ABSTRACT
OBJECTIVE： To optimize and improve the quality standard for Citrus reticulata formula granules. METHODS： Totally 13 batches of C. Reticulata formula granules from 4 different manufacturers were used as trial samples， and qualitative identification of hesperidin and nobiletin in the samples were carried out by TLC according to the method of 2015 edition of Chinese Pharmacopoeia （part Ⅳ）. The quantitative analysis of naringin， hesperidin， hesperetin， nobiletin and tangeretin in C. reticulatae formula granules were conducted by UPLC[The determination was performed on Waters Acquity UPLC BEH C18 column with mobile phase consisted of acetonitrile-0.2％ phosphoric acid aqueous solution （gradient elution）. The detection wavelength was set at 283 nm， and sample size was 3 μL]. RESULTS： The results of TLC showed that in the chromatograms of samples， same color spots were shown in the corresponding positions of the chromatogram of reference substance. The results of UPLC showed， that the linear range of naringin， hesperidin， hesperetin， nobiletin and tangeretin were 0.64-6.44， 15.78-157.80， 0.17-1.66， 2.08-20.85 and 2.04-20.43 μg/mL， respectively （all r≥0.999 2）； the limits of detection were 0.03， 0.33， 0.10， 0.20 and 0.06 μg/mL； the limits of quantitation were 0.07， 1.34， 0.20， 0.60 and 0.22 μg/mL. The average recoveries were 99.4％， 99.6％， 99.7％， 99.7％ and 99.7％ （n＝9）； RSDs of precision （n＝6）， stability （n＝7） and reproducibility （n＝6） tests were all≤2.03％； naringin was detected in only 3 batches of samples from one manufacturer （the content ranged from 0.067 3 to 0.069.6 mg/g）， while the other 4 components were detected in 13 batches of samples （the contents of them ranged 0.646 5-1.728 0， 0.102 6-0.290 5， 0.023 1-0.689 8， 0.018 2-0.270 7 mg/g）. CONCLUSIONS： In this study， the quality standard of C. reticulata formula granules was improved by qualitative and quantitative methods， and the contents of hesperidin， hesperetin， nobiletin and tangeretin were not less than 0.60， 0.10， 0.02 and 0.01 mg/g， respectively.
ABSTRACT
Objective The incidence of cardiovascular and cerebrovascular diseases in postmenopausal women with type 2 diabetes is grim.The study was designed to explore the effect of ω-3 polyunsaturated fatty acids (PUFA) on endothelial function in postmenopausal women with type 2 diabetes. Methods 50 patients admitted to Dingli Medical College of Wenzhou Medical Univer-sity from March 2014 to October 2014 were divided into group A and Group B by random number table .Cross-design of two stages ( I, II) was applied in the investigation .At stage I(3 months ahead of the experiment ), Group A took oral ω-3 PUFA while Group B took placebo .At stage II ( 3 months after the experiment ) , Group B was given oral ω-3 PUFA, while Group A was given placebo .T1 and T3 time was the beginning of the stage I and stage II experiment , while T2 and T4 time was the end of stage I and stage II experiment .At the beginning and end of each stage , detection was made on LDL-C, TG, IL-6, plasminogen activator inhibitor-1 (PAI-1) and endothelium-dependent flow-mediated vasodilation (FMD). Results After the intervention on Group A at stage I , FDM at T2 time was significantly increased compared with that at T 1 time([7.23 ±3.28]% vs [3.62 ±2.13]%, P<0.05), while all the other indexes at T2 time decreased significantly in comparison with T1 time: LDL-C ([2.85 ±0.47]mmol/L vs [3.36 ±0.57] mmol/L), TG([2.41 ±1.06]mmol/L vs [2.96 ±1.12] mmol/L), IL-6([2.83 ± 0.30]ng/L vs [3.42 ±0.32]ng/L), PAI-1 ([7.23 ±3.28]ng/L vs [3.62 ±2.13]ng/L) (P<0.05).After receiving ω-3 PUFA intervention on Group B at stage II , FDM at T4 time was significantly increased compared with that at T 3 time([6.88 ±2.06]% vs [3.60 ±2.18]%, P<0.05), while all the other indexes at T4 time decreased significantly in comparison with T3 time: LDL-C ([3.26 ±0.77]mmol/L vs [3.63 ±0.73] mmol/L), TG([2.28 ±0.94]mmol/L vs [2.77 ±1.25] mmol/L), IL-6([2.91 ± 0.48]ng/L vs [3.30 ±0.52]ng/L), PAI-1 ([45.7 ±24.4]ng/L vs [56.3 ±24.4]ng/L) (P<0.05).Two-period crossover design analysis of variance showed that there was significant difference effect on LDL -C(F=2.754, P=0.019), TG(F=3.115, P=0.011), IL-6(F=1.825, P=0.032), PAI-1(F=2.324, P=0.023) and FMD(F=3.784, P=0.006)between ω-3 PUFA and placebo . Conclusion ω-3 PUFA can improve endothelial function in postmenopausal women with type 2 diabetes , which is of great significance for the primary prevention of cardiovascular disease .