HBDH

α -HYDROXYBUTYRIC DEHYDROGENASE REAGENT KIT（DGKC METHOD） 3. Testing procedure

Sample volume: 6μL Temperature: 37℃
◆Product Name Reagent 1 volume: 240μL Reagent 2 volume: 60μL Main wavelength: 340nm
α-HYDROXYBUTYRIC DEHYDROGENASE REAGENT KIT（DGKC METHOD）
◆Intended Use Change of absorbance
△A/min
This reagent is applied to the invitro quantitive determination of the activity of α-hydroxybutyrate dehydrogenase（α-HBDH）
in serurn or plasma. 300s 60s 60～120s
This method is selective method for type I isoenzyme of lactate dehydrogenase, which is usually used for diseases diagnosis,
such as myocardial infarction and other relevant diseases.
◆Principle
4. Calibration
This method is recommended by Germany Institute of Clinical Chemistry (DGKC). α-HBDH catalyzes α- ketobutyric acid to
It is suggested to use supplementary calibrator: conduct 2 points calibration determination with purified water and the
be α- hydroxybutyric acid, in the process, NADH is oxidized to NAD+, by monitoring the decline rate of the absorbance at
calibrator. After the determination, the instrument will generate calibration curve automatically. When lot number is changed
340nm, α-hydroxybutyrate dehydrogenase activity can be calculated.
or QC is invalid, calibration shall be conducted again. When not using calibrator, Factor K should be inputted.
5. QC
α- HBDH
It is suggested to use QC products produced by Dirui. The laboratory shall establish its own QC area and limit. If QC value is
α- ketobutyric acid + NADH α- hydroxybutyric acid + NAD+
out of control, correction measures shall be taken.
◆Reagent Composition 6. Calculation
Reagent 1 Calculating Method When Using Calibrator:
α- ketobutyric acid 3.75 mmol / L
Buffer 50 mmol / L Sample tube △A/min
Reagent 2 Sample Concentration(U/L)= × Calibrator concentration(U/L)
NADH 0.9 mmol / L calibration tube △A/min
Buffer 50 mmol / L
Note: Ingredient in different lot kits cannot be exchanged. Calculating Method When Not Using Calibrator（340nm）:

◆Storage Conditions and Shelf Life Sample Concentration(U/L) = △A/min×Factor K = △A/min×8199

1. The reagent should be kept at temperature of 2℃~8℃ and sealed in dry place without sunshine. The shelf life is 12 months. It's suggested to use Calibrator for testing. The results obtained with Calibrator are more reliable than those without Calibrator.
2. Under condition of 2℃~8℃, the open vial stability is 30 days.
◆Reference Range
◆Suitable Device At 37℃ 72U/L~182U/L
All semi-auto and auto-chemistry analyzers are suitable. All kinds of parameters of auto-chemistry analyzers are prepared for
The reference range applied is the expected value for this method, which is for reference only. It is recommended for all
reference.
laboratories to do relevant tests to validate such range or establish their own reference range.
◆Sample Requirements
◆Explanation of Results
The most suitable samples are non- hemolyzed serum samples. The plasma samples should adopt heparin anticoagulant.
1. Sample should be diluted by physiological saline if its ΔA/min＞0.200A, and multiply diluting times to calculate the result.
The sample can be stable for 3 days when stored at 2~8℃. The hemolyzed sample could cause the rise of α-HBDH level.
2. α-HBDH determination is only one of the indicators of clinical diagnosis for patients, and clinicians also conduct a
◆Procedure comprehensive diagnosis including body, history diagnosis, as well as other items and diagnostic methods.
1. Reagent Preparation: Both reagent 1 and reagent 2 are liquid reagents and can be used directly.
◆Limit
2. Test conditions
1. The accuracy of results relies on the control of calibration, testing temperature and time.
Temperature 37℃ Reagent 1 volume 240 μL 2. When jaundice is ＞1368μmol/L, hemoglobin is ＞0.5 g/L, triglyceride is ＞5.63mmol/L, ascorbic acid is ＞5.7mmol/L,
Main Wavelength 340 nm Sample volume 6μL the test result will be affected.
Absorbance range 0~2A Reagent 2 volume 60μL ◆Specifications
1. Linearity: up to 1000 U/L
Optical path 1.0 cm Delay time 60 s
2. Blank absorbance A≥1.100
Test mode Rate assay Test time 60~120 s 3. Limit of detection: test normal saline 20 times repeatedly, and the minimum test limit is determined as 8.75U/L by average
+2 times SD.
4. Precision: test two samples with different concentration on the same test system within 20 work days.
 Precision of the same lot number n=20 Precision between days n=20 Photometric
21~26 21~26 20~27 21~31 20~27 24~33 24~36 19~27 19~23 20~28
QC serum point
X ( U/L) SD CV% X ( U/L) SD CV%
Main
340 340 340 340 340 340 340 340 340 340
Sample 1 224.9 5.13 2.28% 230.0 5.11 2.22% wavelength

