Investigating the mechanical effect of the sagittal angle of the cervical facet joint on the cervical intervertebral disc.

Background: Facet tropism is defined as the asymmetry between the left and right facet joints relative to the sagittal plane. Published clinical studies have found that facet tropism is associated with cervical disc herniation. However, the relationship between the facet orientation and the side of cervical disc herniation remains controversial. Therefore, this study used the finite-element technique to investigate the biomechanical effects of the sagittal angle of the cervical facet joints on the cervical intervertebral disc. Objective: The biomechanical effects of the sagittal angle of the cervical facet joint on the cervical disc and facet joint were investigated using the finite-element technique. Methods: The finite-element model was constructed using computed tomography scans of a 26-year-old female volunteer. First, a cervical model was constructed from C3 to C7. The model was verified using data from previously published studies. Second, the facet orientation at the C5-C6 level was altered to simulate different sagittal angles of cervical facet joints. Five models, F70, F80, F90, F100, and F110, were simulated with different facet joint orientations (70°, 80°, 90°, 100°, and 110° facet joint angles at the left side, respectively, and 90° facet joint angles at the right side) at the C5-C6 facet joints. In each model, annular fibres stress and facet cartilage pressure were studied under six pure moments and two combined moments. Results: Comparing the stress of the annulus fibres in flexion combined with right axial rotation and in flexion combined with left axial rotation in the same model, no difference in the maximum stress of the annulus fibres was noted between these two different moments in the F90 model, whereas differences of 12.80%, 8.84%, 14.95% and 33.32% were noted in the F70, F80, F100 and F110 models, respectively. The same trend was observed when comparing the maximum stress of the annulus fibres in each model during left and right axial rotation. No differences in annular fibres stress and facet cartilage pressure were noted among the five models in flexion, extension, lateral bending, left axial rotation, and flexion combined with left axial rotation in this study. However, compared with the F70 model in flexion combined with right axial rotation, the annulus fibres stress of the F80, F90, F100, and F110 models increased by 5.53%, 13.03%, 35.04%, and 72.94%, respectively, and the pressure of the left facet joint of these models decreased by 5.65%, 12.10%, 18.41%, and 25.74%, respectively. The same trend was observed in the right axial moment. Conclusion: Facet tropism leads to unbalanced stress distribution on the annulus fibres at the cervical intervertebral disc. The greater the sagittal angle of the facet joint, the greater the annular fibres stress on this side. We hypothesised that the side with the larger sagittal angle of the facet joint exhibits a greater risk of disc herniation.

[Separation and Quantitative Determination of Four Flavonoids in Tetrastigma hemsleyanum].

OBJECTIVE: To develop a method for separation and quantitative determination of four flavonoids in Tetrastigma hemsleyanum. METHODS: The Separation was performed on a Waters Preparative scale LC-MS autopurification system. Several spectral analyses, such as NMR,HR-MS and UV were used to identify the chemical structures. Quantitative assay was performed on a UHPLC Phenomenex Kinetex C18 column (100 mm x 4.6 mm, 2.6 µm) with the mobile phase consisting of acetonitrile( B) and water containing 0. 1% formic acid (A) in a gradient mode at a flow rate of 0.4 mL/min. The column temperature was at 30 °C and the optimum detection wavelength of DAD was set at 350 nm. RESULTS: Four flavonoids were isolated from the root of Tetrastigma hemsleyanum, which were identified as rutin, isoquercitrin, kaempferol-3-O-rutinoside and astragalin. As to quantitative analysis, a good separation of four flavonoids with in 17.5 min, the linear range of rutin, isoquercitrin, kaempferol-3-O-rutinoside and astragalin was 0.283-11.32 µg/mL (r1 = 0.9998), 0.311-12.44 µg/mL (r2 = 0.9994), 0.277-11.08 µg/mL (r3 = 0.9995) and 0.103-2.06 µg/mL (r4 = 0.9990), respectively. The average recovery (n = 6) was 99.75% (RSD = 2.15%), 98.73% (RSD = 2.58%) ,98. 03% (RSD = 2.23%), and 97.62% (RSD = 1.95%), respectively. CONCLUSION: Four flavonoids are isolated from the root of Tetrastigma hemsleyanum for the first time, and the UHPLC method is simple, rapid and accurate, which can be used for quantitative analysis of multi-component of Tetrastigma hemsleyanum and provide a novel approach for evaluation of the quality of Tetrastigma hemsleyanum comprehensively.

[Rapid simultaneous determination of ten major flavonoids in Tetrastigma hemsleyanum by UPLC-MS/MS].

In this study, a rapid and sensitive analytical method was developed for the determination of 10 major compounds (procyanidin B1, catechin, procyanidin B2, rutin, isoquercitrin, kaempferol-3-O-rutinoside, astragalin, quercitrin, quercetin, and kaempferol) in Tetrastigma hemsleyanum by using ultra-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UPLC-MS/MS) in multiple-reaction monitoring (MRM) mode. UPLC-MS/MS assay with negative ion mode was performed on a Waters CORTECS C18 (2.1 mm x 100 mm, 1.6 µm) with the mobile phase consisting of acetonitrile (A) and 0.1% aqueous formic acid (B) in gradient elution at a flow rate of 0.25 mL · min(-1) and the column temperature was set at 45 °C. Under the optimized chromatographic conditions, good separation for 10 target compounds were obtained including chiral isomer procyanidins B1 and B2 were completely separated within 8.5 min. Satisfactory linearity was achieved with wide linear range and fine determination coefficient (r > 0.996 6), the overall recoveries were ranged from 95.44%-110.40% with the RSD ranging from 2.37%-8.69%. It is the first report about simultaneous analysis of 10 major flavonoids components in Tetrastigma hemsleyanum by using UPLC-MS/MS method, which affords highly sensitive, specific, speedy and efficient method for quality control of Tetrastigma hemsleyanum

[Qualitative and quantitative analysis of major constituents in Tetrastigma hemsleyanum by HPLC-Q-TOF-MS and UPLC-QqQ-MS].

A qualitative analytical method of liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS) was developed for identification of multi-constituents and an analytical method was developed for simultaneously determining 4 major compounds (rutin, isoquercitrin, kaempferol-3-0-rutinoside, and astragalin) in Tetrastigma hemsleyanum Diels et Gilg. The HPLC-Q-TOF-MS assay was performed on a Welch Ultimate XB-C18 column (4.6 mm x 150 mm, 5 microm) with the mobile phase consisting of acetonitrile (A) and water containing 0.1% Formic acid (B) in gradient mode at a flow rate of 0.8 mL x min(-1). The column temperature was at 30 degrees C, and negative ion mode was used for TOF-MS. The UPLC-QqQ-MS assay was performed on a Waters CORTECS C18 (2.1 mm x 100 mm, 1.6 microm) with the mobile phase consisting of acetonitrile (A) and water containing 0.1% formic acid (B) in gradient mode at a flow rate of 0.25 mL x min(-1). The column temperature was at 45 degrees C, and MRM mode was used for QqQ-MS. Based on the retention time and MS spectra, 24 compounds were identified or tentatively characterized by comparing with reference substances or literatures. For quantitative the linear range of 4 detected compounds were good (r > 0.9966), and the overall recoveries ranged from 98.27% to 101.58%, with the RSD ranging from 3.15% to 5.88%. The results indicated that new approach conbined HPLC-Q-TOF-MS and UPLC-QqQ-MS was applicable in qualitative and quantitative quality control of Tetrastigma hemsleyanum.