Nitroglycerin

Molecular formula: C3H5N3O9

Molecular weight: 227.1
CAS Registry No.: 55-63-0

SAMPLE
Matrix: blood
Sample preparation: 3 mL Plasma + 12 mL dichloromethane: ethyl acetate 1:1, shake
mechanically at 250 cycles/min for 5 min, centrifuge at 550 g at 4 for 5 min. Remove the
organic phase and evaporate it to about 20 |xL under a stream of nitrogen at room temperature, inject.
HPLCVARIABLES
Column: 250 X 4 10 |xm Zorbax NH2
Mobile phase: n-Hexane: MeOH 95:5
Flow rate: 5
Injection volume: 20
Detector: Thermal energy analyzer, Thermo Electron Corp. Model 502A, furnace temp
575, argon 15 mL/min, oxygen 25 mL/min, MeOH/dry ice slush bath
CHROMATOGRAM
Retention time: 5.0
Internal standard: nitroglycerin
OTHER SUBSTANCES
Extracted: isosorbide dinitrate, isosorbide mononitrate
KEYWORDS
plasma; nitroglycerin is IS
REFERENCE
Maddock, J.; Lewis, P.A.; Woodward, A.; Massey, RR.; Kennedy, S. Determination of isosorbide dinitrate
and its mononitrate metabolites in human plasma by high-performance liquid chromatography-thermal energy analysis. J.Chromatogr., 1983, 272, 129-136

SAMPLE
Matrix: formulations
Sample preparation: Inject directly.
HPLCVARIABLES
Column: 300 X 3.9 10 jxm pJBondapak C18
Mobile phase: MeCN:MeOH:water 6:6:13
Column temperature: 35
Flow rate: 2.8
Detector: UV 210
CHROMATOGRAM
Retention time: 2.8
KEYWORDS
injections; stability-indicating

REFERENCE
Driver, RS.; Jarvi, E.J.; Gratzer, RL. Stability of nitroglycerin as nitroglycerin concentrate for injection
stored in plastic syringes. Am.J.Hosp.Pharm., 1993, 50, 2561-2563

SAMPLE
Matrix: formulations
HPLCVARIABLES
Column: C18
Mobile phase: MeOH: water 50:50
Flow rate: 1.3
Detector: UV 220
CHROMATOGRAM
Internal standard: isosorbide dinitrate
KEYWORDS
injections; 5% dextrose; stability-indicating
REFERENCE
Pramar, Y; Das Gupta, V.; Gardner, S.N.; Yau, B. Stabilities of dobutamine, dopamine, nitroglycerin
and sodium nitroprusside in disposable plastic syringes. J.Clin.Pharm.Ther., 1991, 16, 203-207

SAMPLE
Matrix: formulations
Sample preparation: Dilute with saline, inject a 10 jxL aliquot.
HPLCVARIABLES
Column: 250 X 4.6 Lichrosorb 10 RP 8
Mobile phase: MeOH: water 50:50
Flow rate: 2
Injection volume: 10
Detector: UV 214
CHROMATOGRAM
Retention time: 5.8
OTHER SUBSTANCES
Simultaneous: isosorbide dinitrate
KEYWORDS
injections; saline
REFERENCE
Martens, H.J.; de Goede, RN.; van Loenen, A.C. Sorption of various drugs in polyvinyl chloride, glass,
and polyethylene-lined infusion containers. Am.J.Hosp.Pharm., 1990, 47, 369-373

SAMPLE
Matrix: formulations
Sample preparation: Weigh out an amount of finely powdered tablets or capsules equivalent to about 25 mg of drug. Add 50 mL buffer, shake for 30 min, add 10 mL MeOH,
make up to 100 mL with buffer, filter (0.45 |xm), inject a 20 JXL aliquot. If the sample
clumps when the buffer is added, agitate with a stirring rod and sonicate. (Buffer was
MeOH:200 mM pH 4.7 ammonium acetate buffer:water 55:10:35.)

HPLCVARIABLES

Guard column: 50 X 6.4 25-37 |xm Whatman Co-Pell ODS

Column: 250 X 4.6 5 |xm Ultrasphere ODS
Mobile phase: MeOH: 200 mM pH 4.7 ammonium acetate buffer .water 55:10:35
Flow rate: 1
Injection volume: 20
Detector: UV 254
CHROMATOGRAM

Retention time: 8
Internal standard: nitroglycerin
OTHER SUBSTANCES

Simultaneous: isosorbide dinitrate, isosorbide mononitrate, pentaerythritol tetranitrate,

saccharin
KEYWORDS

tablets; capsules; nitroglycerin is IS

REFERENCE
Carlson, M.; Thompson, R.D.; Snell, R.P. Determination of isosorbide dinitrate in pharmaceutical products by HPLC. J.Chromatogr.ScL, 1988, 26, 574-578

SAMPLE

Matrix: formulations
HPLCVARIABLES
Column: 150 X 4.6 5 p,m C8 (Altex)
Mobile phase: MeCN: water 40:60
Flow rate: 2.67
KEYWORDS

tablets; mouth rinses

REFERENCE
Noonan, RK.; Benet, L.Z. Incomplete and delayed bioavailability of sublingual nitroglycerin.
Am.J.CardioL, 1985, 55, 184-187

SAMPLE

Matrix: formulations
Sample preparation: 1 mL Solution + 300 |xL isosorbide dinitrate solution, mix, inject an
aliquot.
HPLCVARIABLES

Column: 300 X 3.9 10 |xm jxBondapak phenyl

Mobile phase: MeCN: THF: water 26:10:64
Flow rate: 2
Detector: UV 218
CHROMATOGRAM

