摘要
目的：建立一种利用HPLC法同时测定龟甲胶中14种氨基酸的含量。方法：样品以6 mol·L-1盐酸于110℃水解7 h,以6-氨基喹啉-N-羟基琥珀酰亚胺活化环氨基甲酸酯（AQC）进行衍生化反应,采用Kromasil C18色谱柱（5μm,4.6 mm×250 mm）,柱温37℃,以0.14 mol·L-1三水合乙酸钠（p H 4.85）为流动相A,60%乙腈为流动相B,梯度洗脱,流速1.4 m L·min-1,检测波长248 nm。结果：在一定的浓度范围内,各种氨基酸峰面积和浓度呈良好的线性关系,r值均大于0.999 0;加样回收率（n=9）为94.5%～103.6%;不同样品中14种氨基酸含量范围分别为羟脯氨酸7.08%～8.22%、谷氨酸5.94%～8.21%、丝氨酸2.48%～3.12%、组氨酸0.25%～0.44%、甘氨酸14.35%～16.86%、精氨酸4.78%～5.76%、苏氨酸1.24%～1.75%、丙氨酸4.65%～6.56%、脯氨酸6.78%～8.83%、酪氨酸0.55%～0.84%、缬氨酸0.95%～1.40%、异亮氨酸0.70%～0.95%、亮氨酸1.59%～2.06%、苯丙氨酸1.35%～1.57%。结论：该方法准确、可靠,具有良好的重复性和稳定性,可用于龟甲胶中羟脯氨酸、谷氨酸、丝氨酸、组氨酸、甘氨酸、精氨酸、苏氨酸、丙氨酸、脯氨酸、酪氨酸、缬氨酸、异亮氨酸、亮氨酸、苯丙氨酸的检测。
Objective： To establish an HPLC method for simultaneous determination of 14 kinds of amino acids in tortoise plastron glue. Methods： The sample was hydrolyzed at 110 ℃ for 7 hours in the 6 mol·L- 1hydrochloric acid,and derived with 6-aminoquinolyl-N-hydroxysuccinimidyl-carbamate（ AQC）. HPLC was performed on a Kromasil C18column（ 5 μm,4. 6 mm × 250 mm） with gradient elution of 0. 14 mol·L- 1sodium salt trihydrate（ p H4. 85,mobile phase A） and 60% acetonitrile（ mobile phase B） at the flow rate of 1. 4 m L·min- 1,detected at248 nm. Results： In a certain concentration range,the peak area and the concentration of each amino acid showed a good linear relationship. Correlation coefficient r values were more than 0. 999 0. The recoveries ranged in94. 5%-103. 6%. The content ranges of hydroxyproline,glutamicacid,serine,histidine,glycine,arginine,threonine,alanine,proline,tyrosine,valine,isoleucine,leucine,phenylalanine were 7. 08%-8. 22%,5. 94%-8. 21%,2. 48%-3. 12%,0. 25%-0. 44%,14. 35%-16. 86%,4. 78%-5. 76%,1. 24%-1. 75%,4. 65%-6. 56%,6. 78%-8. 83%,0. 55%-0. 84%,0. 95%-1. 40%,0. 70%-0. 95%,1. 59%-2. 06% and 1. 35%-1. 57% respectively in different samples. Conclusion： The developed method is accurate with high repeatability,which is helpful for the determination of hydroxyproline,glutamicacid,serine,histidine,glycine,arginine,threonine,alanine,proline,tyrosine,valine,isoleucine,leucine,phenylalanine in tortoise plastron glue.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2015年第10期1790-1795,共6页
Chinese Journal of Pharmaceutical Analysis
关键词
龟甲胶
柱前衍生化
水解氨基酸
6-氨基喹啉-N-羟基琥珀酰亚胺活化环氨基甲酸酯
衍生化试剂
高效液相色谱
tortoise plastron glue
pre-column derivatization
hydrolytic amino acids
6-aminoquinolyl-N-hydroxy-succinimidyl-carbamate
derivatization reagent
high performance liquid chromatography
