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The Huo-Xue-Hua-Yu decoction (HXHYD) prescription, which possesses good clinical properties for healing fractures, is made up of 12 types of traditional Chinese medicines (TCMs). According to the drug efficacy of TCM, HXHYD consists of four drug-group prescriptions. This study focused on the type of active mechanism in these drug groups and the type of interaction between them in HXHYD. Surface-enhanced Raman scattering (SERS) spectroscopy was employed to study the effect of the incorporation of calcium hydroxyapatite (CHA), protein and lipid groups on the repair of complete tibial fractures in rats treated with HXHYD and its drug-group prescriptions and without treatment to reveal the efficacy rule of the drug groups in the prescription. The rats were divided into seven groups, each of which was subdivided into three subgroups (evaluated on days 15, 21 and 30 after surgery). The six treatment groups were treated HXHYD, four drug-group prescriptions and Gu-Zhe-Cuo-Shang capsule therapy (treated control). SERS readings were taken at the fractured sites. The results showed that the medical ingredients in HXHYD were not the simple addition of four drug-group prescriptions and the efficacy of HXHYD was stronger than every other drug group prescription because it contained the highest content of CHA and highest carbonate-to-phosphate and phosphate-to-phenylalanine ratios among the treatment groups at all time points in comparison with the fractured control group (no treatment). Thus, the SERS technique has great potential to provide a novel method for effectively and accurately studying the efficacy rule of drug groups in one prescription with the aim to optimize prescriptions.

We have recently applied Raman spectroscopy for biochemical analysis of cell groups treated with 2.5, 5, 7.5 and 10 micromoles per liter of carboplatin and cisplatin, respectively, for apoptosis detection. The continuously decreased intensities of Raman bands assigned to the proteins and deoxyribonucleic acid can be observed as the concentrations of drugs increased, suggesting the cell damage induced by carboplatin and cisplatin. Principal component analysis combined with linear discriminate analysis and support vector machines analysis of the obtained spectral data was used to develop diagnostic algorithms. When cell groups were treated with the same concentration, both principal component analysis combined with linear discriminate analysis and support vector machines analysis executed between control group and groups treated with cisplatin, produce a lower error than those treated with carboplatin, suggesting the more cell damage induced by cisplatin. Among treated groups, an accuracy of 100% is only observed in the group treated with 10 micromoles per liter of cisplatin, demonstrating the apoptosis. Those Statistical analysis results are consistent with the biochemical and Raman spectral analysis. This study indicated that Raman spectroscopy in conjunction with principal component analysis combined with linear discriminate analysis and support vector machines analysis has great potential use in label-free detection of apoptosis in cells and label-free assessment of drug efficacy.