Anticoagulant Citrate Phosphate Dextrose Solution
»Anticoagulant Citrate Phosphate Dextrose Solution is a sterile solution of Citric Acid,Sodium Citrate,Monobasic Sodium Phosphate,and Dextrose in Water for Injection.It contains,in each 1000mL,not less than 2.11g and not more than 2.33g of monobasic sodium phosphate (NaH2PO4·H2O);not less than 24.22g and not more than 26.78g of dextrose (C6H12O6·H2O);not less than 19.16g and not more than 21.18g of total citrate,expressed as citric acid,anhydrous (C6H8O7);and not less than 6.21g and not more than 6.86g of Sodium (Na).It contains no antimicrobial agents.
Prepare Anticoagulant Citrate Phosphate Dextrose Solution as follows:
| Citric Acid (anhydrous) | 2.99g |
| Sodium Citrate (dihydrate) | 26.3g |
| Monobasic Sodium Phosphate (monohydrate;NaH2PO4·H2O) |
2.22g |
| Dextrose (monohydrate) | 25.5g |
| Water for Injection,a sufficient quantity,to make |
1000mL |
Dissolve the ingredients,and mix.Filter the solution until clear,place immediately in suitable containers,and sterilize.
If desired,3.27g of monohydrated citric acid may be used instead of the indicated amount of anhydrous citric acid;23.06g of anhydrous sodium citrate may be used instead of the indicated amount of dihydrated sodium citrate;1.93g of anhydrous monobasic sodium phosphate may be used instead of the indicated amount of monohydrated monobasic sodium phosphate;and 23.2g of anhydrous dextrose may be used instead of the indicated amount of monohydrated dextrose.
Packaging and storage—
Preserve in single-dose containers,of colorless,transparent,Type Ior Type IIglass,or of a suitable plastic material (see Transfusion and Infusion Assemblies and Similar Medical Devices á161ñ).
Labeling—
Label it to indicate the number of mLof Solution required per 100mLof whole blood or the number of mLof Solution required per volume of whole blood to be collected.
Identification—
It responds to the Identificationtest under Dextrose,and to the tests for Phosphate á191ñ,and,when concentrated to one-half its volume,responds to the tests for Citrate á191ñand for Sodium á191ñ.
Bacterial endotoxins á85ñ—
It contains not more than 5.56USP Endotoxin Units per mL.
pHá791ñ:
between 5.0and 6.0.
Chloride á221ñ—
A10-mLportion shows no more chloride than corresponds to 0.50mLof 0.020Nhydrochloric acid (0.0035%).
Other requirements—
It meets the requirements under Injections á1ñ.
Assay for total citrate—
Standard preparations—
Dissolve a suitable quantity of citric acid,previously dried at 90
for 3hours and accurately weighed,in water to obtain a stock solution having a known concentration of about 1.0mg of anhydrous citric acid per mL.Pipet aliquots of 8,9,10,11,and 12mL,respectively,of the stock solution into separate 100-mLvolumetric flasks,dilute with water to volume,and mix.
for 3hours and accurately weighed,in water to obtain a stock solution having a known concentration of about 1.0mg of anhydrous citric acid per mL.Pipet aliquots of 8,9,10,11,and 12mL,respectively,of the stock solution into separate 100-mLvolumetric flasks,dilute with water to volume,and mix.
Assay preparation—
Pipet 10mLof Solution into a 100-mLvolumetric flask,dilute with water to volume,and mix.Pipet 5mLof this solution into another 100-mLvolumetric flask,dilute with water to volume,and mix.
Procedure—
Pipet 1mLof the Assay preparation,each Standard preparation,and water into a suitable test tube.To each tube add 1.3mLof pyridine,and mix by swirling.To one tube at a time add 5.7mLof acetic anhydride,and mix,using a rotary vortex stirrer.Immediately place in a water bath maintained at 31±1.0,and allow the color to develop for 33±1minutes.Determine the absorbance against the reference blank in 2.5-cm cells at 425nm,taking care to measure the absorbance of each solution at the same elapsed time from mixing.Plot the absorbances of the Standard preparationsversus the concentrations,and draw the straight line best fitting the plotted points.From the graph so obtained,calculate the total citrate content,in mg per mL,of the Anticoagulant Citrate Phosphate Dextrose Solution taken by the formula:
,and allow the color to develop for 33±1minutes.Determine the absorbance against the reference blank in 2.5-cm cells at 425nm,taking care to measure the absorbance of each solution at the same elapsed time from mixing.Plot the absorbances of the Standard preparationsversus the concentrations,and draw the straight line best fitting the plotted points.From the graph so obtained,calculate the total citrate content,in mg per mL,of the Anticoagulant Citrate Phosphate Dextrose Solution taken by the formula:
0.2C,
in which Cis the concentration,in µg per mL,of anhydrous citric acid read from the standard curve.
