The reversal effect of albiflorin on multidrug resistance of human ovarian cancer and its mechanism / 中国药理学通报

Aim To study the reversal effect of albiflorin(AL)on multidrug resistance of human ovarian cancer and the potential mechanism. Methods The drug resistance reversal effect of AL on SKOV3/DDP cells was detected by CCK-8 kit,and the effect of AL on P-glycoprotein(P-gp)function was detected by flow cytometry. The effects of AL on MYC,WWP1 and ABCB1 in SKOV3/DDP cells were detected by RT-qPCR and Western blot. The MYC-knockdown SKOV3/DDP cell line was constructed by RNA interference technology,and its drug resistance,P-gp function and related gene and protein expression changes were investigated. Results AL had a drug resistance reversal effect on SKOV3/DDP cells and a concentration-dependent inhibitory effect on P-gp function. The inhibitory effects of AL 25,50 and 100 μmol·L-1 on ABCB1/P-gp,MYC and WWP1 were gradually enhanced. The inhibitory effect of MYCi975,a MYC inhibitor,on ABCB1/P-gp,MYC and WWP1 was stronger than or equivalent to that of AL 100 μmol·L-1 group. After knockdown of MYC in SKOV3/DDP cells,cell drug resistance,P-gp function,and related gene and protein expression were inhibited. Conclusions The drug resistance reversal effect of AL on SKOV3/DDP cells may be related to the inhibition of P-gp function and the expression of ABCB1/P-gp,MYC and WWP1,which provides an experiment base for the development of AL as a drug resistance reversal agent for the clinical treatment of ovarian cancer.

Mechanism of albiflorin in improvement of Alzheimer's disease based on network pharmacology and in vitro experiments / 中国中药杂志

This study aimed to explore the mechanism of albiflorin in the treatment of Alzheimer's disease(AD) based on network pharmacology, molecular docking, and in vitro experiments. Network pharmacology was used to predict the potential targets and pathways of albiflorin against AD, and molecular docking technology was used to verify the binding affinity of albiflorin to key target proteins. Finally, the AD cell model was induced by Aβ_(25-35) in rat pheochromocytoma(PC12) cells and intervened by albiflorin to validate core targets and pathways. The results of network pharmacological analysis showed that albiflorin acted on key targets such as mitogen-activated protein kinase-1(MAPK1 or ERK2), albumin(ALB), epidermal growth factor receptor(EGFR), caspase-3(CASP3), and sodium-dependent serotonin transporter(SLC6A4), and signaling pathways such as MAPK, cAMP, and cGMP-PKG. The results of molecular docking showed that albiflorin had strong binding affinity to MAPK1(ERK2). In vitro experiments showed that compared with the blank group, the model group showed decreased cell viability, decreased expression level of B-cell lymphoma 2(Bcl-2), increased Bcl-2-associated X protein(Bax), and reduced phosphorylation level of extracellular signal-regulated kinase 1/2(ERK1/2) and the relative expression ratio of p-ERK1/2 to ERK1/2. Compared with the model group, the albiflorin group showed potentiated cell viability, up-regulated expression of Bcl-2, down-regulated Bax, and increased phosphorylation level of ERK1/2 and the relative expression ratio of p-ERK1/2 to ERK1/2. These results suggest that the mechanism of albiflorin against AD may be related to its activation of the MAPK/ERK signaling pathway and its inhibition of neuronal apoptosis.

The antidepressant effect of albiflorin based on TSPO and mechanism / 中国药理学通报

Aim To study the antidepressant effects of albiflorin and its relationship with TSPO(translocator protein 18 ku). Methods Mice were divided into eight groups(control group, chronic unpredictable stress group, fluoxetine group, albiflorin low, medium, high dose groups, PK11195 group, PK11195+ albiflorin high dose group)based on the data of the behavioral tests conducted to assess the antidepressant-like effects of albiflorin. After the behavioral tests Western blot and ELISA were conducted to evaluate the TSPO expression, progesterone and allopregnanolone level in hippocampus of mice. Results In the behavioral tests, there were significant differences between the model group and the control group, which indicated that the model was successfully established. The positive drug and albiflorin in different dose groups could reverse the effects of the model group, and PK11195 could reverse the effects of paeoniflorin in high dose group. The results of Western blot and ELISA showed that the TSPO expression, progesterone and allopregnanolone level in the model group significantly decreased. The positive drug and albiflorin groups with different doses could reverse the effects of the model group, and PK11195 could reverse the effects of the high dose group. Conclusions Albiflorin has significant antidepressant and antianxiety effects on CUS mice by TSPO, which provides experimental basis for the further study on the antidepressant effects and mechanism of albiflorin in the future.

