Ampicillin Capsule USP32

.

4903103 4903105 4903106

4903119

4903166

Drug Control I 1 2553

Ampicillin Capsule USP32Ampicillin Penicillin beta lactam ring endocarditis; streptococci, pneumococci, nonpenicillinase-producing staphylococci, Listeria, meningococci; H. influenza, Salmonella, Shigella, E. coli, Enterobacter Klebsiella

Ampicillin capsule 500 mg

Ampicillin capsule USP32

Ampicillin Capsules USP32 Ampicillin Capsules contain an amount of ampicillin (anhydrous or as the trihydrate) equivalent to not less than 90.0 percent and not more than 120.0 percent of the labeled amount of ampicillin (C16H19N3O4S). Packaging and storage Preserve in tight containers. Labeling Label the Capsules to indicate whether the ampicillin therein is in the anhydrous form or is the trihydrate. USP Reference standards Identification Prepare a solution containing about 5 mg of ampicillin per mL, using powder from Ampicillin Capsules, in a mixture of acetone and 0.1 N hydrochloric acid (4:1). Prepare 11 USP Ampicillin RS.

a Standard solution of USP Ampicillin RS to contain 5 mg per mL in the same solvent mixture. Apply separately 2 L of each solution on a thin-layer chromatographic plate (see Chromatography chromatographic silica gel mixture. Place the plate in a suitable chromatographic chamber, and develop the acetone, water, toluene, and glacial acetic acid 621 ) coated with a 0.25-mm layer of

chromatogram in a solvent system consisting of a mixture of (650:100:100:25). When the solvent front has moved about three-fourths of the length of the plate, remove the plate from the chamber, mark the solvent front, and allow to air-dry. Locate the spots on the plate by spraying lightly with a

solution of ninhydrin in alcohol containing 3 mg per mL, and dry at 90 for 15 minutes: the RF value of the principal spot obtained from the solution under test corresponds to that obtained from the Standard solution. Dissolution, Procedure for a Pooled Sample Medium: water; 900 mL. Apparatus 1: 100 rpm. Time: 45 minutes. 711

Standard preparation Dissolve an accurately weighed quantity of USP Ampicillin RS in water to obtain a solution having a known concentration of about L/900 mg per mL, L being the labeled amount, in mg, of ampicillin per Capsule. Procedure Proceed as directed for Procedure in the section AntibioticsHydroxylamine Assay under Automated Methods of

Analysis

16 , using a filtered portion of the solution under

test as the Assay preparation. Calculate the quantity, in mg, of C16H19N3O4S dissolved by the formula: 0.9CP(AU / AS).

Tolerances Not less than 75% (Q) of the labeled amount of C16H19N3O4S is dissolved in 45 minutes. Uniformity of dosage units Water, Method I 905 : meet the requirements. 921 : not more than 4.0% where the

Capsules contain anhydrous Ampicillin, or between 10.0% and 15.0% where the Capsules contain Ampicillin trihydrate. Residual solvents Assay 467 : meet the requirements. (Official January 1, 2007) Standard preparation Prepare as directed for Standard Preparation under Iodometric AssayAntibiotics USP Ampicillin RS.

425 , using

Assay preparation Place not fewer than 5 Capsules in a high-speed glass blender jar containing an accurately measured volume of water, and blend for 4 1 minutes. Dilute an accurately measured volume of this stock solution quantitatively and stepwise with water to obtain an Assay Procedure Proceed as directed for Procedure under Iodometric AssayAntibiotics formula: preparation containing about 1.25 mg of ampicillin per mL. 425 . Calculate the quantity, in

mg, of ampicillin (C16H19N3O4S) in each Capsule taken by the

(T / D)(F / 2000)(B

I),

in which T is the labeled quantity, in mg, of ampicillin in each Capsule; and D is the concentration, in mg per mL, of quantity in each Capsule and the extent of dilution. ampicillin in the Assay preparation on the basis of the labeled

IODOMETRIC ASSAYANTIBIOTICSThe following method is provided for the assay of most of the Pharmacopeial penicillin antibiotic drugs and their dosage forms, for which iodometric titration is particularly suitable. Standard Preparation Dissolve in the solvent specified in the table of Solvents and Final Concentrations a suitable quantity of the USP Reference Standard specified in the individual monograph, previously dried under the conditions specified in the individual monograph and accurately weighed, and dilute quantitatively and stepwise with the same solvent to obtain a

solution having a known concentration of about that specified in the table. Pipet 2.0 mL of this solution into each of two 125-mL glass-stoppered conical flasks. Solvents and Final Concentrations Antibiotic Amoxicillin Ampicillin Ampicillin Sodium Cloxacillin Sodium Cyclacillin Dicloxacillin Sodium Methicillin Sodium Nafcillin Sodium Oxacillin Sodium Penicillin G Potassium Solvent Water Water Buffer No. 1 Water Water Buffer No. 1 Buffer No. 1 Buffer No. 1 Buffer No. 1 Buffer No. 1 Buffer No. 1 Buffer No. 1 Buffer No. 1*