Assay for total phosphate[expressed as monobasic sodium phosphate,monohydrate (NaH2PO4·H2O)]—
1
,2,4-Aminonaphtholsulfonic acid solution—Dissolve 0.5g of 1,2,4-aminonaphtholsulfonic acid in about 150mLof sodium metasulfite solution (3in 20)in a 200-mLvolumetric flask,warming if necessary.Add 5mLof sodium sulfite solution (1in 5),mix,and dilute with sodium metasulfite solution (3in 20)to volume.
Standard preparation—
Dissolve about 0.44g of monobasic potassium phosphate,accurately weighed,in water to make 1000mL,and mix.Pipet 25mLof this solution into a 100-mLvolumetric flask,and dilute with water to volume so as to obtain a solution having a known concentration of about 0.11mg per mLof monobasic potassium phosphate.
Assay preparation—
Dilute 5.0mLof Solution with water to 100mL.
Procedure—
Pipet 5mLof Standard preparation,5mLof Assay preparation,and 5mLof water,to provide a control,into separate 25-mLvolumetric flasks.Treat the contents of each flask as follows.Add 10.0mLof 1Nsulfuric acid,and mix.Add 2.0mLof ammonium molybdate solution (1in 40),and mix.Add 1.0mLof 1,2,4-Aminonaphtholsulfonic acid solution,dilute with water to volume,again mix,and allow to stand for 10minutes at 20to 25.Determine the absorbances of the solutions from the Standard preparationand the Assay preparationagainst the reference solution in 1-cm cells at 660nm,with a suitable spectrophotometer,taking care to measure the absorbance of each solution at the same elapsed time from mixing.Calculate the quantity,in mg,of monobasic sodium phosphate (NaH2PO4·H2O)in each mLof the Solution taken by the formula:
to 25.Determine the absorbances of the solutions from the Standard preparationand the Assay preparationagainst the reference solution in 1-cm cells at 660nm,with a suitable spectrophotometer,taking care to measure the absorbance of each solution at the same elapsed time from mixing.Calculate the quantity,in mg,of monobasic sodium phosphate (NaH2PO4·H2O)in each mLof the Solution taken by the formula:
20.28C(AU/AS),
in which Cis the concentration,in mg per mL,of KH2PO4in the Standard preparation;and AUand ASare the absorbances of the solutions from the Assay preparationand the Standard preparation,respectively.
Assay for dextrose—
Tare a clean,medium-porosity filtering crucible containing several carborundum boiling chips or glass beads.Pipet 50mLof freshly mixed alkaline cupric tartrate TSinto a 400-mLbeaker.Add the boiling chips or glass beads from the tared crucible,45mLof water,and 5.0mLof Solution to the beaker.Heat the beaker and contents over a burner that has been adjusted to cause boiling of the solution to start in 3.5to 4minutes.Boil the solution for 2minutes,accurately timed,and filter immediately through the tared crucible,taking care to transfer all of the boiling chips or glass beads to the crucible.Wash the precipitate with hot water and 10mLof alcohol.Dry the crucible and contents at 110to constant weight.Perform a blank determination,and correct the weight of the precipitate from the sample for any precipitate obtained in the blank.Each mg of cuprous oxide precipitate obtained from the substance under assay is equivalent to 0.496mg of C6H12O6·H2O.
to constant weight.Perform a blank determination,and correct the weight of the precipitate from the sample for any precipitate obtained in the blank.Each mg of cuprous oxide precipitate obtained from the substance under assay is equivalent to 0.496mg of C6H12O6·H2O.
Assay for sodium—
Proceed as directed in the Assay for sodiumunder Anticoagulant Citrate Phosphate Dextrose Adenine Solution.
Auxiliary Information—
Staff Liaison:Radhakrishna S Tirumalai,Scientist
Expert Committee:(BBP)Blood and Blood Products
USP28–NF23Page 165
Phone Number:1-301-816-8339