Chemical Constituents and Pharmacological Effects of Paeoniae Radix Rubra： A Review / 中国实验方剂学杂志

Paeoniae Radix Rubra is a traditional Chinese medicine commonly used in clinical practice， it is mostly wild and widely distributed in different areas of China. In addition， the plant of Paeoniae Radix Rubra also has ornamental value. Modern phytochemical researches showed that the chemical constituents of Paeoniae Radix Rubra were complex. Up to now， more than 300 chemical constituents have been found， mainly including monoterpene glycosides， triterpenoids， flavonoids， tannins， phenolic acids， saccharides， steroids， volatile oils and so on. Among them， the content of monoterpene glycosides was the highest， and the types of volatile oil were the most. Paeoniae Radix Rubra has a wide range of pharmacological effects， exerting different curative effects in multiple systems such as blood， cardiovascular， nervous and digestive system. It can protect myocardial cells and nerve cells， stabilize microcirculation， anti-endotoxin， anti-atherosclerosis， reduce pulmonary hypertension， anti-depression， protect liver， anti-gastric ulcer， anti-tumor， slow down aging， treat Parkinson's syndrome and diabetes and its complications， anti-radiation， anti-inflammatory， anti-virus and so on. Through reviewing the literature on chemical constituents and pharmacological effects of Paeoniae Radix Rubra， it was found that total glycosides and monomers such as paeoniflorin， albiflorin， benzoylpaeoniflorin and gallic acid may be the main active components of Paeoniae Radix Rubra. At present， the research on Paeoniae Radix Rubra mainly focused on monoterpene glycosides， while the research on flavonoids and volatile oil in Paeoniae Radix Rubra was less. It is suggested that research on these two components should be strengthened in the future.

Antidepressant-like effects of albiflorin involved the NO signaling pathway in rats model of chronic restraint stress / 中国天然药物

The depressant-like effects of albiflorin (AF) were studied on stressed chronic restraint stress (CRS) rats. Experimental rats were subjected to immobilization stress for a daily 6 h-restraining in a plastic restrainer for continuous 21 d and were treated with 30 or 15 mg·kg

HPLC fingerprint and simultaneous determination of nine components in Taohong Siwu Decoction of classical prescriptions / 中草药

Objective: To establish chemical fingerprint and multi-components determination of 15 batches of Taohong Siwu Decoction (TSD), and provide reference for the improvement of its quality control. Methods: The separation was performed on Thermo Hypersil Gold C18 column (250 mm × 4.6 mm, 5 μm) for gradient elution with methanol-0.1% phosphoric acid aqueous solution, flow rate 1.0 mL/min, column temperature 30 ℃, and detection wavelength 225 nm. The HPLC fingerprint was established and evaluated by the similarity evaluation system of TCM (version 2012A), and the difference of chemical information between 15 batches of different samples was evaluated by cluster analysis. Furthermore, the content of the nine active components in the sample was determined by HPLC multi-component wavelength switching method, with the partial least squares-discriminant analysis (PLS- DA) by SIMCA 14.1 software to find significant components of the quality between the batches. Results: The HPLC fingerprint of 15 batches of TSD was established. The similarity was greater than 0.96, and 35 common peaks were identified as gallic acid, chlorogenic acid, amygdalin, albiflorin, hydroxysafflor yellow A, paeoniflorin, ferulic acid, senkyunolide I, benzoylpaeoniflorin and ligustilide (corresponding to peaks 2, 8, 9, 13, 14, 15, 16, 25, 31, and 32). The linearity relationships of gallic acid, 5-hydroxymethylfurfural, chlorogenic acid, albiflorin, hydroxysafflor yellow A, paeoniflorin, ferulic acid, verbascoside, and senkyunolide I (r ≥ 0.999 6) were good. The results of content determination respectively were 187.5-344.4, 6.2-154.8, 413.2-459.2, 507.5-923.5, 873.8-1 202.0, 2 122.3-2 782.9, 59.2-121.3, 6.4-26.9, and 38.9-79.6 μg/g, respectively, including higher content of paeoniflorin, hydroxysafflor yellow A, and albiflorin. Furthermore, 15 batches of samples from different origins were classified into three categories. Using PLS-DA analysis, the content determination result showed that paeoniflorin, albiflorin, hydroxysafflor yellow A, and 5-hydroxymethylfurfural were the four components that affected the quality of different batches of TSD. Conclusion: HPLC fingerprint combined with multi-components determination is suitable for quality control and evaluation of TSD preparation.

