Acetaminophen

Chemical Names
N-(4-Hydroxyphenyl)acetamide 4n-Hydroxyacetanilide

Other Names
Paracetamol, Tempra, Tylenol

Form
Acetaminophen

Molecular Formula
C8H9NO2

MW
151.2

CAS
103-90-2

Appearance
Acetaminophen occurs as a white crystalline powder with a slightly bitter taste.

Solubility
Acetaminophen is soluble in boiling water and freely soluble in alcohol.

pKa

Acetaminophen has a pKa of 9.51.

Method 1
Aukunuru et al. described the simultaneous determination of acetaminophen, salicylamide, phenyltoloxamine, and related products by HPLC. A Varian system consisting of a model 9010 pump, a model 9095 autosampler, a model 9050 UV absorbance detector, and a Rainin Dinamax MacIntegrator was used. The stationary phase was a Phenomenex Prodigy C8 column (150 4.6 mm, 5-m particle size). Mobile phase A was 0.1 M phosphate buffer (adjusted to pH 2.7 with phosphoric acid). Mobile phase B was acetonitrile. The mobile phase was linearly delivered from 5% B to 45% B in 17 minutes, followed by 10 minutes for equilibration. The flow rate was 1 mL/min. UV detection was performed at 220 nm. The injection volume was 50 L. Under these conditions, retention times of acetaminophen, salicylamide, and phenyltoloxamine were 5.7, 10.9, and 15.9 minutes, respectively. The stability-indicating nature of the assay was demonstrated by accelerated degradation of the drugs. Drug solutions were prepared in 1 N hydrochloric acid, 1 N sodium hydroxide, or 10% hydrogen peroxide and heated at 60 C. No degradation products interfered with the analysis of drugs. A standard curve for acetaminophen was generated from 0.06 to 300 g/mL. The correlation coefficient was 0.999.

Reference
Aukunuru JV, Kompella UB, Betageri GV. Simultaneous high-performance liquid chromatographic analysis of acetaminophen, salicylamide, phenyltoloxamine, and related products. J Liq Chrom & Rel Technol. 2000; 23: 56578.

Method 2
Hewala described an HPLC method for paracetamol (acetaminophen), guaifenesin, sodium benzoate, and oxomemazine in the presence of degradation products. A Beckman

2ACETAMINOPHEN

Gold system consisting of a model 125 programmable pump, a model 166 programmable UV detector, and a Rheodyne 20-L loop injector was used. The stationary phase was a stainless steel ODS C18 column (250 4.6 mm, 5-m particle size) with a guard column (50 4.6 mm) of the same packing material. Mobile phase A was a mixture of methanol and water (18:82, vol/vol) adjusted to pH 3.9 with phosphoric acid and mobile phase B was a mixture of methanol and water (80:20, vol/vol) adjusted to pH 3.9 with phosphoric acid. Mobile phase A was delivered from 0 to 12 minutes and mobile phase B from 12 to 22 minutes. The flow rate was 1.5 mL/min. UV detection was performed at 235 nm and 0.05 AUFS. Metronidazole was used as an internal standard. A portion of the cough syrup (10 mL) was diluted to 100 mL with methanol, mixed with internal standard solution, and further diluted with methanol to 20 g/mL of metronidazole in the final solution. Under these conditions, retention times for paracetamol, metronidazole, sodium benzoate, guaifenesin, and oxomemazine were about 2.6, 5.5, 8.6, 10.7, and 16.8 minutes, respectively (estimated from the published chromatogram). The method was evaluated to be stability indicating by assaying a mixture of active compounds and possible degradation products. Retention times for possible degradants 4-aminophenol and guaicol were about 3.7 and 7.0 minutes, respectively (estimated from the published chromatogram). A standard curve for acetaminophen was constructed from 5.0 to 30.0 g/mL. The correlation coefficient was 0.9998. Intraday and interday coefficients of variation were 1.06 and 1.86%, respectively.

