§ 436.215 - Dissolution test.

(a) Equipment. Use either Apparatus 1 or 2 as described in the United States Pharmacopeia XXI dissolution test.

(b) Procedure. For each dosage form listed in the table in this paragraph select the appropriate dissolution medium, rotation rate, sampling time, and apparatus, and proceed as set forth in either Apparatus 1 or 2 methodology of the United States Pharmacopeia XXI dissolution test. Determine the amount of drug substance dissolved by performing the assay described in paragraph (c) of this section. The amount of dissolution medium removed for sampling purposes may be disregarded if the amount removed is not more than 15 milliliters. If more than 15 milliliters is removed, then correct for the volume removed.

Dosage formDissolution mediumRotation rate 1Sampling time(s)ApparatusAmoxicillin trihydrate and clavulanate potassium chewable tablets. 900 mL distilled water 75 30 min 2Amoxicillin trihydrate and clavulanate potassium tablets. ......do 75 ......do 2Azithromycin capsules. 900 mL 0.10 M sodium phosphate buffer, pH 6.0, 0.1 mg/mL trypsin 100 45 min 2Bacampicillin hydrochloride tablets. ......do 75 ......do 2Cefadroxil hemihydrate capsules. 900 mL distilled water 100 45 min 1Cefadroxil hemihydrate tablets. 900 mL distilled water 50 30 min 2Cefixime tablets 900 mL 0.05 M potassium phosphate buffer, pH 7.2 100 45 min 1Cefpodoxime proxetil tablets 900 mL pH 3.0 glycine buffer 75 30 min 2Cefprozil tablets. 900 mL purified water 100 45 min 1Cefuroxime axetil for oral suspension900 mL Sorenson's Modified Phosphate Buffer, pH 7.05030 min2Cefuroxime axetil tablets. 900 mL 0.07N hydrochloric acid 55 15 min. and 45 min 2Cephalexin hydrochloride monohydrate tablets. 900 mL distilled water 150 45 min1Cephradine dihydrate capsules. 900 mL 0.12N hydrochloric acid 75 60 min. 2Clarithromycin tablets. 900 mL 0.10 M sodium acetate buffer, pH 5.0 50 30 min. 2Doxycycline hyclate tablets. 900 mL distilled water 75 60 min and 90 min 2Doxycycline monohydrate hydrochloric acid capsules. 900 mL 0.1N hydrochloric acid. 75 60 min 2Erythromycin particles in tablets 900 mL 0.05M potassium phosphate buffer, pH 6.8 75 45 min. 2Loracarbef capsules. 900 mL distilled water. 50 30 min 2Oxytetracycline hydrochloride capsules. 900 mL distilled water 75 30 min and 60 min 2Rifabutin capsules 900 mL 0.01 N hydrochloric acid 100 45 min 1Tetracycline hydrochloride capsules (except 500-mg). ......do 75 ......do 2Tetracycline hydrochloride capsules (500-mg). ......do 75 30 min, 60 min, and 90 min 2Tetracycline hydrochloride tablets. ......do 75 30 min and 60 min 2Vancomycin hydrochloride capsules. 900 mL distilled water. 10045 min 11 Rotation rate of basket or paddle stirring element (revolutions per minute).

(c) Antibiotic drug content—(1) Tetracycline hydrochloride—(i) Preparation of working standard solution. Accurately weigh 20 to 30 milligrams of tetracycline hydrochloride working standard into a suitable-sized volumetric flask. Dissolve and dilute to volume with water. Further dilute an accurately measured portion with distilled water to obtain a known concentration of 0.01 to 0.02 milligram of tetracycline hydrochloride per milliliter.

(ii) Preparation of sample solutions. Dilute an accurately measured portion of the sample with sufficient distilled water to obtain a concentration of 0.01 to 0.02 milligram of tetracycline hydrochloride per milliliter (estimated).

(iii) Procedure. Using a suitable spectrophotometer and water as the blank, determine the absorbance of each standard and sample solution at the absorbance peak at approximately 276 nanometers. Determine the exact position of the absorption peak for the particular instrument used.

