eryth romic in

7/28/2019 Eryth Romic In

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SAMPLEMatrix: bloodSample preparat ion: 200 jxL Plasma or whole blood + 10 jxL 10 |xg/mL oleandomycin +20 |JLL sa tur ate d sodium carbo nate + 1 mL diethyl ether, mix vigorously for 30 s, centri-fuge at 6000 g for 2 min. Remove 750 |xL ether, evaporate to dryness under a stream ofnitrogen at room temperature for 10 min, reconstitute residue in 50 |JLL mobile phase,inject 20 |xL aliquot.

HPLCVARIABLESGuard column: 10 X 4.6 5 jmrn Asahi ODP-50GColumn: 150 X 4.6 5 jxm Asahipak octadecyl polymerMobile phase: MeC N: 50 mM pH 10.5 KH 2PO 4 37:63Flow rate: 1Inject ion volume: 20Detector: E, Shim adzu L-ECD-6A, glassy carbon e lectrode +0.7 2 V ve rsu s an Ag/AgCl ref-erence electrodeCHROMATOGRAMRetention time: 12.1Internal s tandard: oleandomycin (5.7)Limit of detect ion: 100 ng/mLOTHER SUBSTANCESNoninterfering: clenbuterol, diltiazem, dipyridamole, ketotifen, methacholine, orciprena-line, theophyllineKEYWORDSplasma; whole bloodREFERENCEKato, Y.; Yokoyama, T.; Shimokawa, M.; Kudo, K.; Kabe, J.; Mohri, K. Determination of erythromycinin human plasma and whole blood by high-performance liquid chromatography. J.Liq.Chromatogr.,1993, 16, 661-680SAMPLEMatrix: bloodSample preparat ion: 1 mL P las m a + 20 |xL of 100 |xg/mL oleandomycin ph osph ate inethanol + 60 |xL saturated potassium carbonate (final pH 10), mix briefly, add 5 mL t-butyl methyl ether, shake in a reciprocating shaker for 15 min, centrifuge at 800 g for 5min. Remove 4 mL of the upper ether layer and evaporate it to dryness under a streamof nitrogen at room temperature. Wash down tube with 200 JJLL t-butyl methyl ether.Evaporate to dryness again, take up residue in 125 JJLL mobile phase, centrifuge 30 s,

inject aliquot.

ErythromycinMolecular formula: C37H67NO13Molecular weight: 733.9CAS R egistry No.: 114-07-8, 41342-53-4 (ethylsuccinate),96128-89-1 (acistrate), 3521-62-8 (estolate),304-63-2 (gluheptonate), 23067-13-2 (gluheptonate),3847-29-8 (lactobionate), 134-36-1 (propionate),643-22-1 (stearate), 84252-03-9 (stinoprate)


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HPLCVARIABLESGuard column: Alltech Direct Connect packed with ^Bondapak C18Column: 250 X 4.6 5 ixm Ultrasphere C18Mobile phase: MeCN: MeOH: buffer 42:10:48, final pH adjusted to 6.30-6.35 (Buffer was100 mM sodium acetate adjusted to pH 5.0 with 100 mM acetic acid.)Flow rate: 1.20Injection volume: 20Detector: E5 ESA model 5100A, guard cell +0.95 V, detector 1 +0.65 V, detector 2 +0.85VCHROMATOGRAMRetention time: 6Internal standard: oleandomycin phosphate (4)Limit of detection: 250 ng/mLOTHER SUBSTANCESExtracted: anhydroerythromyein, 2'-aeetylerythromycinKEYWORDSplasma; mobile phase recirculatedREFERENCELaakso, S.; Scheinin, M.; Anttila, M. Determination of erythromycin base and 2'-acetylerythromycin inhuman plasma using high-performance liquid chromatography with electrochemical detection.J.Chromatogr., 1990, 526, 475-486

SAMPLEMatrix: bloodSample preparation: 1 mL Plasma + 400 [xL 0.1 M NaOH to adjust pH to 10, mix 30 s,add 5 mL methyl t-butyl ether, vortex 1 min, centrifuge at 3000 rpm for 5 min, evaporateorganic layer to dryness under a stream of nitrogen at 40, dissolve residue in 150 |JLLmobile phase, vortex 2 min, inject 10 |xL aliquot.HPLCVARIABLESColumn: 150 X 4 10 fxm Techopak T-15 C18Mobile phase: MeCN:MeOH:THF:buffer 86:3:3:8 (Buffer was 75 mM sodium acetateadjusted to pH 4.1 with glacial acetic acid.)Flow rate: 1.5Injection volume: 10Detector: E, ESA Model 5100A with ESA model 5020 guard cell, analytical cell + 0.70 V(I) +0.85 V (II), guard cell +0.90 V, 0.5 |xm carbon filtersCHROMATOGRAMRetention time: 10Limit of quantitation: 50 ng/mLKEYWORDSplasmaREFERENCEKokkonen, P.; Haataja, H.; Valttila, S. Determination of 2'-acetylerythromycin and erythromycin inplasma by HPLC using manual and robotic sample preparation. Chromatographia, 1987,24, 680-

