Title: Manual Colorimetric Chloride Reagent
Students: Allison Speer and Donatella Stiers
Team: Team Five
Faculty Mentor: Bill Zundel and Gary NielsenAbstract:
Date: 10-8-04

 A cost effective, manual colorimetric chloride reagent will be prepared and validated for use on the Roche Cobas Mira S Chemistry Analyzer.  Following the Technicon® AutoAnalyzer® protocol, the reagent will be prepared using raw materials purchased from an established supplier and tested in conjunction with commercially prepared standards, controls and reagents.  This project will provide guidance for the reproduction of a working in-house colorimetric chloride reagent for use in the clinical chemistry laboratory.  A double blind study will be performed by collecting 60 random serum samples from two hospital archives with all patient identification removed and utilizing them for pre-use validation. This will include but may not be limited to studies of accuracy, linearity, precision, sensitivity and correlation with the commercially prepared reagent.

Introduction:
Weber State University’s Clinical Laboratory Sciences (CLS) program provides students an invaluable opportunity to practice a broad range of currently applied analytical practices and methods in a working laboratory setting.  As the extent of these opportunities is affected by budgetary constraints, reducing costs where possible may contribute to the quality and variety of activities experienced.   The CLS department recently acquired two Roche Cobas Mira S automated chemistry analyzers capable of, but not limited to, Ion Selective detection of Na+, K+ and Cl- (Roche).  In house production of a chloride reagent grants the student an opportunity to practice clinical chemistry skills while meeting curriculum requirements and potentially reducing the operational and maintenance costs associated with these analyzers.  Team Five will determine the practicability of producing such a reagent (TFCR-05).

Objective:
Team Five is proposing that a quality, colorimetric method chloride reagent for the Cobas Mira S, can be successfully reproduced by students of the CLS department for less than commercially prepared reagents.

Methods:
A manual colorimetric chloride method will be used.  The testing will be done by the Cobas Mira S random analysis analyzer.  A maximum of 60 specimens will be analyzed for chloride levels utilizing a commercially prepared chloride reagent.  The same specimens will again be tested using TFCR-05. Each run will be preceded by a standard, controls (normal and abnormal), and five levels of calibrators.  All of which have a known level of analyte. TFCR-05 will be reproduced as per Technicon® Auto Analyzer® Methodology for mercuric thiocyanate chloride reagent (Technicon).  Assuming the results of the standard, controls and calibrators are accurate; the results of each run will be compared in correlation studies.  This consists of accuracy, linearity, correlation coefficient, within run, run to run, and minimum detection limit.

Materials:
The following items are needed for this research project:

Patient samples
Commercial colorimetric chloride reagent
Controls
Standard
Calibrator
Mercuric thiocyanate
Ferric nitrate
Mercuric nitrate
Nitric acid
Amber bottles
Analyzer cuvettes

References:
Technicon Laboratory Methods File N-56 I/II. Technicon AutoAnalyzer Methodology:  Chloride: Terrytown, New York: Technicon Instruments Corporation, 1970.
Roche Diagnostic Systems. Cobas Mira Plus. New Jersey: Roche, 3rd Edition 1994.