portion of the sample with sufficient distilled water to obtain a solution containing 100 micrograms of amdinocillin per milliliter (estimated).

(ii) Milligrams of amdinocillin per container. Reconstitute the sample as directed in the labeling. Then, using a suitable hypodermic needle and syringe, remove all of the withdrawable contents if it is represented as a singledose container; or, if the labeling specifies the amount of potency in a given volume of the resultant preparation, remove an accurately measured representative portion from each container. Dilute the solution thus obtained with sufficient distilled water to obtain a solution containing 100 micrograms of amdinocillin per milliliter (estimated).

(iii) System suitability requirements—(a) Tailing factor. The tailing factor (T) is satisfactory if it is not more than 2.5 at 5 percent of peak height:

(b) Efficiency of the column. The efficiency of the column (n) is satisfactory if it is greater than 1,500 theoretical plates.

(c) Resolution factor. The resolution factor (R) between the peak for amdinocillin and its nearest eluting impurity is satisfactory if it is not less than 2.5.

(d) Coefficient of variation. The coefficient of variation (SÅ in percent) of five replicate injections is satisfactory if it is not more than 2.0 percent.

If the system suitability parameters have been met, then proceed as described in § 436.353(b) of this chapter.

(iv) Calculations. (a) Calculate the micrograms of amdinocillin per milligram of sample as follows:

m=Percent moisture content of the sample.

(b) Calculate the amdinocillin content of the container as follows:

Milligrams of amdinocillin A, XP,xd per container A, × 1,000

where:

A-Area of the amdinocillin peak in the chromatogram of the sample (at a retention time equal to that observed for the standard);

A-Area of the amdinocillin peak in the chromatogram of the amdinocillin working standard:

in

P-Amdinocillin activity in the amdinocillin working standard solution micrograms per milliliter; and d=Dilution factor of the sample.

(2) Sterility. Proceed as directed in § 436.20 of this chapter, using the method described in paragraph (e)(1) of that section.

(3) Pyrogens. Proceed as directed in § 436.32(a) of this chapter, using a solution containing 40 milligrams of amdinocillin per milliliter.

(4) Moisture. Proceed as directed in § 436.201 of this chapter.

(5) pH. Proceed as directed in §436.202 of this chapter, using an aqueous solution containing 100 100 milligrams of amdinocillin per milliliter.

(6) Crystallinity. Proceed as directed in § 436.203(a) of this chapter.

(7) Identity. Proceed as directed in § 436.211 of this chapter, using a potassium bromide disc containing 1 milligram of amdinocillin in 300 milligrams of potassium bromide, prepared as described in paragraph (b)(1) of that section.

[50 FR 7765, Feb. 26, 1985; 50 FR 10220, Mar. 14, 1985; 50 FR 18243, Apr. 30, 1985; 55 FR 11582, Mar. 29, 1990]

§ 440.3 Amoxicillin trihydrate.

(a) Requirements for certification—(1) Standards of identity, strength, quality, and purity. Amoxicillin trihydrate is the trihydrate form of D(−) a-amino-phydroxybenzyl penicillin. It is so purified and dried that:

(i) Its potency is not less than 900 micrograms and not more than 1,050 micrograms of amoxicillin per milligram on an anhydrous basis. (ii) [Reserved]

(iii) Its moisture content is not less than 11.5 percent and not more than 14.5 percent.

(iv) Its pH in an aqueous solution containing 2 milligrams per milliliter is not less than 3.5 and not more than 6.0.

(v) Its amoxicillin content is not less than 90 percent on an anhydrous basis.

(vi) The acid-base titration concordance is such that the difference between the percent amoxicillin content when determined by nonaqueous acid titration and by nonaqueous base titration is not more than 6. The potencyacid titration concordance is such that the difference between the potency value divided by 10 and the percent amoxicillin content of the sample determined by the nonaqueous acid titration is not more than 6. The potencybase titration concordance is such that the difference between the potency value divided by 10 and the percent amoxicillin content of the sample determined by the nonaqueous base titration is not more than 6.

(vii) It is crystalline.

(viii) It gives a positive identity test for amoxicillin trihydrate.