Sub wavelength 405 405 405 405 405 405 405 405 405 405
Sample 2 398.6 4.64 1.16% 408.9 10.01 2.45%
Reagent R1/T1 240 240 240 240 240 240 240 240 240 240
5. Method comparison: after 200 samples are clinical tested and compared with approved market reagent (x), the relevance
between our reagent (y) and approved market reagent (x) is: y=0.9107x-0.0998，r=0.9755 R2/T2 60 60 0 60 0 60 60 60 60 0

R3/T3 —— —— 60 —— 60 —— —— —— —— 60
◆Standardization Traceability
R4/T4 —— —— 0 —— 0 —— —— —— —— 0
The constant value of calibrator can be traced to the method established by the company.
Normal volume
◆Matters Need Attention of serum sample
6 6 6 6 6 6 6 6 6 6

1. Cautions for Operation

Absorbance
1.1 The product is only for in vitro diagnosis. 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5
limit
1.2 Do not add additive during the test.
Negative Negative Negative Negative Negative Negative Negative Negative Negative Negative
1.3 Volume of reagent and sample can be changed proportionally in accordance with the requirements of instrument. Reaction type
reaction reaction reaction reaction reaction reaction reaction reaction reaction reaction
1.4 The linearity depends on proportion of sample volume and reagent volume and time used for measurement on
-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
auto-chemistry analyzer. Prozone check
limit limit limit limit limit limit limit limit limit limit
1.5 The extinction coefficient of NADH at 334nm and 365nm are 6.18 and 3.40 respectively.
Calibration 2-point 2-point 2-point 2-point 2-point 2-point 2-point 2-point 2-point 2-point
1.6 The reagent cannot be used if it is turbid or water blank absorbance value at 340 nm is less than 1.100A. method linearity linearity linearity linearity linearity linearity linearity linearity linearity linearity
1.7 Unit Conversion: U/L×16.67×10-3=μkat/L
Deflection
2. Cautions for safety check
3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3
2.1 Consider the product as dangerous materials that may cause HIV, HBV, HCV and other infections. To avoid the risk, use
Discreteness
disposable single-use gloves. check
0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1
2.2 Avoid contact with skin, clothes, and eyes. Once in contact with skin or clothes, rinse the contact part with plenty of water,
Sensitivity
and go to see a doctor. -0.07 -0.07 -0.07 -0.07 -0.07 -0.07 -0.07 -0.07 -0.07 -0.07
check
2.3 The samples and waste liquid have potential infectious risk, and the user should manage them according to the laboratory
Blank
safety operation rule, local laws and regulations. horizontal -3.3~3.3 -3.3~3.3 -3.3~3.3 -3.3~3.3 -3.3~3.3 -3.3~3.3 -3.3~3.3 -3.3~3.3 -3.3~3.3 -3.3~3.3
check
◆Reference
Linearity range 9~1000 9~1000 9~1000 9~1000 9~1000 9~1000 9~1000 9~1000 9~1000 9~1000
1. Young DS. et al, Clin. Chem. 1975; 21:5.
2. NCCLS. Interference Testing in Clinical Chemistry; Approved Guideline, 2005.
◆Date of Approval and Revision: 11/2018
◆Packaging Specification
No. Specifications Type

232013202007 R1:4×50mL R2:1×50mL Dirui CS-400/600/800/1200/1300/1600/6400 Package

232013202001 R1:4×80mL R2:4×20mL Dirui CS-240/300 Package

232013202010 R1:2×60mL R2:2×15mL Dirui CS-T Package

232013202011 R1:4×40mL R2:4×10mL Dirui CS-T Package

232013202013 R1:4×150mL R2:1×150mL 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 HBDH HBDH HBDH HBDH HBDH HBDH HBDH HBDH HBDH HBDH

Unit U/L U/L U/L U/L U/L U/L U/L U/L U/L U/L

Method Rate A Rate A Rate A Rate A Rate A Rate A Rate A Rate A Rate A Rate A

Time 20 20 10 10 10 13 10 9 12 9