Q-marker prediction analysis of Paeoniae Radix Alba based on fingerprint and network pharmacology / 中草药

Objective: To explore the potential Q-marker of Paeoniae Radix Alba in Danggui Sini Decoction based on fingerprint and network pharmacology. Methods: The fingerprints of Paeoniae Radix Alba decoction and Danggui Sini Decoction were established, and the law of components transfer was also defined. The "compounds-targets-pathways" network was then established to predict the potential Q-marker of Paeoniae Radix Alba through the network pharmacology. Results: The fingerprints of 15 batches of Paeoniae Radix Alba decoction and 15 batches of Danggui Sini Decoction were established, and the five chromatographic peaks were identified, they were gallic acid, albiflorin, paeoniflorin, 1,2,3,4,6-O-pentagalloylglucose, and benzoyl paeoniflorin. Through the network pharmacology analysis, the potential two active components, eight core targets and 13 key pathways were screened out, which indicated that paeoniflorin and albiflorin were preliminarily predicted to the potential Q-marker of Paeoniae Radix Alba. Conclusion: The analysis and prediction of the Q-marker in this study can provide a reference for the whole control of the Paeoniae Radix Alba quality, which can also provide the basis for the further research on the efficacy-related substance and mechanism of Paeoniae Radix Alba.

Study on application of DESI-MSI in quality control of classical famous prescription Shaoyao Gancao Decoction / 中草药

Objective: Using ultra-high performance liquid chromatography with diode array detection (UHPLC-DAD) and desorption electrospray ionization-mass spectrometry imaging (DESI-MSI) to analyze 15 batches of Shaoyao Gancao Decoction (SGD) substance benchmark and lyophilized powder in order to investigate the advantages of DESI-MSI in quality control of famous classical formulas. Methods: Taking SGD as the research model, fingerprints of the substance benchmark were established by UHPLC-DAD, and the content of index components (paeoniflorin, liquiritin, glycyrrhizic acid) and the yield of dry extract were also investigated. Meanwhile, as the research carrier, the lyophilized powder corresponding to SGD was dissolved in methanol and dotted on qualitative filter paper with quantitative capillary, and fixed it on the slide to make samples. The samples were analyzed on a DESI-MSI system in positive and negative ion mode with methanol-formic acid (1 000:1, flow rate of 3 μL/min) as spray solvent, N2 as spray gas (pressure of 0.5 MPa). The scanning range was m/z 100-1 200, the spatial resolution was 300 μm, the ion source temperature was 120 ℃. Results: DESI-MSI can detect not only the index components of paeoniflorin, liquiritin, glycyrrhizic acid, but also the common peaks of albiflorin. At the same time, DESI-MSI could detect 11 other components from Glycyrrhizae Radix et Rhizoma and Paeoniae Radix Alba, such as licoricesaponin G2, licoricesaponin J2, gallic acid, citric acid, p-hydroxybenzoic acid, and present their relative content visually. The qualitative analysis ability of DESI-MSI was much better than UHPLC-DAD. Conclusion: DESI-MSI can be used as the quality control method for substance benchmark and lyophilized powder and dispensing granules of classical famous formulas with advantages of high sensitivity, strong analytical ability, no complex sample pretreatment, qualitative and relative content analysis of complex samples without reference substance.

HPLC-ELSD fingerprint of Zhenwu Decoction and simultaneous determination of 13 components / 中草药