682


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SAMPLEMatrix: blood, gastric juiceSample preparat ion: Centrifuge plasma or gastric juice at 1200 g for 5 min. 500 |xLPlasma or gastric juice + 500 |JLL pH 11 phosphate buffer (ionic strength 1=1.0) + 50 xL100 |xMoleandomycin in MeCN + 5 mL hex ane : 2-butanol 80 :20, sh ake for 15 min, cen-trifuge at 1200 g for 5 min, freeze in dry ice/acetone. Remove the organic layer and evap-orate it to dryness under a stream of nitrogen, reco nstitute the residue in 300 \xhmobilephase, vortex three times for 1 min, inject a 40 fxL aliquot.HPLC VARIABLESGuard co lumn: 15 X 3.2 7 jxm Brownlee CNColumn: 100 X 4.6 Hypersil C18 BDS base-deactivatedMobile phase: MeCN:2.1 mM N aH 2PO 4:27.1 mM Na2HPO 4 40:30:30Column temperature: 65Flow ra te: 1.2Inject ion vo lum e: 40Detector: E, ESACoulochem Model 5100A, Model 5020 guard cell before injector +1.0 V,model 5011dual electrode analytical cell, screen electrode (detector 1) +0.65 V, sampleelectrode (detector 2) +0.85 V, ESAcarbon filters before guard and analytical cellsCHROMATOGRAMRetent ion t ime: 10.5Internal s tandard: oleandomycin (5)Limit of quant i ta t ion: 20 nM (plasma); 100 nM (gastric juice)KEYWORDSplasma; rugged; pharmacokineticsREFERENCEToreson, H.; Eriksson, B.M.Determination of erythromycin in gastric juice and blood plasma by liquidchromatography and electrochemical detection. J.Chromatogr.B, 1995, 673, 81-89

SAMPLEMatrix: blood, saliva, urineSample preparat ion: Plasma. 2 mL Plasma + 20 |xL 750 |Jig/mL roxithromycin in MeCN+ 5 mL diethyl ether, shake vigorously for 3 min, centrifuge at 900 g at 4 for 5 min.Remove upper layer and evaporate it to dryness under a stream of nitrogen at 45. Re-constitute residue with 100 JJLL MeCN, vortex 5 s, inject 40 |xL aliquot. Urine. 1.5 mLUrine + 100 |xL 750 |xg/mL roxithromycin in saturated K 2HPO 4 + 4 mL diethyl ether,shake vigorously for 3 min, centrifuge at 900 g at 4 for 5 min. Remove uppe r layer andevaporate it to dryness und er a stream of nitrogen at 45. Reconstitute residue with 100jxL MeCN, vortex 5 s, inject 40 |xL aliquot. Saliva. 1.5 mL Saliva + 100 jxL 750 jxg/mLroxithromycin in saturated K 2HPO 4 + 4 mL diethyl ether, shake vigorously for 3 min,centrifuge at 900 g at 4 for 15 min. Remove upper layer and evaporate it to drynessunder a stream of nitrogen at 45. Reconstitute residue with 100 jxL MeCN, vortex 5 s,inject 40 JJLL aliquot.HPLC VARIABLESColumn: Nova-Pak C18Mobile phase: MeCN:MeOH:56 mM sodium acetate buffer 50:4:56, final pH adjusted to7.0 with glacial acetic acidFlow ra te: 1.1Inject ion vo lume: 40Detector: E, Waters M460, +0.9 V versus Ag/AgCl reference electrode


