Acute toxicity study of 77 herbal formulas according to GLP guidelines in rats

Article information

J Korean Med. 2023;44(3):29-38

Publication date (electronic) : 2023 September 1

doi : https://doi.org/10.13048/jkm.23028

Hyeun-kyoo Shin ¹

, Woo-Young Jeon ²

, Chang-Seob Seo ¹

, Mee-Young Lee ³^,

¹KM Science Research Division, Korea Institute of Oriental Medicine

²KM Convergence Research Division, Korea Institute of Oriental Medicine

³Research Infrastructure Team, Korea Institute of Oriental Medicine

Correspondence to: Mee-Young Lee, Ph.D., Research Infrastructure Team, Korea Institute of Oriental Medicine, 1672 Yuseong-daero, Daejeon 34054, Republic of Korea, Tel: +82-42-868-9356, E-mail: cozy11@kiom.re.kr

Received 2023 April 7; Revised 2023 July 13; Accepted 2023 July 14.

Abstract

Objectives

This study aimed to investigate the acute oral toxicity of 77 herbal formulas and performed in male and female Sprague-Dawley (SD) rats as per the guidelines mentioned in Ministry of Food and Drug Safety.

Methods

Each sex of SD rat were administered a single dose (2000 or 5000 mg/kg) of 77 herbal formulas via oral gavage; the control group received vehicle only. After administration, the mortality, clinical signs, gross findings, and body weight were followed up for 15 days.

Results

After administration of 77 herbal formulas, mortality, clinical signs, body weight changes, and gross findings related to the test substances were not observed in both male and female groups.

Conclusion

Our results demonstrate the single-dose oral administration of 77 herbal formulas produced no mortality indicating the approximate lethal dose is greater than 2000 or 5000 mg/kg body weight.

Keywords: Herbal formulas; Acute toxicity; Good Laboratory Practice; approximate lethal dose

Table 1

Acute Oral Toxicity of 28 Kinds of Herbal Formula in Health Insurance

Table 2

Acute Oral Toxicity of 26 Kinds of Herbal Formula in Dispensing Instructions

Table 3

Acute Oral Toxicity of 23 Kinds of the Rest Herbal Formula

References

1. Ministry of Health and Welfare. 2021. PHARMACEUTICAL AFFAIRS ACT (No.18307) https://www.law.go.kr/ .

2. Ministry of Food and Drug Safety. 2023;Regulations on the safety of Pharmaceuticals etc (No. 1848)

3. Ministry of Food and Drug Safety. 2022;Toxicity test standards for pharmaceuticals etc (No.2022–18)

4. Ministry of Food and Drug Safety. 2021;Regulations on notification of product approval for traditional herbal medicine & herbal medicine preparations etc (No. 2021–38)

5. Lee S.D., Kwang S.S.. 1991;A Toxicity Study of Herbal Drugs. The journal of Won kwang Oriental Medicine 1(1):33–38.

6. Ministry of Food and Drug Safety. 2022;Non-clinical test management standard etc (No. 2022–93)

7. Shin H.K., Ha H.K., Seo C.S., Lee M.Y.. 2018. Standard herbal medicine formulary Korea institute of oriental medicine. p. 1–3. 9788959703791.

8. National Institute of Food and Drug Safety Evaluation. 2021;Toxicity test method guideline for foods, etc. Single dose toxicity test method :1–51.

9. Lee S.D.. 1999;Toxic Concept in Oriental Medicine. J Society Prevent Korean Med 3(1):157–172.

10. Chang I.K., Hong N.D.. 1985;Experimental studies on the acute toxicity and the efficacy of Phyllostachys juice. J Int Korean Med 2(1):83–101.

11. Cho S.H., Lee Y.H., Park Y.B.. 1992;Acute Toxicity Study of Aqua-acupuncture of Cervi Cornu Parvum and Ganoderma Lucidum Karsten in Mice. The Journal of Korean Acupuncture & Moxibustion Society 9(1):71–83.

12. Choi M.K., Lee Y.H.. 1992;Studies on the Safety assessment of Cervus elaphus Extract for Aqua-acupuncture. The journal of Kyung Hee Oriental Medicine 15(1):203–229.