Objective: To establish HPLC-ELSD fingerprint of Zhenwu Decoction(ZWD), screen out the signature components of ZWD through chemical pattern recognition, so as to establish the content determination method of ZWD based on this index. Methods: The fingerprint of 16 batches of ZWD was established by HPLC-ELSD method. The similarity evaluation system of traditional Chinese medicine chromatographic fingerprint (2012 Version) was used for similarity evaluation to determine the common peaks and its attribution. Cluster analysis (CA), principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to select the index components of ZWD. Results: The fingerprint of ZWD was established, 38 common peaks were confirmed, and the similarity was > 0.95. The results of CA, PCA and OPLS-DA were consistent and the samples were divided into three categories. Benzoylmesaconine, benzoylaconitine, benzoylhypacoitine, polyporenic acid C, pachymic acid, atractylenolide II, atractylenolide III, oxypaeoniflorin, albiflorin, paeoniflorin and benzoylpaeoniflorin were identified as the 11 index components with significant difference contribution in different batches of ZWD samples. 6-Gingerol and 6-shogaol were the main active components of ginger, so the above 13 components were taken as the index components of ZWD. The chromatographic peak separation degree and linear relationship were good. The average recovery rate was 96.46%-99.80%, RSD ≤ 3.15%. The mass fraction range of benzoylmesaconine, benzoylaconitine, benzoylhypacoitine, polyporenic acid C, pachymic acid, atractylenolide II, atractylenolide III, oxypaeoniflorin, albiflorin, paeoniflorin, benzoylpaeoniflorin, 6-gingerol, 6-shogaol in 16 batches were 283.93-576.86, 25.05-147.39, 62.96-303.37, 31.24-131.27, 9.76-44.04, 32.15-83.55, 76.55-333.13, 17.48-146.61, 456.58-1554.14, 3 322.48-5 590.01, 158.21-556.50, 525.85-582.92 and 68.52-74.73 mg/g, respectively. Conclusion: The fingerprint combined with PCA, CA and OPLS-DA can comprehensively evaluate the quality of ZWD. This method is stable and reliable, providing reference for the quality evaluation.

Comparative analysis of monoterpene glycosides in Paeoniae Rubra Radix Formula Granule from different manufacturers / 中草药

Objective: To establish an HPLC method for simultaneous determination of 15 monoterpene glycosides pyrindyl- paeoniflorin, mudanpioside F, oxyalbiflorin, oxypaeoniflorin, 10-hydroxypaeoniflorin, albiflorin, paeoniflorin, oxypaeonidanin, 4-O- methyl-oxypaeoniflorin, galloylpaeoniflorin, 4-O-methyl-paeoniflorin, albiflorin R1, paeonidanin, benzoyloxypaeoniflorin, and benzoylpaeoniflorin in Paeoniae Rubra Radix Formula Granule, in order to compare the quality of Paeoniae Rubia Radix Formula Granule from different manufacturers and provide the basis for the establishment of a unified quality control method. Methods The separation was performed on an Agilent Zorbax SB-Aq C18 column (250 mm × 4.6 mm, 5 µm), using acetonitrile and potassium dihydrogen phosphate solution (pH 2.8) as the mobile phase at the flow rate of 1.0 mL/min for a gradient elution. The detection wavelength was set at 260 nm. Results: Paeoniflorin, albiflorin, and oxypaeoniflorin were the three main monoterpene glycosides with the highest content in the Paeoniae Rubra Radix Formula Granule. There were extremely significant difference among the contents of 15 monoterpene glycosides in Paeoniae Rubra Radix Formula Granule produced by different manufacturers. The sample of CSPFKL-KRT was remarkable for the highest content of paeoniflorin and oxypaeoniflorin (73.214 mg and 16.935 mg per gram sample, respectively) and the lowest content of albiflorin among all samples (2.343 mg per gram sample), while sample CSPFKL-XLS was remarkable for the lowest content of paeoniflorin and oxypaeoniflorin (26.327 mg and 4.165 mg per gram sample, respectively) and the highest content of albiflorin among all samples (18.893 mg per gram sample). Conclusion: There were extremely significant difference among the contents of main monoterpene glycosides in Paeoniae Rubra Radix Formula Granule produced by different manufacturers, which may affect the clinical use. The establishment of a unified quality standard plays an important role in the quality control of Paeoniae Rubia Radix Formula Granule.

Preliminary discovery of quality marker (Q-marker) of Shenzhiling Oral Liquid based on “fingerprint-efficacy-pharmacokinetics” correlation / 中草药

Objective: To establish a research strategy for discovering quality marker (Q-marker) of Shenzhiling Oral Liquid based on the “fingerprint-efficacy-pharmacokinetics” correlation. Methods: HPLC fingerprints and acetylcholinesterase (AchE) inhibitory activities of 12 batches of Shenzhiling Oral Liquid were analyzed. The correlation analysis between the HPLC fingerprints and AchE inhibitory effects were carried out with orthogonal signal correction-partial least squares regression (OSC-PLSR) method. Combined with network pharmacology, efficacy-related components were determined. By identifying the compounds absorbed and exposed in vivo, pharmacologically active components were determined. Finally, Q-marker could be preliminarily discovered by the comprehensive analysis associated with the integration of efficacy-related components and pharmacologically active ingredients. Results: The results of OSC-PLSR analysis showed that three efficacy-related components were closely related to AchE inhibitory activities. According to mapping the targets of diseases, 61 efficacy-related components were determined. Eleven active compounds in plasma were identified by UHPLC-quadrupole-orbitrap-MS. The Q-markers of Shenzhiling Oral Liquid were liquiritin apioside, albiflorin and azelaic acid preliminarily determined by integrated and comprehensive analysis. Conclusion: The combination of fingerprint-efficacy relationship, network pharmacology and components absorbed in plasma could be an effective way for rapid analysis and discovery of Q-marker in Shenzhiling Oral Liquid, which will be of great significance both to improve the quality control and evaluation, and ensure the safety and effectiveness of traditional Chinese medicines.

