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2) prietest bilirubin total & direct ver. 0prietest .Clinical Chemistry Reagents
BILIRUBIN TOTAL & DIRECT
In vitro diagnostic test kit, for professional use only
Concentration in Sample (mg/dl) = (Abs. of Sample – Abs. of Sample Blank) X 14 (mg/dl)
Concentration in Sample (µmol/L)= (Abs. of Sample – Abs. of Sample Blank) X 239 (µmol/L)
INTENDED USE : Quantitative in vitro determination of concentration of Bilirubin in
CONVERSION FACTOR : mg/dl X 17.1 = µmol/L
serum on photometric systems.
For the calibration of automated photometric systems use of the commercially available calibrator is recommended. QUALITY CONTROL :
CLINICAL SIGNIFICANCE : Red blood cells at the end of their circulating life are broken
To ensure adequate quality, use of the commercially available control sera is down in the reticulo-endothelial systems, mainly the spleen. The resulting haem, once the iron is removed, is then converted to bilirubin (yellow orange bile pigment). Water insoluble bilirubin is called indirect or un-conjugated bilirubin is then released into the MEASURING RANGE:
blood stream where it binds tightly to albumin and is transported to the liver. In the liver The test has been developed to determine Bilirubin concentrations within a measuring un-conjugated bilirubin binds with glucoronic acid (mono and di glucoronides) to form range from 0.05 to 25 mg/dl (0.85 to 427.5 µmol/L). When values exceed higher limit of conjugated bilirubin (direct bilirubin) by the enzyme glucoronyl transferase. Conjugated the range, such samples should be diluted 1 + 1 with NaCl solution (9 g/I) and the result bilirubin or direct bilirubin is excreted via the biliary system into the intestine. multiplied by 2.
Total bilirubin is the sum of un-conjugated bilirubin and conjugated fractions. Bilirubin is SPECIFICITY / INTERFERENCE :
elevated in the hemolysis or lysis of increased breakdown of red blood cells, hepatitis, No interference was observed by Glucose upto 300 mg/dl (16.65 mmol/L), Hemoglobin cirrhosis azo obstruction of the biliary tract e.g. Gallstone. up to 0.5 g/dl (5 g/L) and lipemia up to 600 mg/dl (6.78 mmol/L) of Triglycerides. A list METHOD : Modified Jendrassik & Grof.
of drugs and other interfering substances with Bilirubin determination has been reported PRINCIPLE :
In the determination of total bilirubin, bilirubin is coupled with diazotized sulphanilic acid SENSITIVITY / LIMIT OF DETECTION :
in the presence of caffeine benzoate solution to produce azobilirubin which has The lower limit of detection is 0.05 mg/dl (0.85 µmol/L).
maximum absorbance at 546 nm. Direct Bilirubin in presence of diazotized sulphanilic PRECISION :
acid forms a red colored azo compound in acidic medium which has maximum Intra-assay precision n = 20 absorbance at 546 nm.
R1 : Sulphanilic acid R2 : Sodium Nitrite R4 : Sodium Chloride Preservative & Stabilizer
STORAGE INSTRUCTIONS AND REAGENT STABILITY :
METHOD COMPARISON :
The reagents are stable up to the end of the indicated date of expiry on the vial label, if A comparison between Robonik Prietest Bilirubin (y) and a commercially available test (x) using 20 samples gave following results: stored at 15 to 30 C, protected from light and contamination is avoided. Do not freeze Bilirubin Total Bilirubin Direct
: y = 1.054x - 0.1668 mg/dl Linear Regression : y = 1.0147x +0.3825 mg/dl
WARNINGS AND PRECAUTIONS :
Correlation Coefficient : r = 0.9948 Correlation Coefficient : r = 0.9939
Take the necessary precautions for the use of laboratory reagents. Avoid contact with REFERENCE RANGE :
skin and eyes. If spilled, thoroughly wash affected area with water, flush with plenty of Bilirubin Total Up to 1.1 mg/dl water while disposing. Do not use mouth pipette.