Reference
Hewala II. Stability-indicating HPLC assay for paracetamol, guaiphenesin, sodium benzoate and oxomemazine in cough syrup. Anal Lett. 1994; 27: 7193.

Method 3
Alvi and Castro reported a simultaneous analysis of acetaminophen and hydrocodone bitartrate in a tablet formulation by HPLC analysis. The liquid chromatograph consisted of a Waters model M6000A pump, a model 710B/712B WISP autosampler, a model 490 variable-wavelength detector, and a model 730 data module. The stationary phase was a Waters Nova-Pak C18 Radial-Pak cartridge column (100 8 mm, 4-m particle size) with a Waters C18 guard column. The column temperature was 30 C. The mobile phase consisted of an aqueous phosphate buffer and acetonitrile (84:16, vol/vol), where the phosphate buffer was 0.02 M monobasic potassium phosphate containing 0.2 mL of triethylamine and 0.2 mL of phosphoric acid and adjusted to pH 3.3 with 3 N phosphoric acid. The flow rate was 2.0 mL/min. UV detection was performed at 215 nm and 0.02 AUFS. A portion of powder prepared from ground tablets equivalent to one tablet was weighed, transferred into a 100-mL volumetric flask, mixed with 50 mL of the diluent (0.5 mL of 3 N phosphoric acid in 1000 mL of water), and then shaken for 45 minutes. This mixture was diluted to volume with the diluent, mixed, filtered, and further diluted with the diluent. The injection volume was 10 L. Under these conditions, the retention times of acetaminophen and hydrocodone bitartrate were 2.6 and 5.0 minutes, respectively. The method was evaluated to be stability indicating by assaying a synthetic mixture of drugs and their known degradation products. Retention times for p-aminophenol, hydromorphone hydrochloride, codeine sulfate, and p-chloroacetanilide were 1.6, 2.3, 3.3, and 34.6 minutes, respectively. Samples were also refluxed in 1 N hydrochloric

ACETAMINOPHEN3

acid, 1 N sodium hydroxide solution, or 10% hydrogen peroxide for 4 hours. The decomposition products were separated from their intact drugs. A standard curve for acetaminophen was obtained from 0.2998 to 0.6996 mg/mL. The correlation coefficient was 0.9995.

Reference
Alvi SU, Castro F. A stability-indicating simultaneous analysis of acetaminophen and hydrocodone bitartrate in a tablet formulation by HPLC. J Liq Chromatogr. 1987; 10: 341326.

Method 4
Sisco et al. developed an HPLC method for the simultaneous determination of acetaminophen, codeine phosphate, and sodium benzoate in an elixir formulation. The chromatographic system consisted of a DuPont model 850 high-pressure liquid chromatograph equipped with a DuPont automatic sampler with a 20-L loop, a DuPont column oven, a DuPont 4100 integrator, and a Waters model 440 UV detector. A Hewlett-Packard model 1040A diode-array detector was used for specificity studies of stressed and unstressed samples. The stationary phase was a Waters Bondapak C18 column (300 3.9 mm). The mobile phase consisted of a buffer and methanol (80:20, vol/vol). The buffer contained 2.4 g of 1-butanesulfonic acid sodium salt (0.015 M), 2.04 g of monobasic potassium phosphate (0.015 M), and 2 mL of triethylamine per liter of water. The pH of this solution was adjusted to 4.8 0.1 with dilute phosphoric acid. The flow rate was 2.0 mL/min. UV detection was performed at 214 nm. Samples were diluted 1:40 with distilled water, and the injection volume was 20 L. Under these conditions, retention times for acetaminophen, sodium benzoate, and codeine phosphate were 2.6, 3.8, and 4.7 minutes, respectively. The HPLC method was determined to be stability indicating by accelerated decomposition of acetaminophen. An elixir sample was placed in a clear flint-glass bottle, capped, and stored in a 60 C oven for 2 weeks. Also, a standard solution of acetaminophen and codeine phosphate was spiked with potential degradation products. Retention times for p-aminophenol, codeine N-oxide, and codeinone were 1.8, 6.4, and 8.8 minutes, respectively. Thus, degradation product peaks did not interfere with peaks of acetaminophen, sodium benzoate, and codeine phosphate. A standard curve for acetaminophen was constructed from 0.3 to 0.9 mg/mL (estimated from the published figure). The correlation coefficient was 0.9999.

