Quantitative analysis of the principle soy isoflavones genistein, daidzein and glycitein, and their primary conjugated metabolites in human plasma and urine using reversed-phase high-performance liquid chromatography with ultraviolet detection.

Soy isoflavones are becoming of increasing interest as nutritional agents which can be used to combat osteoporosis and hyperlipidemia, and are also being considered as potential cancer chemopreventive compounds. However, prior to their formulation and distribution as therapeutic agents, thorough pharmacokinetic and toxicological assessment needs to be completed in men and women in a variety of health conditions in order to ensure their therapeutic efficacy and safety. At this time, studies of purified soy isoflavones are possible, and are being designed to fully evaluate the pharmacological utility of these preparations. In support of these studies, quantitative analysis of soy isoflavones in biological fluids can be accomplished with a wide variety of methods and analytical instrumentation. However, the relatively ubiquitous presence of high-performance liquid chromatography with ultraviolet detection (HPLC-UV) in most analytical laboratories, the relative ease of its operation, and the lesser expense of this instrumentation as compared to more sophisticated techniques such as liquid chromatography-mass spectrometry, offers some distinct advantages for its use in pharmacokinetic studies. In this manuscript, the development and validation of an HPLC-UV method for the quantitation of the principal soy isoflavones, genistein, daidzein, and glycitein, and their primary metabolites, in human plasma and urine is described. This analytical approach allows for pharmacologically relevant concentrations of the analytes and their principle metabolites to be detected, and has been validated in close agreement with the US Food and Drug Administration's guidelines for the validation of methods to be used in support of pharmacokinetic studies.

Single-breath measurements of pulmonary oxygen uptake and gas flow rates for ventilator management in ARDS.

Monitoring data in critical care and anesthesiology should be displayed to present a rapid and easily comprehensible definition of the patient's clinical status. A graphic computer display of the analog output of gas flow rates and the O(2) and CO(2) concentrations of respiratory gases profiles the expired breath for an estimation of pulmonary function and gas exchange. An estimate of pulmonary perfusion, cardiac output, and the general adequacy of cardiovascular circulation is obtained from the computer calculation of O(2) uptake and CO(2) elimination, dead space, and alveolar ventilation. Adjunctive data from the spirometric measurements of airway pressures, volumes, and compliance, supplemented by hemodynamic monitoring, aids in the diagnosis of physiologic changes. For > 10 years, we have used this system to monitor patients who are anesthetized, sedated, and receiving mechanical ventilation during anesthesia and surgery, and recently have extended the technique to intensive care areas. Our experience has shown good correlation of changes in the computer-assisted expired breath analysis with coinciding clinical events, including upper airway obstruction, bronchospasm, and alveolar volume/pulmonary capillary blood flow impairment. To demonstrate the use of this system, we describe the ventilator management for a patient with severe ARDS. In this patient, changes in ventilator management, including pressure control ventilation, improved pulmonary O(2) uptake (mean, 18.7 vs 8.5 mL/breath), CO(2) elimination (mean, 17 vs 13 mL/breath), and compliance (mean, 29.7 vs 19.0 mL/cm H(2)O), were compared with intermittent mandatory ventilation.

Intestinal effects of sulfate in drinking water on normal human subjects.

Uncontrolled observations implicate sulfate in drinking water at concentrations exceeding 500-700 mg/liter as a cause of diarrhea, but controlled studies have not been reported. We conducted a controlled study in normal adults to determine the effect of various drinking water sodium sulfate concentrations on bowel function. Ten healthy subjects were given a constant diet and fluid intake. Fluid consisted of 36 ml/kg/day of drinking water of various known sulfate concentrations and 500 ml of other fluid. In a dose-ranging study, four subjects received drinking water with sulfate concentrations of 0, 400, 600, 800, 1000, and 1200 mg/liters for six consecutive two-day periods. In a single-dose study, six other subjects received water with sulfate concentrations of 0 and 1200 mg/liter for two consecutive six-day periods. Stool mass, frequency, and consistency and mouth-to-anus appearance time of colored markers were measured. In the dose-ranging study, the only significant linear trend was decreasing mouth-to-anus appearance time with increasing sulfate concentrations. In the single-dose study, 1200 mg/liter sulfate caused a significant but clinically mild increase in mean stool mass per six-day pool from 621 g to 922 g (P = 0.03). When all 10 subjects were used to compare effects of 0 mg/liter and 1200 mg/liter sulfate, significant differences in stool consistency (P = 0.02) and transit time (P = 0.03) were observed. None of the subjects reported diarrhea or passed more than three stools per day. In 10 normal adult subjects, sulfate in drinking water at a concentration of 1200 mg/liter, which is higher than reported to occur in US municipal water sources, caused a measurable but clinically insignificant increase in stool mass and decrease in stool consistency and appearance time, but no change in stool frequency and no complaint of diarrhea.