Study on fingerprint and gray correlation analysis of Paeoniae Radix Rubra from different habitats / 中草药

Objective: To study the fingerprints of Paeoniae Radix Rubra in different habitats, and determine the content of five chemical components (gallic acid, albiflorin, paeoniflorin, benzoic acid, and benzoyl paeoniflorin) and systematically cluster them. The relationship between origin and content was analyzed by grey correlation degree to provide reference for the quality evaluation of Paeoniae Radix Rubra. Methods: Fingerprints of Paeoniae Radix Rubra from 21 different producing areas were constructed by high performance liquid chromatography. The results were classified by principal component analysis and systematic cluster analysis. The gray correlation degree method was used to process the index components and their relative correlations were calculated. Results: HPLC fingerprints of Paeoniae Radix Rubra from 21 habitats were established, 11 common peaks were confirmed, and five of them (gallic acid, albiflorin, paeoniflorin, benzoic acid, and benzoyl paeoniflorin) were identified. The similarity of Paeonia lactiflora was greater than 0.9, and the similarity of Paeonia veitchii was less than 0.9. It was divided into two categories by principal component analysis combined with cluster analysis. The results of grey correlation analysis showed that the relative correlation (ri) was the largest in Gansu, followed by Ganzi in Sichuan. Conclusion: There is a big difference in the relative yield of Paeoniae Radix Rubra in different habitats. This experiment provides a scientific basis for the quality evaluation of Paeoniae Radix Rubra by fingerprint analysis, principal component analysis combined system cluster analysis and grey correlation analysis method.

Simultaneous Determination of 8 Ingredients in Guizhijiashaoyao Decoction by HPLC / 中国药房

OBJECTIVE： To establish a method for simultaneous determination of albiflorin， paeoniflorin， liquiritin， liquiritigenin， cinnamic acid， cinnamaldehyde， ammonium glycyrrhetate and 6-ginger phenol in Guizhijiashaoyao decoction. METHODS： HPLC method was adopted. The determination was performed on Kromasil C18 column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid solution （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelengths were 235 nm（0-35 min，albiflorin， paeoniflorin， liquiritin）、280 nm（35-65 min，liquiritigenin， cinnamic acid， cinnamaldehyde， ammonium glycyrrhetate and 6-ginger phenol）. The column temperature was 25 ℃， and the sample size was 20 μL. RESULTS： The linear range of albiflorin， paeoniflorin， liquiritin， liquiritigenin， cinnamic acid， cinnamaldehyde， ammonium glycyrrhetate and 6-ginger phenol were 2.125-34.000 μg/mL（r＝0.999 9）， 28.700-459.200 μg/mL（r＝0.999 7）， 3.675-58.800 μg/mL（r＝0.999 7）， 1.235-19.760 μg/mL（r＝0.999 8）， 2.300-36.800 μg/mL（r＝0.999 8）， 0.955-15.280 μg/mL（r＝0.999 7）， 36.000-576.000 μg/mL（r＝0.999 7） and 1.500-24.000 μg/mL（r＝0.999 7）， respectively. The quantitative limits were 0.135， 0.102， 0.096， 0.033， 0.013， 0.023， 0.663， 0.198 μg/mL； the detection limits were 0.041， 0.031， 0.029， 0.010， 0.004， 0.007， 0.201， 0.059 μg/mL. RSDs of precision， stability and reproducibility tests were all lower than 3％ （n＝6）. The recovery rates were 97.47％-100.76％（RSD＝1.33％，n＝6）， 98.15％-103.50％（RSD＝1.82％，n＝6）， 95.65％-100.84％（RSD＝2.38％，n＝6）， 96.75％-100.32％（RSD＝1.31％，n＝6）， 95.88％-102.75％（RSD＝2.52％，n＝6）， 95.63％-100.63％（RSD＝2.00％，n＝6）， 96.78％-100.45％（RSD＝1.35％，n＝6）， 95.71％-100.48％（RSD＝1.80％，n＝6）. CONCLUSIONS： The method is accurate， reliable and exclusive， and suitable for simultaneous determination of 8 ingredients in Guizhijiashaoyao decoction.

