[Characterization and identification of chemical constituents from Schizonepetae Spica based on UHPLC-Q-TOF-MS/MS technique].

The chemical constituents of Schizonepetae Spica were qualitatively analyzed by UHPLC-Q-TOF-MS/MS. An Agilent poroshell 120 SB-C_(18) column(3.0 mm×100 mm, 2.7 µm) was used for gradient elution with 0.1% formic acid water(A)-acetonitrile(B) solution as mobile phase at the flow rate of 0.4 mL·min~(-1) and column temperature of 45 ℃. The data were collected by scanning in positive and negative ion modes, and the compounds were identified by comparison of reference materials and PeakView software. Ninety-seven compounds were identified from Schizonepetae Spica, including 28 flavonoids, 23 phenolic acids, 23 fatty acids, 15 terpenoids, and 8 other compounds. The UHPLC-Q-TOF-MS/MS method established in this study can identify the chemical components of Schizonepetae Spica rapidly, accurately, and comprehensively, and provide a basis for the basic study of pharmacodynamic substances of Schizonepetae Spica.

Discrimination and Characterization of the Volatile Organic Compounds in Schizonepetae Spica from Six Regions of China Using HS-GC-IMS and HS-SPME-GC-MS.

Volatile organic compounds (VOCs) are the main chemical components of Schizonepetae Spica (SS), which have positive effects on the quality evaluation of SS. In this study, HS-SPME-GC-MS (headspace solid-phase microextraction-gas chromatography-mass spectrometry) and HS-GC-IMS (headspace-gas chromatography-ion mobility spectrometry) were performed to characterize the VOCs of SS from six different regions. A total of 82 VOCs were identified. In addition, this work compared the suitability of two instruments to distinguish SS from different habitats. The regional classification using orthogonal partial least squares discriminant analysis (OPLS-DA) shows that the HS-GC-IMS method can classify samples better than the HS-SPME-GC-MS. This study provided a reference method for identification of the SS from different origins.

[Textual research of Schizonepetae Herba and Schizonepetae Spica].

Schizonepetae Herba and Schizonepetae Spica are well-known Chinese herbal medicines for wind dispersing and exterior releasing. Through textual research on Schizonepetae Herba and Schizonepetae Spica, the discrimination of their medicinal parts in history was clarified, and the processing, the property(nature and flavor), meridian tropism, functions, indications, usage, dosage, and the selection of decoction pieces were compared to provide the basis for clinical application. As a result, the whole herb of Schizonepeta tenuifolia was used as medicine in the early records. The aerial part and the dried spike of S. tenuifolia were used as medicines separately in the Song Dynasty, which was recorded in the Atlas of Materia Medica(Ben Cao Tu Jing). Some ancient classics emphasized that only the dried spike could be used as medicine. The separation of Schizonepetae Herba and Schizonepetae Spica meets the different needs of clinical medication and supports the concept of rational development and utilization of Chinese medicine resources. About ten processing methods for Schizonepetae Herba and Schizonepetae Spica have been recorded since ancient times, and raw and charred drugs were the major products. Raw Schizonepetae Herba is required to be used in sections, whereas raw Schizonepetae Spica in clean preparation. Both charred products should avoid scorching. Schizonepetae Herba and Schizonepetae Spica are similar in the property(pungent, bitter, and warm), meridian tropism(lung and liver meridians, as well as qi and blood aspects), and functions(releasing exterior, dispersing wind, regulating and stopping blood, promoting eruption, dispelling sores, promoting digestion, eliminating alcohol effect, etc.), but Schizonepetae Spica is superior in efficacy. For Schizonepetae Herba and Schizonepetae Spica in traditional Chinese medicinal prescriptions, the raw and charred products are similar in usage and dosage, while their focuses in clinical compatibility vary. The raw and charred products of Schizonepetae Herba and Schizonepetae Spica are widely applied clinically. Decoction pieces of different specifications can result in different efficacies and clinical applications, so medication should be performed with caution.

The Potential Transformation Mechanisms of the Marker Components of Schizonepetae Spica and Its Charred Product.

Schizonepetae Spica (SS) is commonly used for treating colds, fevers, bloody stool and metrorrhagia in China. To treat colds and fevers, traditional Chinese medicine doctors often use raw SS, while to treat bloody stool and metrorrhagia, they usually use Schizonepetae Spica Carbonisata (SSC; raw SS processed by stir-frying until carbonization). However, there have been limited investigations designed to uncover the mechanism of stir-fry processing. In the present study, a method combining gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC) was developed for the comprehensive analysis of the chemical profiles of SS and SSC samples. Principal component analysis of the GC-MS data demonstrated that there were 16 significant differences in volatile compounds between the SS and SSC samples. The simultaneous quantification of six nonvolatile compounds was also established based on HPLC, and remarkable differences were found between the two products. These changes were probably responsible for the various pharmacological effects of SS and SSC as well as the observed hepatotoxicity. Finally, the mechanisms could be rationalized by deducing possible reactions involved in the transformation of these marker components. This work reports a new strategy to reveal the chemical transformation of SS during stir-fry processing.