Measurement of lactose consumption reliability and comparison of two methods.

The objective of this study was to assess the reliability of lactose consumption measured with a food-frequency questionnaire and to compare food-frequency data to prospectively recorded data. A self-administered food-frequency questionnaire (FF1) was completed by 295 women ages 38 to 49. Two months later, 75 received the same food-frequency questionnaire (FF2), and 75 received a 1-week diet diary listing 23 commonly eaten lactose-containing foods. Estimated mean intake of lactose was 16.1 g/d with FF1. Intake was slightly (but not significantly) higher at follow-up: Mean difference (95% confidence interval) was 1.4 (-0.8, 3.7) g/d for FF2 versus FF1, and 0.5 g (1.3, 2.3) g/d for the diet diary versus FF1. A moderate level of correlation was found: r = 0.57 between FF1 and FF2, and r = 0.65 between FF1 and the diet diary. Smaller subsets of items (n = 15 and n = 7) could be used to estimate lactose, with 89% and 82% agreement within tertiles, respectively. Since relatively few foods contain substantial amounts of lactose, a limited food-frequency or structured diary may be useful in research studies on the association between lactose consumption and ovarian cancer or other diseases.

Breath-by-breath measurement of oxygen consumption and FIO2-FEO2 with increased oxygen demand.

Continuous on-line breath-by-breath measurement of pulmonary gas exchange was used to monitor the increase in oxygen uptake (VO2) and carbon dioxide excretion (VCO2) induced by the oxidative phosphorylation uncoupling agent 2, 4-dinitrophenol (DNP) in 10 dogs. With incremental doses of DNP totaling 5 mg/kg, the continuously monitored VO2 increased within 2-3 min after the first injection of the drug. VCO2 showed a similar response 4-6 min after the first injection. Temperature increase due to the pharmacological oxidative phosphorylation uncoupling required 20-30 min for a discernible change at this dose. This study also demonstrated a modified and compromised response to the drug in dogs where oxygen delivery was limited by mechanical ventilation.

A pulsed phase measurement ultrasonic flowmeter for medical gases.

Pneumotachometers are used to measure instantaneous flowrate in the respiratory gas streams. The presently available devices suffer from lack of linearity, slow response times, and gas density sensitivity. A new design of an ultrasonic gas flowmeter is presented in this paper: We investigate the acoustic characteristics of ring and piston shaped transducers, and describe a sampling method to avoid the error due to reflection. A microcomputer is used to overcome the 360 degrees detection ambiguity problem associated with phase detection technique. This design has been tested in clinical settings and has been shown to give linear response, independent of gas density, and to have a wide dynamic range.

Automated separation and measurement of lactate dehydrogenase isoenzymes.

We describe a totally automated, computer-controlled system for separating and measuring the activity of lactate dehydrogenase (EC 1.1.1.27) isoenzymes. These isoenzymes in tissue extracts were separated on disposable DEAE-cellulose mini-columns. Resolution was complete except that LD-1 was not resolved from LD-2.

A versatile temperature-controlled reaction cuvet.

A thermostatted reaction cuvet, operating under computerized or manual temperature control in the ultraviolet or visible region, is described and evaluated. The cell was designed for use with the automated chemistry system described earlier [Clin. Chem. 19, 1114 (1973)], but can readily be adapted for other applications. The entire 3-ml cuvet assembly warms from ambient temperature to 37 degrees C in less than 10 min, with plus or minus 0.15 degrees C stability; the long-term stability is plus or minus 0.05 degrees C; temperature recovery time after washing or reagent addition is less than 3 min. The optical pathlength is 10.00 mm and the carryover volume is 18 mu-l.