Troxacitabine prodrugs for pancreatic cancer.

Troxacitabine is a cytotoxic deoxycytidine analogue with an unnatural L-configuration, which is activated by deoxycytidine kinase (dCK). The configuration is responsible for differences in the uptake and metabolism of troxacitabine compared to other deoxynucleoside analogues. The main drawback in the use of most nucleoside anticancer agents originates from their hydrophilic nature, which property requires a high and frequent dosage for an intravenous administration. To overcome this problem several troxacitabine prodrugs modified in the aminogroup with a linear aliphatic chain with a higher lipophilicity were developed. To determine whether these prodrugs have an advantage over Troxacitabine pancreatic cancer cell lines were exposed to Troxacitabine and the lipophilic prodrugs. The addition of linear aliphatic chains to troxacitabine increased sensitivity of pancreatic cancer cell lines to the drug > 100-fold, possibly due to a better uptake and retention of the drug.

In vitro reaction of barbiturates with formaldehyde.

Barbiturates are widely used as sedatives, hypnotics, and antiepileptics, and, when coupled with their narrow therapeutic index, the probability that their use will result in accidental or intentional death is significant. When barbiturates are implicated in a murder or suicide, analysis for their presence is often required. Under certain conditions, barbiturates are quite stable, but conditions found in vivo immediately after death or after embalming may promote barbiturate decomposition. If extensive decomposition occurs, analysis for them may be difficult or impossible. Here, the stability of three representative barbiturates, under conditions that model those likely to prevail in vivo shortly after death and after embalming, have been studied. Solutions of phenobarbital were found to slowly decompose in water over the pH range of approximately 3.5 to 9.5. More rapid decomposition occurred at higher pH, and 2-phenylbutyric acid was the main decomposition product. Formaldehyde (5-20%) accelerated the decomposition rate 3-10-fold such that phenobarbital decomposition could be complete after 30 days. In contrast, pentobarbital decomposed roughly 10 times more slowly and secobarbital did not detectably decompose under any of the conditions studied. Thus, certain barbiturates may partially or completely decompose in vivo after death, especially after embalming, and thus analysis for them may lead to false negatives. However, this work shows that analysis for the parent barbiturate or its predicted decomposition product may provide data that will reduce the likelihood of false negatives.

In vitro reaction of formaldehyde with fenfluramine: conversion to N-methyl fenfluramine.

Embalming is common, and it can create problems for the forensic scientist if a drug has been the cause death and this drug is also reactive toward the embalming fluid. Previous studies have focused on the tricyclic amines nortriptyline and desipramine. In the presence of formaldehyde, a typical component of embalming fluid, either of these two compounds can be rapidly converted to their methylated derivatives amitriptyline and imipramine, respectively. We have begun a larger project designed to determine the reactivity and reactions of a wide range of drugs with formaldehyde. We report here our results from fenfluramine, which, like the tricyclic amines, is reactive towards formaldehyde and is converted into its N-methyl derivative. The rate of conversion is dependent upon pH and formaldehyde concentration. Up to 100% conversion in 24 h was observed. In addition, we have also devised a simplified procedure for monitoring this process that may be useful for others working in this area. Finally, we note that the reactions of fenfluramine studied here and of amines in general with formaldehyde need to be considered when performing postmortem/postembalming forensic analysis.

Activation of AP-1 through the MAP kinase pathway: a potential mechanism of the carcinogenic effect of arenediazonium ions.