Reference
Sisco WR, Rittenhouse CT, Everhart LA, et al. Simultaneous high-performance liquid chromatographic stability-indicating analysis of acetaminophen, codeine phosphate, and sodium benzoate in elixirs. J Chromatogr. 1986; 354: 35566.

Method 5
Wallo and DAdamo developed an HPLC method for the simultaneous assay of hydrocodone bitartrate and acetaminophen in a tablet formulation. A Waters model ALC 204 liquid chromatograph was equipped with a Waters model 710B WISP autosampler, a Schoeffel SF 770 variable-wavelength UV detector, and a Waters data module. The stationary phase was a Waters Bondapak C18 reversed phase column. The mobile phase consisted of 25% methanol and 75% of an aqueous solution containing 0.01 N mono-

4ACETAZOLAMIDE

basic potassium phosphate and 0.05 N potassium nitrate, adjusted to a pH of about 4.5 with 3 N phosphoric acid. The flow rate was about 1.1 mL/min. UV detection was performed at 283 nm and 2.0 AUFS. The injection volume was 13 L. The assay was determined to be stability indicating by spiking the sample with potential degradation products. Degradation product peaks did not interfere with the acetaminophen peak. Retention times for p-aminophenol, hydromorphone, acetaminophen, codeine, hydrocodone, and p-chloroacetanilide were 3.4, 5.2, 5.9, 7.3, 10.0, and 43.3 minutes, respectively. Standard curves were constructed from 5 to 6.5 mg/mL. The correlation coefficient was 0.998.

Reference
Wallo WE, DAdamo A. Simultaneous assay of hydrocodone bitartrate and acetaminophen in a tablet formulation. J Pharm Sci. 1982; 71: 11158.

Method 6
Sena et al. reported an assay of acetaminophen in an effervescent tablet by ion-pair HPLC. A Waters model 6000A pump was equipped with a Valco model CV-6-UHPa-N60 10-L loop injector, a Waters model 440 fixed-wavelength UV detector, and a strip-chart recorder. The stationary phase was a Waters Bondapak phenyl column (300 4 mm, 10-m particle size). The mobile phase was 15% (vol/vol) acetonitrile in 0.005 M tetrabutylammonium phosphate in distilled water, adjusted to pH 7.5 with phosphoric acid or sodium hydroxide. The flow rate was 3.0 mL/min. UV detection was performed at 254 nm and 0.2 AUFS. An acetaminophen tablet was dissolved in 100 mL of a diluent of distilled water and acetonitrile (85:15, vol/vol), diluted by a factor of 200 with the diluent, and filtered. The injection volume was 10 L. Under the described conditions, the retention time for acetaminophen was about 2 minutes (estimated from the published chromatogram). The method was demonstrated to be stability indicating by spiking the acetaminophen solution with its degradation product, p-aminophenol. p-Aminophenol was well resolved from its parent compound. A standard curve for acetaminophen was generated from 10 to 30 g/mL and its correlation coefficient was 0.9998.

Reference
Sena FJ, Piechocki JT, Li KL. Stability-indicating assay of acetaminophen in an effervescent tablet by ion-pair high-performance liquid chromatography. J Pharm Sci. 1979; 68: 14656.

