norge strel

8/17/2019 Norge Strel

1/7

SAMPLE

Matrix: blood

Sample p repa ra ti o n : 1 mL Serum + 3 mL MTBE, vortex for 1 min, centrifuge at 1500

rpm for 5 min. Remove the organic layer and evaporate it to dryness under a stream of

nitrogen at room temperature, reconstitute the residue in 100 jxL MeOH, inject an aliquot.

HPLCVARIABLES

G uar d co lum n: 18 mm long (Brownlee)

Column: 300 X 3.6 10 |xm jxBondapak C18

Mobile ph as e: MeOH: water 80:20

Flow rate: 1

Dete cto r: UV 254; RIA

C H R O M A T O G R A M

Retention time: 5.6

OTHER SUBSTANCES

Extracted: metabolites, norgestimate

KEYWORDS

serum

REFERENCE

Wong, F.A.; Juzw in, S.J.; Tischio, N.S.; Flor, S.C. Determ ination of norgestim ate in serum by a utom ated

high-performance liquid chromatography and subsequent radioimmunoassay. J.Liq.Chromatogr.

1995,28, 1851-1861

SAMPLE

Matrix: bulk

HPLCVARIABLES

Colu mn: 250 X 4 10 |xm LiChrosorb RP-18

Mobile ph as e: MeOH: water 70:30

Flow rate: 1

Injection volume: 25

D etecto r: UV 240


Retention t ime: 9

OTHER SUBSTANCES

Simultaneous: impurities, norethindrone

Interfering: ethinyl estradiol

REFERENCE

Gorog, S.; Hereny i, B. An alysis of stero ids. XXXVIII. The u se of high-perform ance liquid chro m atog raph y

with diode-array UV detection for es tim ating imp urity profiles of steroid drug s. J.Chromatogr. 1987,

400

177-186

Norgestrel

Molecular formula: C

21

H

28

O

2

Molecular weight: 312.5

CAS Registry No :

797-63 -7, 797-64-8 -) form), 6533-00 -2

Previous page

http://81765_13b.pdf/http://81765_13b.pdf/


2/7

SAMPLE

Matrix: culture medium

Sample preparat ion: Extract culture medium twice with 2 volumes of ether, combine the

extracts and evaporate them to dryness, reconstitute with MeOH, inject an aliquot.

HPLCVARIABLES

Column: 300 X 3.9 Techopak 10 C18 (HPLC Technology)

Mobile phase:

MeOH: water 70:30

F low rate: 1.5

Detector:

UV 240; Radioactivity

CHROMATOGRAM

Retent ion t ime: 5

OTHER SUBSTANCES

Extracted:

metabolites, norgestimate

KEYWORDS

tritium labeled

REFERENCE

Wild, M.J.; Rudland, RS.; Back, D.J. Metabolism of the oral contraceptive steroids ethynylestradiol and

norge stima te by norm al (Hum a 7) and m align ant (MCF-7 and ZR-75-1) hu m an b rea st cells in culture.

J.Steroid Biochem.Mol.BioL 1991, 39 5 3 5 - 5 4 3

SAMPLE

Matrix:

formulations

Sample preparat ion: Centrifuge oil formulation at 30° at 2000 rpm for 30 min, filter

(W hatman No. 1 paper), collect the last 4 mL of th e n itra te. D ilute a 10 |xL aliquot to 10

mL with M eCN : w ater 60 :40 containin g 0.3% Tween 80, remove a 2 mL aliquot a nd add

it to 1 mL 3.33 |xg/mL progesterone, vortex for 10 s, inject a 50 |xL aliquot.

HPLCVARIABLES

Column:

300 X 3.9 Novapak C18

Mobile phase: MeCN: water 60:40

Flow rate: 2

Inject ion volume: 50

Detector: UV

248

CHROMATOGRAM

Internal s tandard:

progesterone

KEYWORDS

oils

REFERENCE

Gao, Z.-H.; Shu kla, A. J.; Joh nso n, J.R.; Crowley, W.R. Controlled re leas e of a con tracep tive ste roid from

biodegradable and injectable gel formulations: In vitro evaluation. Ph arm.Res. 1995, 12 857—863

SAMPLE

Matrix:

formulations

Sample preparat ion: Oils. 1 mL Sam ple + 25 mL MeO H-.water 90: 10 , sha ke vigorously

for 5 min, centrifuge, inject a 10

|JLL

aliquot of the supernatant. Tablets. Grind a tablet

to a fine powder, add 25 mL MeOH, sonicate for 5-10 min, filter (0.45 |min), discard first

5 mL of filtrate, inject a 10 |xL aliquot of the remaining filtrate. Suspensions (aqueous).


