Liquid-chromatographic determination of vitamin K1 in plasma, with fluorometric detection.

1986 ◽  
Vol 32 (10) ◽  
pp. 1925-1929 ◽  
Author(s):  
Y Haroon ◽  
D S Bacon ◽  
J A Sadowski
Keyword(s):  
Limit Of Detection ◽  
Lipid Fraction ◽  
Internal Standard ◽  
Reversed Phase ◽  
Phase System ◽  
Fluorometric Detection ◽  
Vitamin K1 ◽  
Crude Lipid ◽  
Zinc Metal ◽  
Liquid Chromatographic

Abstract This assay for phylloquinone (vitamin K1) in plasma requires a single liquid-chromatographic step. Much smaller volumes of plasma (0.5-1.0 mL) are required than in previous assays. Before liquid chromatography, we purified crude lipid extracts by conventional chromatography on silica, then extracted the lipid fraction by dissolving it in an acidic mixture of hexane/acetonitrile (1/4 by vol) containing 70 mmol of zinc chloride per liter. The vitamin K1 was selectively extracted into acetonitrile after being converted to vitamin K1 hydroquinone by addition of zinc metal. This procedure removes greater than 99% of contaminating lipids. We injected the lipid extract directly onto a reversed-phase column after re-converting the vitamin K1 hydroquinone to vitamin K1. Vitamin K1 was quantified by comparison with the internal standard (dihydro-vitamin K1) and detected fluorometrically after post-column "on-line" reduction to the hydroquinone with zinc metal. The lower limit of detection for vitamin K1 in the final reversed-phase system was about 0.05 microgram/L plasma; CVs for replicates were less than 10%. The mean concentration of vitamin K1 in plasma from 22 healthy fasting adults was 0.55 (range 0.09-2.12) micrograms/L.

scholarly journals Liquid Chromatographic Analysis of Vitamin K1 in Milk-Based Infant Formula with Matrix Solid-Phase Dispersion

1999 ◽  
Vol 82 (5) ◽  
pp. 1140-1145 ◽  
Author(s):  
G William Chase ◽  
Ronald R Eitenmiller ◽  
Austin R Long
Keyword(s):  
Infant Formula ◽  
Limit Of Detection ◽  
Solid Phase ◽  
Reversed Phase ◽  
Vitamin K1 ◽  
Matrix Solid Phase Dispersion ◽  
Phase Dispersion ◽  
Limit Of Quantitation ◽  
Liquid Chromatographic Analysis ◽  
Liquid Chromatographic

Abstract A liquid chromatographic method for vitamin K1 in milk-based infant formula is described. The vitamins are extracted from infant formula by matrix solid-phase dispersion and quantitated by reversed-phase chromatography with fluorescence detection. Vitamin K1 is converted to the fluorescent hydroquinone with a postcolumn zinc reductive reactor. The limit of detection is 12 pg, and the limit of quantitation is 38 pg on-column. Linear responses were obtained in the range 0.55-22.1 ng/mL (r2 = 0.9998). Recoveries of vitamin K1 from an analyte-fortified blank material for milk-based infant formula averaged 91.7% (n = 25). The method provides a rapid, specific, and easily controlled assay for vitamin K1 in fortified infant formula.

scholarly journals Determination of Vitamin K1 in Medical Foods by Liquid Chromatography with Postcolumn Reduction and Fluorometric Detection

2000 ◽  
Vol 83 (4) ◽  
pp. 957-962 ◽  
Author(s):  
George M Ware ◽  
G William Chase ◽  
Ronald R Eitenmiller ◽  
Austin R Long
Keyword(s):  
Limit Of Detection ◽  
Solid Phase ◽  
Sodium Bicarbonate Solution ◽  
Reversed Phase ◽  
Mean Value ◽  
Fluorometric Detection ◽  
Vitamin K1 ◽  
Limit Of Quantitation ◽  
Medical Foods

Abstract A liquid chromatographic (LC) method is described for the determination of vitamin K1 in medical foods. The sample is enzymatically digested with lipase and α-amylase and extracted with 1% sodium bicarbonate solution–isopropanol (1 + 1). After C18 solid-phase extraction, vitamin K1 is separated by nonaqueous reversed-phase LC, converted to the hydroquinone by postcolumn zinc reduction, and quantitated by fluorescence detection. The limit of detection is 8 pg (3 σ), and the limit of quantitation is 27 pg (10 σ) on column. Linear response ranged from 0.1 to 1.0 ng vitamin K1 (r = 0.9999). The mean recovery (n = 38) for all spiking levels was 101.6 ± 2.85%. Analysis of Standard Reference Material 1846, Infant Formula, gave a mean value of 0.95 ± 0.088 mg vitamin K/kg (K or K1?)(n = 31) with a coefficient of variation of 9.26.