UPLC-Q-TOF-MS/MS Analysis of Brain and Blood Absorption Components of Buyang Huanwu Tang After Intragastric Administration / 中国实验方剂学杂志

Objective:To analyze and identify the brain and blood absorption components of rats after intragastric administration of Buyang Huanwu Tang(BYHWT). Method:The brain tissue,plasma of normal rats and the cerebral ischemia-reperfusion rats were analyzed by UPLC-Q-TOF-MS/MS.The prototype components in BYHWT were identified according to retention time,accurate relative molecular weight,primary and secondary mass spectrometry data. Result:After the administration of BYHWT,five compounds were found to enter the normal brain tissue through the blood-brain barrier and identified as calycosin-7-glucoside,albiflorin,formononetin-7-O-β-D-glucoside-6″-O-acetyl,safflower yellow A and astragaloside A;two compounds penetrated the blood-brain barrier and entered modeling brain tissue,and they were identified as calycosin-7-glucoside and formononetin-7-O-β-D-glucoside-6″-O-acetyl;seven compounds entered normal plasma and were identified as calycosin-7-glucoside,albiflorin,hydroxysafflor yellow A,et al;three compounds entered model plasma and identified as calycosin-7-O-β-D-glucoside-6″-O-acetyl,6″-O-acetyl-(6αR,11αR)-9,10-dimetho-xypterocarpan-3-O-β-D-glucoside and formononetin-7-O-β-D-glucoside-6″-O-acetyl. Conclusion:BYHWT has different pharmacological material basis in normal and cerebral ischemia-reperfusion rats.

Simultaneous determination of nine components in Paeoniae Radix Rubra by HPLC / 中草药

Objective A high performance liquid chromatographic method (HPLC) was established to simultaneously quantify the paeoniflorin, oxypaeoniflora, albiflorin, benzoyl paeoniflorin, benzoic acid, catechin, paeonol, gallic acid, and 1,2,3,4,6-penta-O- galloy-D-glucose of Paeoniae Radix Rubra. Methods The mobile phase comprised of acetonitrile and water containing 0.1% phosphoric acid. Column temperature was 30 oC, flow rate was 1.0 mL/min, and chromatography was monitored at 230 nm. Results The correlation coefficients between concentration and chromatographic peak area of gallic acid, oxypaeoniflora, catechin, albiflorin, paeoniflorin, 1,2,3,4,6-penta-O-galloy-D-glucose, benzoic acid, benzoyl paeoniflorin, and paeonol were respectively over 0.999 in the ranges of 0.004-1.200, 0.010-1.500, 0.044-0.660, 0.038-1.140, 0.042-2.100, 0.050-1.250, 0.040-0.600, 0.042-1.260, and 0.004-1.080 mg/mL, with good precision, stability, repeatability, and recovery. Conclusion The method is effective, accurate and reproducible, and can be used for the quality control of Paeoniae Radix Rubra.

Rapid determination of paeoniflorin and albiflorin in red peony root by near-infrared spectroscopy / 中草药

Objective To develop a method for the measurement of the content of paeoniflorin and albiflorin in red peony root (the roots of Paeonia lactiflora, RPR) by near-infrared spectroscopy (NIR) for quality rapid assessment. Methods A total of 65 RPR samples were collected from Sichuan, Zhejiang and Anhui of China, and then quantified the content of paeoniflorin and albiflorin by NIR and ultra-high performance liquid chromatography (UPLC). Fifty-three samples were chosen as the calibration set, while the remaining 12 samples were assigned to external validation set. The calibration model of paeoniflorin and albiflorin was further developed by modified partial least squares (MPLS) using UPLC data as reference based on optimized spectral regions, pre-treament of spetrum and the number of principal divisors. The content of paeoniflorin and albiflorin in unknown samples (validation model) was predicted according to the validation set of NIR. Results The standard errors of prediction (SEP) for the contents between the predicted value of NIR method and the estimated value of UPLC method were 1.437 2% for paeoniflorin and 0.784 3% for albiflorin, and their correlation coefficient (R) were 0.990 2 for paeoniflorin) and 0.994 4 for albiflorin in 53 calibration set samples. Moreover, the SEP and R of that in 12 validation set samples were 0.714 5%, 0.988 0% for paeoniflorin, and 0.632 4, 0.994 6 for albiflorin, respectively. The sum of paeoniflorin and albiflorin in RPR cultivated in Sichuan, Anhui, and Zhejiang province in China were 5.49%, 5.33%, and 4.81%, respectively. Conclusion NIR can quantify the amounts of paeoniflorin and albiflorin quickly and simultaneously in RPR for quality rapid assessment. The quality of RPR cultivated in Sichuan, Anhui, and Zhejiang is similar based on the sum of these two bioactive compounds without significant differences.