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CHROMATOGRAMRetention time: 6.0 (erythromycin base), 7.1 (erythromycin B), 34.5 (erythromycin esto-late), 35.5 (erythromycin ethylsuccinate)Internal standard: roxithromycin (14.7)Lim it of det ec tio n: 12.5 ng/mLOTHER SUBSTANCESSimultaneous: 4"-acetylerythromycin, 6-O-methylerythromycinKEYWORDSplasmaREFERENCECroteau, D.; Vallee, F.; Bergeron, M.G.; LeBeI, M. High-performance liquid chromatographic assay oferythromycin and its esters using electrochemical detection. J.Chromatogr., 1987, 419, 205-212SAMPLEMatrix: blood, urineSa m ple p repa ra ti o n : Urine. Centrifuge urine a t 2500 g for 5 min, inject a 100 uX aliquot.Whole blood. 200 |xL Whole blood + 100 |xL 10% EDTA H- 50 |xL 20 |xg/mL josamycin inMeOH: water 50:50, centrifuge to separate plasma. 200 JXL Plasma + 20 |xL saturatedpotassium carbonate + 1 mL MTBE, mix, centrifuge. Remove 800 |xL of the MTBE layerand evaporate it to dryness, reconstitute the residue in 200 |xL MeOH-.water 50:50, injecta 100 |xL aliquot.HPLCVARIABLESColu mn: 250 X 4.6 8 |xm PLRP-S 1000 A (Polymer Labs)Mobile phase : MeCN: t-butanol: 200 mM pH 9.0 phosphate buffer-.water 3:19:5 :73Column temperature: 70Flow rate: 1.5Injection volume: 100Detector: F ex 365 em 450 following post-column extraction. The column effluent mixedwith reagent pumped at 0.7 mL/min and this mixture flow ed through a 1.5 m X 0.5 mmID stainless steel coil. The effluent from the coil mixed with chloroform pumped at 1.5mL/min and this mixture flowed through a 1.5 m X 0.5 mm ID stainless steel coil to asandwich-type phase separator with a 40 (xL groove volume (Vrije Universiteit, Amster-dam). Part of the organic layer was separated and flow ed through the detector a t 0.5mL/min. (Reagent was 5 jxM sodium 9,10-dimethoxyanthracene-2-sulfonate in 100 mMcitric acid.)C H R O M A T O G R A MRetention t ime: 11Internal standard: josamycin (28)Limit of dete ction : 12.5 ng/mL (plasma); 50 ng/mL (urine)OTHER SUBSTANCESExtracted: metabolitesSimultaneous: midecamycin, troleandomycinKEYWORDSplasma; whole blood; post-column extractionREFERENCEKhan, K.; Paesen, J.; Roets, E.; Hoogmartens, J. Analysis of erythromycin A and its metabolites inbiological samples by liquid chromatography with post-column ion-pair extraction. J.Liq.

Chromatogr., 1994, 17, 4195-4213


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SAMPLEMatrix: blood, urineSample preparat ion: Dilute urin e 1:2 with isotonic NaC l. 200 JJLL Plasm a or diluted ur ine+ 100 |JLL water + 600 |JLL pH 9 phosp hate buffer + 3 mL dichloromethane, sh ake for 10min, centrifuge at 2000 g for 5 min. Remove 2.5 mL of the organic layer and evaporateit to dryness und er a stre am of nitrogen, reco nstitute t he resid ue in 50 |xL MeOH , vortexfor 10 s, inject a 15 \xL aliquot.HPLCVARIABLESColumn: 300 X 3.9 10 ^m ixBondapak C18Mobile phase: M eCN :MeO H: 83 mM amm onium acetate 55:2 2:2 3, pH adjusted to 7.5 withacetic acidFlow rate: 1Inject ion volume: 15Detector: E, ESA Coulochem Model 5100A, Model 5020 guard cell 1.0 V (before injector),Model 5010 dual-electrode cell, screen electrode E l + 0.7 V, sam ple electrode E2 +0 .9 V,

0.5 |xm ESA carbon filters placed before guard and analytical cellsCHROMATOGRAMR ete ntio n t ime: 7.0Internal s tandard: erythromycinOTHER SUBSTANCESExtracted: roxithromycinSimul taneous: amitriptyline, clomipramine, disopyramide, erythromycin estolate, eryth-romycin ethylsuccinate, erythromycin stearate, imipramine, josamycin, lidocaine,

