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Rationale: Despite direct immediate intervention and therapy, ST-segment–elevation myocardial infarction (STEMI) victims remain at risk for infarct expansion, heart failure, reinfarction, repeat revascularization, and death. Objective: To evaluate the safety and bioactivity of autologous CD34+ cell (CLBS10) intracoronary infusion in patients with left ventricular dysfunction post STEMI. Methods and Results: Patients who underwent successful stenting for STEMI and had left ventricular dysfunction (ejection fraction≤48%) ≥4 days poststent were eligible for enrollment. Subjects (N=161) underwent mini bone marrow harvest and were randomized 1:1 to receive (1) autologous CD34+ cells (minimum 10 mol/L±20% cells; N=78) or (2) diluent alone (N=83), via intracoronary infusion. The primary safety end point was adverse events, serious adverse events, and major adverse cardiac event. The primary efficacy end point was change in resting myocardial perfusion over 6 months. No differences in myocardial perfusion or adverse events were observed between the control and treatment groups, although increased perfusion was observed within each group from baseline to 6 months ( P P =0.05). At 1 year, 3.6% (N=3) and 0% deaths were observed in the control and treatment group, respectively. Conclusions: This PreSERVE-AMI (Phase 2, randomized, double-blind, placebo-controlled trial) represents the largest study of cell-based therapy for STEMI completed in the United States and provides evidence supporting safety and potential efficacy in patients with left ventricular dysfunction post STEMI who are at risk for death and major morbidity. Clinical Trial Registration: URL: http://www.clinicaltrials.gov . Unique identifier: NCT01495364.

Background: Prior studies suggest an association between naturally high circulating levels of CD34+ cells (CD34) and better outcomes following ischemic events. Pts in the treatment arm of the PreSERVE-AMI study were infused with all CD34 yielded from their bone marrow harvest, resulting in pts with naturally higher CD34 cell counts being infused with a higher cell dose. To determine whether selection bias may have influenced results, we performed an analysis of the relationship between BM CD34 content and outcomes in PreSERVE-AMI. Methods: PreSERVE-AMI is a Phase 2, randomized, double-blind, placebo-controlled trial of autologous, BM derived CD34 (CLBS10) in pts with LV dysfunction (LVEF ≤48% by MRI) ≥4 days post-STEMI. All pts underwent mini BM harvest and were randomized 1:1 to intracoronary infusion of CD34 (minimum dose 10M±20% cells) or placebo (cell diluent). Primary safety endpoints included AEs, SAEs and MACE (CV mortality, HF hospitalization, reinfarction, revascularization). Secondary endpoints included LVEF change from baseline. Regression analyses were performed to assess the influence of BM CD34 content on outcomes. Results: 161 patients were randomized and received intracoronary infusion (Jan 2013-Dec 2014). Mean age was 57±10 and 82% were men. Mean follow-up time was 18 mo, and safety data are accruing. BM CD34 content was not associated with differences in baseline characteristics with the exception of diabetes prevalence being higher in those with higher BM CD34 count (p=0.03). In controls, regression analyses showed no association of BM CD34 content with rates of MACE (p=0.2), SAEs (p=0.4), or change in LVEF (p=0.9). However, in CD34 treated pts, there was a dose-dependent improvement in LV function (p=0.045) and dose-dependent trends in MACE and SAE reduction compared to controls. As previously reported SPECT perfusion improved to a similar degree in control and treated pts. Conclusions: These data suggest that intracoronary infusion of CD34 cells, rather than BM CD34 cell content, is responsible for the benefit observed in the CD34 treatment group. The finding of a dose-response relationship is an important advance for defining the CD34 as an active agent for tissue repair. Future studies will further define CD34 dosimetry.