Acetazolamide
Chemical Names
N-[5-(Aminosulfonyl)-1,3,4-thiadiazol-2-yl]acetamide 5-Acetamido-1,3,4-thiadiazole-2-sulfonamide

ACETAZOLAMIDE5

Other Name
Diamox

Form
Acetazolamide Acetazolamide sodium

Molecular Formula
C4H6N4O3S2 C4H5N4NaO3S2

MW
222.3 244.2

CAS
59-66-5 1424-27-7

Appearance
Acetazolamide is a white to faintly yellowish-white, odorless, crystalline powder. Acetazolamide sodium is a white solid.

Solubility
Acetazolamide is sparingly soluble in water and slightly soluble in alcohol. Acetazolamide sodium is freely soluble in water.

pKa

Acetazolamide has pKa values of 7.4 and 9.1.

Method 1
Allen and Erickson studied the stability of acetazolamide in extemporaneously compounded oral liquids. A Hewlett-Packard series 1050 automated high-performance liquid chromatograph included a multisolvent mixing and pumping system, an autoinjector, a diode-array detector, and a computer with Chem Station software. The stationary phase was a Bakerbond C18 analytical column (250 4.6 mm). The mobile phase contained 950 mL of water, 20 mL of methanol, 30 mL of acetonitrile, and 4.1 g of anhydrous sodium acetate, the pH was adjusted to 4.0 with acetic acid. It was delivered isocratically at 2.0 mL/min. UV detection was performed at 254 nm. Samples were diluted 1:100. The injection volume was 20 L. The retention time for acetazolamide was 3.1 minutes. The HPLC assay was determined to be stability indicating by degrading samples of acetazolamide in water and in various commercial oral vehicles using heat, acid, base, oxidizing agent, and light. A composite chromatogram of acetazolamide after accelerated decomposition showed that degradation product peaks did not interfere with the intact acetazolamide peak. A standard curve was constructed from 50 to 500 g/mL. The intraday and interday coefficients of variation were 1.1 and 1.5%, respectively.

Reference
Allen LV, Erickson MA. Stability of acetazolamide, allopurinol, azathioprine, clonazepam, and flucytosine in extemporaneously compounded oral liquids. Am J Health Syst Pharm. 1996; 53: 19449.

Method 2
Alexander et al. evaluated the stability of acetazolamide 25 mg/mL in a suspension using a stability-indicating HPLC analytical method. The system consisted of a Beckman 110B solvent-delivery system, a Beckman system organizer injector equipped with a 20-L loop, a Beckman 420 system controller programmer, a Knauer 731.710 UV detector, and a Shimadzu C-R3A Chromatopac integrator. The stationary phase was a Whatman ODS-

6ACETYLCHOLINE CHLORIDE

3 C18 Partisil column (250 4.6 mm, 10-m particle size). The mobile phase was 65% 0.02 M phosphate buffer in deionized water and 35% methanol and was delivered isocratically at 1 mL/min. UV detection was performed at 254 nm and 0.04 AUFS. Theophylline 1 mg/mL in methanol was used as an internal standard. Each sample was diluted with the mobile phase. The injection volume was 50 L. Retention times were 4.5 and 5.8 minutes for acetazolamide and theophylline, respectively. The HPLC method was determined to be stability indicating. The acetazolamide suspension was heated for 2 hours under acidic and alkaline conditions. Degradation product peaks did not interfere with either the acetazolamide or the theophylline peak. A calibration curve was obtained by regression analysis of the ratio of acetazolamide peak area to the internal standard peak area versus the acetazolamide concentration from 10 to 100 g per injection.

Reference
Alexander KS, Haribhakti RP, Parker GA. Stability of acetazolamide in suspension compounded from tablets. Am J Hosp Pharm. 1991; 48: 12414.

