Coffee contains cholinomimetic compound distinct from caffeine. I: Purification and chromatographic analysis.

Both regular and decaffeinated coffees were found to have cholinomimetic actions when tested in urethane-anesthetized rats. These actions were distinct from those of caffeine and reversible by atropine. The bioactive fraction was purified from alcoholic extracts of instant decaffeinated coffee by liquid column chromatography and preparative TLC. The purified compound showed similar pharmacological actions as the starting material. Chromatographic behavior was further characterized by analytical TLC and HPLC. Chromatographic analyses of extracts of green coffee beans and roasted ground coffees showed that the cardioactive compound was only present in roasted coffees. Similar analyses of other commonly consumed beverages, including teas and cocoa, showed that this compound was not present in beverages besides coffee.

Cholesterol-raising factor from boiled coffee does not pass a paper filter.

Previous studies have indicated that consumption of boiled coffee raises total and low density lipoprotein (LDL) cholesterol, whereas drip-filtered coffee does not. We have tested the effect on serum lipids of consumed coffee that was first boiled and then filtered through commercial paper coffee filters. Sixty-four healthy volunteers consumed six cups per day of this boiled and filtered coffee for 17 days. Then, they were randomly divided into three groups, which, for the next 79 days, received either unfiltered boiled coffee (lipid content, 1.0 g/l), boiled and filtered coffee (0.02 g lipid/l), or no coffee. Serum total cholesterol levels rose by 0.42 mmol/l (16 mg/dl; 95% confidence interval [CI], 0.14-0.71), LDL cholesterol levels by 0.41 mmol/l (16 mg/dl; 95% CI, 0.16-0.66), and apolipoprotein B levels by 8.6 mg/dl (95% CI, 3.8-13.4) in those who consumed boiled coffee relative to those who consumed boiled and filtered coffee. Responses of triglycerides, high density lipoprotein cholesterol, and apolipoprotein A-I did not differ significantly among these groups. No significant effects on serum lipid levels were found in the boiled and filtered coffee-consuming group compared with those who drank no coffee. In subjects who drank boiled coffee, serum campesterol level, an indicator of cholesterol absorption, remained constant. The serum lathosterol level, an indicator of cholesterol synthesis, increased by 11% (p less than 0.05), but the lathosterol to cholesterol ratio did not change. We propose that paper filters of the type used for drip-filtered coffee retain the lipid present in boiled coffee and in that way remove the hypercholesterolemic factor.(ABSTRACT TRUNCATED AT 250 WORDS)

Determination of food contamination by mineral oil from jute sacks using coupled LC-GC.

Jute fibers are treated with about 5-7% of a high boiling mineral oil fraction ("batching oil") to render them flexible for making fabrics. Foods transported in jute bags are contaminated by this batching oil. A method involving automated on-line LC-GC is described for determining these hydrocarbons in various foods. Complete transfer of the LC fraction to GC is presupposed for obtaining the required sensitivity. Results are given for nuts, coffee, cocoa products, and rice. Contamination ranged between about 5 and 500 ppm.

Synthesis and availability of niacin in roasted coffee.

The coffee bean contains about 1% of trigonelline that is demethylated at temperatures approaching 200 degrees C; it is partially converted into nicotinic acid. This operation is mainly proportional to the severity of dry heat treatment; various other physico-chemical factors also influence the synthesis of niacin during the roasting. The niacin content of weakly roasted commercial coffee is about 10 mg/100 g (American coffee) and it reaches 40 mg in heavy roasted coffees, i.e. Italian coffee. Caffeine-free coffee is lower in niacin than the corresponding raw coffee. The drinking retains 85% of the niacin formed during roasting; it is totally available for the organism and can constitute a noticeable part of the daily supply in niacin.

A randomized study on coffee and blood pressure.