CPT-11 (irinotecan) and mainly its metabolite SN-38 are potent antitumor derivatives of camptothecin. As the active lactone forms of both CPT-11 and SN-38 exist in pH-dependent equilibrium with their respective less potent open-ring hydroxy acid species, the simultaneous monitoring of both forms of both compounds is relevant. CPT-11 and SN-38 derivatives have quite different fluorescence responses. In order to avoid any compromise on the wavelength setting, we developed chromatographic conditions allowing simple automated wavelength setting changes which have been prevented using existing methods involving conventional C18 columns. This was achieved by means of a Symmetry C18 column combined to a gradient elution program using acetonitrile and 75 mM ammonium acetate plus 7.5 mM tetrabutylammonium bromide at pH 6.4. The developed conditions allowed an elution order suitable for a simple automated wavelength change in respect to reliable peak integration. CPT-11 and SN-38 derivatives were detected at lambda ex=362 nm/lambda em=425 nm and lambda ex=375 nm/lambda em =560 nm, respectively. The developed method allowed the detection of amounts less than 3 pg of each derivative injected on column. The method was successfully applied to pharmacokinetic and toxicokinetic studies in rat and dog.

Testing for the equivalence of results generated by different analytical methodology is a common practice in the pharmaceutical sciences. Methodology changes are implemented for both scientific and economic reasons during a scientific study. Thus, the need to demonstrate the appropriateness of considering data generated by distinct methods as part of a single information population arises. This paper describes a rapid and simple approach to the statistical design and interpretation of method comparison experiments. The approach presented is based upon a statistical power calculation technique, a knowledge of the variability associated with the methods to be compared and the criteria for equivalence (the limits within which differences become immeasurable or, for practical purposes, insignificant). Reference tables are included which show necessary sample sizes for comparison experiments for common combinations of these three variables.

An automated system for valve performance testing to determine net weight loss per actuation of pharmaceutical aerosol dosage forms has been developed and is described. The principal element of the system is a novel automatic aerosol actuation device. Details of the validation, comparison to manual methods of analysis and advantages are presented. The automated system has been shown to be a cost-effective, productive and facile alternative to manual testing.

Three test methods for determining in vitro drug release rate from transdermal delivery dosage forms were tested for equivalency of results, ease of implementation and precision. The 'paddle-over-disk' (POD) method is under consideration by the USP as a standarized method for release-rate testing of all transdermal delivery dosage forms. The 'reciprocating disk' (RD) and 'diffusion cell' (DC) methods are both commonly employed throughout the pharmaceutical industry. The three methods were demonstrated to be equivalent in terms of release rate profile (curve shape) and total drug released over the lifetime of the dosage form tested (Transderm-Scop). The precision for the RD method as measured by the mean relative standard deviation over all time points was 4.6%; the precision of the POD method was 5.4% and that for the DC method was 6.7%. Steady-state flux values derived from the POD and RD methods were equivalent ( approximately 4 microg cm(-2) h(-1)) but were approximately 25% greater than the steady-state flux value derived from the DC method ( approximately 3 microg cm(-2) h(-1)). All three methods gave results which were within the specifications of the manufacturer (CIBA-GEIGY). The POD method was the easiest to use on a routine basis, required the least amount of specialized equipment and most resembled the current test methodology for dissolution testing of other dosage forms such as tablets or capsules.

The design and characterization of a nebulizer/spray chamber interface for high performance liquid chromatography-direct current argon plasma spectroscopy are presented. Optimum geometry for this interface was determined by the varying of the chimney tip diameter. Mechanical efficiencies of the interface as a function of nebulizer argon flow are tabulated. The particle velocity distribution within the solvent spray created by the nebulizer and emerging from the interface chimney was measured with the use of laser-Doppler anemometry.

The high-performance liquid chromatographic separation of bis(η 5 -1,2,3,4,5-pentamethylcyclopentadienyl)-titanium, zirconium and hafnium dichlorides was demonstrated on cyano-bonded silica under normal-phase gradient elution conditions. Data reported include capacity factors, separation efficiencies, and compound resolutions. The linearity of detection is shown and detection limits are reported.

