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Five ml of formalin (10%) was added to one sample immediately after collection and this sample was refrigerated until testing, one to five days after collection, the other sample was frozen.
After collection, each blood sample was refrigerated and packed into coolers with ice packs, and at the end of each day shipped by overnight express courier to the Volatile Organics Laboratory at the Centers for Disease Control and Prevention CDCC; Atlanta, GA).
The centrifuge was run at 25°C for 10 minutes at 3,200 g, and the AWF was recovered from the collection cap, the volume was measured and recorded, and the AWF sample was refrigerated prior to assaying performed the same day.
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The serum was then frozen in liquid nitrogen before being stored in a refrigerator – fentanyl and ketamine samples were stored at -4°C whereas propofol samples were refrigerated at -20°C.
All the samples were refrigerated until further use.
Triplicates of blood and serum samples were refrigerated at 4°C until analysis.
All algal samples were refrigerated up to 24 h prior to processing.
Blood samples were refrigerated until the final sample from each participant was collected and then centrifuged at 1500g for 10 min (at 4 °C).
The samples were refrigerated at 4 °C until reaching the laboratory where the collected samples were filtered (Whatman 0.45 µm) and acidified with nitric acid in order to reach a pH below 2.0.
The other samples in the study were sealed and stored under refrigeration (4○C) at IRRI, fumigated for 72 h with Phostoxin, and shipped to the USDA ARS SRRC via courier where the samples were refrigerated until analyzed.
On arrival at the laboratory, mouthwash samples were refrigerated at 4°C or at least 40 minutes, and then centrifuged at 3500 rpm for 5 minutes in a Boeco C-28 centrifuge (Boeckel & Co ,Hamburg, Germany).
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