Final Concentration 1.0 mg per mL 1.25 mg per mL 1.25 mg per mL 1.25 mg per mL 1.0 mg per mL 1.25 mg per mL 1.25 mg per mL 1.25 mg per mL 1.25 mg per mL 2,000 units per mL 2,000 units per mL 2,000 units per mL 2,000 units per mL

Penicillin G Sodium Penicillin V Potassium Phenethicillin Potassium*

Unless otherwise noted, the Buffers are the potassium

Antibiotic

Solvent

*

Final Concentration 81 , except that

phosphate buffers defined in the section Media and Diluents under AntibioticsMicrobial Assays sterilization is not required before use.

Assay Preparation Unless otherwise specified in the individual monograph, dissolve in the solvent specified in the accurately weighed, of the specimen under test, and dilute a known final concentration of about that specified in the glass-stoppered conical flasks. Procedure Inactivation and Titration To 2.0 mL of the Standard Preparation and of the Assay Preparation, in respective flasks, add 2.0 mL of 1.0 N sodium hydroxide, mix by swirling, and allow to stand for 15 minutes. To each flask add 2.0 mL of 1.2 N hydrochloric acid, add 10.0 mL of 0.01 N iodine VS, immediately insert the stopper, and allow to stand for 15 minutes. Titrate with 0.01 N sodium thiosulfate VS. As the endpoint is approached, add 1 drop of starch iodide paste TS, and continue the titration to the discharge of the blue color. table of Solvents and Final Concentrations a suitable quantity, quantitatively with the same solvent to obtain a solution having table. Pipet 2 mL of this solution into each of two 125-mL

Blank Determination To a flask containing 2.0 mL of the Standard Preparation add 10.0 mL of 0.01 N iodine VS. If the Standard Preparation contains amoxicillin or ampicillin, immediately add 0.1 mL of 1.2 N hydrochloric acid.

Immediately titrate with 0.01 N sodium thiosulfate VS. As the endpoint is approached, add 1 drop of starch iodide paste TS, and continue the titration to the discharge of the blue color. Similarly treat a flask containing 2.0 mL of the Assay Preparation. Calculations Calculate the microgram (or unit) equivalent (F) of each mL of 0.01 N sodium thiosulfate consumed by the Standard Preparation by the formula: (2CP) / (B - I),

in which C is the concentration, in mg per mL, of Reference (or units) per mg, of the Reference Standard, B is the

Standard in the Standard Preparation, P is the potency, in g volume, in mL, of 0.01 N sodium thiosulfate consumed in the Blank determination, and I is the volume, in mL, of 0.01 N Calculate the potency of the specimen under test by the formula given in the individual monograph. sodium thiosulfate consumed in the Inactivation and titration.

StandardizationSodium Thiosulfate, Tenth-Normal (0.1 N) Na2S2O35H2O, 24.82 g in 1000 mL 248.19

Dissolve about 26 g of sodium thiosulfate and 200 mg of sodium carbonate in 1000 mL of recently boiled and cooled water. Standardize the solution as follows. Accurately weigh about 210 mg of primary standard potassium dichromate, previously pulverized and dried at 120 for 4 hours, and dissolve in 100 mL of water in a glass-stoppered, and quickly add 3 g of potassium iodide, 2 g of sodium

500-mL flask. Swirl to dissolve the solid, remove the stopper, bicarbonate, and 5 mL of hydrochloric acid. Insert the stopper gently in the flask, swirl to mix, and allow to stand in the dark for exactly 10 minutes. Rinse the stopper and the inner walls of the flask with water, and titrate the liberated iodine with the sodium thiosulfate solution until the solution is yellowish green in color. Add 3 mL of starch TS, and continue the titration until the blue color is discharged. Perform a blank determination. Restandardize the solution as frequently as supported by laboratory stability data. In the absence of such data, restandardize the solution weekly.

