Free Amino Acid Content — Schafer 
51 
mens of Pachygrapsus crassipes live on the floats 
and piling just above the water line. Since the 
area is maintained as a small yacht harbor, the 
floats are scraped occasionally and invariably 
are soon resettled by young forms of the species 
mentioned. 
The conditions at the mouth of the San 
Gabriel River at Alamitos Bay were thoroughly 
investigated and reported by Reish in 1956. 
He reports the primary source of waste dis- 
charge into that region to be the Dow Chemical 
Company, the Santa Fe Springs Waste Disposal 
Company, two domestic sewage disposal plants, 
and the Los Angeles Bureau of Water and 
Power Steam Plant. 
Since specimens of the same species living in 
the same conditions are known to have a con- 
sistent pattern of free amino acids in their mus- 
cle tissues, an investigation of these amino acids 
should indicate whether or not a change from 
a normal to an abnormal metabolism has taken 
place. An analysis of free amino acids was thus 
undertaken by means of two-dimensional paper 
chromatography. 
Muscle tissue only was used for the deter- 
minations. All samples studied were taken from 
individual animals. Pooled samples were not 
used. Specimens were quick-frozen; muscle tis- 
sue from the foot of the abalone and from the 
legs of the crab was excised and subsequently 
lyophilized without thawing. This precaution 
was taken to inhibit the activity of autoenzymes 
and bacterial enzymes. After lyophilization the 
tissue was extracted with cold 70 per cent 
ethanol. Two 50 ml. aliquot portions were used, 
and extraction was permitted for at least 12 
Fig. 2. Chromatogram of free amino acids of Haliotis cracherodii taken from Anacapa Island, a nonpolluted 
