Abstract Determination of biological sex to human remains is a fundamental requirement in anthropological, archeological, and forensic anthropological studies. Sex determination based on morphological criteria is significantly limited in the cases of juvenile remains and adult skeletons in a poor state of preservation. Regular attempts have been made to use alternative techniques to resolve this issue, including analysis of tooth enamel peptides by liquid chromatography/mass spectrometry. Optimization of this method involving acid etching of tooth enamel for 10 min followed by desalting of the products of etching on SDB-RPS StageTips microcolumns and analysis of desalted sample (1/3) by liquid chromatography/mass spectrometry allowed reliable sex determination to fossil remains within a wide range of archeological and biological ages without destructing analyzed teeth. Increasing the duration of enamel etching ensured a 2 to 3-fold increase in the total number of identified peptides and, more importantly, in the number of identified fragments of amelogenin Y isoform specific for male teeth, which facilitated reliable sex determination of fossil remains. The suggested technique was tested with 8 permanent and 15 deciduous teeth of different archaeological age and different degree of preservation. Two amelogenin Y-specific peptide sequences were identified. One of these peptides [SM(+15.99)IRPPYS)] was found in all male-derived samples without exception; the other peptide [IRPPYSS(+79.97)], which contained phosphorylated Ser66 residue, was found only in the enamel from deciduous teeth, which suggests that phosphorylation of Ser66 plays a role in the enamel formation in deciduous teeth.

中文翻译：

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使用来自不同生物世代、考古年龄和埋藏保护程度的牙齿的牙釉质肽组分析优化人类性别测定方法

摘要 确定人类遗骸的生物性别是人类学、考古学和法医人类学研究的基本要求。在保存状况不佳的少年遗骸和成年骨骼的情况下，基于形态学标准的性别确定受到很大限制。已经定期尝试使用替代技术来解决这个问题，包括通过液相色谱/质谱法分析牙釉质肽。该方法的优化包括对牙釉质进行酸蚀刻 10 分钟，然后在 SDB-RPS StageTips 微柱上对蚀刻产物进行脱盐，并通过液相色谱/质谱法对脱盐样品 (1/3) 进行分析，从而可以对化石残骸进行可靠的性别鉴定在广泛的考古和生物年龄范围内，而不会破坏分析过的牙齿。增加牙釉质蚀刻的持续时间确保了鉴定肽的总数增加了 2 到 3 倍，更重要的是，鉴定出男性牙齿特异性牙釉蛋白 Y 异构体的片段数量，这有助于对化石残骸进行可靠的性别确定。建议的技术对不同考古年龄和不同保存程度的 8 颗恒牙和 15 颗乳牙进行了测试。鉴定了两个牙釉蛋白 Y 特异性肽序列。这些肽之一 [SM(+15.99)IRPPYS)] 在所有男性来源的样品中无一例外地被发现；另一种含有磷酸化 Ser66 残基的肽 [IRPPYSS(+79.97)] 仅在乳牙的牙釉质中发现，这表明 Ser66 的磷酸化在乳牙的牙釉质形成中起作用。