Method 3
Parasrampuria et al. studied the stability of acetazolamide sodium in 5% dextrose and 0.9% sodium chloride injections. A Waters ALC 202 high-performance liquid chromatograph was equipped with a Omniscribe recorder and a Schoeffel SF 770 multiplewavelength detector. The stationary phase was a Waters Bondapak nonpolar C18 column. The mobile phase was an aqueous solution containing 0.02 M monobasic potassium phosphate, methanol, and acetonitrile (86:12:2, vol/vol/vol). The flow rate was 2.0 mL/min. UV detection was performed at 265 nm. Sulfamerazine was used as an internal standard for the purpose of calculating the acetazolamide concentrations. The sample injection volume was 20 L. Retention times for acetazolamide and the internal standard were 4.2 and 6.8 minutes, respectively (estimated from the published chromatogram). The HPLC method was determined to be stability indicating by accelerated decomposition of acetazolamide sodium. A 6-mL sample of the acetazolamide solution was mixed with 0.5 mL of 1 N sodium hydroxide and subjected to boiling for 25 minutes. The decomposition product peaks did not interfere with the intact drug peak. This result was confirmed by injecting a sample of the decomposed product onto the column.

Reference
Parasrampuria J, Gupta DV, Stewart KR. Stability of acetazolamide sodium in 5% dextrose or 0.9% sodium chloride injection. Am J Hosp Pharm. 1987; 44: 35860.

Acetylcholine Chloride
Chemical Name
2-(Acetyloxy)-N,N,N-trimethylethanaminium chloride

ACETYLCYSTEINE7

Other Name
Miochol

Form
Acetylcholine chloride

Molecular Formula
C7H16ClNO2

MW
181.7

CAS
60-31-1

Appearance
Acetylcholine chloride occurs as white or off-white crystals or as a crystalline powder.

Solubility
Acetylcholine chloride is very soluble in cold water and alcohol and practically insoluble in ether. It decomposes in hot water or alkalis.

Method
Tao et al. described a simple and rapid HPLC method for quantitating acetylcholine in a lyophilized preparation. The HPLC system consisted of a Waters 6000A pump, a Waters U6K universal loop injector, a Waters 401 refractive index detector, and a SpectraPhysics 4100 microprocessor-controlled data system. The stationary phase was a Waters Bondapak C18 column. The mobile phase was prepared by adding sodium 1-heptanesulfonate (Waters PIC Reagent B-7) to 900 mL of water and adjusting the pH to 4.0 with 6 M ammonium hydroxide. After addition of 50 mL of acetonitrile, the resulting solution was brought to a volume of 1000 mL with additional water. The flow rate was 2.0 mL/min. Samples were diluted with the mobile phase. The injection volume was 50 L. Under these conditions, the retention time for acetylcholine ranged from 7.9 to 8.2 minutes. The method was determined to be stability indicating by heat stressing the sample at 130 C for 19 hours. Degradation product peaks did not interfere with the intact acetylcholine peak. A standard curve was constructed from 50 to 150 g of acetylcholine; the correlation coefficient was 0.9996.

Reference
Tao FT, Thurber JS, Dye DM. High-performance liquid chromatographic determination of acetylcholine in a pharmaceutical preparation. J Pharm Sci. 1984; 73: 13113.

Acetylcysteine
Chemical Names
N-Acetyl-L-cysteine L--Acetamido--mercaptopropionic acid

Other Name
Mucomyst

8ACETYLCYSTEINE

Form
Acetylcysteine Acetylcysteine sodium

Molecular Formula
C5H9NO3S C5H8NNaO3S

MW
163.2 185.2

CAS
616-91-1 19542-74-6

Appearance
Acetylcysteine is a white crystalline powder with a slight acetic odor.

Solubility
Acetylcysteine has an aqueous solubility of 1 in 5. In alcohol the solubility is 1 in 4. It is practically insoluble in chloroform and ether.

