Comprehensive HPLC fingerprint analysis based on a two-step extraction method for quality evaluation of Perilla frutescens (L.) Britt.

Abundant chemical components are key to ensure the evaluation accuracy of fingerprint analysis of traditional Chinese medicines (TCMs). A two-step extraction method combining supercritical fluid extraction (SFE) and water ultrasonic extraction was established for the quality evaluation of Perilla frutescens (L.) Britt. Weakly polar components were extracted under optimal SFE conditions (15% co-solvent (EtOH : n-hexane = 1 : 14, (v/v)), 40 °C, 250 bar, and 30 min), and polar components were subsequently extracted by an ultrasonic step (100% water as solvent, 40 °C, and 45 min). Then, HPLC methods were established, which were validated to be accurate, stable, and reliable. In this work, 25 batches of samples were evaluated and the data were analysed by similarity analysis (SA) and hierarchical cluster analysis (HCA). The similarity values of SFE extracts and aqueous extracts were respectively 0.616-0.999, and 0.252-0.997, proving the importance of the extraction method for the accuracy of the subsequent fingerprint analysis results. For the HCA, 25 samples were divided into two categories (leaves and stems), among which four batches of leaves with less similarity were considered as stems, indicating that quality differences of P. frutescens depending on medicinal parts and origin exist. The two-step extraction method developed in this work has been proved to be suitable for the quality evaluation of TCMs with complex compositions.

Isolation of achiral aliphatic acid derivatives from Piper kadsura using preparative two-dimensional chiral supercritical fluid chromatography.

The separation of structural analogues in natural products has always been one of the challenges in separation science, where supercritical fluid chromatography (SFC) with chiral stationary phases (CSPs) is an unconventional but potential solution. In this study, a preparative two-dimensional chiral SFC (2D cSFC) method that was established with two kinds of CSPs was applied in the isolation of the aliphatic acid derivatives in Piper kadsura (P. kadsura). The RPLC unseparated peaks of two samples A and B of P. kadsura were evenly scattered on the CSP-1 column while they clustered into two groups on the CSP-2 column by SFC. There was impressively complementary selectivity between CSP-1 and CSP-2, which were used for construction of 2D cSFC. The first dimension (1D) separation with CSP-1 fractionated the sample A into six parts by a heart-cutting method and the sample B into nine parts for a comprehensive 2D analysis; then 29 and 71 peaks were respectively found in these parts in the second dimension (2D) separation with CSP-2. Further through 2D preparative separation, 19 high purity components were obtained, and the chemical structures of two of them were confirmed, including a novel unsaturated aliphatic acid compound (8Z,10Z)-12-methoxyheptadeca-8,10-dienoic acid and a known octadecadienoic acid lactone Lactariolide. The 2D cSFC method presented the superiority of separating the achiral compounds of complex samples.

Salvia miltiorrhiza Injection Alleviates LPS-Induced Acute Lung Injury by Adjusting the Balance of MMPs/TIMPs Ratio.

Salvia miltiorrhiza injection (SMI) is a classical traditional Chinese medicine, which plays an active role in the treatment of many diseases such as promoting blood circulation, removing blood stasis, reducing inflammatory reaction, and improving acute lung injury (ALI). Previous studies have shown that matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) are involved in the pathophysiological process of ALI. However, the relationship between SMI and MMPs/TIMPs remains unclear. In this study, Wistar rats were randomly divided into control group (NC), Salvia miltiorrhiza group (SM), lipopolysaccharide group (LPS), and Salvia miltiorrhiza treatment group (Tsm). The four groups were subdivided into four time points (2, 6, 12, and 24 hours), and specimens were collected after animal sacrifice at each time point. Serum TNF-α and IL-6 levels were detected by ELISA. The degree of lung injury was determined by lung tissue hematoxylin-eosin staining, lung wet/dry weight (W/D) ratio, and lung permeability index. The changes in lung MMPs/TIMPs protein and mRNA were detected by Western blot and real-time quantitative PCR. The results showed that rats injected with LPS experience acute lung injury, and the ratio of MMPs/TIMPs in lung tissues increased gradually with time. In the Tsm group, the ratio of MMPs/TIMPs decreased gradually, and likewise, the balance was gradually restored, while indicators related to lung injury were gradually declined. These data suggest that SMI alleviates LPS-induced acute lung injury; this protective effect may be related to regulation of the balance of MMPs/TIMPs ratio.

Isolation, structural elucidation, MS profiling, and evaluation of triglyceride accumulation inhibitory effects of benzophenone C-glucosides from leaves of Mangifera indica L.

Seventy percent ethanol-water extract from the leaves of Mangifera indica L. (Anacardiaceae) was found to show an inhibitory effect on triglyceride (TG) accumulation in 3T3-L1 cells. From the active fraction, six new benzophenone C-glucosides, foliamangiferosides A(3) (1), A(4) (2), C(4) (3), C(5) (4), C(6) (5), and C(7) (6) together with 11 known benzophenone C-glucosides (7-17) were obtained. In this paper, isolation, structure elucidation (1-6), and MS fragment cleavage pathways of all 17 isolates were studied. 1-6 showed inhibitory effects on TG and free fatty acid accumulation in 3T3-L1 cells at 10 µM.

Flavonol glycosides, nigelflavonosides A-F from the whole plant of Nigella glandulifera (Ranunculaceae).

Six new flavonol glycosides, nigelflavonosides A-F (1-6), together with a known compound (7) were isolated from the whole plant of Nigella glandulifera Freyn et Sint (Ranunculaceae). Structure elucidation, especially the localization of the glycosyl or acetyl groups, and complete (1)H- and (13)C-NMR assignments of these compounds were carried out using one- and two-dimensional NMR measurements, including (1)H- and (13)C-NMR, (1)H-(1)H COSY, TOCSY, HMQC, HMBC and NOESY, in addition to HRESI-TOF-MS experiments.

Identification of benzophenone C-glucosides from mango tree leaves and their inhibitory effect on triglyceride accumulation in 3T3-L1 adipocytes.

A 70% ethanol-water extract from the leaves of Mangifera indica L. (Anacardiaceae) inhibited triglyceride (TG) accumulation in 3T3-L1 cells. From the active fraction, seven new benzophenone C-glycosides, foliamangiferosides A (1), A(1) (2), A(2) (3), B (4), C(1) (5), C(2) (6), and C(3) (7), together with five known compounds were isolated and the structures were elucidated on the basis of chemical and physicochemical evidence. The effects of these compounds on TG and the free fatty acid level in 3T3-L1 cells were determined, and the structure-activity relationship was discussed. On the basis of the AMPK signaling pathway, several compounds were found to increase the AMPK enzyme expression and down-regulate lipogenic enzyme gene expression such as SREBP1c, FAS, and HSL.