Quality control for eight components in Guishao Zhenxian Tablets based on quantitative analysis of multi-components with a single-marker / 中草药

Objective To establish a quantitative analysis of multi-components with a single-marker (QAMS) method for the simultaneous determination of oxypaeoniflorin, albiflorin, paeoniflorin, benzoylpaeoniflorin, cinnamic acid, cinnamaldehyde, saikosaponin a, and saikosaponin d in Guishao Zhenxian Tablets (GZT). Methods The separation was performed on a Thermo ODS C18 column (250 mm × 4.6 mm, 5 μm), with the mobile phase consisting of acetonitrile-0.1% phosphate acid solution for gradient elution. The column temperature was 25 ℃, and flow rate was 1.1 mL/min. Using paeoniflorin as internal reference substance, the relative correlation factors of oxypaeoniflorin, albiflorin, benzoylpaeoniflorin, cinnamic acid, cinnamaldehyde, saikosaponin a, and saikosaponin d were calculated and established by HPLC. The results were compared with those obtained by the external standard method to verify the rationality, feasibility, and repeatability of QAMS method. Results Oxypaeoniflorin, albiflorin, paeoniflorin, benzoylpaeoniflorin, cinnamic acid, cinnamaldehyde, saikosaponin a, and saikosaponin d had good relations within the ranges of 2.63-65.75 μg/mL (r = 0.999 7), 9.67-241.75 μg/mL (r = 0.999 8), 13.59-339.75 μg/mL (r = 0.999 5), 1.79-44.75 μg/mL (r = 0.999 4), 2.45-61.25 μg/mL (r = 0.999 1), 7.98-199.50 μg/mL (r = 0.999 7), 2.79-69.75 μg/mL (r = 0.999 3), and 2.51-62.75 μg/mL ( r= 0.999 5), respectively. The recovery rates were 98.64%, 99.33%, 100.02%, 97.42%, 96.95%, 98.75%, 98.21%, and 97.81%, RSDs were 0.89%, 1.19%, 1.00%, 1.33%, 1.51%, 0.99%, 1.43%, and 0.80%, respectively. The relative correlation factors values of oxypaeoniflorin, albiflorin, benzoylpaeoniflorin, cinnamic acid, cinnamaldehyde, saikosaponin a, and saikosaponin d to paeoniflorin were 0.520 9, 1.086 9, 0.476 8, 0.626 1, 0.802 1, 0.713 8, and 0.367 0, respectively. There were no significant difference in assay results between QAMS and the external standard method. Conclusion The QAMS method is feasible and credible, and could be used to determine oxypaeoniflorin, albiflorin, paeoniflorin, benzoylpaeoniflorin, cinnamic acid, cinnamaldehyde, saikosaponin a, and saikosaponin d in Guishao Zhenxian Tablets for the quality control.

Quantitative Study on Simultaneous Determination of Nine Components in Renshen Nüjin Pills Based on QAMS / 中国药学杂志

