Supplementary data

Structural elucidation and anti-osteoporosis activities

of polysaccharides obtained from Curculigo orchioides

Xueqian Wang a, Mengliu Zhang a, Dawei Zhang b, Xinluan Wang c,

Huijuan Cao c, Qian Zhang**a, Chunyan Yan a,*

a School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006,

China

bDepartment of Pharmacology, Guangdong Medical University, Dongguan,

Guangdong 523808, China

cTranslational Medicine R&D Center, Institute of Biomedical and Health

Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of

Sciences, Shenzhen, China

Table S1. Effects of CO70 on bone microarchitecture in the left femur of

ovariectomized (OVX) rats.

Parameters Sham OVX E2 CO70

Conn. Dens (mm-3) 89.50±7.10 45.36±4.69## 51.45±3.52** 51.12±5.69*

SMI -0.27±0.40 1.92±0.23## 1.56±0.22** 1.55±0.21**

BV/TV 0.50±0.07 0.15±0.01## 0.21±0.03** 0.20±0.02**

BS/BV (mm-1) 19.99±2.18 29.54±2.15## 26.19±2.09** 27.61±1.97

Tb.N (mm-1) 4.56±0.15 2.27±0.27## 2.79±0.16** 2.62±0.27*

Tb.Th (mm) 0.099±0.009 0.068±0.003## 0.075±0.004** 0.074±0.006*

Tb.Sp (mm) 0.12±0.01 0.34±0.04## 0.27±0.02** 0.29±0.03*

All values are expressed as the mean ± S.D. ##P < 0.01 versus Sham; *P < 0.05 versus OVX, **P

< 0.01 versus OVX.

Fig. S1. Effects of CO70 on (A) body weight and (B) uterus coefficient of

ovariectomized (OVX) rats. A: The rats underwent adaptive feeding for the first week,

ovariectomy was performed during the second week, and two days after surgery, the

treatment began. B: The uterus was isolated and quickly weighed after the rats were

euthanized. The uterus coefficient was the ratio of uterus weight to body weight. All

values are expressed as the mean ± SD. ##P < 0.01 versus Sham; *P < 0.05 versus

OVX, **P < 0.01 versus OVX.

Fig. S2. The elution curves of CO70

on DEAE-52 (A) and CO70-3 on Sephadex G-75 (B).

Fig. S3. The

HPGPC chromatogram of COP70-3.

Fig. S4. The chromatograms of monosaccharides composition analysis of COP70-3.

A: HPLC analysis of the PMP pre-column derivatives of mixture of standard

monosaccharide. 1: mannose, 2: rhamnose, 3: glucuronic acid, 4: galacturonic acid, 5:

glucose, 6: galactose, 7: xylose, 8: arabinose, 9: fucose. B: HPLC analysis of the PMP

pre-column derivatives of COP70-3. 1: mannose, 2: rhamnose, 3: glucuronic acid, 4:

galacturonic acid, 5: glucose, 6: galactose, 7: arabinose.

Fig. S5. The IR spectrum of COP70-3.

Fig. S6. The gas chromatography of COP70-3 methylation analysis.

The peak at 7.10 min was 1,3,5,6-tetra-O-acetyl-2,4-di-O-methyl-D-galactitol.

The peak at 9.92 min was 1,4,5-tri-O-acetyl-2,3,6-tri-O-methyl-D-mannitol.

The peak at 11.45 min was 1,5-di-O-acetyl-2,3,4,6-tetra-O-methyl-D-glucitol.

The peak at 13.03 min was 1,3,5-tri-O-acetyl-2,4-di-O-methyl-6-deoxy-L-mannitol.

The peak at 13.40 min was 1,5,6-tri-O-acetyl-2,3,4-tri-O-methyl-D-galactitol.

The peak at 16.64 min was 1,3,5,6-tetra-O-acetyl-2,4-di-O-methyl-D-mannitol.

The peak at 18.84 min was 1,5,6-tri-O-acetyl-2,3,4-tri-O-methyl-D-glucose.

The peak at 18.60 min was 1,3,4,5-tetra-O-acetyl-2-O-methyl-L-arabinitol.

The peak at 19.06 min was 1,4,5-tri-O-acetyl-2,3-di-O-methyl-L-arabinitol.

Fig. S7. The standard curve of glucose.

Fig. S8. The chromatograms of monosaccharides composition analysis of CO70.

A: HPLC analysis of the PMP pre-column derivatives of mixture of standard

monosaccharide. 1: mannose, 2: rhamnose, 3: glucuronic acid, 4: galacturonic acid, 5:

glucose, 6: galactose, 7: xylose, 8: arabinose, 9: fucose. B: HPLC analysis of the PMP

pre-column derivatives of CO70. 1: mannose, 2: rhamnose, 3: glucuronic acid, 4:

galacturonic acid, 5: glucose, 6: galactose, 7: xylose, 8: arabinose.