Arenediazonium ions such as those found in the common mushroom Agaricus bisporus have been convincingly demonstrated to be tumorigenic. The specific mechanism of their tumorigenicity remains unclear. It has been shown that arenediazonium ions can be metabolized to aryl radicals, and that reaction of these aryl radicals with DNA produces aryl adducts. These metabolic processes also produce the reactive oxygen species superoxide and hydroxyl radicals which have been implicated in AP-1 activation. To further investigate the mechanism of tumorigenesis by arenediazonium ions, we studied the effect of a representative arenediazonium ion on AP-1 activation and phosphorylation of the signal transduction proteins ERK1, ERK2, JNK, and p38 kinase, both in vitro and in vivo. We also identified the specific radicals produced by spin trapping and ESR analysis. Here, it was found that p-methylbenzenediazonium ion (2a) induced a 16-fold increase in the extent of AP-1 activation at micromolar concentrations, and that this increase coincided with phosphorylation of the signaling kinases ERK1 and -2 and p38 kinase, but not JNK, in JB6 mouse epithelial cells. In vivo studies using AP-1 luciferase reporter-bearing transgenic mice supported the increase in the extent of AP-1 activation in 2a-treated mice over controls, and showed that this effect was different in different tissue types. The antioxidant N-acetylcysteine (NAC), a general antioxidant, showed an inhibitory effect on 2a-mediated AP-1 induction, while aspirin, a hydroxyl radical scavenger, had no effect. Spin trapping studies showed that while NAC suppressed radical formation from 2a, aspirin did not alter radical production from 2a. It appears that 3a, a carbon-centered radical formed from 2a, is responsible for AP-1-induced activation, and therefore, radical species that are not oxygen-centered are also capable of inducing AP-1. These results represent a step toward understanding the mechanism of tumorigenicity of arenediazonium ions.

Estimated content percentages of volatile liquids and fat extractables in ready-to-eat foods.

Content percentages of volatile liquids and fat extractables in 340 samples of ready-to-eat foods were determined gravimetrically. Volatile liquids were determined by drying samples in a microwave oven with a self-contained balance; results were printed out automatically. Fat extractables were extracted from the samples with mixed ethers; extracts were dried and weighed manually. The samples, 191 nonfat and 149 fatty (containing ca 2% or more fat) foods, represent about 5000 different food items and include infant and toddler, ethnic, fast, and imported items. Samples were initially prepared for screening of essential and toxic elements and chemical contamination by chopping and mixing into homogenous composites. Content determinations were then made on separate portions from each composite. Content results were put into a database for evaluation. Overall, mean results from both determinations agree with published data for moisture and fat contents of similar food items. Coefficients of variation, however, were lower for determination of volatile liquids than for that of fat extractables.

Methyl bromide determination in selected foods by headspace technique.

A headspace method used earlier for determining methyl bromide (MB) in assorted nuts and peanut butters has been successfully applied to other foods that could potentially contain traces of this toxic fumigant. The foods tested include 63 off-the-shelf spices and seasonings, 83 table-ready items (grain-based, dried, or highly seasoned), 30 dried fruits and trail mixes, and 38 oil-based items (oil-seeds, cooking oils, or spicy oil-based dressings). Sample headspace gas is produced by blending < or = 50 g sample in 250 +/- 50 mL aqueous solution in a sealed 1000 mL blender cup. After equilibration at 25 degrees C, the headspace is sampled with a gas-tight syringe and injected into a dual column-dual detector gas chromatograph. One determination is made with a 20% OV-101 packed column and a 63Ni electron capture detector (ECD), the other with a GS-Q wide-bore capillary column and a Hall electrolytic conductivity detector (HECD). Of the approximately 200 samples tested, none contained detectable MB residue at a quantitation limit < 100 ng/g sample. All fortified samples yielded MB recovery. Samples were fortified at levels ranging from 78 to 3250 ng MB/g. Recoveries ranged from a mean high of 56% for spices and seasonings to a mean low of 30% for oil-based foods. The overall recovery and CV, including the results from assorted nuts and peanut butters, were 46 and 33%, respectively.

Fumigants and related chemicals in foods: review of residue findings, contamination sources, and analytical methods.

