参苓健体粉指纹图谱和含量测定方法的研究
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摘要
目的:建立高效液相色谱法同时测定参苓健体粉中欧前胡素、异欧前胡素、花椒毒酚、茯苓酸、去氢茯苓酸、去氢土莫酸、白术内酯Ⅰ、白术内酯Ⅱ、白术内酯Ⅲ9个成分的含量,并建立HPLC指纹图谱。方法:采用Agilent Zorbax SB-C_(18)(250 mm×4. 6 mm,5μm)色谱柱,以乙腈-水溶液作为流动相进行梯度洗脱,流速为0. 8 ml·min-1,柱温为30℃,切换波长300 nm、210nm、220 nm及275 nm。以去氢土莫酸为参照峰,用中药色谱指纹图谱相似度评价软件(2004A版)对12批参苓健体粉进行相似度分析。结果:欧前胡素、异欧前胡素、花椒毒酚、茯苓酸、去氢茯苓酸、去氢土莫酸、白术内酯Ⅰ、白术内酯Ⅱ、白术内酯Ⅲ的线性范围分别为4. 15～83. 00μg·ml~(-1)(r=0. 999 9)、5. 06～101. 20μg·ml~(-1)(r=0. 999 8)、6. 88～137. 60μg·ml~(-1)(r=0. 999 7)、8. 87～177. 40μg·ml~(-1)(r=0. 999 9)、4. 03～80. 60μg·ml~(-1)(r=0. 999 5)、7. 32～146. 40μg·ml~(-1)(r=0. 999 8)、3. 84～76. 80μg·ml~(-1)(r=0. 999 7)、3. 81～76. 20μg·ml~(-1)(r=0. 999 8)和5. 37～107. 40μg·ml~(-1)(r=0. 999 6),平均加样回收率分别为98. 9%,99. 3%,99. 8%,100. 8%,101. 1%,100. 3%,101. 3%,100. 5%和99. 9%。12批样品中9个有效成分的含量范围分别在0. 125 8～0. 127 9 mg·g~(-1)、0. 134 7～0. 136 7 mg·g~(-1)、0. 172 6～0. 174 1 mg·g~(-1)、0. 226 6～0. 228 1 mg·g~(-1)、0. 102 6～0. 104 5 mg·g~(-1)、0. 200 7～0. 202 2 mg·g~(-1)、0. 120 0～0. 121 2 mg·g~(-1)、0. 120 4～0. 122 0 mg·g~(-1)和0. 146 0～0. 147 9 mg·g~(-1)。12批参苓健体粉的相似度均在0. 98以上。结论:本研究建立了参苓健体粉的指纹图谱及多指标成分含量的测定方法,为该制剂的质量控制及深入研究奠定了基础。
Objective: To establish a determination method for the characteristic fingerprint of Shenling Jianti powder and to determine imperatorin,isoimperatorin,xanthotoxol,pachymic acid,dehydroporia acid,dehydrotonic acid,atractylodes Ⅰ,atractylodes Ⅱand atractylodes Ⅲ in the preparation simultaneously. Methods: The analysis was carried out on an Agilent Zorbax SB-C_(18) analytical column( 250 mm × 4. 6 mm,5 μm) with gradient elution by acetonitrile-water,and the switching wavelengths were 314 nm,210 nm,220 nm and 275 nm. The column temperature was 30℃ at the flow rate of 0. 8 ml·min-1. Similarity analysis of the 12 batches of preparations was used by Chinese medicine chromatographic fingerprint similarity evaluation software( 2004 A version) with dehydrotonic acid as the reference peak. Results: The linear range of imperatorin,isoimperatorin,xanthotoxol,pachymic acid,dehydroporia acid,dehydrotonic acid,atractylodes Ⅰ,atractylodes Ⅱand atractylodes Ⅲ was 4. 15-83. 00 μg·ml~(-1)( r = 0. 999 9),5. 06-101. 20 μg·ml~(-1)( r = 0. 999 8),6. 88-137. 60 μg·ml~(-1)( r = 0. 999 7),8. 87-177. 40 μg·ml~(-1)( r = 0. 999 9),4. 03-80. 60 μg·ml~(-1)( r =0. 999 5),7. 32-146. 40 μg·ml~(-1)( r = 0. 999 8),3. 84-76. 80 μg·ml~(-1)( r = 0. 999 7),3. 81-76. 20 μg·ml~(-1)( r = 0. 999 8) and5. 37-107. 40 μg·ml~(-1)( r = 0. 999 6),and the average recovery was 98. 9%,99. 3%,99. 8%,100. 8%,101. 1%,100. 3%,101. 3%,100. 5% and 99. 9%,respectively. The contents of 12 batches of preparations were 0. 125 8-0. 127 9 mg·g~(-1),0. 134 7-0. 136 7 mg·g~(-1),0. 172 6-0. 174 1 mg·g~(-1),0. 226 6-0. 228 1 mg·g~(-1),0. 102 6-0. 104 5 mg·g~(-1),0. 200 7-0. 202 2 mg·g~(-1),0. 120 0-0. 121 2 mg·g~(-1),0. 120 4-0. 122 0 mg·g~(-1) and 0. 146 0-0. 147 9 mg·g~(-1),respectively. The similarity of the 12 batches of preparations was greater than 0. 98. Conclusion: The research has established a method for the characteristic fingerprint and multi-component determination of Shenling Jianti powder,which lays foundation for the quality control and further study of the preparation.