Gas-Mediated Intestinal Microbiome Regulation Prompts the Methanol Extract of Schizonepetae Spica to Relieve Colitis.

Intestinal dysbiosis plays an important role in the pathogenesis of colitis (UC). Schizonepetae Herba can achieve anti-inflammatory effects as a medicine and food homologous vegetable. Luteolin, eriodictyol, fisetin, and kaempferol are the main anti-inflammatory active compounds obtained through mass spectrometry from the methanol extract of Schizonepetae Spica (JJSM). JJSM intervention resulted in attenuated weight loss, high disease-activity-index score, colon length shortening and colonic pathological damage in DSS-induced colitis mice. Interestingly, hydrogen sulfide (H2S) was inhibited remarkably, which is helpful to elucidate the relationship between active substance and intestinal flora. Furthermore, JJSM administration improved intestinal flora with down-regulating the abundance of harmful bacteria such as Clostridiales and Desulfovibrio and up-regulating the abundance of beneficial bacteria such as Muribaculaceae and Ligolactobacillus and enhanced the production of SCFAs. It is worth noticing that Desulfovibrio is related to the production of intestinal gas H2S. The elevated levels of Desulfovibrio and H2S will hasten the onset of colitis, which is a crucial risk factor for colitis. The results displayed that JJSM could considerably ameliorate colitis by rebuilding H2S-related intestinal flora, which provides a new therapeutic strategy for Schizonepetae Spica to be utilized as a functional food and considered as an emerging candidate for intestinal inflammation.

Comparisons of the anti-inflammatory, antiviral, and hemostatic activities and chemical profiles of raw and charred Schizonepetae Spica.

ETHNOPHARMACOLOGICAL RELEVANCE: A common view in traditional Chinese medicine (TCM) theory is that "processing can alter the efficacy of crude drugs". The clinical usage of some processed products may have already changed greatly over time during the development of modern scientific analysis. Therefore, the view of "processing can alter the efficacy of crude drugs" should be confirmed by comparative studies. Schizonepetae Spica (SS), a Chinese medicinal herb, is the dried spike of Schizonepeta tenuifolia Briq. It is available in two forms: raw products and charred products (Schizonepetae Spica Carbonisata, SSC; raw SS processed by stir-frying until carbonization). Raw SS is commonly used to treat TCM symptoms that resemble common cold, fever, respiratory tract infection and allergic dermatitis, while SSC has long been used as a remedy for TCM symptoms that resemble bloody stool and metrorrhagia. AIM OF THE STUDY: We aimed to examine whether stir-fry processing alters the anti-inflammatory, antiviral and hemostatic activities of SS and explore the chemical profile behind the potential changes in medicinal properties caused by stir-fry processing. MATERIALS AND METHODS: We used cell models to examine the anti-inflammatory and antiviral effects of raw SS and SSC. The bleeding time of the tail bleeding model and clotting time of the capillary method in mice were used to compare the hemostasis properties of raw SS and SSC. The chemical profiles of SS and SSC were compared using a method combining gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (HPLC/Q-TOF-MS) analysis. RESULTS: The anti-inflammatory effects of SSC were less potent than those of raw SS. Both raw SS and SSC effectively inhibited viral infection in a dose-dependent manner, with IC50 values of 96.30 and 9.73 µg/mL and selectivity index (SI) values of >1.56 and 7.78, respectively. Interestingly, SSC showed more potent antiviral activities than raw SS. Intragastric administration of raw SS and SSC to mice demonstrated that the hemostatic effects of SSC were more potent than those of raw SS. By comparing the volatile chemical profiles of SSC, we found that twenty-nine constituents disappeared and that fifty-four new constituents were formed while the relative contents of five other components decreased and three other components increased. Additionally, the nonvolatile chemical profiles of raw SS and SSC differed, with thirty-two lower peaks and seven higher peaks in SSC than in SS. CONCLUSION: Our study showed that raw SS and SSC support traditional practice for the clinical applications of these two products except for raw SS used for the treatment of viral infection. It is a fascinating challenge to form SSCs with both traditional hemostatic activities and antiviral properties after stir-fry processing. In addition, the volatile and nonvolatile chemical constituents of raw SS changed dramatically during processing. Further studies are warranted to explore whether the change in chemical constituents is in accordance with the purpose of processing.