In the study presented, a simple analytical method for the direct determination of glycine in immunoglobulins by hydrophilic interaction liquid chromatography was developed. The HPLC separation was performed using a SeQuant ZIC-HILIC column (250 mm × 4.6 mm i.d, 5 μm) with the isocratic mobile phase consisting of ammonium formate (20 mM) and acetonitrile (30:70, v/v), and the flow rate set at 0.8 mL/min. UV detection was carried out at a wavelength of 210 nm. The procedure was validated for specificity, precision, linearity, accuracy, limit of detection, limit of quantitation, and robustness. The calibration curve was found to be linear within the concentration range of 1.2–3.6 mg/mL. RSD values for intra-day and inter-day precision were in the range of 0.66 to 1.84%. The limit of quantification and limit of detection were 0.10 mg/mL and 0.03 mg/mL, respectively. The developed chromatographic method was applied for the glycine analysis in various immunoglobulins.

中文翻译：

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建立亲水相互作用高效液相色谱法测定治疗性免疫球蛋白制剂中甘氨酸的方法

在提出的研究中，开发了一种通过亲水相互作用液相色谱法直接测定免疫球蛋白中甘氨酸的简单分析方法。使用SeQuant ZIC-HILIC色谱柱（250 mm×4.6 mm内径，5μm）进行HPLC分离，等度流动相由甲酸铵（20 mM）和乙腈（30:70，v / v）组成），流速设置为0.8 mL / min。在210nm的波长下进行UV检测。验证了该程序的特异性，精密度，线性，准确性，检测限，定量限和稳健性。发现校准曲线在1.2-3.6 mg / mL的浓度范围内是线性的。日内和日间精度的RSD值在0.66％至1.84％的范围内。定量限和检测限分别为0.10 mg / mL和0.03 mg / mL。所开发的色谱方法用于各种免疫球蛋白中的甘氨酸分析。