(iv) Calculation. Determine the total amount of tetracycline hydrochloride dissolved as follows:

ER01JA93.014

(2) Oxytetracycline hydrochloride; preparation of working standard-solution. (i) Accurately weigh 30 milligrams of oxytetracycline-base working standard into a suitable-sized volumetric flask. Add 5 milliliters of 0.1N hydrochloric acid and swirl the flask to dissolve oxytetracycline base. Dilute an accurately measured portion with distilled water to obtain a known concentration of 0.01 to 0.02 milligram of oxytetracycline per milliliter.

(ii) Proceed as directed in paragraphs (c)(1) (ii), (iii), and (iv) of this section except measure the absorbance at the absorption peak at approximately 273 nanometers.

(3) Doxycycline hyclate. Proceed as directed in paragraph (c)(1) of this section, except use the doxycycline working standard.

(4) Bacampicillin hydrochloride. Use the ampicillin working standard as the standard of comparison and assay for ampicillin content by either of the following methods.

(i) Iodometric assay. Proceed as directed in § 436.204 of this chapter, except dilute the working standard to a final concentration of 0.3 milligram of ampicillin per milliliter and use the sample solution as it is removed from the dissolution vessel without further dilution.

(ii) Hydroxylamine colorimetric assay. Proceed as directed in § 442.40(b)(1)(ii) of this chapter, except:

(a) Buffer. In lieu of the buffer described in § 442.40(b)(1)(ii)(b)(2) of this chapter, use the buffer prepared as follows: Dissolve 200 grams of primary standard tris (hydroxymethyl) aminomethane in sufficient distilled water to make 1 liter. Filter before use.

(b) Preparation of the working standard solution. Dissolve and dilute an accurately weighed portion of the ampicillin working standard with sufficient distilled water to obtain a final concentration of 0.3 milligram of ampicillin per milliliter;

(c) Sample solution. Use the sample solution as it is removed from the dissolution vessel without further dilution; and

(d) Calculations. Determine the total amount of ampicillin dissolved as follows:

ER01JA93.409

(5) Cephradine dihydrate—(i) Preparation of working standard solution. Accurately weigh approximately 40 milligrams of cephradine working standard into a suitable-sized volumetric flask. Dissolve and dilute to volume with 0.12N hydrochloric acid. Further dilute with a buffer solution (prepared by dissolving 27.2 grams of sodium acetate trihydrate in a mixture of 12 milliliters of glacial acetic acid and sufficient distilled water to make 2 liters) to obtain a known concentration of 0.01 to 0.03 milligram of cephradine per milliliter.

(ii) Preparation of sample solution. Filter the sample and dilute an accurately measured portion of the filtrate with sufficient buffer solution, described in paragraph (c)(5)(i) of this section, to obtain a concentration of 0.01 to 0.03 milligram of cephradine per milliliter (estimated).

(iii) Proceed as directed in paragraphs (c)(1) (iii) and (iv) of this section, except measure the absorbance at the absorption peak at approximately 262 nanometers.

(6) Amoxicillin trihydrate. Assay for the amoxicillin content as described in § 440.103d of this chapter, except use the sample as it is removed from the dissolution vessel.

(7) Vancomycin hydrochloride. Assay for the vancomycin content as described in § 436.105 of this chapter, except use the sample as it is removed from the dissolution test.

(8) Erythromycin—(i) Preparation of working standard solution. Accurately weigh approximately 140 milligrams of erythromycin working standard into a 250-milliliter volumetric flask and dissolve in 10 milliliters of methyl alcohol. Add water nearly to volume, mix, and allow the solution to cool. Dilute to volume with water and mix. On the day of use, dilute an accurately measured aliquot with water to obtain a known concentration of 0.28 milligram of erythromycin per milliliter (before adjusting for standard potency).

(ii) Preparation of sample solution. Dilute an accurately measured portion of the filtered sample with sufficient 0.05M potassium phosphate buffer, pH 6.8, to obtain a concentration of about 0.28 milligram of erythromycin per milliliter (estimated).