1. Acetone 2. 0.1 HCl 3. 1.2 HCl 4. Toluene 5. Glacial acetic acid 6. Standard Ampicillin 7. USP Ampicillin RS 8. Ninhydrin solution

9. 0.01N SodiumThiosulfat e 10. Potassium iodide 11. sodiumbicarbo

o

Ampicilin capsule 500 mg 20 capsules capsule capsule capsule capsule capsule capsule capsule capsule 20 capsules capsuleNo. 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. . +capsule (g) . capsule (g) 0.6810 0.6440 0.6930 0.6673 0.6914 0.6767 0.7068 0.6817 0.6570 0.6873 0.6862 0.0945 0.0957 0.0946 0.0994 0.0948 0.0983 0.0955 0.0969 0.0962 0.0958 0.0980

12. 13. 14. 15. 16. 17. 18. 19. 20. Total

0.6879 0.6786 0.6645 0.6896 0.6875 0.6983 0.6851 0.6953 0.6623 13.6219

0.0999 0.0979 0.0963 0.0939 0.0956 0.0969 0.0977 0.0950 0.0972 1.9031

capsule capsule capsule capsule 20 capsules capsule 20 capsule 13.6219-1.9031 = 11.6918 g 1 capsule 11.6918/20 = 0.58459 g = 584.59 mg = =

o Identification

Ampicillin 5 mg/mL acetone 0.1 N HCl (4:1)

Spot 2 TLC Mobile phase acetone: water: toluene: glacial acetic acid (650:100:100:25) TLC spray ninhydrin 3 mg/mL 90 Rf

o Standardization

K2Cr2O7 210 mg 100 ml glass-stoppered flask 250 ml KI 3 g, NaHCO3 2 g, HCl 5 ml flask 10 Rinse flask Na2S2O3 starch TS 3 ml

o

Assay Standard preparation

1.25 mg/mL 100 mL

USP Ampicillin RS 125 mg 100 mL volumetric flask 2.0 mL glass-stoppered conical flask 125 mL assay

Assay preparation 1.25 mg/mL 100 mL

Ampicillin capsule Ampicillin 500 mg 584.59 mg Ampicillin 125 mg (584.59 x 125)/500

= 146.15 mg

146.15 mg 100 ml

2 ml glass-stopper conical flask 125 ml assay

Assay

Assay preparation 1.0 N NaOH 2.0 mL swirl 15 1.2 N HCl 2.0 mL 0.01 N Iodine

VS 10.0 mL flask flask 15

0.01 N Na2S2O3

starch iodine paste TS 1

blank

Identification identification thin-layer

chromatography solvent system acetone, water, toluene, and glacial acetic acid 650:100:100:25 ninhydrin in alcohol hot plate 90 C spot sample stardard Rf Std. Sample

Rf standard = 2.4/6.1 = 0.39

Rf sample

= 2.4/6.1

= 0.39 Rf Ampicillin capsule Ampicillin

Result:

Standardization

Standardized Sodium Thiosulfate

Flask 1Weight of K2Cr2O7 Volume of Na2S2O3 after titrate Volume of Na2S2O3 before titrate Normality of Na2S2O3 Average Normality of Na2S2O3 0.2129 g 41.0 mL 0.00 mL 0.1054 N

Flask 20.2160 g 44.20 mL 0.00 mL 0.0997 N

0.1026 N mg of K2Cr2O7

Flask 1; N 49.04 x mL of Na2S2O3 = = 49.04 x 41.0 0.1054 N = 0.2160 0.2129 =

Flask 2;

N

= 0.0997 N Average Normality of Na2S2O3 0.1026 N

49.04 x 44.20

=2

0.1054 + 0.0997=

Result:

Assay

Flask 1 StandardVolume of Na2S2O3 after titrate 1.18

Flask 23.10

Volume of Na2S2O3 before titrate Volume of Na2S2O3 used

1.00 0.15 0.14 0.75 0.00 0.75

3.00 0.13

BlankVolume of Na2S2O3 before titrate Volume of Na2S2O3 used Volume of Na2S2O3 after titrate

Sampletitrate titrate titrate

Volume of Na2S2O3 after Volume of Na2S2O3 after Volume of Na2S2O3 after

0.30 0.00 0.30

1.27 1.00 0.27

0.285 5.00 4.00 1.00

Blanktitrate titrate

Volume of Na2S2O3 after

Volume of Na2S2O3 before Volume of Na2S2O3 used

F

= = = 3404.10

C = P B I

concentration, in mg per mL, of Reference Standard in

the Standard Preparation Standard =

= the potency, in g (or units) per mg, of the Reference volume, in mL, of 0.01 N sodium thiosulfate consumed volume, in mL, of 0.01 N sodium thiosulfate consumed

in the Blank determination =

in the Inactivation and titration

Quantity in mg of Ampicillin in Capsule = = =T = Capsule

each

486.8 mg

the labeled quantity, in mg, of ampicillin in each

D =

the concentration, in mg per mL, of ampicillin in the

Assay preparation on the basis of the labeled quantity in each Capsule and the extent of dilution.

% labeled amount = 97.36 % 500

486.8 x 100

=

Ampicillin capsule 486.8 mg

500 mg 97.36 % monograph %labeled amount 90 120% Ampicillin capsules

antibiotic Penicillins