Extraction and detection of mitragynine in Kratom leaves by high-performance liquid chromatography.

Mitragyna speciosa, also known as Kratom, is an evergreen tree native to Southeast Asia that has been used in traditional medicine to relieve pain. The primary active alkaloid, mitragynine, found in Kratom leaves affects the same µ-receptor as the opioids. The potential effects of Kratom leaves on humans draw international concerns. The purpose of this study is to develop an efficient extraction method for mitragynine from dried Kratom leaves. Mitragynine was extracted from grounded Kratom leaves by liquid-liquid extraction and detected by an HPLC system with a C-18 column. The extracted sample was eluted with a mobile phase of 0.05% formic acid and acetonitrile (50:50, v/v), flow rate = 1.0 mL/min. Mitragynine was detected at 223 nm. The amount of mitragynine in four different strains of Kratom leaves was determined (wt/wt). Intra-day (n = 3) and inter-day(n = 3) studies show excellent precision with RSD < 2.1% and RSD < 3.6% respectively.

Minimal effect of acute caffeine ingestion on intense resistance training performance.

The primary aim of the study was to determine the efficacy of acute caffeine intake to enhance intense resistance training performance. Fourteen resistance-trained men (age and body mass = 23.1 ± 1.1 years and 83.4 ± 13.2 kg, respectively) who regularly consumed caffeine ingested caffeine (6 mg · kg(-1)) or placebo 1 hour before completion of 4 sets of barbell bench press, leg press, bilateral row, and barbell shoulder press to fatigue at 70-80% 1-repetition maximum. Two minutes of rest was allotted between sets. Saliva samples were obtained to assess caffeine concentration. The number of repetitions completed per set and total weight lifted were recorded as indices of performance. Two-way analysis of variance with repeated measures was used to examine differences in performance across treatment and sets. Compared to placebo, there was a small but significant effect (p < 0.05) of acute caffeine intake on repetitions completed for the leg press but not for upper-body exercise (p > 0.05). Total weight lifted across sets was similar (p > 0.05) with caffeine (22,409.5 ± 3,773.2 kg) vs. placebo (21,185.7 ± 4,655.4 kg), yet there were 9 'responders' to caffeine, represented by a meaningful increase in total weight lifted with caffeine vs. placebo. Any ergogenic effect of caffeine on performance of fatiguing, total-body resistance training appears to be of limited practical significance. Additional research is merited to elucidate interindividual differences in caffeine-mediated improvements in performance.

Rapid colorimetric screening test for gamma-hydroxybutyric acid (liquid X) in human urine.

A rapid colorimetric test for the detection of gamma-hydroxybutyric acid (GHB) is described. The ferric hydroxamate test for ester detection has been adapted to detect GHB in human urine samples from a healthy female and a healthy male subject. The assay can be performed within 5 min and with a GHB detection limit of 0.5 mg/ml when 0.3 ml of human urine is used and a GHB detection limit of 0.1 mg/ml when 1 ml of human urine is used. The colored complex indicating the presence of GHB is purple according to the assay conditions. Test results are free from the interference by alcohol, phenolic compounds and other biological chemicals under the assay conditions. In addition, the colorimetric test is free from the potential false-positive test result that could result from physiological concentrations of GHB.

HPLC separation of tetracycline analogues: comparison study of laser-based polarimetric detection with UV detection.

A sensitive and specific high-performance liquid chromatography (HPLC) method based upon laser-based polarimetric detection is developed for the determination of six tetracycline analogues. By interfacing the laser-based polarimeter online with an HPLC system, the specific rotation of each analogue is obtained as compounds elute from the separation system. The six structurally similar tetracycline analogues exhibit significant differences in specific rotations. The experiments suggest that specific rotation can be useful in identifying closely related tetracycline analogues. Linear relationships are found to be in the range of 0.342-0.0043 mg for the tetracycline analogues. Five of the six analogues exhibit excellent linearity (R(2) value >/= 0.99). The polarimetric results are compared with UV detection. The HPLC-laser-based polarimetric detection instrument is able to quantitate the studied tetracycline analogues with high precision, accuracy, and sensitivity, which make it useful for the development of a standard method for the determination of tetracyclines in biological specimens. The performance of the HPLC-polarimetric system for the analysis of tetracyclines in a biological matrix is evaluated. The selectivity of polarimetric detection provides a distinct advantage in the analysis of tetracycline analogues in milk. The HPLC-polarimetric system provides a rapid and sensitive technique that involves minimal sample cleanup and pretreatment for the analysis of tetracyclines in milk.