(5) pH. Proceed as directed in §436.202 of this chapter, using an aqueous suspension containing 50 milligrams per milliliter.

(6) Identity. Proceed as directed in § 436.211 of this chapter, using the sample preparation as described in paragraph (b)(2) of that section.

[49 FR 25847, June 25, 1984; 49 FR 40006, Oct. 12, 1984, as amended at 55 FR 11583, Mar. 29, 1990]

§ 442.52 Cefotetan.

(a) Requirements for certification—(1) Standards of identity, strength, quality, and purity. Cefotetan is (6R,7S)-4-[[2carboxy-7-methoxy-3-[[(1-methyl-1Htetrazol-5-yl)thio]methyl]-8-oxo-5-thia1-azabicyclo[4.2.0]oct-2-ene-7-yl]-carbamoyl]-1,3-dithietane-2,α-malonamic acid. It is so purified and dried that:

(i) Its potency is not less than 950 micrograms and not more than 1,030 micrograms of cefotetan activity per milligram on the anhydrous basis.

(ii) Its moisture content is not more than 2.5 percent.

(iii) It gives a positive identity test for cefotetan.

(2) Labeling. It shall be labeled in accordance with the requirements of § 432.5 of this chapter.

(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, and identity.

(ii) Samples, if required by the Director, Center for Drug Evaluation and Research: 10 packages each containing approximately 500 milligrams.

(b) Tests and methods of assay—(1) Potency. Proceed as directed in § 436.216 of this chapter, except use the resolution test solution to determine resolution in lieu of the working standard solution. Perform the assay at ambient temperature, using an ultraviolet detection system operating at a wavelength of 254 nanometers, a column packed with microparticulate (3 to 10 micrometers in diameter) reversed phase packing material such as octadecyl hydrocarbon bonded silicas, a flow rate not exceeding 2.0 milliliters per minute, and a known injection volume of between 10 and 20 microliters.

Reagents, working standard solution, sample solution, resolution test solution, system suitability requirements, and calculations are as follows:

(i) Reagents-(A) Diluting solution. Mix water:methanol:acetonitrile (90:5:5).

(B) Mobile phase. Mix 0.1M phosphoric acid:glacial acetic

acid:methanol:acetonitrile

(1700:100:105:105). Filter through a suitable filter capable of removing particulate matter greater than 0.5 micron in diameter. Degas the mobile phase just prior to its introduction into the chromatograph.

(ii) Preparation of working standard, sample, and resolution test solutions—(A) Working standard solution. Accurately weigh approximately 50 milligrams of the cefotetan working standard into a 250-milliliter volumetric flask containing 12.5 milliliters of methanol. Swirl the flask for several minutes, then add 12.5 milliliters of acetonitrile. Swirl the flask until the cefotetan is dissolved. Dilute to volume with water to obtain a solution containing approximately 200 micrograms of cefotetan per milliliter. Mix well. Protect the working standard solution from light.

(B) Sample solution. Dissolve an accurately weighed portion of the sample with sufficient diluting solution described in paragraph (b)(1)(i)(A) of this section to obtain a concentration of approximately 200 micrograms of cefotetan per milliliter.

(C) Resolution test solution. Place 10 milliliters of the working standard solution in a stoppered flask containing a few milligrams of magnesium carbonate. Close the flask and sonicate for 10 minutes. If the solution is not slightly turbid, add more magnesium carbonate and repeat sonication. Filter the turbid solution through a 0.5-micron filter and use within 2 hours. As this solution stands, the tautomer concentration increases.

(iii) System suitability requirements— (A) Tailing factor. The tailing factor (T) is satisfactory if it is not more than 1.3 at 10 percent of peak height in lieu of 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. The resolution (R) between the peak for cefotetan and its tautomer is satisfactory if it is not less than 2.0.

(D) Coefficient of variation. The coefficient of variation (S,in percent) of five replicate injections is satisfactory if it is not more than 2.0 percent. If the system suitability requirements have been met, then proceed as described in § 436.216 (b) of this chapter. Alternate chromatographic conditions are acceptable provided comparable system suitability requirements are met. However, the sample preparation described in paragraph (b)(1)(ii)(B) of this section should not be changed.

Ps

in

= Cefotetan activity in the cefotetan working standard solution micrograms per milliliter;

= Volume of flask used to dilute standard; and

V1 = Volume of sample diluted.

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

(3) Identity. Proceed as directed in § 436.211 of this chapter using the potassium bromide discs prepared as described in § 436.211(b)(1) of this chapter or the mineral oil mull prepared as described in § 436.211(b)(2) of this chapter. [59 FR 26940, May 25, 1994, as amended at 60 FR 33712, June 29, 1995]

§ 442.53a Sterile cefotetan disodium.

(a) Requirements for certification—(1) Standards of identity, strength, quality, and purity. Sterile cefotetan disodium is a white to off-white lyophilized powder. It is so purified and dried that:

(1) If the cefotetan disodium is not packaged for dispensing, its potency is not less than 830 micrograms and not more than 970 micrograms of cefotetan

per milligram on the anhydrous basis. If the cefotetan disodium is packaged for dispensing, its potency is not less than 830 micrograms and not more than 970 micrograms of cefotetan per milligram on the anhydrous basis and also, each container contains not less than 90 percent and not more than 120 percent of the number of milligrams of cefotetan that it is represented to contain.

