Vitamin B2 interference with TDx drugs-of-abuse assays.

Kunsman GW, Levine B, Smith ML.

Bexar County Medical Examiner's Office, San Antonio, TX, USA.

Migraine is a headache condition found in significant frequency in the general population. One recent study has shown that riboflavin, Vitamin B2, is an effective prophylactic treatment for this headache condition. One subject in a recent study conducted by the Division of Forensic Toxicology, Armed Forces Institute of Pathology (AFIP) was taking 200 mg of riboflavin twice daily for the prevention of migraine headaches. When that subject's urine was tested using Abbott TDx drugs-of-abuse assays a number of tests resulted in a MX BKG error and all samples had BLK I values greater than those observed with normal urine specimens. The MX BKG error occurs when the BLK I value is greater than the upper limit determined by the manufacturer for a particular assay. High BLK I values may result if the specimen being analyzed contains a fluorophore that will compete with the fluorescein-labeled antibody used in the assay. This error serves as a notification that an interfering substance may be present and the assay is not performing according to manufacturer-specifications. Upon termination of riboflavin therapy the subject's BLK I values began to decrease within 60 h of the last 200 mg dose. A second subject began chronic riboflavin use to confirm this interferent effect. Elevated BLK I values resulted within 3 h of a single 200 mg dose and MX BKG errors occurred 1 h after a second 400 mg dose. No false negative results were noted with either subject (both subjects used butalbital and the first subject also used hydrocodone and diazepam during the study), suggesting that riboflavin is not an adulterant. Riboflavin use, however, does interfere with the TDx DAU assays and may result in quantitative values being determined which are of questionable validity in the face of an elevated BLK I value or may result in only an MX BKG error and no quantitative value reported. It is unclear if the interfering fluorophore is simply riboflavin itself or a combination of riboflavin and its metabolic products. Results obtained on urine samples collected from individuals using prophylactic riboflavin for migraine prevention and analyzed by TDx may be of questionable validity. Such samples may require analysis utilizing another immunoassay technique that does not employ a fluorescein-labeled antibody.

Source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=&dopt=Abstract butalbital fioricet barbiturate




Use of a statistically designed experimental approach to optimize the propylketal derivatization of barbiturates.

Kushnir MM, Urry FM.

ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, UT 84108, USA.

The derivatization of barbiturates with dimethylformamide dipropylacetal and dimethylformamide diisopropylacetal is studied with respect to the optimization of reaction recovery and reliability. A second-order orthogonal experimental design is utilized in order to obtain regression equations for the reaction recovery dependence on the derivatization solution composition, incubation temperature, and time for amobarbital, butalbital, pentobarbital, phenobarbital, and secobarbital. Regression equations for the effect of incubation temperature and time on the derivative recovery and the optimum conditions for derivatization recoveries are obtained. Differences in the phenomena of the derivative formation are evaluated between the two derivatizing reagents and the barbiturates. Based on the analysis of the obtained equations, it is concluded that the dipropylketal derivative of barbiturates is superior in comparison with diisopropylketal when considering the milder conditions of the reaction, absence of sudden changes in the recovery with a variation in the derivatization parameters, and reliability for the simultaneous testing of the barbiturates. A method for the routine testing of the barbiturates by gas chromatography-mass spectrometry in urine specimens is included.

Source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=&dopt=Abstract butalbital fioricet barbiturate




Isotopic analogues as internal standards for quantitative analyses of drugs and metabolites by GC-MS--nonlinear calibration approaches.

Whiting TC, Liu RH, Chang WT, Bodapati MR.

Department of Justice Sciences, University of Alabama at Birmingham, 35294-2060, USA.