3/7

Make up 5 mL to 50 mL with MeOH, filter (0.45 jjim), discard first 5 mL of filtra te, inject

a 10 |xL aliquot of the remaining filtrate.

HPLCVARIABLES

Column: 250 X 4.6 5 |xm Zorbax ODS

Mobile phase:

MeOH: water 75:25

Flow rate: 1.5


Detector: UV

240

CHROMATOGRAM

Retention time: 5.9

Limit of detection:

5 |xg/mL

OTHER SUBSTANCES

Simultaneous:

aspirin, benzyl alcohol, benzyl benzoate, boldenone, caffeine, caluster6ne,

cortisone, dehydroepiandrosterone (UV 210), fluoxym esterone, formebolone, mesterolone

(UV 210), methandriol (UV 210), methenolone acetate, methyltestosterone, mibolerone,

nandrolone acetate, nandrolone propionate, norethandrolone, norethindrone, norethin-

drone acetate, oxandrolone (UV 210), oxymetholone, stanozolol, testolactone, testosterone

acetate, testosterone propionate, trenbolone acetate

Interfering:

ethisterone, methandrostenolone, nandrolone, testosterone

KEYWORDS

oils;

tablets; suspensions

REFERENCE

Walters, M.J.; Ayers, R.J.; Brown, D.J. Analysis of illegally distributed anabolic steroid products by

liquid chromatography with identity confirmation by mass spectrometry or infrared spectrophotom-

etry J.Assoc.OffAnaLChem. 1990, 73 904-926

SAMPLE

Matrix:

microsomal incubations, mucosal fluid

Sample preparation:

Mucosal fluid . Extract 1 mL mucosal fluid twice with 5 mL diethyl

ether, evaporate extracts to dryness, resuspend residue in 100 |xL MeOH, inject an ali-

quot. Microsomal incubations. Extract 2.5 mL microsomal preparation with 5 mL diethyl

ether, proceed as before.

HPLCVARIABLES

Guard column:

on-line guard column

Column:

100 X 8 ixBondapak radial compression module

Mobile phase: MeOH: water 70:30

Flow rate: 1.5

Injection volume:

100

Detector: UV 240

CHROMATOGRAM

Retention time: 6

OTHER SUBSTANCES

Simultaneous:

3-ketonorgestimate, 17-deacetylnorgestimate, norgestimate

REFERENCE

Madden, S.; Back, D.J. Metabolism of norgestimate by human gastrointestinal mucosa and liver micro-

somes in vitro. J.Steroid Biochem.Mol.Biol. 1991, 38 4 9 7 - 5 0 3


4/7

SAMPLE

Matrix: solutions

HPLC VARIABLES

Column: 50 X 4.6 5 |xm Supelcosil LC-18

Mobile phase: MeOH: THF: water 10:20:70

Flow rate: 2


Detector: UV

220

CHROMATOGRAM

Retention time:

5.5

OTHER SUBSTANCES

Simultaneous:

ethinyl estradiol, norethindrone, norethindrone acetate, norethynodrel

acetate

REFERENCE

Supelco Catalog Supelco Inc. Bellefonte PA

1996, p. A130

SAMPLE

Matrix: solutions

Sample preparation:

Inject an aliquot of a 10 |xg/mL solution.

HPLC VARIABLES

Column:

150

X 4.6

Supelco

ODS

Mobile phase: MeCN: water 25:75 containing 14 mM cyclodextrin

Column temperature: 0

Flow rate: 1 3


Detector: UV 240

CHROMATOGRAM

Retention time: 16 D +)), 18 D -))

KEYWORDS

chiral

REFERENCE

Lamparczyk, H.; Zarzycki, RK.; Nowakowska, J. Effect of temperature on separation of norgestrel en-

antiomers

by

high-performance liquid chromatography. J.Chromatogr.A

1994,

668 413-417

SAMPLE

Matrix: solutions

Sample preparation: Direct injection.

HPLCVARIABLES

Column:

250

X 4.6 10

m Partisil

C 18 ODS 3

Mobile phase: MeCN-.water 50:50

Flow rate: 2

Detector: UV 243

CHROMATOGRAM

Retention time: 6.0


5/7

KEYWORDS

see also J.Pharm.Sci. 1989, 78, 477

REFERENCE

Catz, P.; Friend, D.R. In vitro evaluations of transdermal levonorgestrel.

D rug D es.Deliv.

