EXPERIMENTAL METHODS 



Apparatus 



All atomic absorption analyses were made using a Perkin-Elmer atomic 

 absorption unit (Model 360) coupled to a Perkin-Elmer heated graphite 

 atomizer (Model No. HGA-2100). The weight measurements of the worms 

 were made with a Perkin-Elmer microbalance (Auto balance Model No. 

 AD-2Z). Freeze-drying of the worms was accomplished using a Virtis (Model 

 No. 10-145MR-BA) lyophilizer. Low temperature ashing of the samples was 

 done with a L.F.E. (Model LTA-505) low temperature asher. 



Reagents and Materials 



Ultrex HNO3 was used throughout the analytical elution and dissolution 

 procedures. Copper standards were made up from a stock solution of ALPHA 

 atomic absorption standard copper. All 2/5 dram polyethylene snap-cap vials 

 were acid washed in concentrated HNO3 for two days, soaked in demineralized 

 water for two days, and finally rinsed five times with copious quantities of 

 demineralized water. The vials were allowed to air dry in a class-100 clean 

 bench. 



Procedure 



The 61 polychaete specimens used in this study were raised in the 

 laboratory. Complete details of the methods to raise the worms are given by 

 Pesch and Morgan (2). Live polychaete samples were removed from the 

 seawater tanks with the aid of a nylon brush and rinsed in control seawater for 

 approximately one minute, and then placed in precleaned polyethylene vials 

 (1.2 ml capacity) fitted with snap-caps. The samples were frozen and then 

 freeze-dried for 24 hours. The freeze-dried worms were then weighed. The 

 average weight was 8.7 ± 4.7 mg. One ml of 5 percent HNO3 was added to the 

 worms in their respective vials. The sample vials were capped and allowed to 

 stand at room temperature for two days. The worms were then transferred 

 from the first extraction vial (A) to precleaned tared vials (B) with the aid of a 

 teflon fiber. The tared vials (B) containing the wet acid leached worms were 

 again weighed. One ml of 5 percent HNO3 was added to the B vials. The (B) 

 vials were capped and allowed to stand at room temperature for two days. The 

 worms were again transferred to pretared vials (C) and weighed wet. The 

 worms were then freeze-dried and weighed again. During these transfer steps 

 care was taken so that the worms did not disintegrate. The insoluble worm 

 carcasses were then destroyed by low temperature ashing. The freeze-dried 

 worms were inserted into teflon beakers (10 ml capacity) and ashed for 24 

 hours at the following conditions: O9 flow 50 cc/min; and RF power of 50 



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