Lovastatin

Molecular formula: C24H36O5

Molecular weight: 404.6
CAS Registry No.: 75330-75-5

SAMPLE
Matrix: bile, blood
Sample preparation: Prepare a 1 mL Bond-Elut C2 SPE cartridge by washing with 1 mL
MeOH and 1 mL buffer. Plasma. 500 |xL Plasma + 500 jxL buffer, keep in an ice water
mixture, add 100 jxL MeCN: water 60:40, add 50 |JLL 5 |xg/mL methyl mevinolinic acid in
MeCN: water 60:40, mix, add to the SPE cartridge, wash twice with 1 mL buffer, wash
twice with 1 mL MeCN: buffer 20:80, elute with 400 |JLL MeCN: water 75:25, inject a 30
|xL aliquot of eluant. Bile. Inject a 10 |xL aliquot directly. (Buffer was 100 mM pH 7.2
potassium phosphate buffer.)
HPLCVARIABLES
Column: 50 X 4.6 3 |xm Sepralyte C18
Mobile phase: MeCN: 50 mM ammonium phosphate + 10 mM phosphoric acid. Isocratic
50:50 (plasma). Gradient 20:80 to 75:25 over 10 min (bile).
Flow rate: 1.5
Injection volume: 30 (plasma), 10 (bile)
Detector: UV 238
CHROMATOGRAM
Retention time: 4.95
Internal standard: methyl mevinolinic acid (3.50)
Limit of detection: 25 ng/mL
OTHER SUBSTANCES
Simultaneous: mevinolinic acid
KEYWORDS
plasma; human; rat; dog; SPE
REFERENCE
Stubbs, R.J.; Schwartz, M.; Bayne, W.F. Determination of mevinolin and mevinolinic acid in plasma and
bile by reversed-phase high-performance liquid chromatography. J.Chromatogr., 1986, 383, 438-443

SAMPLE
Matrix: bile, microsomal incubations
Sample preparation: Microsomal incubations. Extract 10 mL microsomal incubations
with 30 mL ethyl acetate: acetone 2:1, centrifuge. Remove organic layer and dry it over
anhydrous sodium sulfate, filter, evaporate under vacuum, reconstitute in n-propanol,
inject an aliquot. Bile. Acidify to pH 3.5 with 20 mM formic acid, extract with toluene:
n-propanol 85:15, extract with n-butanol saturated with water, combine extracts, evaporate under vacuum, reconstitute in n-propanol, inject an aliquot.
HPLCVARIABLES
Column: 300 X 10 |jJBondapak ODS

Mobile phase: Gradient. MeCN: water from 30:70 to 90:10 over 30 min.
Flow rate: 3
Detector: UV 238
CHROMATOGRAM

Retention time: 26
OTHER SUBSTANCES

Simultaneous: metabolites
REFERENCE
Vyas, K.R; Kari, RH.; Pitzenberger, S.M.; Halpin, R.A.; Ramjit, H.G.; Arison, B.; Murphy, J.S.; Hoffman,
W.F.; Schwartz, M.S.; UIm, E.H. Biotransformation of lovastatin. I. Structure elucidation of in vitro
and in vivo metabolites in the rat and mouse. Drug Metab.Dispos., 1990, 18, 203-211

SAMPLE

Matrix: microsomal incubations

Sample preparation: 1 mL Microsomal incubation + 1 mL acetone + 2 mL ethyl acetate,
extract, centrifuge. Remove the organic layer and dry it over anhydrous sodium sulfate,
concentrate under a stream of nitrogen. Reconstitute in 200 |xL n-propanol, inject 20-50
juiL aliquot.
HPLC VARIABLES

Column: 250 X 4.6 5 jxm Ultrasphere ODS

Mobile phase: Gradient. MeCN: 5 mM formic acid from 30:70 to 90:10 over 30 min.
Flow rate: 1
Injection volume: 20-50
Detector: UV 238
CHROMATOGRAM

Retention time: 25
OTHER SUBSTANCES

Simultaneous: metabolites
REFERENCE
Vyas, K.R; Kari, RH.; Prakash, S.R.; Duggan, D.E. Biotransformation of lovastatin. II. In vitro metabolism by rat and mouse liver microsomes and involvement of cytochrome P-450 in dehydrogenation
of lovastatin. Drug Metab.Dispos., 1990, 18, 218-222

SAMPLE

Matrix: solutions
HPLCVARIABLES

Column: 250 X 4.6 5 ynn Supelcosil LC-DP (A) or 250 X 4 5 fxm LiChrospher 100 RP-8 (B)
Mobile phase: MeCN:0.025% phosphoric acid:buffer 25:10:5 (A) or 60:25:15 (B) (Buffer
was 9 mL concentrated phosphoric acid and 10 mL triethylamine in 900 mL water, adjust
pH to 3.4 with dilute phosphoric acid, make up to 1 L.)
Flow rate: 0.6
Injection volume: 25
Detector: UV 229
CHROMATOGRAM

Retention time: 10.49 (A), 14.79 (B)