(iii) Procedure. Transfer 5.0-milliliter aliquots of the working standard solution and sample solution to 25-milliliter volumetric flasks and treat as follows: Add 2.0 milliliters of water, allow to stand for 5 minutes with intermittent swirling. Add 15.0 milliliters of 0.25N sodium hydroxide, dilute to volume with sufficient 0.05M potassium phosphate buffer, pH 6.8, and mix. Heat to 60 °C for 5 minutes and allow to cool. Using a suitable spectrophotometer and a blank (prepared as per the procedure above except that 2.0 milliliters of 0.5N sulfuric acid is substituted for the 2.0 milliliters of water) for each solution, determine the absorbance of each working standard and sample solution at the absorbance peak at approximately 236 nanometers. Determine the exact position of the absorption peak for the particular instrument used.

(iv) Calculation. Proceed as directed in paragraph (c)(1)(iv) of this section.

(9) Cefuroxime axetil tablets and powder for oral suspension—(i) Preparation of working standard solution—(a) Cefuroxime axetil tablets. Accurately weigh approximately 60 milligrams of cefuroxime axetil working standard into a suitable-sized volumetric flask. Dissolve in 5 milliliters of methanol and dilute to volume with 0.07N hydrochloric acid to obtain a known concentration equivalent to 0.01 to 0.02 milligram of cefuroxime activity per milliliter.

(b) Cefuroxime axetil for oral suspension. Accurately weigh approximately 15 milligrams of cefuroxime axetil working standard into a 100-milliliter volumetric flask. Dissolve in 5 milliliters of methanol and dilute to volume with Sorenson's Modified Phosphate Buffer, pH 7.0 (4.2 grams of sodium dihydrogen orthophosphate dihydrate and 14.3 grams of hydrogen disodium orthophosphate dodecahydrate per liter of water).

(ii) Preparation of sample solution—(a) Cefuroxime axetil tablets. Filter through a 0.45-micron filter and dilute an accurately measured portion of the filtrate with sufficient 0.07N hydrochloric acid to obtain a concentration equivalent to 0.01 to 0.02 milligram of cefuroxime activity per milliliter (estimated).

(b) Cefuroxime axetil for oral suspension. Filter the sample through an 8-micron filter. A coarse prefilter may be used to prevent clogging. Use the filtrate solution without further dilution.

(iii) Procedure—(a) Cefuroxime axetil tablets. Using a suitable spectrophotometer and 0.07N hydrochloric acid as the blank, determine the absorbance of each standard and sample solution at the absorbance peak at approximately 280 nanometers. Determine the exact position of the absorption peak for the particular instrument used.

(b) Cefuroxime axetil for oral suspension. Using a suitable spectrophotometer and Sorenson's Modified Phosphate Buffer, pH 7.0 (4.2 grams of sodium dihydrogen orthophosphate dihydrate and 14.3 grams of hydrogen disodium orthophosphate dodecahydrate per liter of water) as the blank, determine the absorbance of each standard and sample solution at the absorbance peak at approximately 280 nanometers. Determine the exact position of the absorption peak for the particular instrument used.

(iv) Calculations. Determine the total amount of cefuroxime activity dissolved as follows:

ER01JA93.015

(10) Cefixime—(i) Preparation of working standard solution. Accurately weigh approximately 25 milligrams of cefixime working standard into a 500-milliliter volumetic flask. Wet the powder with 0.5 milliliters of methanol, and dilute to volume with 0.05 M potassium phosphate buffer, pH 7.2 (prepared by dissolving 6.8 grams of monobasic potassium phosphate in distilled water to a volume of one liter. The pH is adjusted to 7.2 with 1.0N NaOH). Sonicate to assure dissolution and mix.

(ii) Preparation of sample solution. Forty-five minutes after the beginning of the rotation, withdraw and filter a portion of the solution. For the 400-milligram tablets, pipet 10.0 milliliters of the filtered sample solution into a 100-milliliter volumetric flask. For the 200-milligram tablets, pipet 10.0 milliliters of the filtered sample into a 50-milliliter volumetric flask. Dilute to volume with 0.05 M postassium phosphate buffer, pH 7.2.

