Matskási István (szerk.): A Magyar Természettudományi Múzeum évkönyve 93. (Budapest 2001)

Csapó, J., Bernert, Zs. , Csapó, Zs. , Pohn, G. , Csapó-Kiss, Zs. , Költő, L. , Szikossy, I. ; Némethy, S.: Introduction of amino acid racemisation based age estimation into paleoanthropological [sic] research

Table 2. Methods for age estimation of adults Methods for age estimation Transparency of root and periodontosis Degree of abrasio of the teeth Ectocranial ossification of sutures Endocranial ossification of sutures Changes of surface of the faciès symphyseos ossis pubis Changes of surface of sternal extremity of the rib IDENTIFICATION OF D- AND L-ENANTIOMERS Apparatus - D- and L-amino acids were identified by a high efficiency LaChrom Hitachi Merck liquid Chromatograph. The apparatus was built up from a LaChrom D-7000 System Manager device, a LaChrom L-7100 HPLC pump, a LaChrom L-7300 column thermostat, a LaChrom L-7200 Autosampler, a LaChrom L7400 programmable UV detector and a LaChrom 7480 fluorescent detector. Amino acid enantiomers were separated after o-phtaldialdehyd (OPA) and 2,3,4,6-tetra-O-acetil-l -tio-ß-D-glycopyranosid (TATG) derivatisation in a non-chiral column aided by gradient elution. Chemicals - Acetonitril and methanol were purchased from Rathburn (Walkerburn, U.K.), amino acid standards, OPA and TATG were purchased from Sigma (St. Louis, Mo.). Elution puffers were produced from mono- and dinatrium-hydrogen-phosphat. Derivatisation - Reaction was executed in 120 urn micro phials put into 1.8 cm

3 volume ampoule with teflon covered internal covers and caps. The automatic sample feeder was programmed to mix the sample (free amino acids or protein hydrolysate distilled in nitrogen stream) dissolved in 90 ml of boric puffer (0.4M; pH:9.5) with 15 ml of reagent (8 mg of OPA and 44 mg of TATG dissolved in 1 cm

3 of methanol). After this the solution was thoroughly mixed by repeated sucking up and releasing back, then it was left to rest for 6 minutes. The injecting device was carefully washed through and then we injected 25 mm of this reaction compound into the column of analyser. When finishing injection the system was three times flushed through by a 70:30 ratio 100 ml acetone water mixture. Separation and determination of enantiomers - Enantiomers were separated by reversed phase chromatography according to the method of ElNARSSON et al.