1990 6 4 9 -

60

SAMPLE

Matrix: solutions

HPLCVARIABLES

Column: 250 X 4.6 10 jim Partisil ODS-3 C18

Mobile phase: MeCN-.water 50:50

Flow rate: 2

D etecto r: UV 243


Retention time: 6.0

REFERENCE

Friend, D.R.; C atz, P.; Heller, J.; O kagaki, M. Tran sderm al delivery of levonorgestrel IV : Ev aluation of

membranes.

J.Pharm.Sci.

1989

78

477-480

SAMPLE

Matrix: solutions

HPLCVARIABLES

Guard column: Chromsep reverse phase guard column (Chrompack)

Column: 100 X 3 5 |xm Chromspher glass column

M obile ph as e: MeCN: water 35:65

Flow rate: 0.4

Detec tor: UV 247


Retention time: 20

OTHER SUBSTANCES

Sim ul tane ou s: nandrolone, trenbolone (at UV 340)

REFERENCE

Haa snoot, W.; Schilt, R.; Ha m ers , A.R.; Huf, F.A.; Farjam , A.; Fre i, R.W.; B rin km an , U.A. Dete rm ina tion

of p-19-nortestosterone a nd its metab olite a-19-nortestosterone in biological samples at th e sub pa rts

per billion level by high-performance liquid chromatography with on-line immunoaffinity sample

pretreatment. J.Chromatogr 1989 489 157-171

SAMPLE

Matrix: solutions

HPLCVARIABLES

Colum n: 250 X 4.6 10 jxm Nucleosil C18

Mobile pha se : MeCN:THF: water 12.9:22.4:64.7

Flow rate: 1

D etecto r: UV 240


6/7


Retention t ime: 13

OTHER SUBSTANCES

Sim ul tane ous : estrone, ethinyl estradiol, mestranol, norethindrone, norethindrone acetate

REFERENCE

Gazdag, M.; Szepesi, G.; Szeleczki, E. Selection of high-performance liquid chromatographic methods in

pharmaceutical analysis. I. Optimization for selectivity in reversed-phase chromatography.

J.Chromatogr.

1988

454

8 3 - 9 4

SAMPLE

Matrix: solutions

HPLCVARIABLES

Co lumn: 250 X 4.6 5

\xm

LiChrosorb Si 60

Mobile phase : Hexane: dioxane: isopropanol 95 : 3:2

Flow rate: 1

D etec tor: UV 254


Retention t ime: 13

OTHER SUBSTANCES

Sim ul tane ou s: estrone, ethinyl estradiol, mestranol, norethindrone, norethindrone acetate

KEYWORDS

normal phase

REFERENCE

Gazdag, M.; Szepesi, G.; Fabian-Varga, K. Selection of high-performance liquid chro ma tographic m eth-

ods in pharmaceutical analysis. II. Optimization for selectivity in normal-phase systems.

J.Chromatogr.

1988

454 9 5 - 1 0 7

SAMPLE

Matrix: solutions

HPLCVARIABLES

Colum n: 250 X 4.6 jmBondapak C18

M obile phase: Dioxane: water 50:50 (Caution Dioxane is a carcinogen )

Flow rate: 1.4

D etec tor: UV 254

OTHER SUBSTANCES

Simultaneous: norgestimate

REFERENCE

Killinger, J.; Ha hn, D.W.; Phillips, A.; Hetyei, N.S.; McGuire, J.L. T he affinity of norgestim ate for u ter ine

progestogen receptors and its direct action on the uterus.

Contraception

1985,

32

311-319

SAMPLE

Matrix: tissue

Sa m ple p re pa ra ti o n : Incubate endometrial tissue with buffer, remove tissue, extract me-

dium twice with 2 volumes of diethyl ether, evaporate to dryness, reconstitute in a small

volume of MeOH, inject an aliquot.


7/7

HPLC VARIABLES

Column: 300 X 3.9 Technopak 10 C18

Mobile phase: MeOH: water 70:30

Flow rate: 1.5

Detector: UV

240

CHROMATOGRAM

Retention time:

OTHER SUBSTANCES

Simultaneous:

metabolites, norgestimate

KEYWORDS

endometrial tissue

REFERENCE

Wild,

M.J.;

Rudland,

RS.;

Back, D.J. M etabolism of the oral contraceptive steroids ethyny lestradiol,

norgestimate and 3-ketodeso gestrel by a hu m an endo me trial cancer cell line (HEC-IA) and endo-

metrial tissue in

vitro. J.Steroid Biochem .Mol.BioL

1993,

45

407-420

norge strel

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