Improved liquid-chromatographic method for determination of serum cortisol.

1979 ◽  
Vol 25 (7) ◽  
pp. 1293-1296 ◽  
Author(s):  
P M Kabra ◽  
L L Tsai ◽  
L J Marton
Keyword(s):  
Limit Of Detection ◽  
Internal Standard ◽  
Serum Cortisol ◽  
Peak Height ◽  
Reversed Phase ◽  
Column Temperature ◽  
Coefficients Of Variation ◽  
Liquid Chromatographic Method ◽  
Liquid Chromatographic

Abstract We describe a specific and precise method for measuring concentrations of cortisol in serum or plasma by liquid chromatography. Cortisol, together with an internal standard, equilenin, is extracted from 1 mL of serum or plasma and analyzed isocratically on a reversed-phase column with a mobile phase of acetonitrile/phosphate buffer (30/70, by vol.), at a flow rate of 2.0 mL/min. The eluted cortisol is detected by its absorption at 254 nm and quantitated by peak height measurements. Each analysis requires no longer than 15 min at the optimum column temperature of 50 degrees C. The lower limit of detection for cortisol is about 2 ng/sample for a standard solution; sensitivity is routinely 5 micrograms/L of serum. Analytical recoveries exceeded 95%, with good day-to-day precision (coefficients of variation between 4 and 7%). Of more than 50 drugs and steroids tested for possible interference, only the steroids cortisone, prednisone, and prednisolone may interfere with the analysis of cortisol.

Liquid Chromatographic Determination of Residual Isocyanate Monomers in Plastics Intended for Food Contact Use

1995 ◽  
Vol 78 (3) ◽  
pp. 711-718 ◽  
Author(s):  
Andrew P Damant ◽  
Sue M Jickells ◽  
Laurence Castle
Keyword(s):  
Limit Of Detection ◽  
Internal Standard ◽  
Standard Addition ◽  
Chromatographic Determination ◽  
Reversed Phase ◽  
Good Precision ◽  
Food Packages ◽  
Food Package ◽  
Liquid Chromatographic Determination ◽  
Liquid Chromatographic

Abstract A liquid chromatographic (LC) method was developed for the analysis of 10 isocyanates in polyurethane articles and laminates intended for food use. Residual isocyanates are extracted by dichloromethane with concurrent derivatization by 9-(methylaminomethyl)anthracene. The resultant derivatives are analyzed by reversed-phase LC with fluorescence detection. Separation of the isocyanates was studied and optimized. Quantitation uses 1-naphthyl isocyanate as internal standard and standard addition to the food package. Validation demonstrated the method to have good precision (± 2–5%) and recovery (83–95%) for samples spiked with isocyanates at 0.1 mg/kg. The limit of detection was 0.03 mg/kg. Analysis of 19 commercial polyurethane or laminate food packages demonstrated that the method was not prone to interferences. Residues of diphenylmethane-4,4′-diisocyanate were detected in 5 packages and ranged from 0.14 to 1.08 mg/kg.

Sensitive and Selective Liquid-Chromatographic Assay of Fluoxetine and Norfluoxetine in Plasma with Fluorescence Detection After Precolumn Derivatization

1992 ◽  
Vol 38 (9) ◽  
pp. 1756-1761 ◽  
Author(s):  
R F Suckow ◽  
M F Zhang ◽  
T B Cooper
Keyword(s):  
Fluorescence Detection ◽  
Limit Of Detection ◽  
Internal Standard ◽  
Reversed Phase ◽  
Pharmacokinetic Studies ◽  
Precolumn Derivatization ◽  
Ionization Mass ◽  
Ionization Mass Spectroscopy ◽  
Positive Chemical Ionization ◽  
Liquid Chromatographic