Bilirubin Direct Up to 0.25 mg/dl WASTE MANAGEMENT :
0 to 1 day < 8.0 mg/dl Please refer to local regulation requirements.
1 to 2 days < 12 mg/dl REAGENT PREPARATION :
3 to 5 days < 16 mg/dl The reagents are ready-to-use. Above 5 days 0.3 to 1.2 mg/dl (5.13 to 20.52 µmol/L) MATERIAL REQUIRED BUT NOT PROVIDED :
It is recommended that each laboratory should assign its own normal range. NaCl solution 9 g/l, General laboratory equipment, Analyser / Photometer, pipettes LITERATURE :
Nuttall, K.L., Klee, G.G., Analytes of haemoglobin metabolism-Porphyrins, Iron and SPECIMEN :
Bilirubin, Tietz Fundamentals of Clinical Chemistry, 5 Ed., Burtis, C. A. & Ashwood, Serum free from hemolysis. E.R. (W.B. Saunders eds. Philadelphia USA). Storage: Specimen should be protected from bright light as bilirubin is photo labile.
Tietz, N.W., Clinical guide to laboratory tests. 3 Ed., (W.B. Saunders eds. Specimen may be stored refrigerated for 3 days. Philadelphia USA). ASSAY PROCEDURE : 1 - Total Bilirubin End Point (Differential)
Vassault, A., et al., Protocole de validation de techniques. (Document B, stade 3), Ann. Application sheets for automated systems are available on request.
Biol. Clin.,. (1986) Young DS. Effects of drugs on Clinical Lab. Tests, 4 ed. AACC Press, 1995.
Room Temperature End Point, Differential prietest TOUCH Bring all the contents of the kit to Room Temperature prior to use.
Name : T-BIL , Mod : DIFF Read absorbance of Sample against Sample Blank.
Pri.: 546 , Sec.: 0 Label the test tube as Sample Blank, Sample for every patient sample. Control Blank and Temp: 30C , KF : 1.000 control sample for every control sample. Pipette into respective test tube the reagent, Vol : 500ul , Unit : mg/dl sample, control sample as per the table given below : Sample Blank / Control Blank Std : N , Factor: 14 QCABH = *QCABL = * Mix, let stand at room temperature for 5 minutes and read the absorbance of sample Rgnt. Linearity : 25 against the sample blank.
* Indicates user definable parameter.
Concentration in Sample (mg/dl) = (Abs. of Sample – Abs. of Sample Blank) X 14 (mg/dl) Concentration in Sample (µmol/L)= (Abs. of Sample – Abs. of Sample Blank) X 239 (µmol/L) NA Implies Not Applicable Reagent Linearity ASSAY PROCEDURE : 2 - Direct Bilirubin End Point (Differential)
NOTE : Instrument application is similar for Direct Bilirubin Test except name of the test &
Normal / Reference range. "Assay procedure 2" to be followed for Direct Bilirubin Test.
Room Temperature prietest is the Trade Mark of ROBONIK (INDIA) PVT.LTD., for Clinical Chemistry Reagents. End Point, Differential prietest TOUCH is the Trade Mark of ROBONIK (INDIA) PVT. LTD., for Biochemistry Analyser. Read absorbance of Sample against sample blank.
Label the test tube as Sample Blank, Sample for every patient sample. Control Blank and Manufactured and Marketed by:
control sample for every control sample. Pipette into respective test tube the reagent, ROBONIK (INDIA) PVT. LTD.,
sample, control sample as per the table given below : A-374, TTC Industrial Area, Mahape, Navi Mumbai - 400 710, INDIA.
Sample Blank / Control Blank Tel. No.: + 91 (22) 67829700 An ISO 9001 : 2008 Certified Company
Fax. No.: + 91 (22) 67829701 An ISO 13485 : 2003 Certified Company
Orders : firstname.lastname@example.org Queries : email@example.com Website : www.robonik.in Mix, let stand at room temperature for EXACTLY 5 minutes and read the absorbance of
For in vitro diagnostic use Consult Instructions for use REF Catalogue Number sample against the sample blank. Manufacturer's Address Date of Manufacture
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