Method 1
Anaizi et al. determined the stability of acetylcysteine in an extemporaneously prepared ophthalmic solution using an HPLC method. The HPLC system included a Perkin-Elmer model 250 binary LC pump, a model LC 290 UV detector, a model ISS 100 autoinjector, an Epson computer workstation, and a Perkin-Elmer model 0258-0195 reduced-activity Pecosphere C18 column (330 4.6 mm, 3-m particle size). The mobile phase consisted of 50 mM phosphate buffer in HPLC-grade water (adjusted to pH 3.0 with phosphoric acid) and was delivered isocratically at 3.0 mL/min. UV detection was performed at 210 nm and 0.01 AUFS. DL-Phenylalanine 10 g/mL in HPLC-grade water was used as an internal standard. Samples were diluted with water and then the internal standard solution. The injection volume was 10 L. Under these conditions, retention times for acetylcysteine and phenylalanine were 0.5 and 1.2 minutes, respectively. To demonstrate the stability-indicating nature of the assay, solutions of acetylcysteine 100 mg/mL were adjusted to pH 0.9, 8.2, and 12.3 with 10 N sodium hydroxide and 10 N hydrochloric acid and were placed in a 60 C oven for 7 days. The pH was then adjusted and solutions were diluted before injecting onto the column. Degradation product peaks did not interfere with the intact acetylcysteine peak. A standard curve was generated from 5 to 40 g/mL; the correlation coefficient was 1.000. The intraday and interday coefficients of variation were less than 1.88 and 1.22%, respectively.

Reference
Anaizi NH, Swenson CF, Dentinger PJ. Stability of acetylcysteine in an extemporaneously compounded ophthalmic solution. Am J Health Syst Pharm. 1997; 54: 54953.

Method 2
Fawcett et al. investigated the stability of acetylcysteine 10% eyedrops stored in lowdensity polyethylene eyedrop bottles at various temperatures and under in-use conditions. The HPLC system included a Shimadzu LC-5A pump, a Rheodyne injector, a Shimadzu SPD-6AV variable-wavelength detector, and a Hitachi D-2500 chromatointegrator. The stationary phase was a Shandon Hypersil-ODS C18 column (100 4.6 mm, 5-m particle size). The mobile phase was 50 mM potassium dihydrogen phosphate solution. The flow rate was 1.5 mL/min. UV detection was performed at 214 nm. DL-Phenylalanine was used as an internal standard. Samples were diluted 1:200 with 0.05% sodium metabisulfite solution. The injection volume was 20 L. Under these conditions, the retention times of acetylcysteine,

ACYCLOVIR9

the decomposition product (N,Nn-diacetylcystine), and the internal standard were 1.4, 2.4, and 4.6 minutes, respectively. The stability-indicating nature of the assay was demonstrated by the accelerated decomposition of the drug. Acetylcysteine was oxidized by hydrogen peroxide 30%. The decomposition product, N,Nn-diacetylcystine, did not interfere with the analysis of the drug. A standard curve for acetylcysteine was constructed from 0.25 to 0.75 mg/mL. The correlation coefficient of the curve was 0.999.

Reference
Fawcett JP, Woods DJ, Hayes P, et al. The stability of acetylcysteine eyedrops. Aust J Hosp Pharm. 1993; 23: 1821.

Acyclovir
Chemical Names
2-Amino-1,9-dihydro-9-[(2-hydroxyethoxy)methyl]-6H-purin-6-one 9-[(2-Hydroxyethoxy)methyl]guanine

Other Names
Aciclovir, Zovirax

Form
Acyclovir Acyclovir sodium

Molecular Formula
C8H11N5O3 C8H10N5NaO3

MW
225.2 247.2

CAS
59277-89-3 69657-51-8

Appearance
Acyclovir and acyclovir sodium occur as white crystalline powders. Acyclovir has solubilities of 1.3 mg/mL in water at 25 C and 0.2 mg/mL in alcohol. Acyclovir sodium has a solubility of greater than 100 mg/mL in water at 25 C.

Solubility

pKa

Acyclovir has pKa values of 2.27 and 9.25.