(ii) It is sterile.

(iii) It is nonpyrogenic.

(iv) Its moisture content is not more than 1.5 percent.

(v) Its pH in an aqueous solution containing 100 milligrams of cefotetan disodium per milliliter is not less than 4.0 and not more than 6.5.

(vi) It gives a positive identity test for cefotetan.

(2) Labeling. It shall be labeled in accordance with the requirements of § 432.5 of this chapter.

(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, sterility, pyrogens, moisture, pH, and identity.

(ii) Samples, if required by the Director, Center for Drug Evaluation and Research:

(a) If the batch is packaged for repacking or for use in the manufacture of another drug:

(1) For all tests except sterility: 10 packages, each containing approximately 500 milligrams.

(2) For sterility testing: 20 packages, each containing approximately 300 milligrams.

(b) If the batch is packaged for dispensing:

(1) For all tests except sterility: A minimum of 10 immediate containers.

(2) For sterility testing: 20 immediate containers, collected at regular intervals throughout each filling operation.

(b) Tests and methods of assay-(1) Potency. Proceed as directed in § 436.216 of this chapter, except use the resolution test solution to determine resolution in lieu of the working standard solution. Perform the assay at ambient temperature, using an ultraviolet detection system operating at a wavelength of 254 nanometers, a column

packed with microparticulate (3 to 10 micrometers in diameter) reversed phase packing material such as octadecyl hydrocarbon bonded silicas, a flow rate not exceeding 2.0 milliliters per minute, and a known injection volume of between 10 and 20 microliters. Reagents, working standard solution, sample solution, resolution test solution, system suitability requirements, and calculations are as follows:

(i) Reagents-(a) Diluting solution. Mix water:methanol:acetonitrile (90:5:5). (b) Mobile phase. Mix 0.1M phosphoric acid:glacial acetic

acid:methanol:acetonitrile

(1700:100:105:105). Filter through a suitable filter capable of removing particulate matter greater than 0.5 micron in diameter. Degas the mobile phase just prior to its introduction into the chromatograph.

(ii) Preparation of working standard, sample, and resolution test solutions—(a) Working standard solution. Accurately weigh approximately 50 milligrams of the cefotetan working standard into a 250-milliliter volumetric flask containing 12.5 milliliters of methanol. Swirl the flask for several minutes, then add 12.5 milliliters of acetonitrile. Swirl the flask until the cefotetan is dissolved. Dilute to volume with water to obtain a solution containing approximately 200 micrograms of cefotetan per milliliter. Mix well. Protect the working standard solution from light.

(b) Sample solutions—(1) Product not packaged for dispensing (micrograms of cefotetan per milligram). Dissolve an accurately weighed portion of the sample with sufficient diluting solution described in paragraph (b)(1)(i)(a) of this section, to obtain a concentration of approximately 200 micrograms of cefotetan per milliliter.

(2) Product packaged for dispensing. Determine both micrograms of cefotetan per milligram of the sample and milligrams of cefotetan per container. Use separate containers for preparation of each sample solution as described in paragraphs (b)(1)(ii)(b)(2) (i) and (ii) of this section.

(i) Micrograms of cefotetan per milligram. Dissolve an accurately weighed portion of the sample with sufficient diluting solution described in paragraph (b)(1)(i)(a) of this section, to ob

tain a concentration of approximately 200 micrograms of cefotetan per milliliter.

(ii) Milligrams of cefotetan 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. Further dilute an aliquot of the solution thus obtained with sufficient diluting solution described in paragraph (b)(1)(i)(a) of this section, to obtain a concentration of approximately 200 micrograms of cefotetan per milliliter.

(c) Resolution test solution. Place 10 milliliters of the working standard solution in a stoppered flask containing a few milligrams of magnesium carbonate. Close the flask and sonicate for 10 minutes. If the solution is not slightly turbid, add more magnesium carbonate and repeat sonication. Filter the turbid solution through a 0.5-micron filter and use within 2 hours. As this solution stands, the tautomer concentration increases.

(iii) System suitability requirements(a) Tailing factor. The tailing factor (T) is satisfactory if it is not more than 1.3 at 10 percent of peak height in lieu of 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. The resolution (R) between the peak for cefotetan and its tautomer is satisfactory if it is not less than 2.0.

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

If the system suitability requirements have been met, then proceed as described in § 436.216(b) of this chapter. Alternate chromatographic conditions are acceptable provided comparable system suitability requirements are met. However, the sample preparation described in paragraph (b)(1)(ii)(b) of this section should not be changed.

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

A, Area of the cefotetan peak in the chromatogram of the cefotetan working standard;

P-Cefotetan activity in the cefotetan working standard solution in 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(b) of this chapter, using a solution containing 50 milligrams of cefotetan 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 milligrams of cefotetan disodium per milliliter.