Public concern over chemical residues in foods increased in the United States during the early 1980s. Potentially hazardous levels of ethylene dibromide (EDB), a relatively non-volatile fumigant, were detected in several finished grain-based products by governmental food-monitoring laboratories. As a result, the U.S. Environmental Protection Agency banned the use of EDB as a fumigant in 1983. Commercial fumigators then began using more of the highly volatile chemicals such as methyl bromide and phosphine. These chemicals are less likely to leave residues on stored crops than the previously used fumigants such as EDB, chloroform, and carbon tetrachloride. However, trace residues of many pest-control fumigants and related industrial chemicals are currently found in assorted foods. This contamination may come from the original fumigation of stored crops, or from the industrial chemicals occurring in the environment and in food processing chains. No potential health problem is indicated at this time. Yet scientists continue to uncover the sources of this chemical contamination, and to develop better methods to monitor foods for it. They also seek better ways to protect foodstuffs from pests prior to human consumption.

Recovery of 1,3-, 2,3-dichloropropenes, 1,2-dibromo-3-chloropropane, and o-, p-dichlorobenzenes from fatty and non-fat foodstuffs by liquid extraction technique.

Food samples including fatty, non-fatty, grain-based, and nongrain-based types were fortified with the following five nematocides and fumigants: 1,3-dichloropropene, 2,3-dichloropropene, 1,2-dibromo-3-chloropropane, o-dichlorobenzene, and p-dichlorobenzene. Then, depending on sample consistency and type, the samples were diluted in, or extracted with organic solvent such as isooctane. A few of the high-fat extracts were passed through Florisil to remove excess fat or endogenous interferences. Analysis of the initial or cleaned up extracts was done by gas chromatography (GC) at 90 degrees C. The dichloropropenes were determined on 20% OV-101 columns with electron-capture and Hall electroconductivity detectors. The dichlorobenzenes and 1,2-dibromo-3-chloropropane, which elute beyond 30 min on the above columns, were determined on 5%-loaded columns using the same detectors. All five analytes were recovered from these techniques. Mean analyte recovery following a direct dilution or extraction was 83%, and following the Florisil cleanup step, was 52%. In 1986, a fumigant survey of about 200 foodstuffs by using this overall technique gave no findings of the five compounds studied here.

A displacement pump procedure to load extracts for automated gel permeation chromatography.

Automated gel permeation chromatography (GPC) effectively separates lipids from pesticides in sample extracts that contain fat. Using a large syringe to load sample extracts manually onto GPC models having 5 mL holding loops is awkward, slow, and potentially hazardous. Loading with a small-volume displacement pump, however, is convenient and fast (ca 1 loop every 20 s). And more importantly, the analyst is not exposed to toxic organic vapors because the loading pump and its connecting lines do not leak in the way that a syringe does.

Determination of pesticide residues in food with a 6% cyanopropylphenyl capillary column.

A small-diameter 6% cyanopropylphenyl column is studied for its suitability for determining pesticides in food. Repeatability and linearity are satisfactory, and the column is capable of separating residue combinations that are known not to separate on methyl silicone columns. At 150 degrees C or 130 degrees C, the column satisfactorily separates five by-products of tecnazene, a growth regulator and sprout suppressant found in potatoes, and four by-products of quintozene, a soil and seed fungicide found in peanut products.

Rapid determination of fumigant and industrial chemical residues in food.

