention relates to a reagent and a preparation method for measuring serum components, in particular to a reagent and a preparation method for measuring cholesterol in low-density lipoprotein in serum, which can be widely used in the technical fields of medicine and biochemistry. Background technique [0002] The positive correlation between serum (serum) low-density lipoprotein cholesterol (LDL-C) and the incidence of cardiovascular and cerebrovascular diseases has been confirmed by epidemiology and related clinics. The detection of low-density lipoprotein cholesterol can also be widely used in relevant clinical diagnosis and treatment, and As one of the risk indicators of abnormal lipid metabolism. [0003] There are known ultracentrifugation and electrophoresis and the Friedewald formula method recommended by the "National Clinical Inspection Operation Regulations" (page 281 of the second edition) compiled by the Ministry of Health of the Peopl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31
Inventor 王贤理蒙凯蔡其浩
Owner 浙江伊利康生物技术有限公司
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