The effects of coffee on blood pressure and heart-rate and the mediating effect of two common brewing methods, were studied in a randomized trial in 107 young, normotensive adults. After a three-week run-in period, subjects were randomly assigned to one of three groups, receiving either (1) 4-6 cups filtered coffee per day, (2) 4-6 cups boiled coffee per day, or (3) no coffee at all for a period of nine weeks. Because all participants consumed filtered coffee before the trial, the group continuing on filtered coffee was considered as the reference group. Both systolic (SBP) and diastolic blood pressure (DBP) decreased in the abstinence group. Compared to the filter group, only the fall in SBP after 9 weeks was statistically significant, -6.1 mmHg (95% confidence limits -10.8, -1.4). After adjustment for SBP at baseline and body weight change during the study, the observed reduction decreased, to -3.4 mmHg (-7.1, 0.3). The patterns for SBP and DBP were remarkably similar in the groups using either filtered coffee or boiled coffee. After 9 weeks of boiled coffee, mean changes from baseline for SBP and DBP were 0.4 mmHg (-3.7, 4.5) and -0.1 mmHg (-3.4, 3.2), compared to the filter group. The heart rate showed a slight, non-significant decrease in the abstinence group. In conclusion, these findings suggest that abstinence from coffee for a period of several weeks may slightly reduce blood pressure in young normotensive subjects.

Analysis of hydrogen peroxide in freshly prepared coffees.

Coffee has been shown unequivocally to be genotoxic in vitro, but no genotoxicity has been seen in vivo testing. Since the in vitro genotoxicity appears to be dependent on hydrogen peroxide, it is important to know whether hydrogen peroxide is present in prepared coffee and whether it is being formed during the in vitro testing. We have devised a procedure to measure hydrogen peroxide in prepared coffee using disposable reversed-phase columns to decolourize the coffee and retain its catechols. Hydrogen peroxide was assayed in the eluate from the columns by two chromogenic methods: horseradish-peroxidase-mediated oxidation of phenol red and non-enzymatic oxidation of iodide. Three brands of brewed and instant coffee prepared in the manner recommended to the consumer were studied. Although six of the twelve preparations of coffee contained 3-29 microM-hydrogen peroxide, in the other six none could be detected. Sampling, batch, and aging effects may contribute to the variability in the testing, but there is no indication of greater than 100 microM-hydrogen peroxide levels in freshly prepared coffees, as reported in the literature using other methods. Hydrogen peroxide did form slowly in prepared coffee as the beverage became oxygenated, but it formed quickly if the coffee was diluted by addition to an oxygen-containing solution at neutral pH and then incubated at 37 degrees C. These results strongly suggest that adventitious formation of hydrogen peroxide is a confounding factor in the analytical and in vitro genotoxicological testing of coffees.

Mass spectrometric confirmation of the presence of N-nitrosopyrrolidine in instant coffee.

Traces of N-nitrosopyrrolidine (NPYR) may occur in some samples of both instant coffee and fine-ground roasted coffee. The identity of NPYR in 2 samples of instant coffee was confirmed by mass spectrometry as well as by liquid chromatography-thermal energy analysis. A 2-step cleanup procedure, involving fractionation on basic alumina followed by gradient elution on reverse-phase C18 cartridge, is described that allows full-scan mass spectrometric confirmation of NPYR in tested samples.

Caffeine as main interfering compound in radioimmunoassay of aflatoxin B1 in coffee samples.

The content of caffeine in coffee extracts prepared for radioimmunoassay of aflatoxin B1 was determined by gas chromatography. The extracts from coffee beans and decaffeinated coffee contained 1.76-4.60 and 0.71-0.85 g caffeine/kg, respectively. These concentrations of caffeine caused false results in radioimmunoassay of aflatoxin B1 in the range 1.0-2.8 micrograms/kg for coffee beans and 0.3-0.4 micrograms/kg for decaffeinated coffee.

A study of the contamination by ochratoxin A of green and roasted coffee beans.

A study was performed to evaluate the contamination by ochratoxin A in coffee beans. Twenty-nine samples of green coffee were collected from large lots of material by representative sampling. The analyses of green coffee samples showed a significantly high contamination percentage (58%) ranging from 0.2 to 15 micrograms/kg. Naturally and artificially contaminated samples were roasted at different operation times (5-6 min) to verify the percentage of destruction of the mycotoxin. The percentage ranged from 48% to 87% and from 90% to 100% in artificially and naturally contaminated samples respectively. The beverages prepared from artificially contaminated coffee using the most common types of coffee makers showed no residues of ochratoxin A.

Contribution of coffee aroma constituents to the mutagenicity of coffee.

