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1.江西中医药大学,南昌 330004
2.中国中医科学院 中药研究所,北京 100700
3.天津中医药大学,天津 301617
4.北京城市学院,北京 100083
严林,在读博士,从事中药制剂研究,E-mail:3104483640@qq.com
刘德文,副研究员,硕士生导师,从事中药制剂与中药炮制研究,E-mail:dwliu@icmm.ac.cn;
仝燕,研究员,硕士生导师,从事中药制剂研究,E-mail:tongyan1012@sohu.com
纸质出版日期:2023-12-05,
网络出版日期:2023-04-28,
收稿日期:2023-02-23,
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严林,欧则民,王艳静等.基于UPLC-Q-TOF-MS的泽泻盐制前后血清药物化学比较分析[J].中国实验方剂学杂志,2023,29(23):122-130.
YAN Lin,OU Zemin,WANG Yanjing,et al.Comparative Analysis of Serum Pharmacochemistry of Alismatis Rhizoma Before and After Salt Processing Based on UPLC-Q-TOF-MS[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(23):122-130.
严林,欧则民,王艳静等.基于UPLC-Q-TOF-MS的泽泻盐制前后血清药物化学比较分析[J].中国实验方剂学杂志,2023,29(23):122-130. DOI: 10.13422/j.cnki.syfjx.20230866.
YAN Lin,OU Zemin,WANG Yanjing,et al.Comparative Analysis of Serum Pharmacochemistry of Alismatis Rhizoma Before and After Salt Processing Based on UPLC-Q-TOF-MS[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(23):122-130. DOI: 10.13422/j.cnki.syfjx.20230866.
目的
2
基于超高效液相色谱-四极杆-飞行时间质谱法(UPLC-Q-TOF-MS)对泽泻、盐泽泻水提液灌胃给药后大鼠血清中的移行成分进行比较分析。
方法
2
SD大鼠随机分为空白组、泽泻组(10 g·kg
-1
)和盐泽泻组(10 g·kg
-1
),每组3只,给药组分别灌胃给予泽泻、盐泽泻水提液,空白组灌胃等体积饮用水,早晚各1次,连续3 d,末次给药60 min后,眼眶采血,制备血清样品。采用ACQUITY UPLC BEH C
18
(2.1 mm×50 mm,1.7 μm)色谱柱,流动相乙腈(A)-0.1%甲酸水溶液(B),梯度洗脱(0~10 min,10%~50% A;10~27 min,50%~95% A;27~27.1 min,95%~10% A;27.1~30 min,10% A),流速0.3 mL·min
-1
,正离子模式下采集数据,扫描范围
m
/
z
100~1 200。在自建泽泻化学成分库的基础上借助UNIFI 1.9.2软件对泽泻、盐泽泻水提液及给药血清、空白血清的总离子流图和二级质谱碎片信息进行比对,推导泽泻、盐泽泻可能的入血移行成分及其裂解规律,并计算各化学成分炮制前后的响应强度比。
结果
2
在给予泽泻水提液大鼠血清中分析推测得到20个成分,包括5个原型成分和15个代谢产物;在给予盐泽泻水提液大鼠血清中分析推测得到14个成分,包括5个原型成分和9个代谢成分;其中两者共有成分13个,包括5个原型成分和8个代谢成分。5个原型成分分别为16-氧代泽泻醇A、24-乙酰泽泻醇A、泽泻醇A、泽泻醇B、泽泻醇C,代谢产物主要涉及Ⅰ相代谢(氧化反应)和Ⅱ相代谢(葡萄糖醛酸化反应);盐泽泻组大鼠血清中原型成分16-氧代泽泻醇A、泽泻醇B、泽泻醇C的响应强度升高,而代谢成分种类及响应强度则普遍降低。
结论
2
泽泻盐制后可能通过减缓萜类成分代谢速率从而促进原型成分吸收入血,可为泽泻、盐泽泻的药效物质基础研究提供了数据支撑。
Objective
2
Based on serum pharmacochemistry and ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) the transitional components in the serum of rats after intragastric administration of water extract of Alismatis Rhizoma(AR)and salt-processed Alismatis Rhizoma(SAR) were compared.
Method
2
SD rats were randomly divided into blank group
AR group(10 g·kg
-1
) and SAR group(10 g·kg
-1
)
3 rats in each group
the administration groups were given AR and SAR aqueous extracts by gavage
respectively
and the blank group was given an equal volume of drinking water by gavage once in the morning and once in the evening
for 3 consecutive days. Sixty minutes after the last administration
blood was collected from the eye orbits
and the serum samples were prepared. The serum samples were prepared on an ACQUITY UPLC BEH C18 column(2.1 mm×50 mm
1.7 μm) with the mobile phase of acetonitrile(A)-0.1% formic acid aqueous solution(B) in a gradient elution(0-10 min
10%-50% A; 10-27 min
50%-95%A; 27-27.1 min
95%-10% A; 27.1-30 min
10%A)
the data were collected at a flow rate of 0.3 mL·min
-1
in positive ion mode with a scanning range of
m
/
z
100-1 200. Based on the self-constructed chemical composition library of AR
the total ion flow diagrams and secondary MS fragmentation information of the aqueous extracts of AR and SAR
as well as the administered serum and the blank serum
were compared with each other by UNIFI 1.9.2
so as to deduce the possible blood-migrating constituents and their cleavage patterns in the aqueous extracts
and the response intensity ratios of each chemical component were calculated before and after processing.
Result
2
A total of 20 components
including 5 prototypical components and 15 metabolites
were analyzed and deduced from the serum of rats given aqueous extract of AR. And 14 components
including 5 prototypical components and 9 metabolites
were analyzed and deduced from the serum of rats given aqueous extract of SAR. Of these
13 components were common to both of them
including 5 prototypical components and 8 metabolites. The 5 prototypical components were 16-oxoalisol A
alisol A 24-acetate
alisol A
alisol B and alisol C. The metabolites were mainly involved in phase Ⅰ metabolism(oxidation) and phase Ⅱ metabolism(glucuronidation). There was a big change in the intensity of response of the common components before and after salt-processing
and the response intensities of the prototypical components
16-oxoalisol A
alisol B and alisol C
were elevated
while the type and response intensity of metabolites were generally decreased
and it was hypothesized that the metabolic rate of terpenoids might be slowed down after salt-processing of AR
so that the blood-migrating constituents could participate in the metabolism of the body more in the form of prototypes.
Conclusion
2
Salt-processing of AR may promote the absorption of prototypical components into the blood by slowing down the metabolic rate of terpenoids
which can provide support for the research on material basis of AR and SAR.
泽泻中药炮制血清药物化学萜类裂解规律代谢产物超高效液相色谱-四极杆-飞行时间质谱法(UPLC-Q-TOF-MS)
Alismatis Rhizomaprocessingserum pharmacochemistryterpenoidfragmentation patternmetaboliteultra-high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS)
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