(2) Labeling. In addition to the labeling requirements of § 432.5 of this chapter, each package shall bear on its outside wrapper or container and the immediate container the following statement: "For use in the manufacture of nonparenteral drugs only".

(3) Requests for certification; samples. In addition to complying with the requirements of § 431.1 of this chapter, each such request shall contain:

(i) Results of tests and assays on the batch for potency, moisture, pH, amoxicillin content, concordance, crystallinity, and identity.

(ii) Samples required: 12 packages, each containing approximately 500 milligrams.

(b) Tests and methods of assay—(1) Potency. Use any of the following methods; however, the results obtained from the iodometric assay shall be conclusive:

(i) Microbiological agar diffusion assay. Proceed as directed in §436.105 of this chapter, preparing the sample for assay as follows: Dissolve an accurately weighed portion of the sample in sufficient sterile distilled water to give a stock solution containing 1.0 milligram of amoxicillin per milliliter (estimated). Further dilute an aliquot of the stock solution with solution 3 to the reference concentration of 0.1 microgram of amoxicillin per milliliter (estimated).

(ii) Iodometric assay. Proceed as directed in § 436.204 of this chapter, except in paragraph (d) of that section, add 3 drops of 1.2N hydrochloric acid to both the sample and working standard solutions after the addition of 0.01N iodine solution.

(iii) Hydroxylamine colorimetric assay. Proceed as directed in § 436.205 of this chapter.

(2) [Reserved]

(3) Moisture. Proceed as directed in § 436.201 of this chapter.

(4) pH. Proceed as directed in § 436.202 of this chapter, using an aqueous solution containing 2 milligrams per milliliter.

(5) Amoxicillin content. Proceed as directed in § 436.213 of this chapter using both the titration procedures described in paragraphs (e) (1) and (2) of that section. Calculate the percent amoxicillin content as follows:

(i) Acid titration.

Percent amoxicillin content

where:

A=Milliliters of lithium methoxide reagent used in titrating the sample. B=Milliliters of lithium methoxide reagent used in titrating the blank.

m=Percent moisture content of the sample. Calculate the difference between the potency and the amoxicillin content as follows:

Difference

Potency in micrograms per milligram 10

percent amoxicillin content

(ii) Base titration.

where:

Percent amoxicillin content

titration and by nonaqueous base titration is not more than six. The potencyacid titration concordance is such that the difference between the potency value divided by 10 and the percent ampicillin content of the sample determined by the nonaqueous acid titration is not more than six. The potency-base titration concordance is such that the difference between the potency value divided by 10 and the percent ampicillin content of the sample determined by the nonaqueous base titration is not more than six.

(vii) It is crystalline.

(viii) It gives a positive identity test for ampicillin.

(2) Labeling. In addition to the labeling requirements prescribed by § 432.5(b) of this chapter, each package shall bear on its outside wrapper or container and the immediate container the following statement, "For use in the manufacture of nonparenteral drugs only".

(3) Requests for certification; samples. In addition to complying with the requirements of § 431.1 of this chapter, each such request shall contain:

(i) Results of tests and assays on the batch for potency, loss on drying, pH, ampicillin content, concordance, crystallinity, and identity.

(ii) Samples required: 10 packages, each containing approximately 300 milligrams.

(b) Tests and methods of assay—(1) Potency. Assay for potency by any of the following methods; however, the results obtained from the microbiological agar diffusion assay shall be conclusive.

(i) Microbiological agar diffusion assay. Proceed as directed in §436.105 of this chapter, preparing the sample for assay as follows: Dissolve an accurately

weighed portion of the sample in sufficient sterile distilled water to give a stock solution containing 0.1 milligram of ampicillin per milliliter (estimated). Further dilute an aliquot of the stock solution with 0.1M potassium phosphate buffer, pH 8.0 (solution 3), to the reference concentration of 0.1 microgram of ampicillin per milliliter (estimated).

(ii) Iodometric assay. Proceed as directed in § 436.204 of this chapter, except in paragraph (d) of that section, add 3 drops of 1.2N hydrochloric acid to both the sample and working standard solutions after the addition of 0.01N iodine solution.