spiramycinKEYWORDSplasma; erythromycin is ISREFERENCEDemotes-Mainaird, RM.; Vingon, GA.; Jarry, C.H.; Albin, H.C. Micro-method for the determination ofroxithromycin in human plasma and urine by high-performance liquid chromatography using elec-trochemical detection. J.Chromatogr., 1989, 490, 115-123SAMPLEMatrix: blood, urineSample preparat ion: Prew ash a 1 mL C18 Bondelut C18 SPE cartridge with 3 mL MeCNand 3 mL water. 1 mL Serum or urine + 0.25 mL (0.50 mL for urine) 6-12 |Jig/mL ole-andomycin pho sph ate in w ater, add 1 mL MeCN , vortex 1 min, centrifuge at 1600 g for5 min, add to 8 mL water, load onto the SPE cartridge, wash with 5 mL water, wash with5 mL MeCN.water 1:1, suck dry, elute with two 0.5 mL aliquots of MeCN:50 mM pH6.30 phosphate buffer. Dry under vacuum in a rotary vacuum centrifuge, reconstitute in20 |JLL water, vortex 1 min, add 25 |JLL MeC N, vortex 1 m in, centrifuge at 1600 g for 1min, inject 3-5 JJIL aliquot of upper layer.HPLC VARIABLESGuard column: Waters Guard-Pak with Anatech 40-60 |xm glass beadsColumn: 150 X 3.9 Novapak C18Mobile phase: M eCN : 50 mM pH 6.30 pho sph ate buffer 30:70Column tem peratu re: 35Flow rate: 1Injection volume: 3-5Detector: E, Metrohm 656 with a glassy carbon electrode, 1.15 V versus Ag/AgCl referenceelectrode; also UV at 200 nm with LOD 250-1000 ng/mL (J.Pharm.Sci. 1985, 74, 1126-

1128)


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CHROMATOGRAMRetention time: 6.3Internal standard: oleandomycin phosphate (4.4)Limit of detection: 100 ng/mLOTHER SUBSTANCESSimultaneous: anhydroerythromycinKEYWORDSserum; SPE; stability-indicatingREFERENCEStubb s, C ; H aigh, J.M .; Kanfer, I. A stability-indicating liquid chroma tographic me thod for the analy sisof erythromycin in stored biological fluids using amperometric detection. J.Liq.Chromatogr., 1987,10, 2547-2557SAMPLEMatrix: formulationsHPLCVARIABLESColumn: 250 X 4.6 5 jim Bakerbond C18Mobile phase: MeCN: MeOH: 200 mM ammonium acetate: water 45 :10:10:25 , pH 6.25FIo1W rate: 1Detector: UV 215CHROMATOGRAMRetention time: 7.46 (erythromycin lactobionate)KEYWORDSinjections; saline; water; stability-indicatingREFERENCEStiles, M.L.; Allen, L.V., Jr .; P rince , S.J. Stability of various antibiotics kept in an insulated pouch during

administration via portable infusion pump. Am.J.Health-Syst.Pharm., 1995, 52, 7074SAMPLEMatrix: formulationsSample preparation: Weigh out material corresponding to ca. 250 mg erythromycin ethyl-succinate, add 10 mL acetone, sonicate 5 min, centrifuge at 2500 g for 5 min, dilute a 6mL aliquot of supernatant to 10 mL with 200 mM pH 6.5 tetrabutylammonium hydrogensulfate:200 mM pH 6.5 phosphate buffer:water 12.5:7.5:80.

HPLC VARIABLESColumn: 250 X 4.6 10 |xm RSiI LL C18 (RSL-Bio-Rad)Mobile phase: MeCN: 200 mM pH 6.5 tetrabutylamm onium hydrogen sulfate (adjust pHwith NaOH):200 mM pH 6.5 phosphate buffer:water 42.5:5:5:47 .5Column temperature: 35Flow rate: 1.5Injection volume: 20Detector: UV 215CHROMATOGRAMRetention time: 24 (erythromycin A ethylsuccinate), 8 (erythromycin A)