13. Lee H.J.. 1993;Study for the Toxicity of Kinds of Ginseng Radix Aqua-acupuncture Extract. The Journal of Korean Acupuncture & Moxibustion Society 10(1):167–173.

14. Nam Y.S., Lee Y.H.. 1996;Studies on the Safety Assessment of Red-ginseng Radix Extract Solution for Herb-acupuncture. J korean Medicine 17(1):478–493.

15. Park M.R.. 1995. Studies on the Acute oral toxicity of Sojeokbaekchoolsan in mice Taejeon university. p. 1–39.

16. Kim Y.C., Lee J.H., Lee H.J.. 1997;Studies on oral toxicity of Inchinchunggan-tang in mice. The journal of Kyung Hee Oriental Medicine 20(1):57–89.

17. Yong K.H.. 1985;Prospect of good laboratory practice in Korea. Korean J. of Toxicology 1(1):43–48.

18. Park G.L.. 2005;Meaning and perspective of GLP. The Monthly Technology & Standards 39:93–96.

19. Ma J.Y., Huang D.S., Lee N.H., Ha H.K., Yu Y.B., Shin H.K.. 2008;Acute Toxicity Study on Sipjeondaebo-tang in Rats. Korean J. Oriental Physiology & Pathology 22(5):1192–1195. (364)

20. Ha H.K., LeeJ K., Lee H.Y., Seo C.S., Kim J.H., et al. 2010;Evaluation of safety of the herbal formula Ojeok-san: acute and sub-chronic toxicity studies in rats. J Ethnopharmacol 131(2):410–416. https://doi.org/10.1016/j.jep.2010.07.011 .

21. Ha H.K., Lee J.K., Lee H.Y., Koh W.S., Seo C.S., Shin H.K.. 2011;Safety Evaluation of Yukmijihwang-tang: Assessment of Acute and Subchronic Toxicity in Rats. Evid Based Complement Alternat Med 2011:672136. https://doi.org/10.1155/2011/672136 .

22. Shin I.S., Kim J.H., Ha H.K., Seo C.S., et al. 2010;Acute Toxicity Study on Gumiganghwal-tang (Jiuweiqianghuo-tang) in Sprague-Dawley Rats. The Korean Journal of Oriental Medical Prescription 18(1):79–85.

23. Yoo S.R., Jeong S.J., Lee M.Y., Shin H.K., Seo C.S., Ha H.K.. 2018;Single Oral Acute Toxicity of Banhasasim-Tang and Its Antiobesity Effect on Diet-Induced Obese Mice and 3T3-L1 Adipocytes. Evidence-Based Complementary and Alternative Medicine 2018:3865434. https://doi.org/10.1155/2018/3865434 .

24. Lee M.Y., Seo C.S., Kim J.Y., Shin H.K.. 2015;Evaluation of a water extract of So-CheongRyong-Tang for acute toxicity and genotoxicity using in vitro and in vivo tests. BMC Complementary and Alternative Medicine 15:235. https://doi.org/10.1186/s12906-015-0737-x .

25. Lim H.S., Lee M.Y., Seo C.S., Shin I.S., et al. 2011;Single Oral Dose Toxicity Evaluation of Leejung-tang, a Korean Traditional Herbal Formula, in Crl:CD (SD) rats. The Journal of Korean Oriental Medicine 32(3):18–24.

26. Shin I.S., Lee M.Y., Seo C.S., Lim H.S., Ha H.K., Shin H.K.. 2012;ijin-tang, an Oriental Herbal Formula Reduces Ethanol-induced Acute Gastric Injury in Rats. J Korean Soc Appl Biol Chem (55):197–204. https://doi.org/10.1007/s13765-012-1173-y .

27. Ma J.Y., Huang D.S., You Y.B., Ha H.K., Shin H.K.. 2007;Acute Toxicity Study on Palmul-tang (Bawu-tang) in Mice. Kor. J. Herbology 22(2):13–16.