OBJECTIVE: To establish a quantitative analysis of multi-components by single-marker (QAMS) method for simultaneous determination of senkyunolide H, senkyunolide I, senkyunolide A, ligustilide, oxypaeoniflorin, albiflorin, paeoniflorin, benzoylpaeoniflorin and α-cyperone in Renshen Nüjin pills, in order to provide basis for studying its quality standards.METHODS: The analysis of the methanol extract of Renshen Nüjin pills was performed on Agilent Zorbax SB C18 column (4.6 mm×250 mm,5 μm), with mobile phase composed of methanol-acetonitrile (2∶1)-0.1% phosphoric acid solution at a flow rate of 0.9 mL•min-1 in gradient elution mode. The column temperature was maintained at 30 ℃, and the detection wavelengths were set at 280, 230 and 242 nm. Paeoniflorin was selected as the internal standard, and the relative correlation factors (RCF) of senkyunolide H, senkyunolide I, senkyunolide A, ligustilide, oxypaeoniflorin, albiflorin, paeoniflorin, benzoylpaeoniflorin and α-cyperone were determined by HPLC. The accuracy and feasibility of the method were validated by comparing the results of QAMS method and external standard method.RESULTS: The standard curves of senkyunolide H, senkyunolide I, senkyunolide A, ligustilide, oxypaeoniflorin, albiflorin, paeoniflorin, benzoylpaeoniflorin and α-cyperone had good linear relationship in the ranges of the tested concentrations. The precision, stability and repeatability complied with the requirements of methodology. The recoveries were 97.38%, 98.16%, 98.84%, 99.63%, 97.04%, 99.14%, 100.04%, 96.93% and 98.48%, RSDs were 1.38%, 1.18%, 0.97%, 0.86%, 1.68%, 1.30%, 0.57%, 1.32% and 1.19%, respectively. No significant differences were observed in the determination results by QAMS method and external standard method.CONCLUSION: The QAMS method can be used for the content determination and quality control of the nine components in Renshen Nüjin pills.

Enhancement of exposure and reduction of elimination for paeoniflorin or albiflorin via co-administration with total peony glucosides and hypoxic pharmacokinetics comparison / 中国药理学与毒理学杂志

OBJECTIVE Paeoniflorin (PF) and albiflorin (AF) are the major active components of total peony glucosides(TPG)from Paeonia lactiflora Pal,which have many biological activities such as anti-inflammatory, antioxidation and anti-hypertension effects. The drug-drug pharmacokinetic interaction among PF,AF and TPG,the pharmacokinetic comparisons of AF between hypoxia and normoxia,the transport of AF cross the blood-brain barrier cell model and the transport of AF/PF/TPG cross Caco-2 cell model were investigated.METHODS A highly sensitive and rapid UPLC-MS method with multiple-reaction monitoring(MRM)scanning via electrospray ionization(ESI)source operating both in the positive and negative ionization mode was successfully developed and validated for simultaneous quantitation of PF and AF in rat plasma after an oral administration of PF,AF and TPG. RESULTS The validated and developed UPLC-MS/MS method was successfully applied to simultaneously determine the AF and PF concentration in rat plasma and investigate pharmacokinetic interactions after a single intragastrical ad-ministration of PF,AF,co-administration of PF with AF and TPG,respectively.The elimination of both PF co-administered with AF and PF in TPG were slower than those for PF alone and the distribution in the tissues was wider.The combination of PF with AF or TPG could significantly increase the values of the AUC, MRT and t1/2of the drug PF, and reduce the values of CL of PF. From a comparison of the main pharmacokinetic parameters among AF alone, AF combined with PF and AF in TPG, the values of the MRT and t1/2of AF in TPG were greater than that of AF alone,and there were statistically signifi-cant differences in these parameters(P<0.05,P<0.01).It was also noticed that AUC and Cmaxof PF in hypoxia rats were significantly decreased compared with that of normaxia rats, suggesting that there was a decreased exposure of PF in rats under hypoxia. The multiple active components in TPG may lead to DDIs between some P-gp substrates. CONCLUSION The clinical performance of total peony glucosides would be better than that of single constitute. The outcomes of the study are expected to serve as a basis for development of clinical guidelines on total peony glucosides usage.

Simultaneous determination of five constituents in Baishao Formula Granules by HPLC / 中成药

AIM To establish an HPLC method for the simultaneous content determination of five constituents in Baishao Formula Granules (Paeoniae Radix Alba).METHODS The analysis of aqueous extract of this drug was performed on a 30 ℃ thermostatic Phenomenex C18 column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.05％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 230 nm.RESULTS Paeoniflorin,albiflorin,paeoniflorin sulfonate,gallic acid and benzoic acid showed good linear relationships within the ranges of 0.020-0.639 mg/mL (r =0.999 8),0.005-0.172 mg/mL (r =0.999 9),0.020-0.652 mg/mL (r =1.000 0),0.003-0.097 mg/mL (r =0.999 8),0.002-0.058 mg/mL (r =0.999 7),whose average recoveries were 99.1％,98.3％,98.6％,98.1％ and 99.5％ with the RSDs of 1.86％,1.37％,1.69％,1.46％ and 2.26％,respectively.The contents of various constituents in twenty-seven batches of samples demonstrated obvious differences.CONCLUSION We should pay attention to Paeoniae Radix Alba in Baishao Formula Granules due to its unstable quality.