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KEYWORDSpowders; tabletsREFERENCECachet, T.; Lannoo, P.; Paesen, J.; Janssen, G.; Hoogmartens, J. Determination of erythromycin ethylsuccinate by liquid chromatography. J.Chromatogr, 1992, 600, 9 9 - 1 0 8SAMPLEMatrix: solutionsHPLCVARIABLESColumn: 250 X 4.6 Zorbax C8Mobile phase: MeC N: 200 mM pH 6.5 tetrame thylam m onium pho sph ate: 200 mM pH 6.5ammonium phosp hate: water 35:2 0:5:4 0Column temperature: 35F lo w rate : 1.5Inject ion volum e: 20Detector: UV 215CHROMATOGRAMRetent ion t ime: 12KEYWORDSbetter results with aged columnsREFERENCECachet, T.; Quintens, I.; Roets, E.; Hoogmartens, J. Improved separation of erythromycin on aged re-versed-phase columns. J.Liq.Chromatogr., 1989, 12, 2171-2201 ANNOTATED BIBLIOGRAPHYZierfels, G.; Petz, M. [Fluorimetric determination of erythromycin residues in foods of animal originafter derivatization with FMOC and HPLC separation]. Z.Lebensm.Unters.Forsch., 1994, 198, 3 0 7 -312J ane cek, M.; Quilliam, M.A.; Bailey, M.R.; N orth, D.H. De term ination of erythromycin A by liquid chro-matography and electrochemical detection, with application to salmon tissue. J.Chromatogr, 1993,619, 63-69 [electrochemical detection; fish; tissue; SPE; column temp 40; LOD 100 ng/g]Paese n, J .; Calam , D.H.; Miller, J.H.McB .; Raiola, G.; Rozanski, A.; Silver, B.; Hoo gm artens, J . Collab-orative study of the analysis of erythromycin by liquid chromatography on wide-pore poly(styrene-divinylbenzene). J.Liq.Chromatogr, 1993,16, 152 9-15 44 [column temp 70; simultaneous impurities]Cachet, T.; Quintens, L; Paesen, J.; Roets, E.; Hoogmartens, J. Improved separation of erythromycin onaged reversed-phase columns. II. J.Liq.Chromatogr, 1991, 14, 1203-1218Paesen, J. ; Roets, E.; Hoogm artens, J . L iquid chrom atography of erythromycin A and related substanceson poly(styrene-divinylbenzene). Chromatographia, 1991, 32, 162-166Stubbs, C ; Kanfer, I. A stability-indicating high-performance liquid chrom atographic a ssay of erythro-mycin estolate in pharmaceutical dosage forms. Int.J.Pharm., 1990, 63, 113-119Araman, A.; Temiz, D.; Guven, K.C. Stability studies of erythromycin in simulated gastric mediumstudied by high-performance liquid chromatography. Ada Pharm.Turc, 1988, 30, 3 7 - 4 2Croteau, D.; Bergeron, M .G.; LeBeI, M. Pharm acok inetic adv antag es of erythromycin estolate over ethyl-succinate as determined by high-pressure liquid chromatography. Antimicrob.Agents Chemother,1988, 32, 5 6 1 - 5 6 5Grgurinovich, N.; Matthews, A. Analysis of erythromycin and roxithromycin in plasma or serum byhigh-performance liquid chromatography using electrochemical detection. J.Chromatogr, 1988, 433,298-304


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Haataja, H.; Kokkonen, P. Determination of 2'-acetylerythromycin and erythromycin in human tonsiltissue by HPLC with coulometric detection. J.Antimicrob.Chemother., 1988, 21, 6 7 - 7 2Stubbs, C; Kanfer, I. High-performance liquid chromatography of erythromycin propionyl ester anderythromycin base in biological fluids. J.Chromatogr., 1988, 427, 93-101 [extracted erythromycin,erythromycin propionate; oleandomycin (IS); electrochemical detection; column temp 35; serum;urine; SPE; pharmacokinetics; LOQ 250 ng/mL]Cachet, T.; Kibwage, LO.; Roets, E.; Hoo gm artens, J.; V anderhae ghe, H. O ptimization of the sepa rationof erythromycin and related substances by high-performance liquid chromatography. J.Chromatogr.,1987, 409, 91-100 [column temp 35; simultaneous impurities, erythromycin A, erythromycin B,erythromycin C]Geria, T.; Hong, W.H.; Daly, R.E. Improved high-performance liquid chrom atographic assay of erythro-mycin in pharmaceutical solid dosage forms. J.Chromatogr., 1987, 396, 191-198Nilsson, L.G.; Walldorf, B.; Paulsen, O. Determination of erythromycin in hu m an plasma , using columnliquid chromatography with a polymeric packing material, alkaline mobile phase and amperometricdetection. J.Chromatogr., 1987, 423, 189-197Kibwage, 1.0.; Roets, E.; Hoogmartens, J.; Vanderhaeghe, H. Separation of erythromycin and related

substan ces by high-performance liquid chrom atography on poly(styrene-divinylbenzene) packin g ma-terials. J.Chromatogr., 1985, 330, 275-286Stubbs, C; Haigh, J.M.; Kanfer, I. Determination of erythromycin in serum and urine by high-perfor-mance liquid chromatography with ultraviolet detection. J.Pharm.ScL, 1985, 74, 1126-1128Du thu, G.S. Assay of erythromyc in from h um an serum by high performance liquid chrom atograph y withelectrochemical detection. J.Liq.Chromatogr., 1984, 7, 1023-1032 [serum; electrochemical detection;also josamycin, oleandomycin, tylosin]Chen, M.L.; Chiou, W.L. Analysis of erythromycin in biological fluids by high-performance liquid chro-matography with electrochemical detection. J.Chromatogr, 1983, 278, 9 1 - 1 0 0Tsuji, K.; Kane, M.P. Improved high-pressure liquid chromatographic method for the analysis of eryth-romycin in solid dosage forms. J.Pharm.ScL, 1982, 71, 1160-1164

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