Method 1
Dubhashi and Vavia developed an HPLC method for the quantitation of acyclovir in pharmaceutical dosage forms. The instrument consisted of a Jasco PU-980 intelligent pump with a 20-L fixed-loop injector and a Jasco UV-975 intelligent detector. The stationary phase was a Jasco Finepak SiL C18 column (250 4.6 mm, 5-m particle size). The mobile phase was 5% (vol/vol) acetonitrile in 0.01 M monobasic potassium phosphate aqueous buffer solution (pH 4.8). The flow rate was 1.0 mL/min. UV detection was performed at 255 nm and 0.16 AUFS. A sample of gel (about 100 mg) was accurately weighed, transferred into a 10mL volumetric flask, and extracted in distilled water by ultrasonication for 30 minutes;

10ACYCLOVIR

the flask was then filled to the 10-mL mark with water. The aqueous layer was collected, diluted with water to a nominal acyclovir concentration of about 100 g/mL, filtered, and assayed. For ointment samples, extraction of acyclovir was performed the same way as for the gel sample. The resulting mixture was centrifuged at 5000 rpm for 30 minutes. The lower, clear aqueous layer was collected and assayed. The injection volume was 20 L. Under these conditions, the retention time for acyclovir was about 11 minutes. The analytical method was evaluated to be stability indicating by accelerated decomposition of acyclovir. An acyclovir solution (2.5 mg/2.5 mL) was mixed with 10 mL of 1 N sodium hydroxide solution or 1 N sulfuric acid, diluted with 10 mL of water, heated to boiling for 10 minutes using a hot plate, cooled to room temperature, neutralized using 1 N sulfuric acid or 1 N sodium hydroxide solution, and assayed. The peak of the intact drug was resolved chromatographically from peaks of the degradation products. A linear relationship between the acyclovir concentration and peak area was obtained from 50 to 200 g/mL. The correlation coefficient was greater than 0.9985. The limit of detection was 75 ng and the limit of quantification was 250 ng.

Reference
Dubhashi SS, Vavia PR. Stability indicating reverse phase HPLC method for acyclovir. Indian Drugs. 2000; 37: 4648.

Method 2
Zhang et al. investigated the stability of acyclovir sodium 1, 7, and 10 mg/mL in 5% dextrose injection and 0.9% sodium chloride injection. The liquid chromatograph was a Waters model LC Module 1 Plus equipped with a multisolvent-delivery pump, a UV detector, and an autosampler. The stationary phase was a Waters Bondapak C18 column (300 3.9 mm, 10-m particle size). The mobile phase was 5 mM sodium acetate in water adjusted to pH 3.0 with acetic acid. The flow rate was 1.45 mL/min. UV detection was performed at 254 nm and 0.5 AUFS. Samples were appropriately diluted with HPLC-grade water. The injection volume was 20 L. Under these conditions, the retention time for acyclovir was 8.6 minutes. The analytical method was determined to be stability indicating by accelerated decomposition of acyclovir. Boiling the acyclovir sodium in 0.1 N hydrochloric acid for 5 minutes led to a 50% loss in peak area for intact acyclovir and the formation of a new peak at about 4.2 minutes. The degradation product peak did not interfere with the intact acyclovir peak. Calibration curves for acyclovir were constructed from a linear plot of peak area versus concentration from 0.025 to 0.150 mg/mL; the correlation coefficient was greater than 0.9999. The intraday and interday coefficients of variation were 0.8 and 0.9%, respectively.

Reference
Zhang Y, Trissel LA, Martinez JF, et al. Stability of acyclovir sodium 1, 7, and 10 mg/mL in 5% dextrose injection and 0.9% sodium chloride injection. Am J Health Syst Pharm. 1998; 55: 5747.

Method 3
Gupta et al. studied the chemical stability of acyclovir sodium in 5% dextrose and 0.9% sodium chloride injections. The chromatograph was a Waters ALC 202 liquid