[Product Name] Tri-potassium EDTA

[Formula] EDTAK3·2H2O

[Package Specification] Plastic bottle, 500g/bottle

[Performance] White crystal powder, odorless, soluble in water, easy to absorb moisture, molecular weight of 442.56, pH value of 7.3±1 (5% aqueous solution, g/ml,), solubility in water ≥60% (g/ml, at 25℃). Stable at room temperature. The main content of tri-potassium EDTA ≥99.0%. See the product test report for other indicators.

[Application] This product is specially used for in vitro anticoagulation of blood, and is an anticoagulant additive for clinical blood test. It is used for blood sample pretreatment (anticoagulation treatment) in the process of clinical blood collection and test.

[Scope of application] EDTA salt can protect the cellular components of the blood, does not affect the white blood cell count and size, has the least effect on the red blood cell morphology, and can inhibit the aggregation of platelets, suitable for general hematological tests. Except for the isolation and examination of platelets, it is not suitable for coagulation test and platelet kinetic energy test, nor for the determination of calcium, potassium, sodium, iron, alkaline phosphatase, creatine kinase and leucine amino peptidase and PCR test.

The international committee for standardization of hematology (icsh) recommends the preferred anticoagulant for blood counts and classification of tripotassium EDTA. Compared with dipotassium EDTA, it has higher solubility and faster anticoagulation rate.   
[Method of use] the product is dissolved with deionized water (distilled water), prepared into a solution with a certain concentration and content, and then added into the blood collection tube.  

I. Solution preparation method and procedure:

1. Preparation: electronic balance (Accurate to1/10,000), volumetric bottle (100ml, 200ml, 500ml, 1000ml, as required), beaker, deionized water (distilled water), glass rod, washing bottle, etc.

2. Weighing: accurate weighing of certain pre-determined tripotassium EDTA powder by electronic balance, accurate to 1%;  

3. Dissolution: This product is dissolved with distilled water and deionized water. Pour the tripotassium EDTA powder into the volumetric bottle or beaker with part of distilled water (deionized water) before stirring. Carefully pour the powder attached to the mouth and wall of the volumetric bottle or beaker into the bottle or cup with the washing bottle. Especially pay attention to the volumetric bottle and beaker to the cleaning of the volumetric flask and beaker without impurities, oil stains and oxides. Please refer to the relevant contents in the operator's Manual for the cleaning of volumetric flask and beaker. 

When dissolving with beaker, pay special attention to the control of water consumption in the process of dissolving, and reserve enough volume for EDTA tripotassium on the later cleaning glass rod, beaker and other tools, otherwise it will cause over capacity and reduce tripotassium concentration.

4. Constant volume: After all tripotassium EDTA is dissolved, the eye and the scale line of the volumetric flask shall be kept horizontal. Use the washing bottle to slowly add deionized water along the wall of the volumetric flask mouth, and wait until the concave liquid level bottom and the scale line are on the same line.

5. Sufficient mixing: Seal the opening of the volumetric flask with the volumetric cork, hold the upper part of the volumetric flask with one hand and press the cork with fingers, hold the bottom of the volumetric flask with another hand, and mix it upside down carefully for 5-8 times.

6. Inspection and confirmation: Carefully check the key factors such as the weight and volume of the tripotassium EDTA, recalculate the tripotassium EDTA content, and confirm by the supervisor. 
II. Adding method:

1. Adding with machine:

(1) Cleaning: Before using the machine, it is necessary to carefully clean the containers and delivery pipes on the machine, rinse with deionized water, then clean the containers and pipes with 80% alcohol for 3 times, then clean the containers and pipes with more than 99% ethanol, and dry the alcohol with hot air. Ensure that no impurities, water adhere to the tube and wall, avoid contamination or dilution of tripotassium EDTA.

(2)Adjusting machine: Determine the amount or volume of tripotassium EDTA to be added according to the planned volume of blood tubes, adjust the flow regulator of the machine to the appropriate value, add 10-100 tubes, and check whether the added dose meets the requirements. The admissible error of solution addition is ±10%. 

2. Add manually:

(1) Clean containers and tools: Cleaning containers and pipettes for repacking tripotassium EDTA solution with the same method as that of the machine.

(2) Distribute tripotassium solutions and tools: divide the tripotassium solutions into cleaned containers and distribute them to the operators together with the cleaned and adjusted pipettes. Note that the scale of the pipette should be adjusted to ensure that the volume of added solution meets the specified requirements.

(3) Adding solution: use a pipette to add the tripotassium solution into the blood collection tube. Confirm the quantity of each operator. In the process, random inspection shall be conducted at any time. The allowable error range of adding amount is ± 10%.

III. Drying: it can be dried by hot air blowing, drying oven, drying room, etc. The drying temperature of the hot air is 40-80 ℃ (the drying temperature of the plastic test tube is subject to the temperature that the plastic can withstand, usually no more than 40 ℃), and the drying time is 10-300 seconds. Air drying oven and drying room shall be dried at 40-80 ℃ for 2-6 hours. The drying time is related to the volume, drying temperature and blowing speed of the Tripotassium solution. Special attention should be paid to the speed of the air blast, so as not to cause the three potassium solution to be blown out of the blood tubes.