(iii) Procedure. Proceed as directed in paragraphs (c)(1) (iii) and (iv) of this section, except measure the absorbance of the peak at approximately 320 nanometers using 0.05 M potassium phosphate buffer, pH 7.2 as the blank.

(11) Cephalexin hydrochloride monohydrate. Assay for cephalexin activity of the cephalexin hydrochloride monohydrate as directed in § 442.28 of this chapter, and use U.S.P. dissolution apparatus 1 (10 mesh basket). Use the sample as it is removed from the dissolution vessel.

(12) Doxycycline monohydrate. Proceed as directed in paragraph (c)(1) of this section, except use the doxycycline standard.

(13) Clarithromycin. Proceed as directed in § 452.50(b)(1) of this chapter except:

(i) Dissolution medium. Instead of the mobile phase described in § 452.50(b)(1)(i) of this chapter, use 0.10 M sodium acetate buffer prepared as follows: Weigh 13.6 grams of sodium acetate trihyrate into a container sufficient to hold 1 liter of solution. Dissolve the salt in 750 milliliters of distilled water. Adjust the pH of the solution to 5.0±0.05 with glacial acetic acid. Dilute to 1,000 milliliters with distilled water.

(ii) Preparation of the standard and sample solutions—(a) Standard solution. Dissolve (with shaking or sonication) an accurately weighed portion of the clarithromycin working standard, in sufficient methanol to obtain a solution having a known concentration of approximately 625 micrograms per milliliter of clarithromycin. Quantitatively transfer and dilute an aliquot of this solution with mobile phase (described in § 452.50(b)(1)(i) of this chapter) and mix to obtain a solution of known concentration of approximately 125 micrograms per milliliter of clarithromycin.

(b) Sample solution. Use the sample solution as it is removed from the dissolution vessel after diluting and mixing with mobile phase (described in § 452.50(b)(1)(i) of this chapter) 1:2 for the 250-milligram tablet and 1:4 for the 500-milligram tablet.

(c) Calculations. Determine the total amount of clarithromycin activity dissolved as follows:

ER01JA93.016

(14) Azithromycin. Proceed as directed in § 452.60(b)(1) of this chapter, except:

(i) Dissolution medium. Dissolve 85.2 grams of sodium phosphate dibasic and dilute to volume with ultrapure deionized or high-performance liquid chromatographic-grade water in a stoppered 2-liter graduated cylinder. Dilute this entire solution in an appropriate, suitably sized container with 4 liters of ultrapure deionized or high-performance liquid chromatographic-grade water. Adjust the pH to 6.0±0.05 with concentrated hydrochloric acid (about 40.5 milliliters). Add 600 milligrams of trypsin and mix well.

(ii) Preparation of the standard and sample solutions—(a) Standard solution. Accurately weigh approximately 15 milligrams of the azithromycin working standard into a 50-milliliter volumetric flask. Add 25 milliliters of the dissolution medium and sonicate briefly. Dilute to volume with dissolution medium and mix well. Pipet 2.0 milliliters of this solution into a 25-milliliter volumetric flask and dilute to volume with the mobile phase described in § 452.60(b)(1)(i) of this chapter. Pipet 4.0 milliliters of this solution into a 25-milliliter volumetric flask and bring to volume with the mobile phase.

(b) Sample solution. Filter the sample solutions through a 0.45-micron filter before use. Pipet 2.0 milliliters of the filtered aliquot into a 25-milliliter volumetric flask and dilute to volume with the mobile phase described in § 452.60(b)(1)(i) of this chapter. Pipet 4.0 milliliters of this solution into another 25-milliliter volumetric flask and bring to volume with the mobile phase. The solution is stable at room temperature for 24 hours.

(c) Calculations. Determine the percent of azithromycin dissolved as follows:

ER01JA93.017 ER01JA93.018

(15) Cefprozil. Proceed as directed in § 442.80(b)(1) of this chapter except:

(i) Sample solutions. Filter the sample solutions through a 0.45-micron filter before use. Use the sample solution as it is removed from the dissolution vessel without further dilution for the 250-milligram tablet; prepare the sample solution for the 500-milligram tablet by diluting a 5-milliliter aliquot of the filtered solution to volume in a 10-milliliter volumetric flask with distilled water.

