REFERENCE

Kruger, RB.; Albrecht, C.RDe V.; Jaarsveld, RR Use of guanidine hydrochloride and ammonium sulfate
in comprehensive in-line sorption enrichment of xenobiotics in biological fluids by high-performance
liquid chromatography. J.Chromatogr., 1993, 612, 191-198

SAMPLE
Matrix: blood, CSF, peritoneal fluid, synovial fluid, tissue, urine
Sample preparation: Condition a C18 SPE cartridge (J.T. Baker) with 1 mL MeOH, 1 mL
water, and 1 mL 100 mM pH 4.5 acetate buffer. CSF, peritoneal fluid, serum, synovial
fluid, tissue. Homogenize (TenBroeck tissue grinder) endometrial tissue with saline, cen-
trifuge at 510 g, remove supernatant. Centrifuge fluids at 510 g. 1 mL Sample + 1 mL
100 mM acetate buffer, mix, add to the SPE cartridge, wash with 1 mL water, elute with
1 mL MeCN:MeOH: 100 mM acetate buffer 12.5:12.5:75, add 12.5 ng ormetoprim, inject
an aliquot. Urine. 1 mL Urine + 1 mL MeCN, centrifuge at 510 g for 10 min. Remove 25
IxL of the supernatant and add it to 1 mL 100 mM acetate buffer, add to the SPE cartridge,
wash with 1 mL water, elute with 1 mL MeCN: MeOH: 100 mM acetate buffer 12.5:
12.5:75, add 12.5 ng ormetoprim, inject an aliquot.

HPLCVARIABLES
Column: C18 (Rainin)
Mobile phase: MeCN: MeOH: 50 mM phosphate buffer 12.5:12.5:75, pH 3.0
Flow rate: 1.5
Detector: UV 280

CHROMATOGRAM
Retention time: 6
Internal standard: ormetoprim (8)
Limit of detection: 15 ng/mL

OTHER SUBSTANCES
Extracted: sulfamethoxazole

KEYWORDS
serum; endometrium; SPE; horse; pharmacokinetics

REFERENCE
Brown, M.P.; Gronwall, R.; Castro, L. Pharmacokinetics and body fluid and endometrial concentrations
of trimethoprim-sulfamethoxazole in mares. Am.J.Vet.Res., 1988, 49, 918-922

SAMPLE
Matrix: blood, CSF, urine
Sample preparation: 50 |xL Serum, urine, or CSF + 50 |xL 100 |xg/mL antipyrine in
MeOH, vortex for 15 s, centrifuge at 10000 g for 2 min, inject a 20 |xL aliquot of the
supernatant.

HPLCVARIABLES
Guard column: 37-50 |xm fxBondapak C18
Column: 300 X 3.9 10 |xm jxBondapak C18
Mobile phase: MeOH: buffer 35:65 (Buffer was 97% 67 mM KH2PO4 + 3% 67 mM
Na2HPO4 and the pH was adjusted to 3.5 with phosphoric acid.)
Flow rate: 1
Injection volume: 20
Detector: UV 225

CHROMATOGRAM
Retention time: 4.6