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No. 10 (PC 10)
Direct analysis of chemical substances in biofluids without sample pretreatment
Ritsuko Wakayama 1 , Yasushi Iwasaki 1 , Motoaki Kamachi 1 , Natsumi Sawa 2 , Katsuko Hara 2 ,
Yutaka Komiyama 2
1 Showa Denko K.K, Japan, 2 Department of Clinical Sciences and Laboratory Medicine, Kansai Medical
University Takii hospital, Japan
Sample pretreatment is often a time consuming and laborsome step which can be a restricting factor for
the rapid analysis. This is especially true for the analysis of drugs in biofluids containing many
interfering matrices. Therefore, we developed a direct analysis method for drugs in biofluids, using a
polymer-based reversed-phase HPLC column which has a unique feature of retaining hydrophilic target
compounds while excluding large biological matrices prior to the elution of targets. Following
conditions were used. Column; Shodex ODP2 HP-4D (4.6 mm ID x 150 mm L) and ODP2 HP-4E (4.6
mm ID x 250 mm L) from Showa Denko K.K. Eluent; 0.1% TFA/CH3CN = 93/7. Flow rate; between
0.3 and 0.6 mL/min. Injection volume; from 5 to 10 uL. Detector; UV with wavelength between 215 and
272 nm. Column temperature; 40°C. Standards were prepared in the eluent. Optimal conditions for each
target and biofluids varied slightly. For the identification purpose, each peak were fractionated, and then
injected directly to a MS/MS. The components of typical cold medicines were detected without the
influence of biological matrices. Blood serum, urine, and gastric juice samples were collected from
suspected drug-poisoned patients transported to the emergency and critical care medicine. Several
components in cold medicine were detected in one patient’s serum: 44.1 ug/mL acetaminophen, 11.0
ug/mL dihydrocodein phosphate, and 22.6 ug/mL aspirin. A small amount of salicylamide and salicylic
acid was also found. Theophylline and risperidone were detectable in other patients’ serum samples. In
one case of accidental insecticide poisoning, imidacloprid was detected in the patient’s gastric juice.
The presented method using the ODP2 HP column provides a simple and rapid analysis of drugs in
biofluids without the necessity of complex sample pretreatment. This method is applicable for the
diagnosis and treatment of patients who are poisoned or overdosed counter medicine.
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