Furosemide was found to be a potent diuretic and saluretic drug comparable to

other substances. But animal experiments and studies in volunteers showed an

effect at doses at which other diuretic drugs were no longer effective. Thus the
name “high-ceiling diuretic” was coined.
The discovery of sildenafil, a highly selective inhibitor of phosphodiesterase-5
(PDE-5), was the result of research on chemical agents that might be useful in the
treatment of coronary heart disease. Initial clinical studies on sildenafil were not
promising with respect to its anti-anginal potential. However, the incidental
discovery of its anti-impotence effect led to its approval for the treatment of
erectile dysfunction.
Drug Discovery and Evaluation, Pharmacological Assays, 3rd Ed - Vogel HG
(Ed) – 2008.

Diuretics are drugs used in certain pathological conditions to eliminate somatic

fluids by promoting renal excretion of water and salts (53). Chemically, they are
heterogenous compounds that present different pharmacological properties and,
accordingly, are classified into several different groups. The groups of loop
diuretics and thiazide diuretics are the most important in veterinary practice (54).
The former group includes three compounds (furosemide, ethacrynic acid, and
bumetadine), but only furosemide has been approved for use in cattle (55).
Included in the latter group are chlorothiazide, hydrochlorothiazide, and
trichlormethiazide.
Furosemide and thiazide diuretics (Fig. 8.4) have been approved for use in dairy
cattle for treatment of postparturient edema of the mammary gland and associated
structures (56). Furosemide and hydrochlorothiazide are administered
intramuscularly or intravenously at a dosage of 500 and 125–250 mg/animal,
respectively. Chlorothiazide and trichlormethiazide are administered orally at
dosage of 2000 and 200 mg/animal, respectively.
Unauthorized use of these diuretics, or the failure to follow label indications for
approved use in the cattle, could lead to unacceptable residues in meat and milk
destined for human consumption. While there are no official tolerances for these
drugs in milk, the Food and Drug Administration (FDA) has established safe
levels that range from 7 ppb for trichlormethiazide, to 10 ppb for furosemide, and
67 ppb for the other thiazides (56). Administration of diuretics is associated with
potential toxic effects such as bone marrow depression, hyperbilirubinemia,
altered carbohydrate metabolism, and elevated levels of urea, uric acid, and sugar.
Furosemide is a strongly acidic o-chlorosulfonamide compound that includes an
additional carboxyl group that differentiates it from the weakly acidic thiazides.
Pharmacokinetic studies showed that 30 min after oral administration of 20 mg
furosemide/kg bw in dogs, 22.73 ppb was the maximum plasma concentration
attained. The oral bioavailability of the compound was estimated at approximately
77%. Furosemide is extensively bound to plasma proteins (91%). In dogs, the
elimination half-life of furosemide was found to be 1.42 h after oral dosing, and
1.13 h after intravenous dosing. Excretion of furosemide was rapid and proceeded
primarily through kidney, mostly in form of the parent drug.
Following oral administration of radiolabeled furosemide, excretion was reported
to be almost complete within 3 days in rats (96–98%) and dogs (98–99%). Rat
urine contained 40–50% of the parent drug, 30% 4-chloro-5- sulfamoyl-
anthranilic acid, and four unidentified metabolites that accounted for the rest of
the administered radioactivity. In contrast, urine of dog and monkey contained
85% unmetabolized furosemide, 7% 4-chloro-5-sulfamoyl-anthranilic acid, and
the remainder was due to unidentified metabolites. Following intramuscular
injection of 5 mg furosemide/kg bw in cattle, the half-life for plasma elimination
was estimated at 4.3 h. In contrast, the half-life of furosemide in cattle was
reported to be less than 1 h following intravenous administration.
Residue depletion studies in lactating cows given an intramuscular injection of 5
mg furosemide/kg bw showed that residues could be detected in milk for at least
24 h after treatment. The half-life in milk was estimated to be 3 h. When cows
were administered three intramuscular injections of 1.5 mg furosemide/kg bw per
day, milk contained 660 ppb at 7 h after dosing.
Diuretics are therapeutic agents used in certain pathological conditions to
eliminate bodily fluids. Furosemide and the thiazide diuretics, chlorothiazide,
hydrochlorothiazide, and trichlormethiazide are approved for use in dairy cattle
for treatment of postparturient edema of the mammary gland and associated
structures. The potential misuse of these diuretic drugs in cattle could lead to
unacceptable residues in meat or milk destined for human consumption.
Therefore, analytical methods sufficiently sensitive to monitor residue
concentration levels in foods are valuable in preventing unapproved use of
diuretics.
In determining diuretic residues in foods, it is often necessary to know their
physicochemical characteristics. In general, the diuretics are soluble in methanol,
ethanol, and water with the exception of hydrochlorothiazide that is insoluble in
water (552, 553). Furosemide, which is a strongly acidic o-chlorosulfonamide
compound, is the least stable among these diuretics. Its degradation proceeds with
both a hydrolysis and a photochemical oxidation process. The major product
generated is 4-chloro-5-sulfamoylanthranilic acid, which is further converted into
4-chloro-5-sulfoanthranilic acid. Acid hydrolysis of the furosemide also gives 4-
chloro-5-sulfamoylanthranilic acid and furfuryl alcohol. Chlorothiazide,
hydrochlorothiazide, and trichlormethiazide are all characterized by two
ultraviolet absorbance maxima at 225 and 270 nm, whereas furosemide exhibits a
natural fluorescence with excitation and emission wavelengths at 272 and 410 nm,
respectively.
Extensive literature reviews (554, 555) have indicated that almost all reported
analytical methods for the analysis of diuretics employ liquid chromatography.
Most of these methods are limited, however, to assaying diuretics in urine and
plasma. With the exception of a liquid chromatographic method for the
determination of furosemide, another one for chlormethiazide, and a third method
for chlorothiazide and hydrochlorothiazide residues in bovine milk, no
chromatographic method has been reported in the literature for assaying diuretics
in meat (Table 29.17)
The furosemide extraction procedure was later examined for potential application
in the analysis of thiazide diuretics in milk. Since this procedure could not provide
sufficiently clean extracts for thiazides, additional acidic and basic extraction
procedures were evaluated (557). Thus, milk was deproteinized with
trichloroacetic acid, phosphoric acid, or potassium dihydrogen phosphate and
centrifuged. The supernatants were extracted with ethyl acetate, evaporated to
dryness, reconstituted in mobile phase, and analyzed by liquid chromatography.
The recoveries in most cases were low and widely variable. Basic extraction, on
the other hand, with sodium bicarbonate/potassium carbonate mixture or
potassium monohydrogen phosphate followed by extraction with ethyl acetate
also gave poor recoveries in most cases. It appears that a significant degradation
of chlorothiazide occurred under the basic conditions.
Drug Residues in Foods-Botsoglou-2000.pdf