About 40 coffee aroma constituents belonging to the classes of dicarbonyls, sulphur-containing compounds, furfuryls, N-heterocyclics and others were systematically evaluated in three Ames tester strains. Only aliphatic dicarbonyl compounds showed notable direct mutagenic activity, which mainly affected 'base-pair substitution' in Ames tester strains TA100 and TA102. Very weak effects were also seen with some N-heterocyclics, mainly affecting frameshift tester strain TA98 upon metabolic activation. However, it was shown that these N-heterocyclics do not contribute substantially to the mutagenicity in coffee. The hydrogen peroxide and methylglyoxal contents of coffee were determined up to 26 hr after preparation. Their concentrations tended to decrease whereas mutagenic activity decreased significantly with time in tester strains TA100 and TA102. It is concluded that several highly labile coffee constituents contribute to the bacterial mutagenicity and also that the synergism between hydrogen peroxide and methylglyoxal is not the main factor. The absence of coffee mutagenicity/carcinogenicity in rodents with these highly reactive coffee aroma compounds can be explained in part by detoxification of microsomal enzyme systems.

Application of high performance liquid chromatography to the analysis of some non-volatile coffee components.

High performance liquid chromatography (HPLC) was applied to the analysis of caffeine, trigonelline, nicotinic acid and sucrose in Arabica and Robusta coffee. Green and roasted coffee samples were used in this study and the degradation of sucrose and trigonelline, with the formation of nicotinic acid, was followed during roasting. Caffeine did not undergo significant degradation with only 5.4% being lost under severe roasting. Sucrose was degraded rapidly during processing with light roasting producing a 97% loss and dark roasting degrading it completely. Loss of trigonelline was strongly dependent on the degree of roasting, being higher in the Robusta coffee. Trigonelline degradation was associated with nicotinic acid formation both in the Arabica and Robusta coffees as a consequence of the roasting process. Trigonelline and sucrose were determined simultaneously by partition chromatography and detection with the mass detector. Determination of caffeine was carried out using reversed phase chromatography and nicotinic acid by ion-pair reversed phase chromatography. Detection in both cases was achieved using an ultraviolet detector at 272 nm or 254 nm, respectively. HPLC showed adequate precision and accuracy for routine analyses. In addition, the methods used were more rapid and simple than traditional procedures. HPLC appears to be a suitable technique for quality control in the coffee industry, and for fundamental investigation on the mechanisms involved in the roasting process.

Application of high performance liquid chromatography to the analysis of some non-volatile coffee components

High performance liquid chromatography (HPLC) was applied to the analysis of caffeine, trigonelline, nicotinic acid and sucrose in Arabica an Robusta coffee. Green and roasted coffee samples were used in this study and the degradation of sucrose and trigonelline, with the formation of nicotinic acid, was followed during roasting. Caffeine did not undergo significant degradation with only 5.4% being lost under severe roasting. Sucrose was degraded rapidly during processing with light roasting producing a 97% loss and dark roasting degrading it completely. Loss of trigonelline was strongly dependent on the degree of roasting being higher in the Robusta coffee. Trigonelline degradation was associated with nicotinic acid formation both in the Arabica and Robusta coffees as a consequence of the roasting process. Trigonelline and sucrose were determined simultaneously by partition chromatography and detection with the mass detector. Determination of caffeine was carried out using reversed phase chromatography and nicotinic acid by ion-pair reversed phase chromatography. Detection in both cases was achieved using an ultraviolet detector at 272nm or 254nm, respectively. HPLC showed adequate precision and accuracy for routine analyses. In addition, the methods used were more rapid and simple than traditional procedures. HPLC appears to be a suitable technique for quality control in the coffee industry, and for fundamental investigation on the mechanisms involved in the roasting process (AU) s

Genotoxicity testing of Maillard reaction products.

Since the development of short-term genotoxicity tests such as the Ames assay, the mutagenicity of Maillard reaction products has been tested extensively. Some products have exhibited strong activity. For example, one of the earliest studies demonstrated some mutagenic activity in a dichloromethane extract of a D-glucose/ammonia Maillard model system. Many researchers have attempted to pinpoint the principal chemical(s) of mutagenicity of the Maillard products using various sugar-amino acid browning model systems over last two decades. However, no mutagenic individual Maillard product has been isolated and identified. Nitrite has been also used as a reactant in browning reaction model systems, primarily to investigate the formation of potentially mutagenic or carcinogenic N-nitroso compounds. Recently some potent mutagens isolated from pyrolyzed amino acids or proteins have begun to receive attention as Maillard reaction products.