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Absorption of NPH-insulin from subcutaneous tissue: a methodological study in pigs.


To elucidate the validity of the indirect method of following insulin absorption from the subcutaneous tissue, i.e. external counting of injected 125-I insulin, the disappearance of 125-I NPH insulin from subcutaneous tissue was measured by a biotelemetric method (external measurement of radioactivity) and by direct determination of radioactivity and insulin in subcutaneous tissue extracted one to 300 min. after injection of 8 units 125-I NPH porcine insulin, 5 mm beneath the surface of the skin in 6 anaesthesized pigs. Furthermore, the appearance of insulin in arterial plasma was measured after inhibiting the endogenous insulin secretion of the pigs by epinephrine and propanolol. The disappearance of radioactivity measured continuously for 5 hours was of first order kinetics with t1/2=462+/-36 min. (mean+/-S.E.M.). The half-life of the injected depot of radioactivity (t1/2) demonstrated a large intra- as well as interindividual variation as seen in diabetics. In tissue extracts a highly significant (2P less than 0.001) correlation was found between radioactivity and insulin (r=0.93), indicating the radioactivity to be representative of insulin. During gel chromatography on Sephadex G 50 M only one peak of radioactivity was seen in all tissue extracts. 300 min. after NPH-insulin injection, however, the specific radioactivity of insulin was significantly higher (2P less than 0.05) than in tissue extracts sampled one min. after NPH insulin injection, indicating a slight accumulation of non-immunoreactive insulin degradation products. A fair correlation (r=0.75, 2 P less than 0.05) was seen between the disappearance of externally measured radioactivity or insulin from tissue extracts and the appearance of insulin in plasma. It is concluded that the insulin absorption coefficient is a relevant and biologically sensible expression of insulin absorption.

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