The Perfect Safety Guide for Vacutainer Blood Collection

Phlebotomy using vacutainers is by far one of the most preferred method for drawing blood samples for laboratory testing. Vacutainers are small and sterile glass containers (sometimes made of high-grade plastic). The vacuum inside these containers allow for multiple samples to be collected with a single puncture or needle prick.


Colour Coding of Vacutainers

Vacutainer blood collection tubes are coated with additives (such as anti-coagulants or preservative substances) to help preserve the blood sample. Now since these additives may be different for the type of test to be performed on the blood samples, these vacutainer tubes are colour coded (stopper colour) to help the phlebotomist identify the right container for the right type of test. There is a universal standard protocol used for these stopper colours and the order of blood draw has to be maintained to procure a viable sample.


Order of Draw

The standardised Order of Draw refers to the sequence in which the vacuum blood collection tubes must be used to collect blood samples. Since a single needle prick is used to draw the blood additives may be carried on from one sample to the next, thus contaminating samples. Using the order of draw prevents this contamination and makes the blood sample viable.

Order of Draw




Colour Code






Blood Culture Bottle

Anticoagulant and Growth Media

Blood culture for microorganisms

Light Blue Stopper

Sodium Citrate

Coagulation Studies, D-Dimer, INR

Red Stopper

No Additive

Viral Serology, Endocrinology, Vitamin D

Gold Stopper

Clot Activator and Gel

Hepatitis, CMP, BMP, U&E, LFT

Dark Green Stopper

Sodium Heparin

HLA typing, ammonia, insulin

Purple Stopper



Pink Stopper


Blood cross match, direct antiglobulin

Royal Blue Stopper


Heavy metals testing, toxicology

Gray Stopper

Sodium Fluoride

Glucose, Lactate

Yellow Stopper

Acid citrate Dextrose A

HIV testing, Tissue typing

White Stopper




For effective blood sample collection, it is very important that the phlebotomist understand the order or draw and also the number of inversions that each tube requires to maintain the integrity of the blood collection tubes and tests. The light blue tube, for example, requires to be inverted 3-4 times while the red tube 5 times and the purple tube 8 - 10 times. This is a very important part of the blood collection processes. The tubes must be labelled immediately after collection. Vacutainers are far better than single syringe draws due to the low risk of hemolysis.


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