应用UPLC-MS/MS观测6种中药对尿酸盐体外溶解度的影响＊

目的 建立水基质溶液中尿囊素含量的测定方法,观测6种治疗痛风常用中药对尿酸盐体外溶解度的影响。方法 采用超高效液相色谱-串联质谱（UPLC-MS/MS）法定量分析9组溶液中尿酸盐溶解后产生的尿囊素含量：实验组溶剂为6种治疗痛风常用中药的超声水提取溶液,对照组溶剂为超纯水、5%碳酸氢钠溶液和尿酸酶溶液。结果 尿囊素的色谱在1.40 min出峰,在0.1-100 ng·mL^-1范围内线性关系良好（r² = 0.9995）,平均加标回收率为102.36%（n = 6）,RSD为2.33%。尿囊素浓度与溶液pH值无线性相关;溶解时间24 h,土茯苓、黄柏和海金沙组尿囊素浓度与超纯水组均无统计学差异（P > 0.05）;溶解时间24 h,山慈菇、苍术和大黄组尿囊素浓度均高于超纯水组（P < 0.05）;溶解时间24 h,山慈菇组尿囊素浓度高于5%碳酸氢钠组（P < 0.05）;尿酸酶组的尿囊素浓度在1 h和24 h溶解时间内均为9组中最高（P < 0.05）。通过尿酸盐溶液中分解产生的尿囊素含量估算尿酸盐的溶解度（1 h,24 h）,以1h超纯水组的溶解度为100%：超纯水组（100%,121%）,山慈菇组（279%,463%）,土茯苓组（69%,172%）,海金沙组（21%,97%）,苍术组（105%,400%）,大黄组（146%,311%）,黄柏组（65%,74%）,尿酸酶组（8247%,12454%）和5%碳酸氢钠组（63%,285%）。结论 本方法可灵敏、准确地测定尿酸盐在水基质中药溶液中溶解后产生的尿囊素含量。山慈菇、苍术和大黄对尿酸盐的体外溶解度有一定促进作用,但未能达到临床溶解痛风石的要求。土茯苓、黄柏和海金沙对MSU的溶解无促进作用。

Effect of Six Herbs on the Solubility of Urate: An In-Vitro Study Using UPLC-MS/MS

Objective To establish a method for the determination of allantoin content in aqueous matrix solution and to observe the effects of 6 traditional Chinese medicines commonly used in the treatment of gout on the in vitro solubility of urate.Methods The content of allantoin in 9 groups was quantitatively analyzed by UPLC-MS /MS. The solvent of the experimental group was 6 kinds of traditional Chinese medicines commonly used in the treatment of gout, and the solvent of the control group was ultra-pure water, 5% sodium bicarbonate solution and uricase solution.Results The chromatographic peak of allantoin was 1.40 min, and the linear relationship was good in the range of 0.1-100 ng·mL^-1 (r² = 0.9995). The average standard recovery was 102.36% (n = 6), and the RSD was 2.33%. There was no linear correlation between allantoin concentration and pH value (P > 0.05). In 24 h, there was no statistically significant difference between the allantoin concentration of Rhizoma Smilacis Glabrae, Cortex Phellodendri and Spora Lygodii groups and that of the ultra-pure water group (P > 0.05). In 24h, the allantoin concentration of Pseudobulbus Cremastrae seu Pleiones, Rhizoma Atractylodis and Dahuang was higher than that of the ultra-pure water group (P < 0.05). In 24 h, the allantoin concentration of Pseudobulbus Cremastrae seu Pleiones group was higher than that of 5% sodium bicarbonate group (P < 0.05). In both 1 h and 24 h, the allantoin concentration of uricase group was the highest among all groups (P < 0.05). The solubility in other groups were calculated based on allantoin concentration as follow (1 h-24 h), the urate solubility of ultra-pure water group was defined as 100%: Ultra-pure water (100%, 121%), Pseudobulbus Cremastrae seu Pleiones group (279%, 463%), Rhizoma Smilacis Glabrae group (69%, 172%), Spora Lygodii group (21%, 97%), Rhizoma Atractylodis group (105%, 400%), Radix et Rhizoma Rhei group (146%, 311%), Cortex Phellodendri group (65%, 74%), Uricase group (8247%, 12454%) and 5% sodium bicarbonate group (63%, 285%).Conclusion This method is sensitive and accurate for determination of the content of allantoin produced after urate is dissolved in aqueous Chinese medicine solution. In vitro solubility of urate was improved by Pseudobulbus Cremastrae seu Pleiones, Rhizoma Atractylodis and Radix et Rhizoma Rhei, but they failed to meet the standard of clinical dissolution of gout. The dissolution of MSU could not be promoted by Rhizoma Smilacis Glabrae, Cortex Phellodendri and Spora Lygodii.