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Journal of Chinese Integrative Medicine ›› 2010, Vol. 8 ›› Issue (5): 438-447.doi: 10.3736/jcim20100507

• Original Experimental Research • Previous Articles     Next Articles

Preclinical safety evaluation of the aqueous acetone extract of Chinese herbal formula Modified Huo Luo Xiao Ling Dan

Yin Fan Arthura,b, Lixing Laoa, Rui-xin Zhanga, An-nan Zhoua,b,Brian M. Bermana   

  1. a Center for Integrative Medicine, School of Medicine, University of Maryland, Baltimore, MD 21201, USA
    b McLean Center for Complementary and Alternative Medicine, PLC. Vienna,VA 22182,USA
  • Received:2009-12-10 Accepted:2010-03-30 Online:2010-05-20 Published:2010-06-15
  • Contact: Yin Fan Arthur E-mail:ArthurFan@ChineseMedicineDoctor.US


To investigate the safety of oral administration of Modified Huo Luo Xiao Ling Dan (HLXLD), a compound traditional Chinese herbal medicine.

The toxicological information of HLXLD and its individual constituent herbs was searched in cintcm or TCMlars (, PubMed (MEDLINE), Chinese Herbal Medicine (1999) and WHO Monographs on Selected Medicinal Plants (Vol. Ⅰ — Ⅲ). Single-dose acute toxicity was assessed by using the highest possible dosage. Motor function test was used to determine whether the herbal formula might cause motor impairment. Nine-day HLXLD repeat-dose sub-chronic toxicity/adverse effects, and 42-day chronic toxicity/adverse effects in rats were also assessed.

The literature searches showed that HLXLD and its eleven ingredient herbs had no side/adverse effects listed in the traditional Chinese medicine literature. Under the dosages proposed in the formula, the HLXLD formula had no side/adverse effects according to MEDLINE, Chinese Herbal Medicine and WHO Monographs on Selected Medicinal Plants. The studies in rats showed: (1) in single-dose acute toxicity assessment, the maximal feasible single oral dose, 9.20 g/kg HLXLD, showed no significant effect on clinical signs, or body weight and mortality over a 14-day period in rats; (2) during motor function test, nine-day repeat-dose of daily HLXLD treatment at 4.60 g/kg did not cause motor impairment; (3) in nine-day HLXLD repeat-dose sub-chronic toxicity/adverse effects assessment, there were no noticeable abnormal behavioral changes or obvious adverse reactions and signs in complete Freund's adjuvant inflamed rats (highest observed dosage: 4.60 g/kg), and no noticeable adverse effects were observed during, or 14 days after nine-day treatment at 4.60 g/kg in non-inflamed rats; (4) during 42-day chronic toxicity/adverse effects assessments, no noticeable abnormal behavioral changes, no obvious adverse reactions and signs were observed in normal rats administered with HLXLD at a dose of 2.30 g/kg and the values of serum biochemistry and histopathology were in normal range.

Both existing information and animal data support that Modified HLXLD is a safe herbal product for clinical application.

Key words: Chinese herbal formula, Modified Huo Luo Xiao Ling Dan, Safety, Literature investigation, Acute toxicity tests, Motor function, Sub-chronic toxicity, Chronic toxicity tests, Rats


No. Chinese herb name in pinyin [Botanic name] Plant part Human
dose (g)*
Oral administration acute toxicity**
(1) Ruxiang [Boswellia carterii Birdw.] Gum-resin 15 17.8% N/A
(2) Qianghuo [Notopterygium incisum Ting ex H. T. Chang.] Rhizome and root 12 9.5% MFD: Water exact 12 g/kg
(3) Danggui [Angelica sinensis (Oliv.) Diels.] Root 12 11.6% N/A
(4) Baishao [Paeonia lactiflora Pall.] Root, rootlets and
12 10.0% MFD: 6 g/kg (methanol extract)
(5) Gancao [Glycyrrhiza uralensis Fisch.] Root 12 9.7% N/A***
(6) Yanhusuo [Corydalis yanhusuo W.T.Wang] Root 12 1.8% LD50: 100 g/kg
(7) Danshen [Salvia miltiorrhiza Bge.] Root 12 1.8% N/A***
(8) Chuanxiong [Ligusticum chuanxiong S.H. Qiu.] Root 12 9.2% MFD: 6 g/kg (methanol extract)
(9) Qinjiao [Gentiana macrophylla Pall.] Root 12 10.0% LD50: 18.96 g/kg (water extract); 17.36 g/kg (ethanol extract)
(10) Guizhi [Cinnamomum cassia Presl.] Twig 15 4.1% MFD: 50 g/kg (water extract)
(11) Duhuo [Angelica pubescens Maxim.] Root 12 14.4% N/A
Total 11 herbs 138 100% N/A



Group n ALT (IU/L) AST (IU/L) Total protein (g/dL) Albumin (g/dL) Globulin (g/dL)
Baseline 20 44.45±1.23 114.20±2.95 6.32±0.06 3.63±0.03 2.69±0.04
End 20 69.44±2.99** 153.06±8.43** 8.02±0.22** 4.50±0.12** 3.51±0.11**
Baseline 10 45.70±2.28 115.50±5.76 6.43±0.13 3.70±0.06 2.73±0.08
End 10 65.20±5.21** 146.70±11.62 7.79±0.20** 4.47±0.10** 3.32±0.10**


Group n LDH (IU/L) CK (IU/L) BUN (mg/dL) Creatinine (mg/dL)
Baseline 20 1 619.20±92.88 931.05±99.26 0.47±0.04 19.15±1.84
End 20 937.05±119.33** 1 094.68±290.13 0.51±0.02 25.31±0.67**
Baseline 10 1 693.10±167.63 883.40±85.69 0.59±0.09 22.5±3.15
End 10 926.30±107.45** 630.90±143.90 0.52±0.02 24.9±1.39
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