Hemastix Presumptive Test for Blood

1. Principle:
1.1. Hemastix are reagent strips originally designed for use in testing for blood in urine. This quick and easy test
has been found to be applicable as a presumptive test for the presence of blood. The test strips contain
diisopropylbenzene dihydroperoxide and 3,3’,5,5’-tetramethylbenzidine. The test is based on the
peroxidase-like activity of hemoglobin which has the ability to cleave oxygen molecules from H2O2 and
catalyzes the reaction from the reduced colorless form of3,3’,5,5’- tetramethylbenzidine to the oxidized
colored form. The resulting color ranges from orange t o g re en . Very h i g h c on ce nt r at i o ns o f bl oo d
m ay c a u s e t h e c ol or development to continue to blue.
2. PREPARATIONS
2.1. Deionized Water
2.2. Hemastix
3. MINIMUM STANDARDS & CONTROLS
3.1. Positive control (known blood stain)
3.2. Negative control
4. PROCEDURE OR ANALYSIS
4.1. Check the kit tag no. and lot number and document the same in Crime Scene Detail Form
(CSPM/WS/L4/002).
4.2. Check the expiry date of the kit.
4.3. Direct Sampling Approach
4.3.1. Apply one drop of distilled/ deionized water to the pad end of the test strip, positive control or negative
control.
4.3.2. Rub the damp pad onto the questioned blood stain for not more than 5-10 sec.
4.3.3. Note color change within 60 seconds.
4.4. Indirect Sampling Approach
4.4.1. Moist a small piece of filter paper or polyester tipped applicator with distilled water
4.4.2. Once removed any water excess, rub the pointed end gently against the questioned stain, positive
control or negative control.
4.4.3. Press slightly the reactive pad of the strip on the transferred material on the filter paper or the polyester
tipped applicator for not more than 5–10 s.
4.4.4. Note color change within 60 seconds.
4.5. On-scene QC Check
4.5.1. Observe positive control and negative control for any color change.
4.5.2. The test results will be consistent if
4.5.2.1. A color change from orange to green or blue is observed in positive control within 60 seconds.
4.5.2.2. No color change is observed in negative control within 60 seconds.
4.5.3. The test results will be inconsistent if
4.5.3.1. No color change from orange to green or blue is observed in positive control within 60 seconds.
4.5.3.2. Any color change from orange to green or blue is observed in negative control within 60 seconds.
4.5.4. The QC check is documented in Crime Scene Detail Form (CSPM/WS/L4/002).
4.5.5. If the results are inconsistent, do not proceed for test interpretation with the same kit. If the kit is not
previously expired, the root cause is undertaken on Corrective & Preventive Action Request Form
(QSP/QF/L4/015) and Root Cause Analysis (RCA) Form (QSP/QF/L4/068).
4.6. Test Interpretation
4.6.1. A color change from orange to green or blue indicates a positive result.
4.6.2. No color change indicates a negative result. A negative result means there is no blood present or is
below the limit of detection of the test.
4.6.3. The test results are documented in Case Notes (CSPM/WS/L4/007).
4.7. SAFETY CONSIDERATIONS
4.7.1. Eating, drinking and smoking should be prohibited in areas where this material is handled, stored and
processed.
4.7.2. CSIs should wash hands and face before eating, drinking and smoking.
4.7.3. Store in original container protected from direct sunlight in a dry, cool and well-ventilated area.
5. REFERENE:
5.1. https://static.training.nij.gov/lab-manual/Linked%20Documents/Protocols/pdi_lab_pro_2.17.pdf
5.2. De Vittori E, Barni F, Lewis SW, Antonini G, Rapone C, Berti A. Forensic application of a rapid one-step
tetramethylbenzidine-based test for the presumptive trace detection of bloodstains at the crime scene and in
the laboratory. Forensic Chemistry. 2016 Nov 1; 2:63-74.
6. Related Documents:
6.1. Crime Scene Detail Form (CSPM/WS/L4/002).
6.2. Case Notes (CSPM/WS/L4/007)
6.3. Corrective & Preventive Action Request Form (QSP/QF/L4/015)
6.4. Root Cause Analysis (RCA) Form (QSP/QF/L4/068)