ABSTRACT
Objective:To develop an UPLC-MS/MS method for the determination of paeoniflorin, tetramethylpyrazine, nobiletin, paeonol, amygdalin and ligustilide in Gexia-Zhuyu Decoction. Methods:Isocratic elution was carried out with mobile phase consisting of methanol- 4 mM ammonium formate. The separation was performed on Waters XTerra MS C18 (2.1 mm × 50 mm, 3.5 μm), and the mass spectrometer was operated in the positive and negative ionization electrospray (ESI) mode using multiple monitoring (MRM) for analysis of six components. Quantitative analysis by external standard method. The precursor to product ion transitions monitored for paeoniflorin, tetramethylpyrazine, nobiletin, paeonol, amygdalin and ligustilide were m/z 525.0→449.1, 137.1→55.1, 403.0→373.0, 167.2→149.4, 456.5→323.3 and 191.1→91.1, respectively. Results:Paeoniflorin, tetramethylpyrazine, nobiletin, paeonol, amygdalin and ligustilide were all analyzed exactly, the linear ranges were 0.000 2-0.012 8, 0.000 8-0.051 2, 0.000 1-0.006 4, 0.000 2-0.012 8, 0.000 6-0.038 4, 0.000 1- 0.006 4 ng, respectively. The r were 0.999 3, 0.997 5, 0.999 6, 0.992 6, 0.995 5 and 0.999 1, respectively. The recoveries of six analytes ranged from 98.00% to 105.3% and the relative standard deviations ( RSD) were all below 2.47%. Conclusions:This method was sensitive, which could be applied for the determination of paeoniflorin, tetramethylpyrazine, nobiletin, paeonol, amygdalin and ligustilide in Gexia-Zhuyu Decoction.
ABSTRACT
Objective To observe the effect of Ziyin-Jianghuo formula in treatment of Yin deficiency generating intrinsic heat type of the levels of monoamine neurotransmitters and estrogens in rats, and investigate its intervention in the neuroendocrine system. Methods There were 7 groups, which were sham operation group, model control group, estrogen tablet group, Gengnian capsule group, Ziyin-Jianghuo formula low, medium and high dose groups. Castration was performed by castration (extraction of ovaries) plus hot traditional Chinese medicine. The rats in the treatment group were given the above drugs 24 hours after the last administration of the hot Chinese medicine, once a day for 30 consecutive days. The sham operation group and the model control group were given an equal volume of purified water by simultaneous intragastric administration. The serum estradiol (E2), luteinizing hormone (LH), follicle stimulating hormone (FSH), prolactin (PRL) levels,uterine coefficient and monoamine neurotransmitters dopamine (DA), norepinephrine (NE), serotonin (5-HT), 5-hydroxyindoleacetic acid (5-HIAA) content in the hypothalamus were detected after 12 hours of the last treatment. Results Compared with the model group, the levels of serum FSH (4.39 ± 0.22 IU/L, 2.89 ± 0.91 IU/L, 2.84 ± 0.98 IU/L vs. 5.51 ± 0.24 IU/L), LH (14.48 ± 0.24 IU/L, 11.46 ± 0.33 IU/L, 5.28 ± 1.31 IU/L vs. 15.02 ± 0.37 IU/L) in the low, middle and high doses of Ziyin-Jianghuo formula significantly decreased (P<0.05), the levels of serum E2 (39.84 ± 6.08 pmol/L, 48.65 ± 6.77 pmol/L, 64.96 ± 7.97 pmol/L vs. 33.16 ± 4.62 pmol/L) significantly increased (P<0.05). The content of DA, 5-HT, 5-HIAA in the hypothalamus and the 5-HT/NE (0.48 ± 0.02, 0.43 ± 0.03, 0.27 ± 0.02 vs. 0.67 ± 0.02), 5-HIAA/5-HT (1.74 ± 0.09, 1.71 ± 0.10, 1.80 ± 0.17 vs. 2.00 ± 0.10) in the low, middle and high doses of Ziyin-Jianghuo formula significantly decreased ( P<0.05), the content of NE (663.34 ± 9.81 ng/kg, 695.94 ± 10.54 ng/kg, 790.76 ± 16.35 ng/kg vs. 602.95 ± 13.24 ng/kg) in the hypothalamus in the low, middle and high doses of Ziyin-Jianghuo formula significantly increased (P<0.05). The levels of serum PRL (10.16 ± 1.26 μg/L, 7.22 ± 1.26 μg/L vs. 14.80 ± 1.64 μg/L) in the middle and high doses of Ziyin-Jianghuo formula significantly decreased (P<0.05). Conclusions The Ziyin-Jianghuo formula has a significant positive regulation effect on the neuroendocrine system of menopausal Yin deficiency generating intrinsic heat type of rats, and this process is dose-dependent and can improve a series of symptoms caused by autonomic dysfunction.