OTHER SUBSTANCES
Also analyzed: acebutolol, acepromazine, acetaminophen, acetazolamide, acetophenazine,
albuterol, alprazolam, amitriptyline, amobarbital, amoxapine, antipyrine, atenolol, atropine, azatadine, baclofen, benzocaine, bromocriptine, brompheniramine, brotizolam,
bupivacaine, buspirone, butabarbital, butalbital, caffeine, carbamazepine, cetirizine,
chlorcyclizine, chlordiazepoxide, chlormezanone, chloroquine, chlorpheniramine,
chlorpromazine, chlorpropamide, chlorprothixene, chlorthalidone, chlorzoxazone, cimetidine, cisapride, clomipramine, clonazepam, clonidine, clozapine, cocaine, codeine, colchicine, cyclizine, cyclobenzaprine, dantrolene, desipramine, diazepam, diclofenac, diflunisal,
diltiazem, diphenhydramine, diphenidol, diphenoxylate, dipyridamole, disopyramide, dobutamine, doxapram, doxepin, droperidol, encainide, ethidium bromide, ethopropazine,
fenoprofen, fentanyl, flavoxate, fluoxetine, fluphenazine, flurazepam, flurbiprofen, fluvoxamine, furosemide, glutethimide, glyburide, guaifenesin, haloperidol, homatropine, hydralazine, hydrochlorothiazide, hydrocodone, hydromorphone, hydroxychloroquine, hydroxyzine, ibuprofen, imipramine, indomethacin, ketoconazole, ketoprofen, ketorolac,
labetalol, levorphanol, lidocaine, loratadine, lorazepam, loxapine, mazindol, mefenamic
acid, meperidine, mephenytoin, mepivacaine, mesoridazine, metaproterenol, methadone,
methdilazine, methocarbamol, methotrexate, methotrimeprazine, methoxamine, methyldopa, methylphenidate, metoclopramide, metolazone, metoprolol, metronidazole, midazolam, moclobemide, morphine, nadolol, nalbuphine, naloxone, naphazoline, naproxen,
nifedipine, nizatidine, norepinephrine, nortriptyline, oxazepam, oxycodone, oxymetazoline, paroxetine, pemoline, pentazocine, pentobarbital, pentoxifylline, perphenazine,
pheniramine, phenobarbital, phenol, phenolphthalein, phentolamine, phenylbutazone,
phenyltoloxamine, phenytoin, pimozide, pindolol, piroxicam, pramoxine, prazepam, prazosin, probenecid, procainamide, procaine, prochlorperazine, procyclidine, promazine, promethazine, propafenone, propantheline, propiomazine, propofol, propranolol, protriptyline, quazepam, quinidine, quinine, racemethorphan, ranitidine, remoxipride, risperidone,
salicylic acid, scopolamine, secobarbital, sertraline, sotalol, spironolactone, sulfinpyrazone, sulindac, temazepam, terbutaline, terfenadine, tetracaine, theophylline, thiethylperazine, thiopental, thioridazine, thiothixene, timolol, tocainide, tolbutamide, tolmetin,
trazodone, triamterene, triazolam, trifluoperazine, triflupromazine, trimeprazine, trimethoprim, trimipramine, verapamil, warfarin, xylometazoline, yohimbine, zopiclone
KEYWORDS
some details of plasma extraction
REFERENCE
Koves, E.M. Use of high-performance liquid chromatography-diode array detection in forensic toxicology.
J.Chromatogr.A, 1995, 692, 103-119

SAMPLE
Matrix: solutions
Sample preparation: Centrifuge at 2000 rpm, inject an aliquot.
HPLCVARIABLES
Column: 150 X 4.6 5 |xm Hypersil ODS
Mobile phase: MeCN: water: triethylamine: glacial acetic acid 500:500:1:1
Column temperature: 30
Flow rate: 2
Detector: UV 238
CHROMATOGRAM
Retention time: 14.4 (4.4 hydroxy acid form)
Limit of detection: 10 ng/mL
Limit of quantitation: 25 ng/mL

REFERENCE
Serajuddin, A.T.; Ranadive, S.A.; Mahoney, E.M. Relative lipophilicities, solubilities, and structurepharmacological considerations of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors pravastatin, lovastatin, mevastatin, and simvastatin. J.Pharm.Sci., 1991, 80, 830-834

SAMPLE

Matrix: tissue
Sample preparation: Homogenize in 4 mL water, add 4 mL ice-cold acetone, centrifuge at
3000 rpm for 15 min, extract twice with an equal volume of hexanes. Evaporate extracts
under vacuum, reconstitute in n-propanol, inject an aliquot.
HPLCVARIABLES

Column: 300 X 7 Hamilton PRP-I

Mobile phase: Gradient. MeCN: 5 mM formic acid from 30:70 to 90:10 over 30 min.
Flow rate: 3
Detector: UV 238
CHROMATOGRAM

Retention time: 19
OTHER SUBSTANCES
Simultaneous: metabolites
KEYWORDS

liver
REFERENCE
Vyas, K.P.; Kari, RH.; Pitzenberger, S.M.; Halpin, R.A.; Ramjit, H.G.; Arison, B.; Murphy, J.S.; Hoffman,
W.F.; Schwartz, M.S.; UIm, E.H. Biotransformation of lovastatin. I. Structure elucidation of in vitro
and in vivo metabolites in the rat and mouse. Drug Metab.Dispos., 1990, 18, 203-211