目的: 建立采用亲水作用色谱（HILIC）测定复方硫酸软骨素滴眼液中尿囊素含量的测定方法。方法: 采用丙基酰胺键合硅胶色谱柱（Venusil HILIC,4.6 mm×250 mm,5 μm）,以乙腈 - 水（98∶2）为流 动相进行等度洗脱,流速 1.0 mL·min-1,柱温 30 ℃,检测波长 191 nm;同时采用紫外可见分光光度法对尿囊素含量测定结果进行验证。结果: 尿囊素与复方硫酸软骨素滴眼液中其他组分可以完全分离,尿囊素在 0.010 4~0.148 4 mg·mL-1 的浓度范围内线性关系良好（r=1.000）;精密度试验的 RSD 为 0.25%,平均回收率 为 98.7%（n=9）,RSD 为 0.69%;紫外可见分光光度法与亲水作用色谱法对 3 批样品的尿囊素测定结果基本一致。结论: 本方法灵敏、准确,专属性好,可用于复方硫酸软骨素滴眼液中尿囊素的含量测定。
Objective: To establish a new method for the determination of allantoin content in compound chondroitin sulfate eye drops using hydrophilic interaction liquid chromatography（HILIC）. Methods: The Venusil HILIC column（4.6 mm×250 mm,5 μm）was adopted for isocratic elution with acetonitrile-water（98∶2）as the mobile phase. The flow rate was 1.0 mL·min-1,the column temperature was 30 ℃ and the detection wavelength was 191 nm. Ultraviolet and visible spectrophotometry was used to verify the results of allantoin content. Results: Allantoin could be completely separated with other components in compound chondroitin sulfate eye drops. The calibration curve was linear in the concentration range of 0.010 4-0.148 4 mg·mL-1 with the correlation coefficient（r）of 1.000. The relative standard deviation（RSD）of the precision test was 0.25%. The average recovery was 98.7% （n=9）and the RSD was 0.69%. The three batches results of allantoin using HILIC method were in accordance with the results of UV-VIS spectrophotometry. Conclusion: This method is sensitive,accurate and specific. It can be used to determine the content of allantoin in compound chondroitin sulfate eye drops.
[1] 章茹,万义玲,张定亨.尿囊素的性质与应用[J].南昌大学学报(工科版),1997,19(2):81
ZHANG R,WAN YL,ZHANG DH.The properties and application of allantoin[J].J Nanchang Univ(Eng Sc Edit),1997,19(2):81
[2] MADRAZO-JIMENEZ M,RODRIGUEZ-CABALLERO A SERRERA-FIGALLO MA,et al.The effects of a topical gel containing chitosan,0.2% chlorhexidine,allantoin and despanthenol on the wound healing process subsequent to impacted lower third molar extraction[J].Med Oral Patol Oral Cir Bucal,2016,21(6):e696
[3] MANCA ML,MATRICARDI P,CENCETTI C,et al.Combination of argan oil and phospholipids for the development of an effective liposome-like formulation able to improve skin hydration and allantoin dermal delivery[J].Int J Pharm,2016,505(1-2): 204
[4] FLORENTINO IF,SI LVA DP B,GAL DINO PM,e t al.Antinociceptive and anti-inflammatory effects of Memora nodosa and allantoin in mice[J].J Ethnopharmacol,2016,186:298
[5] SEAN CS.Martindale:The Complete Drug Reference Edited 38[M].London:Pharmaceutical Press,2014:1693
[6] 金鹏飞,吴学军,邹定,等.HPLC-DAD 同时测定复方尿维氨滴眼液中 3 种活性成分的含量[J].中国药学杂志,2009,44(21):1669
JIN PF,WU XJ,ZOU D,et al.Simultaneous determination of the content of three active ingredients in compound urovitamin eye drops by HPLC-DAD[J].Chin Pharm J,2009,44(21):1669
[7] 许辉,陈锋.HPLC 法测定缩泉丸中尿囊素含量[J].湖南中医杂志,2016,32(12):159
XU H,CHEN F.Determination of allantoin in Shuoquan pills by HPLC[J].Hunan J Tradit Chin Med,2016,32(12):159
[8] 朱群英,张亚锋,苏超男,等.HILIC-HPLC 双波长法测定山药中尿囊素和腺苷的含量[J].西北药学杂志,2018,33(3):54
ZHU QY,ZHANG YF,SU CN,et al.Determination of allantoin and adenosine in Yam by HILIC-HPLC dual wavelength method[J]. Northwest Pharm J,2018,33(3):54
[9] ASMIN A,DE RECHTER S,PETER J,et al.Simultaneous determination of allantoin and adenosine in human urine using liquid chromatography-UV detection[J].J Chromatogr B,2018,1096: 201
[10] 胡翮,李昂.反相高效液相色谱法测定保健品纯怀山药粉中尿囊素的含量[J].药物分析杂志,2017,37(8):1520
HU H,LI A.Determination of allantoin in health care products pure Huai yam powder by reversed-phase high performance liquid chromatography[J].Chin J Pharml Anal,2017,37(8):1520
[11] 中华人民共和国药典 2020 年版.一部[S].2020:1853 ChP 2020.Vol Ⅰ[S].2020:1853
[12] 文庆,聂平,丁野,等.HILIC-HPLC 法测定山药中尿囊素的含量[J].中南药学,2014,12(2):169
WEN Q,NIE P,DING Y,et al.Determination of aAllantoin in yam by HILIC-HPLC method[J].Cent South Pharm,2014,12(2): 169
[13] 刘应蛟,楚世峰,袁志鹰,等.HPLC 法同时测定山药饮片、麸炒及土炒山药中尿囊素、腺苷和苯丙氨酸的含量[J].时珍国医国药,2019,30(3):568
LIU YJ,CHU SF,YUAN ZY,et al.Simultaneous determination of allantoin,adenosine and phenylalanine in yam slices,bran-fried and soil-fried yam by HPLC[J].Lishizhen Med Mater Med Res,2019, 30(3):568
[14] 吴颖. 亲水作用色谱-高效液相色谱法测定纯淮山药粉中尿囊素的含量[J].医疗装备,2016,29(20):84
WU Y.Determination of allantoin in pure yam powder by hydrophilic interaction chromatography-high performance liquid chromatography[J].Med Equip,2016,29(20):84
[15] 徐露露,陈路晓,骆宜,等.亲水作用色谱在天然产物分离分析中的应用进展[J].药物分析杂志,2016,36(9):1516
XU LL,CHEN LX,LUO Y,et al.Application progress of hydrophilic interaction chromatography in separation and analysis of natural products[J].Chin J Pharm Anal,2016,36(9):1516
[16] 张玉婵,黄奇建,江志华.溶液pH 值对尿囊素稳定性的影响[J].今日药学,2014,24(5):343
ZHANG YC,HUANG QJ,JIANG ZH.The effect of solution pH on the stability of allantoin[J].Pharm Today,2014,24(5):343