Tripotassium EDTA anticoagulants are usually best in solution, but some users require dried EDTA tripotassium tubes.

[Dosage] the recommended anticoagulant dosage range in the industry standard of vacuum blood collection is 1.5-2.2mg tripotassium EDTA per ml of blood. The anticoagulant effect of micro blood collector is the same. The company recommends 1.8-2.0mg of Tripotassium EDTA to anticoagulate 1ml blood for one-time venous vacuum blood collection tube, and 18mg of anticoagulation 1ml blood for peripheral blood anticoagulation.

Example table: tripotassium EDTA solution preparation and common additives scale

Volume of blood collection tube(ml)	2	3	4	5	6	7	8
Adding amount of Tripotassium EDTA(mg)	3.0-4.0	4.5-6.0	6.0--8.0	7.5--10.0	9.0--12.0	10.5-14.0	12.0-16.0
Volume of additive solution amount(ul)	20	30	40	50	60	70	80
Concentration of tripotassium EDTA(g/l)	150-200	150-200	150-200	150-200	150-200	150-200	150-200

Instructions:

• The dosage of tripotassium EDTA was calculated according to 15-20mg per ml of blood;
• The main content of tripotassium shall be calculated at 100%;
• In the actual production process, the added dose of tripotassium can be controlled by 1.5-2.2mg/ml blood. The main content of tripotassium is subject to the actual data in the inspection report. When the main content is less than 99% and the control value of added dosage is 1.5mg/ml, the main content error should be considered.  

4. Calculation method:

Concentration (g / L) = Weight of Tripotassium EDTA (mg) / volume of additive (ul).

Pay attention to the conversion of units. mg and ul are converted into g and ml at the same time. The percentage obtained is the grams of Tripotassium added in 100ml of water. In the end, ml needs to be converted into l by multiplying the molecular denominator by 10 at the same time. The weight of Tripotassium added in each liter of water is obtained.

[Precautions]

The key to the preparation of the whole blood test specimen is to fully and completely anticoagulant, so it is necessary to reverse 5-8 times as soon as possible after blood collection to prevent the formation of small blood clots. Especially when the ambient temperature of blood collection is higher than 25 ℃, the mixing of blood and EDTA must be timely and sufficient, otherwise it will easily lead to blood coagulation or local coagulation.

The anticoagulation of EDTA salt is not irreversible, so the test should be completed within 24 hours after blood collection in EDTA anticoagulation test tube, otherwise the test result may be wrong. The preservation time of EDTA salt anticoagulant samples was 48 hours at 4 ℃ and no more than 24 hours at 23 ℃. Platelet count can be preserved for 24 hours at 4 ℃.

Anticoagulation requires sufficient amount of EDTA, but excessive amount of EDTA may cause changes in the morphology of blood cells, so it is particularly important to control the amount of EDTA tripotassium.

EDTA salt can inhibit platelet aggregation, but it cannot completely prevent platelet aggregation. A small number of patients showed lower than normal platelets in routine blood test, which may be caused by pseudoscientification of platelets. In this case, sodium citrate anticoagulant could be used for platelet test verification. This condition has nothing to do with the quality of EDTA anticoagulant, but has more to do with the patient's blood characteristics and clinical test operation.

EDTA Tripotassium can be used in combination with sodium fluoride and separation gel. When EDTA Tripotassium / sodium fluoride is used in combination, 1.2-2.0mg Tripotassium and 2-4mg sodium fluoride can be added to each ml of blood. When EDTA Tripotassium / separation gel is mixed, the dosage of Tripotassium is the same, and the amount of separation gel added to each tube is 0.8-1.2g. The anticoagulant effect, silicification of the tube wall, centrifugation conditions and the quality of the separation gel will affect the blood separating effect when they are used together with the separation gel. It is s ested that high quality plasma samples can be obtained by using the separation gel produced by our company.

It is recommended to use γ - ray irradiation at a dose of 8-25kgy. The irradiation dose can be determined by the initial colony number.

[Warning and Prevention]

When impurities, foreign matters, abnormal smell, color, serious moisture absorption and expiration date occur in Tripotassium EDTA, please do not use.

Do not use Tripotassium EDTA solution in case of turbidity.

This product is a chemical reagent with strong corrosiveness. It is forbidden to touch it directly. When splashing on the skin, wash with plenty of water. Splashing into eyes can be washed with plenty of water immediately. In case of emergency, it should be sent to hospital for examination and treatment immediately.

[Storage Conditions] The product can be sealed and stored in a cool and dry place, away from light and moisture. The aqueous solution of this product can be sealed and stored in sterile state at room temperature. It is recommended that the aqueous solution of this product be prepared and used immediately, and not stored in solution state for a long time.

[Validity] 3 years

Desheng is an established blood test reagent company with 14 years of R&D and production experience. It has a lot of research on blood collection additives. Enterprises or individuals in need of Tripotassium EDTA can directly contact 15071057538(Wechat same number) . We look forward to your calls, to join hands in creating a better tomorrow!