An analytical method was developed for the determination of famotidine, potential degradates, and preservatives in several pharmaceutical formulations. The method employs reversed-phase chromatography on a silica column with a methanol—phosphate solution as the mobile phase. The influence of the concentration of phosphate and organic modifier are discussed. Accuracy and precision for this method along with assay data from different formulations of famotidine are presented. Two different commercial silica columns were tested with this method. Chromatographic differences related to the surface area and pore size of the silica are discussed.

A novel method for the determination of timolol in pharmaceutical products has been developed and is described. The method employs high-performance liquid chromatography (HPLC) on silica dynamically modified with the cetyltrimethylammonium cation to quantitate the analyte. The use of this type of reversed-phase HPLC system for timolol determinations results in improved quality of chromatography, especially in terms of peak tailing and peak efficiency, in comparison to chromatography on bonded-phase silica. Column-to-column as well as manufacturer-to-manufacturer reproducibility for this separation on silica columns has been obtained and is better in our hands than that encountered with bonded-phase column packings. The method has been shown to be linear for the compounds studied, comparably accurate and precise to bonded-phase methods and specific for timolol in a variety of pharmaceutical formulations. Under the conditions specified, timolol can be successfully separated from its three known potential degradates. Possible explanations of the primary retention mechanisms for the analytes are offered.

The high-performance liquid chromatographic (HPLC) behavior of homologous titanaindene complexes has been investigated. Various polar (silica, cyano-bonded, phenyl-bonded) and non-polar (C18-bonded) adsorbents were employed in this study for normal-phase and reversed-phase HPLC, respectively. Data reported include capacity factors (k′), separation efficiencies (N), tailing factors (T) and compound resolutions (R). Normal-phase separation of a titanaindene structural isomer pair on a phenyl-bonded silica column is demonstrated.

The loss of available iodine from povidone-iodine solution stored in contact with two different polyetherpolyurethane foams was monitored as a function of time and temperature. Statistical evaluation of the results for the four temperatures studied [ambient (25 degrees), 30, 45 and 55 degrees C] indicated the chemical equivalence of the two foams as storage and delivery systems for povidone-iodine solution in terms of solution stability. In addition, application of a first-order kinetic model to the data produced an acceptable fit. An Arrhenius-type evaluation of the resulting rate constants yielded a linear relationship which was shown to be useful for predicting loss of available iodine under ambient temperature conditions of storage.

A flow injection (FI) method has been developed to determine simultaneously a drug with an ultraviolet chromophore, lovastatin, and butylated hydroxyanisole (BHA) in a tablet. The system involves ultraviolet (UV) absorbance detection for the drug and oxidative amperometric electrochemical detection for BHA. The method has been shown to be reproducible for routine determinations with an accuracy of +/-1% for lovastatin and +/-4% for BHA. Precision for both analytes was approximately +/-1% (RSD). The use of FI compared with HPLC for these determinations has led to a dramatic decrease in the time required per assay. The method with UV detection has been shown to be specific for the drug in the presence of potential autoxidation products as well as BHA and its oxidation products. The use of electrochemical detection in series allows the method to be simultaneously specific for low levels of BHA (40 microg/tablet) in the presence of its oxidation products as well as the drug and its potential autoxidation products.

The high-performance liquid chromatographic (HPLC) analysis of dopazinol, an anti-Parkinsonian drug, using either ultraviolet (UV) absorbance detection at 282 nm or oxidative amperometric electrochemical detection at the glassy carbon electrode (+0.92 V vs. Ag/AgCl) is described and evaluated. The influence of such factors as mobile phase composition, pH and column temperature on the detection of dopazinol using both detectors is reported. The HPLC analysis of samples resulting from the development and evaluation of a pharmaceutical formulation demonstrated a precision of less than 2.5% (R.S.D.) in the working ranges of the UV and electrochemical detectors. The limits of detection were 100 ng/ml for the UV detector and 1 ng/ml for the electrochemical detector. The higher sensitivity of the latter makes its use appropriate for the analysis of samples with low concentrations of dopazinol. Studies to compare the UV and electrochemical detectors for the routine analysis of dopazinol in pharmaceutical formulations are presented.