Abstract We determined fluoxetine (Prozac) and its major metabolite norfluoxetine in plasma by liquid chromatography with fluorescence detection. After liquid-liquid extraction from 1 mL of plasma, the extract was derivatized at room temperature with dansyl chloride, and the highly fluorescent derivatives were chromatographed with a reversed-phase C18 column and a mobile phase of phosphate buffer and acetonitrile. Dansylated fluoxetine, norfluoxetine, and the internal standard were eluted in less than 14 min with no interference from endogenous material. The calibration curve was linear over the concentration range 25-800 micrograms/L with inter- and intra-assay imprecision (CV) of less than 10%. Validity of the assay was checked by comparing results for 110 patients' samples with those by a liquid-chromatographic method with ultraviolet detection (r = 0.993 for fluoxetine, 0.957 for norfluoxetine). The identity of the dansylated derivatives was verified by positive chemical ionization mass spectroscopy. The lower limit of detection was approximately 3 micrograms/L. Because no major antidepressant, neuroleptic, or respective drug metabolites interfere with the quantification of fluoxetine and norfluoxetine, this is a useful procedure for pharmacokinetic studies and in clinical settings.

Simultaneous liquid-chromatographic determination of 12 common sedatives and hypnotics in serum.

1978 ◽  
Vol 24 (4) ◽  
pp. 657-662 ◽  
Author(s):  
P M Kabra ◽  
H Y Koo ◽  
L J Marton
Keyword(s):  
Serum Proteins ◽  
Limit Of Detection ◽  
Internal Standard ◽  
Chromatographic Determination ◽  
Reversed Phase ◽  
Acetonitrile Solution ◽  
Column Temperature ◽  
Liquid Chromatographic Determination ◽  
Liquid Chromatographic

Abstract We present a method for simultaneously determining 12 hypnotics and sedatives (primidone, methyprylon, phenobarbital, butabarbital, butalbital, ethchlorvynol, pentobarbital, amobarbital, phenytoin, glutethimide, secobarbital and methaqualone) in 200 microliter of serum. Serum proteins are precipitated with an acetonitrile solution containing 5-(4-methylphenyl)-5-phenylhydantoin, the internal standard. The drugs are eluted from a reversed-phase column with a mobile phase consisting of an acetonitrile/phosphate buffer, at a flow rate of 3.0 ml/min. The eluted drugs are detected by their absorption at 195 nm; their quantities are estimated from their peak heights. Each analysis requires no longer than 30 min at the optimum column temperature of 50 degrees C. The lower limit of detection for all of these drugs is less than 10 ng/sample for drug standard. A sensitivity of 1.0 mg/liter of serum is attained routinely for each of the drugs. Analytical recoveries for the 12 drugs varied from 94 to 112%, with good day-to-day precision (CV between 3.8 and 10.4%). Of more than 35 drugs tested for possible interference, only ethotoin interferes with the analysis of phenobarbital.

Liquid-chromatographic determination of alpha- and gamma-tocopherols in erythrocytes, with fluorescence detection.

1983 ◽  
Vol 29 (10) ◽  
pp. 1840-1842 ◽  
Author(s):  
J Lehmann ◽  
H L Martin
Keyword(s):  
Mobile Phase ◽  
High Performance ◽  
Limit Of Detection ◽  
Internal Standard ◽  
Chromatographic Determination ◽  
High Performance Liquid Chromatographic ◽  
Reversed Phase ◽  
Liquid Chromatographic Determination ◽  
Liquid Chromatographic

Abstract We have adapted to erythrocytes a method for the determination of alpha- and gamma-tocopherols in plasma and platelets. Erythrocytes (50 microL) were extracted with methanol containing tocol (internal standard) and pyrogallol. Tocopherols were partitioned into chloroform, washed, and injected in methanol onto a reversed-phase (C18) "high-performance" liquid-chromatographic column. The mobile phase was methanol/water (99/1 by vol) at a flow rate of 2 mL/min and detection was with a "high-performance" spectrophotofluorometer. The limit of detection for either tocopherol is 0.10 microgram/mL of packed cells. Analytical recoveries ranged from 93 to 104%. Some values for tocopherols in human erythrocytes are presented.

Improved liquid-chromatographic determination of mexiletine, an antiarrhythmic drug, in plasma.