Internal standard: isosorbide dinitrate

KEYWORDS

injections; 5% dextrose; saline

REFERENCE
Thompson, M.; Smith, M.; Gragg, R.; Soliman, K.F. Stability of nitroglycerin and dobutamine in 5%
dextrose and 0.9% sodium chloride injection. Am.J.Hosp.Pharm., 1985, 42, 361-362

SAMPLE
Matrix: formulations
Sample preparation: Powder tablets, weigh out a portion equivalent to 3 mg erythrityl
tetranitrate, add to 10 mL 75 jJig/mL nitroglycerin in MeOH, sonicate for 2 min, shake
mechanically for 30 min, filter, inject an aliquot
HPLCVARIABLES
Guard column: 40 X 4.6 ixBondapak C18/Corasil
Column: 300 X 3.9 10 |xm jxBondapak C18
Mobile phase: MeOH: water 40:60
Flow rate: 1
Injection volume: 20
Detector: UV 220
CHROMATOGRAM
Retention time: 14
Internal standard: nitroglycerin
OTHER SUBSTANCES
Simultaneous: erythrityl tetranitrate, isosorbide dinitrate, pentaerythritol tetranitrate
KEYWORDS
tablets; nitroglycerin is IS
REFERENCE
Olsen, CS.; Scroggins, H.S. High-performance liquid chromatographic determination of the nitrate esters isosorbide dinitrate, pentaerythritol tetranitrate, and erythrityl tetranitrate in various tablet
forms. J.Pharm.Scl, 1984, 73, 1303-1304

SAMPLE
Matrix: formulations
Sample preparation: Dilute with water, inject an aliquot.
HPLC VARIABLES
Column: 300 X 3.9 ixBondapack phenyl
Mobile phase: MeCN: THF: water 26:10:64
Flow rate: 2
Injection volume: 10-40
Detector: UV 218
CHROMATOGRAM
Retention time: 10
Internal standard: isosorbide dinitrate (6)
KEYWORDS
inj ections; stability-indicating
REFERENCE
Baaske, D.M.; Carter, J.E.; Amann, A.H. Rapid and accurate stability-indicating assay for nitroglycerin.
J.Pharm.ScL, 1979, 68, 481-483

SAMPLE

Matrix: solutions
Sample preparation: Add 10 g 10% nitroglycerin solution in lactose to 125 mL MeOH,
sonicate for 5 min, shake mechanically for 30 min, dilute to 200 mL with MeOH, let
undissolved lactose settle, filter through paper. Dilute 5 mL to 250 mL with mobile phase,
inject a 20 |xL aliquot.
HPLCVARIABLES

Column: 250 X 4.6 5 |xm Ultrasphere ODS C18

Mobile phase: MeCN: water 65:35
Flow rate: 1
Injection volume: 20
Detector: UV 230
CHROMATOGRAM

Retention time: 6
Internal standard: nitroglycerin
OTHER SUBSTANCES

Simultaneous: pentaerythritol tetranitrate

KEYWORDS

collaborative study; nitroglycerin is IS

REFERENCE
Carlson, M. Liquid chromatographic determination of pentaerythritol tetranitrate in Pharmaceuticals:
collaborative study. J.Assoc.Off.Anal.Chem., 1990, 73, 693-697

SAMPLE

Matrix: solutions
HPLCVARIABLES

Column: 150 X 4.6 5 |xm Supelcosil LC-8

Mobile phase: MeOH: water 50:50
Flow rate: 1.5
Detector: UV 210
CHROMATOGRAM

Retention time: 4.5-5

REFERENCE
Roberts, M.E.; Mueller, K.R. Comparisons of in vitro nitroglycerin (TNG) flux across yucatan pig, hairless mouse, and human skins. Pharm.Res., 1990, 7, 673-676

SAMPLE

Matrix: solutions
HPLCVARIABLES

Column: 33 X 4.6 3 u.m C18 (Perkin-Elmer)

Mobile phase: MeCN: MeOH: water 24:24:52
Flow rate: 3.5
Detector: UV 210

CHROMATOGRAM
Retention time: 0.6
Limit of quantitation: 1 |xg/mL
OTHER SUBSTANCES
Simultaneous: degradation products
REFERENCE
Severin, G. Rapid high-performance liquid chromatographic procedure for nitroglycerin and its degradation products. J.Chromatogr., 1985, 320, 445-449

ANNOTATED BIBLIOGRAPHY
Carlson, M.; Thompson, R.D. Determination of pentaerythritol tetranitrate in Pharmaceuticals by high
performance liquid chromatography. J.Liq.Chromatogr., 1988, 11, 2603-2620 [simultaneous pentaerythritol tetranitrate; tablets; capsules; nitroglycerin is IS]
Torok, L; Paal, T.; Koszegine Szalai, H.; Keseru, P. [High performance liquid chromatographic assay and
determination of the even distribution of active ingredients in nitroglycerin and isosorbide dinitrate
tablets]. Ada Pharm.Hung., 1985, 55, 154-162
Baaske, D.M.; Karnatz, N.N.; Carter, J.E. High-performance liquid chromatographic assay for partially
nitrated glycerins in nitroglycerin. J.Pharm.Sci., 1983, 72, 194-196
Gelber, L.; Papas, A.N. Validation of high-performance liquid chromatographic methods for analysis of
sustained-release preparations containing nitroglycerin, isosorbide dinitrate, or pentaerythritol tetranitrate. J.Pharm.Sci., 1983, 72, 124-126
Yu, W.C.; Goff, E.U. Determination of vasodilators and their metabolites in plasma by liquid chromatography with a nitrosyl-specific detector. Anal.Chem., 1983, 55, 29-32 [TEA detection; plasma; SPE;
extracted isosorbide dinitrate, isosorbide mononitrate, pentaerythritol tetranitrate; LOD 0.1 ng]