Prilocaine and Epinephrine Injection
»Prilocaine and Epinephrine Injection is a sterile solution prepared from Prilocaine Hydrochloride and Epinephrine with the aid of Hydrochloric Acid in Water for Injection,or a sterile solution of Prilocaine Hydrochloride and Epinephrine Bitartrate in Water for Injection.The content of epinephrine does not exceed 0.002percent (1in 50,000).Prilocaine and Epinephrine Injection contains the equivalent of not less than 95.0percent and not more than 105.0percent of the labeled amount of prilocaine hydrochloride (C13H20N2O·HCl)and the equivalent of not less than 90.0percent and not more than 115.0percent of the labeled amount of epinephrine (C9H13NO3).
Packaging and storage—
Preserve in single-dose or in multiple-dose,light-resistant containers,preferably of Type Iglass.
Labeling—
The label indicates that the Injection is not to be used if its color is pinkish or darker than slightly yellow or if it contains a precipitate.
USP Reference standards á11ñ—
USP Prilocaine Hydrochloride RS.USP Epinephrine Bitartrate RS.USP Endotoxin RS.
Color and clarity—
Using the Injection as the Test solution,proceed as directed for Color and clarityunder Epinephrine Injection.
Identification—
B:
The chromatogram of the Assay preparationobtained as directed in the Assay for prilocaine hydrochlorideexhibits a major peak for prilocaine,the retention time of which corresponds to that exhibited in the chromatogram of the Standard preparationobtained as directed in the Assay for prilocaine hydrochloride.
C:
The chromatogram of the Assay preparationobtained as directed in the Assay for epinephrineexhibits a major peak for epinephrine,the retention time of which corresponds to that exhibited in the chromatogram of the Standard preparationobtained as directed in the Assay for epinephrine.
Bacterial endotoxins á85ñ—
It contains not more than 0.9USP Endotoxin Unit per mg of prilocaine hydrochloride.
pHá791ñ:
between 3.3and 5.5.
Other requirements—
It meets the requirements under Injections á1ñ.
Assay for prilocaine hydrochloride—
Mobile phase,Standard preparation,Resolution preparation,andChromatographic system—
Proceed as directed in the Assayunder Prilocaine Hydrochloride Injection.
Assay preparation—
Transfer an accurately measured volume of Injection,equivalent to about 200mg of prilocaine hydrochloride,to a 50-mLvolumetric flask,dilute with Mobile phaseto volume,and mix.
Procedure—
[NOTE—Use peak areas where peak responses are indicated.]Separately inject equal volumes (about 10µL)of the Assay preparationand the Standard preparationinto the chromatograph,record the chromatograms,and measure the responses for the major peaks.Calculate the quantity,in mg,of prilocaine hydrochloride (C13H20N2O·HCl)in each mLof the Injection taken by the formula:
50(C/V)(rU/rS),
in which Cis the concentration,in mg per mL,of USP Prilocaine Hydrochloride RSin the Standard preparation,Vis the volume,in mL,of Injection taken,and rUand rSare the prilocaine peak responses obtained from the Assay preparationand the Standard preparation,respectively.
Assay for epinephrine—
Mobile phase—
Mix 50mLof glacial acetic acid and 930mLof water,and adjust with 1Nsodium hydroxide to a pHof 3.40.Dissolve 1.1g of sodium 1-heptanesulfonate in this solution,add 1.0mLof 0.1Mdisodium ethylenediaminetetraacetate,and mix.Mix about 9volumes of this solution with 1volume of methanol.Filter through a membrane filter (1µm or finer porosity),and degas.Make adjustments if necessary (see System Suitabilityunder Chromatography á621ñ).
Standard preparation—
Dissolve an accurately weighed quantity of USP Epinephrine Bitartrate RSin Mobile phaseto obtain a solution having a known concentration of about 9µg of epinephrine bitartrate per mL.Pipet 10mLof this solution into a 50-mLvolumetric flask,dilute with Mobile phaseto volume,and mix to obtain a Standard preparationhaving a known concentration of about 1.8µg of epinephrine bitartrate per mL.
Assay preparation—
Transfer an accurately measured volume of Injection,equivalent to about 50µg of epinephrine,to a 50-mLvolumetric flask,dilute with Mobile phaseto volume,and mix.
Chromatographic system
(see Chromatography á621ñ)—The liquid chromatograph is fitted with a 30-cm ×3.9-mm stainless steel column packed with packing L1,and is equipped with an electrochemical detector held at a potential of +650mV,a controller capable of regulating the background current,and a suitable recorder,and it is operated at a temperature between 20
and 25maintained at ±1.0of the selected temperature.The flow rate is about 1mLper minute.Chromatograph the Standard preparationas directed for Procedure:the relative standard deviation of the peak responses of successive injections of the Standard preparationis not more than 1.5%.
and 25maintained at ±1.0of the selected temperature.The flow rate is about 1mLper minute.Chromatograph the Standard preparationas directed for Procedure:the relative standard deviation of the peak responses of successive injections of the Standard preparationis not more than 1.5%.
Procedure—
Separately inject equal volumes (about 20µL)of the Assay preparationand the Standard preparationinto the chromatograph,record the chromatograms,and measure the responses for the major peaks.Calculate the quantity,in µg,of epinephrine (C9H13NO3)in each mLof the Injection taken by the formula:
(183.21/333.30)50(C/V)(rU/rS),
in which 183.21and 333.30are the molecular weights of epinephrine and epinephrine bitartrate,respectively,Cis the concentration,in µg per mL,of USP Epinephrine Bitartrate RSin the Standard preparation,Vis the volume,in mL,of Injection taken,and rUand rSare the peak responses obtained from the Assay preparationand the Standard preparation,respectively.
Auxiliary Information—
Staff Liaison:Karen A Russo,Ph.D.,Scientist
Expert Committee:(PA1)Pharmaceutical Analysis 1
USP28–NF23Page 1618
Phone Number:1-301-816-8379