Amino acid end-group in commercial heparin. I. Determination of the number of amino and hydroxyl end groups in heparin

Three commercial heparins of different molecular weights and anticoagulant activities were dinitrophenylated (DNP) with 2,4-dinitrofluorobenzene under aqueous conditions. The absorption spectra observed for DNP heparins in a 1% NaHCO₃ solution consisted of the two absorption spectra characteristic of DNP—amino (N—DNP) and DNP—hydroxyl (O—DNP) groups. The number of N—DNP, O—DNP, and (N + O)—DNP groups were determined as well as the number of N—DNP groups per heparin chain; different values (i.e., 0.16, 0.17, and 0.55) for the latter were obtained with the three heparins. Further calculations showed that two of the heparins had approximately two (N + O)—DNP groups per chain (i.e., 2.1 and 1.8) whilst the third sample, considered to be a “cruder” heparin, had a value of 1.4.