Analysis of benzalkonium chloride and its homologs: HPLC versus HPCE.

Benzalkonium chloride (BAK) is a mixture of alkylbenzyldimethylammonium chloride homologs with n-C,2H25, n-C,4H29, and n-C16H33 comprising a major portion of the alkyl groups present. An analytical method for BAK must differentiate and quantitate the homologs in the BAK mixture. Reversed-phase high performance liquid chromatography (HPLC) separates compounds based on their affinity for a nonpolar column, which is a direct correlation to the compounds' polarity. High performance capillary electrophoresis (HPCE), however, separates compounds in an electric field according to their charge and size. The BAK homologs are suitable for separation by either of these methods because their polarity and sizes differ significantly. The HPLC method employed a mobile phase of 60% acetonitrile and 40% 0.1 M sodium acetate buffer pH 5 pumped at 1.0 ml min(-1), a 4.6 x 250 mm cyano column with 5 microm packing, and UV detection at 254 nm. The HPCE method utilized a run buffer of 30% acetonitrile and 70% 0.05 M sodium phosphate pH 3.06, a 50 microm x 20 cm open silica capillary, 7.5 kV electric field and UV detection at 214 nm. Both HPLC and HPCE demonstrated good linearity in the range of 0.025 to 0.8 mg ml(-1) with r2 values of approximately 0.99. The HPLC method produced good separation of the homolog peaks with a total analysis time of 25 min. HPCE run time was less than 5 min and demonstrated good separation of the three homologs. The HPLC method, however, was superior to HPCE in the areas of sensitivity and precision. The HPLC has been extensively used in the routine quantitation and qualitation of benzalkonium chloride concentrations in various products; however, long analysis times make this method inefficient. The HPCE method produced comparable results to the HPLC method but with much shorter analysis times. An HPCE analysis method, as presented here, may prove to be a much more useful and efficient method for the analysis of benzalkonium chloride and its homologs.

Stability of Two Concentrations of Heparin Sodium Prefilled in CADD-Micro* Pump Syringes.

The purpose of this study was to determine the stability of two concentrations (1000 units/mL and 40,000 units/mL) of heparin sodium when individually prefilled in CADD-Micro pump syringes (Sims Deltec, Inc., St Paul MN) and stored at near-body (30 deg C) temperature for extended periods of time, up to 30 days. Three syringes were prepared for each sample period and each concentration. Following aseptic filling, the syringes were capped and stored under controlled temperatures for the duration of the study. The contents of each of three syringes per sample set were expelled individually into screw-cap, glass sample vials at designated sample times. Samples were immediately stored at -70 deg C until the time of analysis. The results of the study indicate that, when prefilled in CADD-Micro pump polypropylene syringes and maintained at 30 deg C for up to 30 days, both concentrations of heparin sodium studied retained greater thatn 95% of the intact drug.

Stability of ramipril in water, apple juice, and applesauce.

The stability of ramipril in water, in apple juice, and in applesauce was studied. The contents of a single capsule each of ramipril 1.25, 2.5, and 5 mg were mixed in glass beakers with 120 mL of deionized and filtered water, apple juice, or applesauce. Each mixture was apportioned into 10 120-mL amber polyethylene terephthalate (PET) containers. Five of the containers in each set were stored at 23 degrees C, and samples were taken at 0, 1, 2, 6, 12, and 24 hours. The other five containers were stored at 3 degrees C, and samples were taken at 4, 8, 12, 24, and 48 hours. The samples were analyzed for ramipril concentration by stability-indicating high-performance liquid chromatography (HPLC). The quantity of drug remaining in the PET container after "administration" was determined by mixing the contents of single 5-mg ramipril capsules with 60 mL of apple juice, pouring the mixture into a waste receptacle, rinsing the PET container three separate times with 10 mL of water, and analyzing the pooled fluid from these rinses for ramipril concentration by HPLC. Under no condition did the percentage of ramipril remaining drop below 90%. No peaks for degradation products appeared in the chromatograms. The mean +/- S.D. quantity of ramipril remaining in the PET containers after draining was 0.3 +/- 0.3% for the apple juice. Ramipril from 1.25-, 2.5-, and 5-mg capsules mixed in water, in apple juice, and in applesauce was stable for 24 hours at 23 degrees C and for 48 hours at 3 degrees C.

RNA polymerase II transcription factor SIII. II. Functional properties and role in RNA chain elongation.

A novel transcription factor that stimulates synthesis of accurately initiated transcripts by mammalian RNA polymerase II has been identified and purified to apparent homogeneity from rat liver extracts (Bradsher, J. N., Jackson, K. W., Conaway, R. C., Conaway, J. W. (1993) J. Biol. Chem. 268, 25587-25593). This factor, which we designate SIII, has a native molecular mass of approximately 140 kDa and is composed of three polypeptides of 110, 18, and 15 kDa. In this report, we demonstrate that SIII stimulates promoter-specific transcription by increasing the overall rate of RNA chain elongation by RNA polymerase II. Results of pulse-chase experiments indicate that SIII does not need to be present during preinitiation complex formation or transcription initiation in order to stimulate promoter-specific transcription. In addition, SIII is able to stimulate the rate of RNA chain elongation by RNA polymerase II during transcription of double stranded oligo(dC)-tailed templates. Taken together, these findings indicate that the factor exerts its activity directly on the elongation complex.