A gas chromatographic (GC) method is described for the determination of 22 fumigant and industrial chemical residues in a variety of foods. The fumigants and industrial chemicals determined are methyl bromide, methylene chloride, carbon disulfide, chloroform, 1,1-dichloroethane, ethylene dichloride, methyl chloroform, carbon tetrachloride, methylene bromide, propylene dichloride, 2,3-dichloropropene, trichloroethylene, 1,3-dichloropropylene, 1,1,2-trichloroethane, chloropicrin, ethylene dibromide, tetrachloroethylene, propylene dibromide, 1,1,2,2-tetrachloroethane, p-dichlorobenzene, o-dichlorobenzene, and 1,2-dibromo-3-chloropropane. Except for the latter three, the fumigants are determined at 90 degrees C on 3.6 m 20% loaded OV-101 columns with electron-capture and Hall-electroconductivity detectors. The other 3 compounds (o-dichlorobenzene, p-dichlorobenzene, and 1,2-dibromo-3-chloropropane), which elute beyond 30 min on the above columns, are determined at 90 degrees C on 1.8 m 5% loaded OV-101 columns with the same detectors. The ng/g-level fortifications have an overall mean analyte recovery of 70% and a coefficient of variation of 40%. The variety of foods examined includes both fatty and nonfatty food types (e.g., off-the-shelf cooked and uncooked grain-based items, dairy products, fresh and canned fruits and vegetables, and meats). Samples are extracted and cleaned up according to fat content and food type. Samples containing less than 71% fat are extracted by using an aqueous: nonaqueous shakeout (20% acetone solution under isooctane). Most extracts (isooctanes) are analyzed directly. Extracts from samples containing from 21 to 70% fat (e.g., ground beef, pecans, and corn chips) are cleaned up further on micro-Florisil columns to remove excess fat. A few other samples containing more than 71% fat or oil (e.g., butter, salad dressing, and vegetable oil) are diluted directly in isooctane and, depending on the degree of dilution, can be cleaned up further on micro-Florisil columns. Also, clear beverages (e.g., soda and tea) are extracted directly with isooctane. These extraction and cleanup techniques were tested on 231 different table-ready foods. Three-hundred incurred residues of 10 different fumigants were found in 138 items examined; 93 items had no detectable residues. The main advantage of the method is rapid semiquantitative determination of multiple fumigants from all food types.

Determining multifumigants in whole grains and legumes, milled and low-fat grain products, spices, citrus fruit, and beverages.

Whole grain and legumes, milled and low-fat products, spices, citrus fruit, and dry beverage ingredients are leached with purified, acidified acetone-water solutions. Portions of these leachates are then back-extracted with purified isooctane. Liquid beverages are directly extracted with the isooctane. Six to 10 microL of each isooctane extract is then screened for 11 fumigant residues by gas chromatography (GC) using electron-capture and Hall electroconductivity detectors, and dual 20% OV-101 columns. Further confirmation of residue identity is done on 20% OV-225/20% OV-17 (2.5 + 1 mixed-bed) and 10% SP-1000 columns. The analytes determined include methyl bromide, methylene chloride, carbon disulfide, chloroform, ethylene dichloride, methyl chloroform, carbon tetrachloride, trichloroethylene, chloropicrin, ethylene dibromide, and tetrachloroethylene, using mixed-component reference solutions. Average recoveries from fortified grain range from 25 to 85%; methyl bromide and chloropicrin were recovered the least. Recoveries from the other kinds of food samples range from 43 to 111%. Advantages of this procedure are (1) clean sample extracts, (2) ppb detection limits, (3) residue stability, (4) relative speed, quality control, and safety of the analysis, and (5) results which gave an accurate picture of residual fumigants in grain and food products.

Gas chromatographic determination of fumigant residues in stored grains, using isooctane partitioning and dual column packings.

A gas chromatographic (GC) procedure for determining fumigants in grains was developed. Fumigants were leached from grain samples with the official AOAC method using acetone-water (5 + 1). They were then partitioned from the leachate with isooctane, yielding a dry, stable extract that was analyzed by GC. Fortified sample recoveries ranged from 90 to 100%. Two GC columns were used, 20% OV-101 and 20% OV-225/20% OV-17 (2 + 1). These columns gave dissimilar retention profiles and baseline resolution for the 7 fumigants investigated: chloroform, ethylene dichloride, carbon tetrachloride, trichloroethylene, chloropicrin, ethylene dibromide, and tetrachloroethylene. Further tests showed that grain samples could be screened for fumigant residues by direct injection of the acetone-water leachates obtained using the AOAC method.