(ii) Calculations. Determine the total percent of cefprozil dissolved as follows:

ER01JA93.019 ER01JA93.020

(16) Loracarbef—(i) Preparation of the working standard solution. Accurately weigh approximately 110 milligrams of the loracarbef working standard into a suitable-sized volumetric flask. Dissolve and dilute to volume with water. Further dilute an accurately measured portion with distilled water to obtain a known concentration of 0.02 milligram of loracarbef activity per milliliter.

(ii) Preparation of sample solutions. Dilute an accurately measured portion of the sample with sufficient distilled water to obtain a concentration of 0.02 milligram of loracarbef activity per milliliter (estimated).

(iii) Procedure. Using a suitable spectrophotometer and water as the blank, determine the absorbance of each standard and sample solution at the absorbance maximum at approximately 260 nanometers. Determine the exact position of the absorbance maximum for the particular instrument used.

(iv) Calculations. Determine the total amount of loracarbef dissolved as follows:

ER01JA93.021

(17) Cefadroxil hemihydrate. Proceed as directed in paragraph (c)(1) of this section, except use the cefadroxil working standard and measure the absorbance at the absorption peak of approximately 264 nanometers.

(18) Rifabutin—(i) Preparation of the working standard solution. Accurately weigh approximately 45 milligrams of the rifabutin working standard into a suitable-sized volumetric flask. Dissolve and dilute to volume with 0.01N hydrochloric acid (prepared by diluting 5.0 milliliters of hydrochloric acid (37 percent) to 6 liters with distilled water) to obtain a concentration of approximately 13 micrograms rifabutin activity per milliliter.

(ii) Preparation of sample solutions. Forty-five minutes after the beginning of the rotation, withdraw a 10-milliliter aliquot from the vessel. Dilute a 2-milliliter portion of the sample to 25 milliliters with 0.01N hydrochloric acid.

(iii) Procedure. Using a suitable spectrophotometer and 0.01N hydrochloric acid as the blank, determine the absorbance of each standard and sample solution at the absorbance maximum at approximately 280 nanometers. Determine the exact position of the absorbance maximum for the particular instrument used.

(iv) Calculations. Determine the total amount of rifabutin dissolved as follows:

ER01JA93.022

(19) Cefpodoxime proxetil—(i) Dissolution fluid:0.04 molar glycine buffer, pH 3.0—(A) Stock solution. Dissolve 54.5 grams of glycine (aminoacetic acid) and 42.6 grams of sodium chloride in about 500 milliliters of deionized water in a 1-liter volumetric flask. Add cautiously, and with swirling, 14.2 milliliters of concentrated hydrochloric acid. Cool to room temperature. Dilute to volume with deionized water and mix. Check the pH of the solution obtained by diluting 50 milliliters of the stock solution to 900 milliliters with deionized water. The pH should be 3.0±0.1. If necessary, adjust the pH of the stock solution with 50 percent sodium hydroxide or concentrated hydrochloric acid. Recheck that the pH of the working solution is 3.0±0.1.

(B) Working solution. Dilute 50 milliliters of stock solution to 900 milliliters with deionized water.

(ii) Preparation of the working standard solutions. Accurately weigh approximately 28 milligrams for the 100-milligram tablets and 56 milligrams for the 200-milligram tablets of the cefpodoxime proxetil working standard and dissolve in 10 milliliters of methanol. Dilute to 200 milliliters with dissolution fluid. Prepare fresh daily.

(iii) Sample solutions. Filter the sample solutions through a 0.45-micron filter before use. Use the sample solution as it is removed from the dissolution vessel without further dilution.

(iv) Procedure. Using a suitable spectrophotometer and water as the blank, determine the absorbance of each standard and sample solution at the absorbance peak at approximately 259 nanometers. Determine the exact position of the absorption peak for the particular instrument used.

(v) Calculations. Determine the percent of label dissolved as follows:

Percent dissolved = (Asam/Astd) X (Cs/L) X V X P X F1

(d) Evaluation. Use the dissolution acceptance table and interpretation in the United States Pharmacopeia XXI.