Biological monitoring of hospital pharmacy personnel occupationally exposed to cytostatic drugs: urinary excretion and cytogenetics studies.

For evaluation of the risk borne by hospital pharmacy personnel exposed to antineoplastic agents, the incorporation of cyclophosphamide, ifosfamide, and platinum-containing drugs was quantified by the determination of urinary concentrations. In addition, the induction of micronuclei (MN) and sister-chromatid-exchange (SCE) rates in peripheral blood lymphocytes were studied for correlation with the urinary excretion of cytostatic drugs. Cyclophosphamide and ifosfamide were determined in 24-h urine samples using gas chromatography with electron capture (detection limit 2.5 micrograms/l). Voltammetric analysis enabled the determination of platinum concentrations of 4 ng/l. Heparinized blood (20 ml) was drawn and lymphocytes were cultured for MN and SCE studies. In all, 13 hospital pharmacists and pharmacy technicians regularly involved in the preparation of cytostatic drugs participated in this investigation (7 persons represent a follow-up group). All subjects applied standard safety precautions, including the use of a vertical laminar air-flow hood, protective gowns, and latex gloves. On the day of urine sampling an average of 4,870 mg cyclophosphamide, 5,580 mg ifosfamide, and 504 mg platinum-containing drugs were handled. The excretion of 5 and 9 micrograms cyclophosphamide/l urine was measured in two samples, respectively. An elevated level of urinary platinum was found in one pharmacist (22.3 ng/g creatinine) in comparison with a nonexposed control group. Mean frequencies of MN and SCE did not differ significantly between the drug exposed group and control group. The employees who had incorporated chemotherapeutic agents were part of the follow-up group and, thus, particularly cautious and sensitive to a possible hazard. The results emphasize the necessity of improving personal protection of hospital pharmacy personnel occupationally exposed to cytostatic drugs and support the importance of biological monitoring. In an ongoing project in our department the sources of contamination are being investigated parallel to biological monitoring so as to determine critical situations and improve personal protection.

Biological monitoring of cyclophosphamide and ifosfamide in urine of hospital personnel occupationally exposed to cytostatic drugs.

The occupational exposure of 21 nurses and pharmacy personnel from eight hospitals to cyclophosphamide and ifosfamide was determined by quantifying the amount of the drugs handled and by measuring the urinary excretion of the unmetabolised substances. Preparing antineoplastic drugs for intravenous treatment was the major task of all study participants. Twenty four hour urine was collected on days when cyclophosphamide and/or ifosfamide were mixed, on average 3900 mg cyclophosphamide and/or 5900 mg ifosfamide. The analyses were performed by gas chromatography with electron capture, detection limit 2.5 micrograms/24 hour urine. Despite standard safety precautions, including a vertical laminar air flow safety cabinet and gloves, cyclophosphamide was detected in 12 of 31 and ifosfamide in four of 21 urine samples on days when the drugs were handled. Excretion of cyclophosphamide ranged from 3.5 to 38 micrograms/24 h (mean 11.4 micrograms/24 h) urine, ifosfamide from 5 to 12.7 micrograms/24 h (mean 9 micrograms/24 h) urine. Based on an excretion rate of 11.3% unmetabolised cyclophosphamide, the average amount excreted corresponded to an uptake of 101 micrograms cyclophosphamide. For ifosfamide the mean quantity incorporated was 20 micrograms assuming that 45% of the drug was excreted. Pertaining to the doses handled, the uptake of cyclophosphamide and ifosfamide was estimated to be approximately 0.0025% and 0.0004% respectively. Despite time-consuming purification procedures, gas chromatographic analysis is a suitable method for monitoring personnel occupationally exposed to cyclophosphamide and ifosfamide and is a major contribution to the evaluation of potential health risks of exposed personnel.

Urinary platinum in hospital personnel occupationally exposed to platinum-containing antineoplastic drugs.

Urinary platinum levels of 21 nurses and hospital pharmacy personnel occupationally exposed to platinum containing antineoplastic drugs were determined in 24-h urine by voltammetric analysis after UV photolysis. All study participants applied standard safety measures, including a vertical laminar air-flow cabinet and latex gloves. The amount of platinum-containing drugs prepared for intravenous application ranged from 40-3260 mg/day. Urinary platinum was detected in 9 of 52 urine samples collected on days when platinum-containing drugs were mixed (limit of determination 4 ng/l). In comparison with a non-exposed control group, elevated urinary platinum levels were found in one pharmacist (35 ng/g creatinine) and one pharmacy technician (28 ng/g creatinine). The pharmacist's urinary platinum remained elevated after 2 days without occupational exposure to antineoplastic drugs. The urinary platinum level of the pharmacy technician dropped considerably after several weeks without handling cytostatic drugs. Voltammetric detection of urinary platinum is a highly sensitive method suitable for biological and environmental monitoring.

Comparison of cytolytic and proliferative activities of human gamma delta and alpha beta T cells from peripheral blood against various human tumor cell lines.

Peripheral blood lymphocytes (PBLs) from healthy donors were more than 95% enriched for gamma delta T cells. V gamma 9/V delta 2 was the predominant T-cell subset and represented more than two thirds of the isolated gamma delta T cells. When activated by surface-immobilized anti-TCR delta 1 monoclonal antibody (MoAb) during 5 days of culture, gamma delta T cells demonstrated a significant fourfold to 15-fold stronger cytolytic activity against various tumor cell lines than did PBL polyclonal alpha beta T cells analogously activated by surface-immobilized anti-CD3 MoAbs. Blocking experiments with specific MoAbs demonstrated that the cytolytic activity of gamma delta T cells was non-major histocompatibility complex (MHC) restricted and did not involve the gamma delta T-cell antigen receptor (TCR) but that cytolytic activity was dependent on LFA-1 beta/ICAM1 interaction. The proliferative responses of gamma delta and alpha beta T cells to Ovcar-3 tumor cells and irradiated allogeneic PBLs were inhibited by anti-HLA-A,B,C MoAb. Our findings suggest that gamma delta T cells activated via the TCR have a significant advantage in non-MHC-restricted lysis of various tumor cell lines over PBL alpha beta T cells stimulated analogously with anti-CD3, which may be important in terms of applicability of activated gamma delta cells for adoptive immunotherapy of metastatic cancers.