Objective: To establish a determination method for the characteristic fingerprint of Shenling Jianti powder and to determine imperatorin,isoimperatorin,xanthotoxol,pachymic acid,dehydroporia acid,dehydrotonic acid,atractylodes Ⅰ,atractylodes Ⅱand atractylodes Ⅲ in the preparation simultaneously. Methods: The analysis was carried out on an Agilent Zorbax SB-C_(18) analytical column( 250 mm × 4. 6 mm,5 μm) with gradient elution by acetonitrile-water,and the switching wavelengths were 314 nm,210 nm,220 nm and 275 nm. The column temperature was 30℃ at the flow rate of 0. 8 ml·min-1. Similarity analysis of the 12 batches of preparations was used by Chinese medicine chromatographic fingerprint similarity evaluation software( 2004 A version) with dehydrotonic acid as the reference peak. Results: The linear range of imperatorin,isoimperatorin,xanthotoxol,pachymic acid,dehydroporia acid,dehydrotonic acid,atractylodes Ⅰ,atractylodes Ⅱand atractylodes Ⅲ was 4. 15-83. 00 μg·ml~(-1)( r = 0. 999 9),5. 06-101. 20 μg·ml~(-1)( r = 0. 999 8),6. 88-137. 60 μg·ml~(-1)( r = 0. 999 7),8. 87-177. 40 μg·ml~(-1)( r = 0. 999 9),4. 03-80. 60 μg·ml~(-1)( r =0. 999 5),7. 32-146. 40 μg·ml~(-1)( r = 0. 999 8),3. 84-76. 80 μg·ml~(-1)( r = 0. 999 7),3. 81-76. 20 μg·ml~(-1)( r = 0. 999 8) and5. 37-107. 40 μg·ml~(-1)( r = 0. 999 6),and the average recovery was 98. 9%,99. 3%,99. 8%,100. 8%,101. 1%,100. 3%,101. 3%,100. 5% and 99. 9%,respectively. The contents of 12 batches of preparations were 0. 125 8-0. 127 9 mg·g~(-1),0. 134 7-0. 136 7 mg·g~(-1),0. 172 6-0. 174 1 mg·g~(-1),0. 226 6-0. 228 1 mg·g~(-1),0. 102 6-0. 104 5 mg·g~(-1),0. 200 7-0. 202 2 mg·g~(-1),0. 120 0-0. 121 2 mg·g~(-1),0. 120 4-0. 122 0 mg·g~(-1) and 0. 146 0-0. 147 9 mg·g~(-1),respectively. The similarity of the 12 batches of preparations was greater than 0. 98. Conclusion: The research has established a method for the characteristic fingerprint and multi-component determination of Shenling Jianti powder,which lays foundation for the quality control and further study of the preparation.
