Simultaneously quantitative analysis of peptides and chemical components in Cervus and Cucumis polypeptide injection (Songmeile®) using reversed phase liquid chromatography-hydrophilic interaction liquid chromatography-tandem mass spectrometry.

Co-occurrences of peptides and chemical components are usually observed in complicated matrices. Notably, those traditional Chinese medicine prescriptions (TCMPs) contain both plant and animal ingredients. It is still challenging to simultaneously monitor peptides and chemical components attributing to their different liquid chromatographic (LC) and mass spectrometric (MS) behaviors. Herein, efforts were made to pursue an eligible approach enabling simultaneous determination of peptides and chemical components in a TCMP namely Cervus and Cucumis polypeptide injection (CCPI, Songmeile®) that is prepared from the acid hydrolytic peptide-enriched extract of Sika deer (Cervus nippon Temminck) bone and the aqueous extract of muskmelon (Cucumis melo L.) seeds. Reversed phase liquid chromatography and hydrophilic interaction liquid chromatography were serially connected (RPLC-HILIC) to achieve comprehensive retention and separation. Sensitive detection was accomplished with selected reaction monitoring (SRM) mode, and multiply charged and singly charged ion transitions were defined for peptides and chemical components, respectively. Inter-batch variations of CCPI were evaluated in an authentic compound-independent manner. In particular, online energy-resolved MS was proposed to gain optimal parameters for five targeted peptides after that CCPI peptidome was profiled using nanoLC-LTQ Orbitrap Velos Pro MS. A so-called universal metabolome standard (UMS) sample was built for calibration curve construction and subsequently applied to acquire the quasi-contents of all 31 analytes, including five peptides and 26 chemical components, in ten batches of CCPI (CCPI1-CCPI10). The quantitative dataset revealed mild fluctuation for the quasi-content profiles of analytes-of-interest within different batches. More importantly, RPLC-HILIC-SRM is a promising method to fully address the demands of simultaneous measurement of peptides and chemical components in complicated matrices, and it might be a robust analytical tool for in-depth quality evaluation of CCPI as well as other TCMPs.

[Two new polypeptides from extract of deer bone extract].

Two new polypeptides were isolated and purified from the extract of deer bone (constitutive part of Cucumis and Cervus polypeptide injection) by various column chromatography including C4 300Å and Sephadex G-50, as well as semipreparative HPLC. Their N-terminal amino acid sequences were identified by De Novo sequencing on the basis of MALDI-TOF-MS data and Explorer™ software. The N-terminal amino acid sequences of polypeptides were identified as NH2-Gly-Pro-Val-Gly-Pro-Thr-Gly-Pro-Val-Gly-Ala-Ala-Gly-Pro-Ser-Gly-Pro-Asp (Mei18 peptide, 1) and NH2-Ala-Gly-Pro-Ala-Gly-Pro-Leu-Gly-Pro-Leu-Gly-Pro-Leu-Gly-Pro-Leu-Gly-Pro-Pro-Asp-Ser-Try-Asp (Mei23 peptide, 2), respectively. Mei18 and Mei 23 peptides are new polypeptides.

[Clinical analysis of cervus and cucumis polypeptide injection for lumbar vertebrae diseases based on real world hospital information system].

To analyze the clinical drug use characteristics and medication rules of cervus and cucumis polypeptide injection (CCP injection) for the patients with lumbar vertebra diseases. Based on the electronic medical data of 6 990 patients with CCP injection for lumbar vertebra diseases from 25 first-class hospitals over the whole country from 2004, statistical analysis was conducted for their general information, diagnostic information, orders information, laboratory test information and treatment results. The results showed that the age distribution was highest in patients aged 45 to 64 (3 883, 55.55%); protrusion of lumbar intervertebral disc was most common among lumbar vertebra diseases (5 519, 82.50%); and TCM syndrome mainly included hemostasis. The daily dose was most between 8-12 mL, with a usual treatment course of 1-3 d. Commonly used western medicines in combined drug use included hypertonic dehydrating agent, antibiotics and nutrition nerve medicines, and the commonly used traditional Chinese medicines in combined drug use included the drugs with promoting blood circulation to remove blood stasis, kidney-tonifying and bone-building agents, and formula for clearing heat and removing toxicity, etc. The most commonly used western medicines are methylamine, adenosine, lidocaine injection, mannitol, et al, and the most commonly used Chinese medicines included Jintiange capsules, Qianggu capsules, et al. It is suggested that CCP injection for lumbar vertebrae diseases is mainly used in middle-aged and elderly patients, and is frequently used in combination with kidney-tonifying and bone-building agents, drugs with promoting blood circulation to remove blood stasis, analgesic agents and antibiotics.

[Clinical analysis of Cervus and Cucumis Polypeptide injection based on real world hospital information system].

To analyze the clinical application of Cervus and Cucumis Polypeptide injection in the real world, in order to define the characteristics of clinical drug use and correlation, and provide reference for risk management and further study for Cervus and Cucumis Polypeptide injection. Descriptive analysis and association rules analysis were performed on 37 721 cases using Cervus and Cucumis Polypeptide injection in 26 hospitals nationwide. Cervus and Cucumis Polypeptide injection were mostly adopted by patients aged between 45 and 64(39.84%); mainly used to treat fracture patients in clinic(17 362 cases, 33.97%); 12 mL(41.81%) was the commonest dosage. And the course of treatment mainly lasted for 1-3 days(28 467 cases, 76.26%), which was basically consistent with the description of package insert. In clinic, traditional Chinese medicines, such as blood activating and stasis removing agents and Bushen Zhuanggu agents, were frequently combined with it(rule support degree of 19.38%). Such western medicine as antibiotics and nutritional drugs were frequently combined with it(rule support 39.9%). The main single combined medicine were vitamin C(13 202 cases, 35%), and Jintiange capsule(7 285 cases, 19.31%). The commonly used combined drug pairs were Hulisan capsule and Jintiange capsule (rule support 4.458%), phenobarbital and ceftazidime azole oxazoline(rule support degree of 10.62%). Cervus and Cucumis Polypeptide injection is mainly adopted by elderly patients in clinic, used to treat fracture patients, and often combined with blood activating and stasis removing agents, Bushen Zhuanggu agents, antibiotics, and nutritional medicine to enhance fracture healing. In clinical application, attention shall be paid to drug safety of elderly patients and types of combined medicines and their interaction, so as to prevent adverse reactions.