(6) Identity. The high-performance liquid chromatogram of the sample determined as directed in paragraph (b)(1) of this section, compares quali

tatively to that of the cefotetan working standard.

[51 FR 20263, June 4, 1986, as amended at 52 FR 35912, Sept. 24, 1987; 55 FR 11583, Mar. 29, 1990]

§ 442.54 Cefpodoxime proxetil.

(a) Requirements for certification—(1) Standards of identity, strength, quality, and purity. Cefpodoxime proxetil is (±)1-hydroxyethyl(+)-(6R,7R)-7-[2-(2amino-4-thiazolyl)glyoxylamido]-3(methoxymethyl)-8-oxo-5-thia-1azabicyclo[4.2.0]oct-2-ene-2carboxylate,72-(Z)-(O-methyloxime), isopropyl carbonate (ester). It is so purified and dried that:

(i) Its potency is not less than 690 micrograms and not more than 804 micrograms of cefpodoxime activity per milligram, on an anhydrous basis.

(ii) The ratio of its R-epimer to total cefpodoxime is not less than 0.5 and not more than 0.6.

(iii) Its moisture content is not more than 3 percent.

(iv) It gives a positive identity test. (2) Labeling. It shall be labeled in accordance with the requirements of § 432.5 of this chapter.

(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 cefpodoxime potency, isomer ratio, moisture, and identity.

(ii) Samples, if required by the Director, Center for Drug Evaluation and Research: 10 packages, each containing approximately 500 milligrams.

(b) Tests and methods of assay—(1) Potency. Proceed as directed in § 436.216 of this chapter, using a suitable thermostatted column heating mechanism to maintain a column temperature of 40 °C, an ultraviolet detection system operating at a wavelength of 254 nanometers, a 15 centimeter X 4.6 millimeter (i.d.) column packed with microparticulate (5 micrometers in diameter) reversed phase packing material such as octadecyl silane bonded to silicas, a flow rate of 0.8 milliliter per minute, and a known injection volume of 2 microliters. The retention time for

the S-epimer is approximately 22 minutes and the retention time for Repimer is approximately 28 minutes. The internal standard (propylparaben) has a retention time of 34 minutes. Mobile phase, dilution solvent, resolution solution, internal standard solution, working standard and sample solutions, system suitability requirements, and calculations are as follows:

(1) Mobile phase. The mobile phase consists of 420 milliliters of methanol, 580 milliliters of deionized water, and 230 milligrams of L-histidine hydrochloride. The pH is adjusted to 2.5±0.1 using 2N sulfuric acid. The mobile phase must be at room temperature for a correct pH measurement. The methanol concentration may be adjusted to achieve comparable retention times from column to column. Increasing methanol reduces retention times. Filter the mobile phase through a suitable filter capable of removing particulate matter 0.5 micron in diameter and degas it just before its introduction into the chromatograph.

(ii) Dilution solvent. Prepare a solvent for dilution by thoroughly mixing 495 milliliters of deionized water, 495 milliliters of acetonitrile, and 10 milliliters of acetic acid in an appropriate container.

(iii) Resolution solution. Prepare a 1 milligram per milliliter solution of any bulk containing ANTI-A in dilution solvent. Use this solution to determine the resolution between ANTI-A and the later-eluting drug epimer (R-epimer). Alternately, the resolution factor can be determined between the R and S isomers.

(iv) Internal standard solution. Prepare a solution of propylparaben in dilution solvent at a concentration of 10 milligrams per milliliter.

(v) Preparation of working standard solutions. Accurately weigh weigh approxi

the

mately 42 milligrams of cefpodoxime proxetil working reference standard add 3 milliliters of internal standard solution and 25 milliliters of dilution solvent. The standard solution is stable for at least 48 hours. Refrigeration is not recommended.

(vi) Sample solution. Accurately weigh approximately 42 milligrams of the sample, add 3 milliliters of internal standard and 25 milliliters of dilution solvent. The sample solution is stable for at least 48 hours. Refrigeration is not recommended.

(vii) System suitability requirements— (A) Asymmetry factor.The asymmetry factor (A,) is satisfactory if it is not less than 0.8 and not more than 1.1 for the R-epimer of cefpodoxime peak.

(B) Efficiency of the column. The absolute efficiency (h,) is satisfactory if it is not more than 5 for the R-epimer peak.

(C) Resolution factor. The resolution factor (R) between the peak for ANTIA and the peak for the R-epimer is satisfactory if it is not less than 1.3. Alternately, the resolution factor (R) between the peak for the R-epimer and the peak for the S-epimer of cefpodoxime is not less than 11.

(D) Coefficient of variation (Relative standard deviation). The coefficient of variation (SRin percent of 5 replicate injections) is satisfactory if it is not more than 2 percent.

(E) Capacity factor (k'). The capacity factor (k') for the R-epimer of cefpodoxime is satisfactory if it is not less than 10.4 and not more than 15.6.

(F) If the system suitability parameters in this paragraph (b)(1)(iv) have been met, then proceed as described in § 436.216(b) of this chapter.

(viii) Calculations. Calculate the micrograms of cefpodoxime proxetil per milligram of sample on an anhydrous basis as follows: