High-pressure liquid chromatographic method for analysis of cephalosporins

Signs, Steven A.; File, Thomas M.; Tan, James S.

doi:10.1128/aac.26.5.652

Cited by 65 publications

(29 citation statements)

References 14 publications

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“…Two methods introduced for cefaclor were based on protein precipitation. In the first method, the supernatant was directly quantified by RP-HPLC after precipitation with cold methanol-0.1 M sodium acetate (65). Recovery was excellent, and the detection limit was 0.2 Fkg/ml.…”

Section: Methodsmentioning

confidence: 99%

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Recent analytical methods for cephalosporins in biological fluids

Toothaker¹,

Wright²,

Pachla³

1987

Antimicrob Agents Chemother

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Since 1980, RP chromatography has been the principal analytical technique used for cephalosporins. This technology offers selectivity, accuracy, and ease of use. Most of the methods rely on protein precipitation and, to a lesser extent, solid-phase isolation or extraction procedures. The proper selection of a method depends on the analytical constraints imposed by the overall objective of the study. For example, pharmacokinetic datum interpretation mandates that the method be validated and provide specific and accurate results. LC is the preferred technique, since it not only meets these specifications but may also distinguish between the drug and metabolites. Those chromatographic methods which quantify several different cephalosporins are not desirable for pharmacokinetic datum interpretation, since accuracy and precision are usually compromised in order that many different drugs may be quantified in a single analysis. The proper selection of sample preparation method is dependent on the presence of potential interferences and the acceptable lower limit of quantitation. Protein precipitation methods offer ease of sample preparation but may suffer from nonselectivity. Solid-phase isolation and extraction procedures may increase selectivity and improve the limit of quantitation. Although LC provides specific and accurate results, clinical laboratories may prefer to use the less specific methods for therapeutic drug monitoring. In this case, microbiological, enzymatic, and fluorimetric methods offer improved sample throughput but less specificity. However, these methods should not be used for drugs that may have a low margin of safety or if the patient is on multiple-antibiotic therapy. Future methods may involve incorporating solid-phase isolation columns to enhance the specificity of chromatographic, microbiological, enzymatic, and fluorescence methods. Advancements in microbore column technology may allow improvements in the selectivity and sensitivity of LC methods. Many investigators prefer to use simple protein precipitation procedures for sample preparation because of sample throughput constraints. However, advances in robotic sample preparation may allow the more cumbersome solid-phase isolation or extraction techniques to be used to improved sample throughput and specificity.

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Section: Methodsmentioning

confidence: 99%

“…Two RP-HPLC methods were reported for cephaloridine in plasma or urine during this review period (36,65). One of the methods was applied to rat urine samples contaminated with feces and diet pellets (36).…”

Section: Methodsmentioning

confidence: 99%

Recent analytical methods for cephalosporins in biological fluids

Toothaker¹,

Wright²,

Pachla³

1987

Antimicrob Agents Chemother

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“…When the gasket is placed with the Millipore filter in contact with body tissues, the filter allows the free diffusion of interstitial fluid into the chamber but is impermeable to bacteria. The 6.64 x 106 CFU/ml (n = 30) as determined by the plating of serial dilutions in brain heart infusion agar. The concentration of E. coli averaged 1.34 x 107 CFU/ml (n = 12).…”

Section: Methodsmentioning

confidence: 99%

Canine model for the simultaneous measurement of antibiotic levels in tissues and bacterial killing rate

Wagner¹,

Salstrom²,

File³

et al. 1986

Antimicrob Agents Chemother

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Antibiotic levels in serum are.commonly used to guide ahtibiotic therapy. The (7) and Raebtirn (5) studied interstitial fluid antibiotic levels in the abraded forearms of human volunteers. Animal models in which subcutaneous tissue cages were used to measure the antibiotic levels in interstitial fluid have been described (2, 4). There is no model, however, which allows measurement of the in vivo killing of bacteria along with a determination of antibiotic levels in tissues.We developed a canine model that allows measuremnent of the antibiotic levels in interstitial fluid and the bacterial killing rate by using a modified Sykes-Moore chamber. This model can be used to study the penetration of antibiotics into body tissues and to compare the in vivo efficacy of antibiotics.MATERIALS AND METHODS Adult mongrel dogs of either sex ranging in weight from 14 to 34 kg were used. They were placed under general anesthesia by induction with sodium pentothal, orotracheally intubated, and maintained with intravenous sodium pentothal, pentobarbital, and lactated Ringer solution with 5% glucose. The dogs were routinely maintained under general anesthesia for 9 h without difficulty.Each animal was placed in the supine position, and its abdomen was shaved and prepped with povidone-iodine solution. Oblique abdominal skin incisions were made bilaterally. The external oblique muscle belly was freed by blunt and sharp dissection from the surrounding tissues with the origin and insertion of the muscle left intact. Bleeding from the skin and subcutaneous tissue was meticulously con-* Corresponding author. trolled with electrocautery. This dissection produced four potential spaces in the tissue planes: between the skin and anterior surface of the external oblique muscle and between the internal oblique muscle and the posterior surface of the external oblique muscle, bilaterally. These pockets were used to place the modified Sykes-Moore chamber in intimate contact with the muscle belly.Our Sykes-Moore chambers consist of two stainless steel rings with a glass cover slip, a self-sealing rubber gasket, a membrane filter (pore size, 0.22 ,um; Millipore Corp., Bedford, Mass.), a surface area of 3.14 cm2, and a volume of 0.8 ml (Fig. 1). The rubber gasket allows sampling of the chamber fluid. When the gasket is placed with the Millipore filter in contact with body tissues, the filter allows the free diffusion of interstitial fluid into the chamber but is impermeable to bacteria. The chambers were assembled and then steam autoclaved at 121°C under 15 lb (ca. 6.8 kg) of pressure for 15 min.The chambers were inoculated with clinical isolates of either Staphylococcus aureus or Escherichia coli immediately before being placed into the dog. The bacteria had been grown for 18 h in Trypticase soy broth (BBL Microbiology Systems, Cockeysville, Md.). A 1:100 dilution of bacteria in phosphate-buffered saline (PBS) at a pH of 7.4 was prepared, and 0.8 ml of this suspension was injected into the chamber through the rubber gasket. The concentration of S...

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“…Cephalosporin antibacterial used in the treatment of infections due to susceptible Gram-positive and Gram-negative bacteria, including infections of the abdomen, bones and joints, CNS, skin and skin structure, genito-urinary tract (including gonorrhoea) and respiratory tract, in gynaecological infections, and in early Lyme disease [2,3]. Literature survey revealed that several HPLC [4][5][6][7][8][9][10][11], HPTLC [12] and spectrophotometric methods [13][14][15][16][17][18] have been used for determination of Cefotaxime Sodium. Paracetamol is chemically 4-hydroxy acetanilide.…”

Section: Introductionmentioning

confidence: 99%