1984 ◽  
Vol 30 (2) ◽  
pp. 319-322 ◽  
Author(s):  
W Mastropaolo ◽  
D R Holmes ◽  
M J Osborn ◽  
J Rooke ◽  
T P Moyer
Keyword(s):  
Methylene Chloride ◽  
Limit Of Detection ◽  
Internal Standard ◽  
Chromatographic Determination ◽  
Peak Height ◽  
Reversed Phase ◽  
Internal Standard Peak ◽  
Chromatographic Procedure ◽  
Liquid Chromatographic

Abstract In this improved reversed-phase liquid-chromatographic procedure for determination of mexiletine in plasma, mexiletine and an internal standard, chlorodisopyramide, are extracted with methylene chloride from 0.5 mL of serum or plasma; the extract is then concentrated and injected onto a C18 chromatographic column. Mexiletine in the column effluent is detected by monitoring absorbance at 210 nm. It is quantified by use of mexiletine-internal standard peak-height ratios. The relation between this ratio and mexiletine concentration is linear from 0.1 to 5.0 mg/L. The lower limit of detection is about 50 micrograms/L. At a mexiletine concentration of 2.0 mg/L in serum, intrarun precision (CV) is 2.9% and inter-run precision is 5.9%; at 0.5 mg/L, these CVs are 5.7% and 9.6%, respectively. Analytical recovery of added mexiletine in serum is 68-88%. Therapeutic concentrations of some commonly administered drugs in patients' specimens did not interfere. In serum from 38 patients receiving mexiletine for cardiac arrhythmia, concentrations measured by this method correlated with therapeutic efficacy.

scholarly journals Liquid Chromatographic Determination of Amphotericin B in Different Pharmaceuticals

2002 ◽  
Vol 85 (1) ◽  
pp. 15-19 ◽  
Author(s):  
Louis P Lue ◽  
Susan T Hadman ◽  
Ales Vancura
Keyword(s):  
Amphotericin B ◽  
Limit Of Detection ◽  
Fungal Infections ◽  
Internal Standard ◽  
Pharmaceutical Formulations ◽  
Chromatographic Determination ◽  
Reversed Phase ◽  
Drug Of Choice ◽  
Limit Of Quantitation ◽  
Liquid Chromatographic

Abstract Amphotericin B (AmB) is one of the most potent antifungal agents and the drug of choice in the treatment of serious fungal infections. A liquid chromatographic (LC) method was developed to determine AmB in pharmaceutical formulations for injection, tissue culture, cream, and lotion. μBondapak C18 reversed-phase column and a simple mobile phase consisting of acetonitrile–water–acetic acid (40 + 54 + 6, v/v) was used. The flow rate was 1.8 mL/min and the effluent was monitored at 405 nm. The developed LC method uses piroxicam as an internal standard and has a limit of detection of 10 ng/mL, a limit of quantitation of 30 ng/mL, and the assay is linear from 0.01 to 100 μg/mL. AmB and piroxicam elute with retention times of 12.4 and 4.0 min, respectively, and the resolution between AmB and piroxicam was 10.6. In comparison with the official United States Pharmacopeia microbial assay for AmB, this LC method is more rapid, selective, sensitive, and offers positive identification.

Liquid-chromatographic determination of tobramycin in serum with spectrophotometric detection.

1983 ◽  
Vol 29 (4) ◽  
pp. 672-674 ◽  
Author(s):  
P M Kabra ◽  
P K Bhatnagar ◽  
M A Nelson ◽  
J H Wall ◽  
L J Marton
Keyword(s):  
Limit Of Detection ◽  
Internal Standard ◽  
Chromatographic Determination ◽  
Reversed Phase ◽  
Water Soluble ◽  
Trinitrobenzenesulfonic Acid ◽  
Column Temperature ◽  
Derivatizing Agent ◽  
Liquid Chromatographic

Abstract We describe a simple, precise, accurate, and specific liquid-chromatographic procedure for determination of tobramycin in 50 microL of serum. Tobramycin and the internal standard (sisomicin) are quantitatively converted into their trinitrophenyl derivatives by reaction with a water-soluble derivatizing agent (2,4,6-trinitrobenzenesulfonic acid) at 70 degrees C for 30 min. The derivatives are extracted from the crude reaction mixture by using a reversed-phase Bond-Elut C18 column, and separated on a reversed-phase octyl column with a mobile phase consisting of an acetonitrile/phosphate buffer (70/30 by vol) at a flow rate of 3.0 mL/min. The eluted compounds are detected at 340 nm, and quantified from their peak areas. Chromatography is complete in less than 4.5 min at the optimum column temperature of 50 degrees C. The lower limit of detection for tobramycin is less than 0.2 mg/L. Analytical recoveries for tobramycin varied from 94 to 99%, linearity extended to 25 mg/L, and day-to-day precision (CV) was between 4.6 and 5.1%. Numerous drugs and antibiotics tested do not interfere. Results correlate well (r greater than 0.95) with those by radioimmunoassay and EMIT.

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