Hutchinson James

Rapid method for the simultaneous measurement of nicotine and cotinine in urine and serum by gas chromatography–mass spectrometry

A simple, sensitive, and rapid gas chromatographic-mass spectrometric method is described for the simultaneous detection and quantitation of nicotine and its metabolite, cotinine, in urine and serum. The analytes and their respective deuterated internal standards were extracted by liquid-liquid extraction coupled to centrifugation and evaporation. The detection limit of the assay was 0.16 ng/ml for both nicotine and cotinine. The limit of quantitation for each analyte was 1.25 ng/ml.

Analgesic responses to intrathecal morphine in relation to CSF concentrations of morphine-3,β-glucuronide and morphine-6,β-glucuronide

Abstract This study was performed to determine whether variations in analgesic responses to intrathecal morphine could be explained by cerebrospinal fluid (CSF) concentrations of morphine metabolites. Twenty-four CSF samples were collected at the beginning, middle and end of treatment periods in seven cancer patients with pain of malignant origin. CSF concentrations of morphine-3,β-glucuronide (M3G) and morphine-6,β-glucuronide (M6G) metabolites were measured by gas chromatography/mass spectrometry. Analgesic responses to morphine were estimated concurrent with CSF collection using a visual analog scale representing percentages of pain relief. Effective analgesia was defined as ≥ 75% pain relief. CSF concentration of M3G and M6G in the 24 samples were 722 ± 116 ng/ml and 699 ± 158 ng/ml, respectively. CSF samples were categorized into two groups: (1) those collected during effective analgesia (N = 14), and (2) those collected during ineffective analgesia (N = 10). M6G levels detected in group 1 samples (effective analgesia) were significantly greater than those found in group 2 samples (ineffective analgesia) (978 ± 243 ng/ml vs 309 ± 68 ng/ml, P M3G M6G ratios were not significant. It is concluded that CSF M6G may be indicative of effectiveness of analgesia in cancer patients subjected to intrathecal morphine.

Relationship between peripheral blood lymphocytes and their functional capacity in sarcoidosis

Abstract Sarcoidosis is a disease of unknown etiology characterized by granuloma formation in tissues and anergy to delayed hypersensitivity. In 34 sarcoidosis patients and 16 normal persons, T cells from peripheral blood were quantitated and their functional capacity assessed by sheep red blood cell (SRBC) rosette assay and mitogen response, respectively. Effect of Kveim-Siltzbach suspension (KSS) on rosette formation and its ability to induce blast formation were also examined. Normal spleen cell suspension (NSS) was used as control. The results show that 84% of patients had suppressed T-cell numbers, while 79% had suppression of both active (rosettes formed immediately on incubation with SRBC) and total (rosettes formed after a 1-hr incubation on ice) T cells. Mean active was 30% in sarcoidosis patients vs 44% in controls, while mean total was 48% in sarcoidosis patients vs 64% in controls, while mean total was 48% in sarcoidosis patients vs 64% in control subjects. Suppressed mitogen response to phytohemagglutinin was seen in only 34% of patients and of this number, 90% had decreased T-cell numbers (active and total), suggesting that T-cell suppression does not necessarily reflect functional abnormality in these cells; however, the reverse is true. KSS caused no blastogenesis or change in rosette formation when incubated with lymphocytes from sarcoid patients (KSS active 36%; total 48%: NSS active 34%; total 48%). Either the cells are insensitive to KSS stimulation or sensitive cells have migrated to sites of granuloma formation.