In order to achieve accurate quantitation of drugs and metabolites (analytes) in complex matrices, 2H- (and less commonly 13C-) labeled analogues of the analytes are now routinely adapted as the internal standards (IS) using linear calibration models to fit data generated by selected ion monitoring gas chromatography-mass spectrometry (GC-MS) protocols. In this study, the effects of cross-contribution (contribution of the IS to the intensity of the ion designated for the analyte and vice versa) on the linearity of the calibration data are examined. Nonlinear approaches that may address this problem are also studied. Two ion pairs (one with least and one with significant cross-contribution) from each of the following analyte/IS pairs are used as the exemplar systems for this study: butalbital/13C4-butalbital, butalbital/2H5-butalbital, secobarbital/13C4-secobarbital, and secobarbital/2H5-secobarbital. Analyte/IS ion intensity ratios of a series of standard solutions are correlated with the analyte/IS concentration ratios using one-point, multiple-point (unweighted and weighted) linear, and hyperbolic functions. The one-point calibration approach produces excellent calibration results in treating data derived from ion pairs with no significant cross contribution. In cases where significant cross-contribution exists, results derived from the one-point approach show, as expected, significant deviations at both ends of the concentration range. With the cross-contribution phenomenon accounted for, the hyperbolic calibration model is clearly more effective in fitting calibration data at both the lower and higher analyte concentration ends, thus significantly lowering the detection limit and extending the calibration range to a higher level. However, the calibration range cannot be extended indefinitely. At the low concentration end, noise-to-signal ratio and the cross-contribution of the IS to the intensity of the ion designated for the analyte, however insignificant, will incrementally reduce the quality of the observed ion intensity and intensity ratio data. At the high concentration end, detection saturation and the cross-contribution of the analyte to the intensity of the ion designated for the IS, however insignificant, will incrementally decrease the "slope" of the calibration curve. Thus, acceptable sensitivity (increase in analyte/IS ion-pair intensity ratio per unit increase in analyte concentration) of the calibration curve will become the limiting factor.

Source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=&dopt=Abstract butalbital fioricet barbiturate




Quantitative design of optimal analgesic combination of acetaminophen, caffeine, and butalbital.

Zheng QS, Wang XW, Gui CQ, Sun RY.

Laboratory of Structure Biology, School of Life Science, University of Science and Technology of China, Hefei 230021, China. zhengqs mail.ahwhptt.net.cn

AIM: To quantitatively seek an optimal analgesic combination of acetaminophen (Ace), butalbital (Bul), and caffeine (Caf), and to characterize the pharmacodynamics of interaction among the three drugs. METHODS: The models of acute inflammatory pain in carrageenin-injected rats were applied to measure the vocalization threshold to paw pressure. Six groups with different ratios and doses were set to seek an optimal combination of Ace, Caf, and Bul, analyzed by the weighted modification method. Based on the ratio and doses in the optimal combination, four continuous doses were set to analyze the interactions of therapeutic effects by the reflection method. The interaction of the acute toxicity was evaluated by the parameter method. RESULTS: According to the degree of importance to the combined analgesic effect, Ace > Caf > Bul; Ace showed a significant dose-response relationship, whereas in Caf and Bul, this relationship was not apparent. A new combination was obtained by the theoretical analysis and confirmed further by experimentation. Namely, at a ratio of 8.6:1:1.5 Ace + Caf + Bul (240 + 28 + 42 mg/kg, ig) was an optimal combination. Both Caf and Bul had a synergism to Ace, but Caf was a stronger synergist in contrast to Bul. Such synergism increased the therapeutic effects in the range of Ace 153.6 - 300 mg/kg combined with Caf 17.9 - 35 mg/kg and Bul 26.8 -5 2.5 mg/kg (8.6:1:1.5). However, the dose of Ace + Caf + Bul at 300 + 35 + 52.5 mg/kg resulted in sedation in rats. The peak latency was approximately 1 h for all four continuous doses, but the peak amplitude was dose-related, and the duration of the therapeutic effect was less than 2 h. The acute toxicity of the three drugs in combination remained the same. CONCLUSION: Ace + Caf + Bul at a dose of (240 + 28 + 42) mg/kg (ig) results in an optimal combination. The therapeutic window of the combination is located in the range of Ace (153.6 - 240 mg/kg)+Caf (17.9 - 28 mg/kg)+Bul (26.8 - 42 mg/kg) (8.6:1:1.5). Caf has a stronger synergism with Ace than Bul.

Source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=&dopt=Abstract butalbital fioricet barbiturate