引文
1刘东方,赵丽娜,李银峰,等.中药指纹图谱技术的研究进展及应用[J]中草药,2016,47(22):4085-4094
2杨方良,张晶,孙国祥,等.中药组方指纹图谱研究方法和思路[J].色谱,2016,34(7):715-725
3杨柳,李婷,范彦博,等.妇科洗剂的HPLC指纹图谱研究[J].中国药师,2017,20(3):463-465
4原卫生部药品标准[S].中药成方制剂.第8册.1992:107
5桂元,薛承斌,詹继东,等.HPLC法测定参苓健体粉中去氢土莫酸、去氢茯苓酸、茯苓酸、白术内酯Ⅲ和白术内酯Ⅰ[J].现代药物与临床,2016,31(9):1331-1334
6陈琴华,余飞,王红梅,等.白术内酯Ⅰ、Ⅱ、Ⅲ对炎性巨噬细胞细胞因子表达的影响[J].中国药师,2017,20(12):2112-2116
7梁学清,李丹丹,黄忠威.茯苓药理作用研究进展[J].河南科技大学学报(医学版),2012,30(2):154-156
8季晓,宣槐斌,黄宝康.明党参活性成分及药理作用研究进展[J].药学实践杂志,2015,(2):102-105
9董凤彩.白术不同化学成分的药理作用[J].中医临床研究,2015,7(14):28-29
10李羾,何丹,杨常成.高效液相色谱法测定白术中白术内酯Ⅰ、Ⅱ和Ⅲ的含量[J].中南药学,2014,12(1):70-73
11步达,姚晓,郑紫婷,等.HPLC法测定明党参组织培养物和栽培品中香豆素成分的含量[J].天然产物研究与开发,2014,26(10):1638-1643
12李习平,庞雪,周逸群.不同加工方法对茯苓及茯苓皮中茯苓酸含量的影响[J].中国药师,2015,18(9):1453-1455
13王坚,徐菁,李鹏,等.HPLC法同时测定补益资生丸中白术内酯Ⅲ、白术内酯Ⅰ、去氢土莫酸、猪苓酸C和茯苓酸[J].中成药,2015,37(8):1718-1722
14车爽,李清,霍艳双,等.波长转换RP-HPLC法同时测定茯苓不同部位中5种三萜酸含量[J].药学学报,2010,45(4):494-497
2杨方良,张晶,孙国祥,等.中药组方指纹图谱研究方法和思路[J].色谱,2016,34(7):715-725
3杨柳,李婷,范彦博,等.妇科洗剂的HPLC指纹图谱研究[J].中国药师,2017,20(3):463-465
4原卫生部药品标准[S].中药成方制剂.第8册.1992:107
5桂元,薛承斌,詹继东,等.HPLC法测定参苓健体粉中去氢土莫酸、去氢茯苓酸、茯苓酸、白术内酯Ⅲ和白术内酯Ⅰ[J].现代药物与临床,2016,31(9):1331-1334
6陈琴华,余飞,王红梅,等.白术内酯Ⅰ、Ⅱ、Ⅲ对炎性巨噬细胞细胞因子表达的影响[J].中国药师,2017,20(12):2112-2116
7梁学清,李丹丹,黄忠威.茯苓药理作用研究进展[J].河南科技大学学报(医学版),2012,30(2):154-156
8季晓,宣槐斌,黄宝康.明党参活性成分及药理作用研究进展[J].药学实践杂志,2015,(2):102-105
9董凤彩.白术不同化学成分的药理作用[J].中医临床研究,2015,7(14):28-29
10李羾,何丹,杨常成.高效液相色谱法测定白术中白术内酯Ⅰ、Ⅱ和Ⅲ的含量[J].中南药学,2014,12(1):70-73
11步达,姚晓,郑紫婷,等.HPLC法测定明党参组织培养物和栽培品中香豆素成分的含量[J].天然产物研究与开发,2014,26(10):1638-1643
12李习平,庞雪,周逸群.不同加工方法对茯苓及茯苓皮中茯苓酸含量的影响[J].中国药师,2015,18(9):1453-1455
13王坚,徐菁,李鹏,等.HPLC法同时测定补益资生丸中白术内酯Ⅲ、白术内酯Ⅰ、去氢土莫酸、猪苓酸C和茯苓酸[J].中成药,2015,37(8):1718-1722
14车爽,李清,霍艳双,等.波长转换RP-HPLC法同时测定茯苓不同部位中5种三萜酸含量[J].药学学报,2010,45(4):494-497