(iii) Hydroxylamine colorimetric assay. Proceed as directed in § 436.205 of this chapter.

(2) [Reserved]

(3) Loss on drying. Proceed as directed in § 436.200(a) of this chapter.

(4) pH. Proceed as directed in § 436.202 of this chapter, using an aqueous solution containing 10 milligrams per milliliter.

(5) Ampicillin content. Proceed as directed in § 436.213 of this chapter, using both the titration procedures described in paragraphs (e) (1) and (2) of that section. Calculate the percent ampicillin content as follows:

(i) Acid titration.

micrograms of ampicillin per milligram on an anhydrous basis.

(ii) [Reserved]

(iii) Its loss on drying is not less than 12 percent and not more than 15 percent.

(iv) Its pH in an aqueous solution containing 10 milligrams per milliliter is not less than 3.5 and not more than 6.0.

(v) Its ampicillin content is not less than 90 percent on an anhydrous basis.

(vi) The acid-base titration concordance is such that the difference between the percent ampicillin content when determined by nonaqueous acid titration and by nonaqueous base titration is not more than 6. The potencyacid titration concordance is such that the difference between the potency value divided by 10 and the percent ampicillin content of the sample determined by the nonaqueous acid titration is not more than 6. The potency-base titration concordance is such that the difference between the potency value divided by 10 and the percent ampicillin content of the sample determined by the nonaqueous base titration is not more than 6.

(vii) It is crystalline.

(viii) It gives a positive identity test for ampicillin trihydrate.

(2) Labeling. In addition to the labeling requirements prescribed by § 432.5(b) of this chapter, this drug shall be labeled "ampicillin" and each package shall bear on its outside wrapper or container and the immediate container the following statement "For use in the

manufacture of nonparenteral

drugs only".

(3) Requests for certification; samples. In addition to complying with the requirements of §431.1 of this chapter, each such request shall contain:

(i) Results of tests and assays on the batch for potency, loss on drying, pH, ampicillin content, concordance, crystallinity, and identity.

(ii) Samples required: 10 packages each containing approximately 300 milligrams.

(b) Tests and methods of assay—(1) Potency. Use any of the following methods; however, the results obtained from the microbiological agar diffusion assay shall be conclusive.

as

an

(i) Microbiological agar diffusion assay. Proceed as directed in §436.105 of this chapter, preparing the sample for assay follows: Dissolve accurately weighed portion of the sample in sufficient sterile distilled water to give a stock solution containing 0.1 milligram of ampicillin per milliliter (estimated). Further dilute an aliquot of the stock solution with 0.1M potassium phosphate buffer, pH 8.0 (solution 3), to the reference concentration of 0.1 microgram of ampicillin per milliliter (estimated).

(ii) Iodometric assay. Proceed as directed in § 436.204 of this chapter, except in paragraph (d) of that section, add 3 drops of 1.2N hydrochloric acid to both the sample and working standard solutions after the addition of 0.01N iodine solution.

(iii) Hydroxylamine colorimetric assay. Proceed as directed in § 436.205 of this chapter.

(2) [Reserved]

(3) Loss on drying. Proceed as directed in § 436.200(a) of this chapter.

(4) pH. Proceed as directed in § 436.202 of this chapter, using an aqueous solution containing 10 milligrams per milliliter.

(5) Ampicillin content. Proceed as directed in § 436.213 of this chapter, using both the titration procedures described in paragraphs (e) (1) and (2) of that section. Calculate the percent ampicillin content as follows:

(i) Acid titration.

Percent ampicillin content=[(A-B) (normality of lithium methoxide reagent) (349.4) (100) (100)]/[(Weight of sample in milligrams) (100-m)],

where:

A-Milliliters of lithium methoxide reagent used in titrating the sample;

B-Milliliters of lithium methoxide reagent used in titrating the blank; m=Percent moisture content of the sample. Calculate the difference between the potency and the ampicillin content as follows: Difference (Potency in micrograms per milligram/10) - percent ampicillin content. (ii) Base titration.

Percent ampicillin content=[(A−B) (normality of perchloric acid reagent) (349.4) (100) (100)]/[(Weight of sample in milligrams) (100-m)],

where: