<sup>125</sup>
            I Radioimmunoassay of Netilmicin

Stevens, Paul; Young, Lowell S.; Hewitt, William L.

doi:10.1128/aac.11.4.768

Cited by 11 publications

(3 citation statements)

References 8 publications

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“…DISCUSSION Netilmicin in plasma or body fluids can be analyzed by microbiological assay (7). A radioimmunoassay for netilmicin has also been reported (10). Although the forner assay is simple and inexpensive and the latter is quite sensitive, they have some disadvantages in common with assays using biological agents or substances derived from biological sources (5a).…”

Section: Resultsmentioning

confidence: 99%

High-Pressure Liquid Chromatographic Assay of Netilmicin in Plasma

Peng¹,

Jackson²,

Chiou³

1977

Antimicrob Agents Chemother

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A high-pressure liquid chromatographic method for the quantitative determination of netilmicin in plasma was developed. The procedures involve acetonitrile protein precipitation, methylene chloride extraction, and dansylation to form the fluorescent dansyl derivative of netilmicin, which is extracted into ethyl acetate and chromatographed on a reverse-phase column with aqueous acetonitrile as the mobile phase. A good linear relationship between peak height measurements and netilmicin concentrations was found. This method is sensitive and reproducible; a netilmicin concentration as low as 0.5 Ag/ml can be measured with only 0.1 ml of plasma sample. The results of assays ofplasma or serum samples by this high-pressure liquid chromatographic method correlate well with those obtained by microbiological assays.Netilmicin is a new semisynthetic derivative of sisomicin. Its antimicrobial spectrum and activity are similar to those of other structurally related aminoglycosidic antibiotics such as gentamicin, sisomicin, and tobramycin (5,6,9). Although compared with gentamicin, netilmicin demonstrated lower nephrotoxicity in animals (3, 4) and less variation in peak levels after intramuscular administration in humans (7), it has potential for plasma level variations and oto-and nephrotoxicity. We wish to report a high-pressure liquid chromatographic method for determining plasma or body fluid levels of netilmicin. Such an assay could be useful in dosage regimen adjustments, in maintaining therapeutic levels in plasma, in averting untoward reactions, and in obtaining useful pharmacokinetic data. Similar benefits have been derived from the assays of gentamicin in plasma (2, 8). MATERIALS AND METHODSNetilmicin was obtained from Schering Corp., Bloomfield, N.J. 5-Dimethylamino-1-naphthalene sulfonyl chloride (dansyl chloride) was purchased from Sigma Chemical Co., St. Louis, Mo. Glassdistilled acetonitrile, methylene chloride, and ethyl acetate were from Burdick and Jackson Laboratory, Muskegon, Mich. Sodium carbonate, sodium bicarbonate, and phosphoric acid were reagent grade from Fisher Scientific Co., Fair Lawn, N.J.Sample preparation. Portions (0.1 ml) of plasma samples were diluted with 0.9 ml of phosphate buffer (0.1 M, alkalinized with sodium hydroxide to pH 11), placed in culture tubes (13 by 100 mm) containing 2.5 ml of acetonitrile, blended in a Vortex mixer, and centrifuged. The supernatant solutions were poured into new tubes, blended in a Vortex mixer with 2 ml of methylene chloride, and centrifuged at 2,000 rpm for about 1 min. Portions (0.5 ml) of the separated aqueous (upper) layer were transferred to new, screw-cap culture tubes containing 4 mg of dansyl chloride in 0.3 ml of acetonitrile.These tubes were closed and incubated in a water bath at 75°C for 5 min. The reaction mixtures were cooled in ice-water, and 0.5 ml of ethyl acetate and 6 ml of carbonate buffer (containing 0.5 M sodium carbonate and bicarbonate, pH 9.5) were added. After Vortex mixing and centrifugation, 1-to 5-,l portions of the organic (u...

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Section: Resultsmentioning

confidence: 99%

High-Pressure Liquid Chromatographic Assay of Netilmicin in Plasma

Peng¹,

Jackson²,

Chiou³

1977

Antimicrob Agents Chemother

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“…Early detection of prostate cancer through clinical and novel bioanalytical platforms will impact survival rates, 9 hence the need to find improved diagnostic tools with high selectivity, sensitivity and stability. To date, several antibody-based methods such as radioimmunoassays, 10 enzyme-linked immunosorbent assays (ELISA), 11 surface plasmon resonance (SPR) immunoassays, 12 and fluorescence immunoassays 13 have been used to detect this biomarker. Limitations of these methods include high cost, technical complexity, limitations of laboratory testing, a lack of portable equipment and the role of interferences.…”

Section: Introductionmentioning

confidence: 99%

A NiFe PBA/AuNPs nanocomposite sensitive immunosensor for electrochemical detection of PSA

Hua,

Zhao,

Gui

et al. 2024

Anal. Methods

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“…Numerous assays have been developed for aminoglycosides (4,8,9,11,16,18). Radioenzymatic assays (REA) with gentamicin adenyl transferase is one technique used for the quantitation of gentamicin (17).…”

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confidence: 99%

Radioenzymatic assays for aminoglycosides with kanamycin 6'-acetyltransferase

Weber¹,

Smith²,

Opheim³

1985

J Clin Microbiol

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To facilitate the rapid and accurate quantitation of parenterally administered aminoglycosides, we defined the optimum conditions (pH, duration of incubation, and cofactor concentrations) to permit radioenzymatic assays with kanamycin acetyltransferase. The accuracy in quantitating tobramycin, netilmicin, kanamycin, and amikacin at concentrations in the therapeutic range was greater than 90%, with a mean recovery of 102.8%. The mean of the interassay coefficient of variation was 7.8%. Typical standard curves at six different concentrations resulted in a correlation coefficient (r value) of greater than 0.99 for each aminoglycoside. The radioenzymatic assay correlates well with the bioassay (tobramycin and netilmicin) and radioimmunoassay (amikacin and kanamycin); the correlation coefficient is greater than 0.90 for all. We conclude that the radioenzymatic assay utilizing kanamycin 6'-acetyltransferase is feasible for all commercially available parenterally administered aminoglycosides.

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¹²⁵ I Radioimmunoassay of Netilmicin

Abstract: We report a radioimmunoassay for the new semisynthetic aminoglycoside netilmicin with a sensitivity of 480 pg per tube and a correlation coefficient of 0.94 between this radioimmunoassay and a microbioassay for measurement of netilmicin in serum.

Cited by 11 publications

References 8 publications

High-Pressure Liquid Chromatographic Assay of Netilmicin in Plasma

High-Pressure Liquid Chromatographic Assay of Netilmicin in Plasma

A NiFe PBA/AuNPs nanocomposite sensitive immunosensor for electrochemical detection of PSA

Radioenzymatic assays for aminoglycosides with kanamycin 6'-acetyltransferase

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125 I Radioimmunoassay of Netilmicin

Abstract: We report a radioimmunoassay for the new semisynthetic aminoglycoside netilmicin with a sensitivity of 480 pg per tube and a correlation coefficient of 0.94 between this radioimmunoassay and a microbioassay for measurement of netilmicin in serum.

Cited by 11 publications

References 8 publications

High-Pressure Liquid Chromatographic Assay of Netilmicin in Plasma

High-Pressure Liquid Chromatographic Assay of Netilmicin in Plasma

A NiFe PBA/AuNPs nanocomposite sensitive immunosensor for electrochemical detection of PSA

Radioenzymatic assays for aminoglycosides with kanamycin 6'-acetyltransferase

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¹²⁵ I Radioimmunoassay of Netilmicin