28. Shin I.S., Kim J.H., Ha H.K., Huang D.S., Shin H.K.. 2010;tudy on Safety of Pyungwi-san in Sprague-Dawley Rats. Journal of Physiology & Pathology in Korean Medicine 24(3):426–429.

29. Lee M.Y., Seo C.S., Kim Y.B., Shin H.K.. 2015;Safety assessment of Guibi-tang: Subchronic toxicity study in Crl:CD SD rats. Regulatory Toxicology and Pharmacology 73(2):485–493. https://doi.org/10.1016/j.yrtph.2015.09.030 .

30. Shin I.S., Kim J.H., Ha H.K., Seo C.S., et al. 2010;Acute Toxicity Study on Bangpungtongsung-san (Fangfengtongsheng-san) in Sprague-Dawley Rats. The Journal of Korean Oriental Medical Ophthalmology & Otorhinolaryngology & Dermatology 23(1):111–117.

31. Ma J.Y., Huang D.S., You Y.B., Ha H.K., Shin H.K.. 2007;Acute Toxicity Study on Sagunja-tang (Sijunzi-tang) in ICR Mice. The Journal of Korean Medicine 28(2):200–204.

32. Ma J.Y., You Y.B., Ha H.K., Huang D.S., Shin H.K.. 2007;Acute Toxicity Study on Samul-tang (Siwu-tang) in Mice. The Korean Journal of Oriental Medical Prescription 15(2):113–117.

33. Kim S.J., Lee M.Y., Shin I.S., Seo C.S., Huh J.I., et al. 2010;ingle Dose Acute Toxicity of Ssanghwa-tang in Crl: CD (SD) Rats. Kor. J. Herbology 26(2):39–43.

34. Jeon W.Y., Lee M.Y., Shin I.S., Lim H.S., Shin H.K.. 2012;Protective Effects of the Traditional Herbal Formula Oryeongsan Water Extract on Ethanol-Induced Acute Gastric Mucosal Injury in Rats. Evidence-Based Complementary and Alternative Medicine 2012:438191. https://doi.org/10.1155/2012/438191 .

Article information Continued

Table 1

Acute Oral Toxicity of 28 Kinds of Herbal Formula in Health Insurance

Herbal formula	Reference Herbal formula	Extract code (KE)	Single-serving size (g)	Yield of extract (%)	GLP organizations	Human equivalent dose for rats (mg/kg)	ALD (mg/kg)
Gamisoyo-san	B	57	31.870	26.80	KIT	882.6	> 5,000
Galgeun-tang	U	02	44.000	12.60	KIT	572.9	> 2,000
Gumiganghwal-tang²²⁾	B	16	37.500	22.76	KIT	882.0	> 2,000
Gungha-tang	B	58	21.270	20.14	KIT	442.7	> 5,000
Naeso-san	B	52	41.250	11.98	KIT	510.6	> 5,000
Daesiho-tang	B	61	50.640	25.62	KIT	1340.6	> 5,000
Daehwajung-eum	B	62	37.520	21.38	KIT	828.9	> 5,000
Banhabaekchulcheonma-tang	B	11	43.750	17.60	KIT	795.7	> 5,000
Banhasasim-tang²³⁾	D	38	37.500	14.99	KIT	580.9	> 5,000
Banhahubag-tang	B	72	26.500	11.66	NIKOM	319.3	> 5,000
Bojungikgi-tang	B	12	22.875	25.40	KIT	600.4	> 2,000
Bulhwangeumjeonggi-san	D	55	33.750	12.48	KIT	435.2	> 5,000
Samso-eum	D	28	41.250	18.60	KIT	792.8	> 2,000
Samchulgeonbi-tang	D	06	43.500	24.50	KIT	1101.3	> 2,000
Saengmaek-san	B	40	15.000	26.58	KIT	412.0	> 5,000
Sosiho-tang	B	26	35.625	22.94	KIT	844.5	> 2,000
Socheongryong-tang²⁴⁾	B	13	37.500	21.70	KIT	840.9	> 2,000
Yeongyopaedog-san	D	59	35.690	23.00	KIT	848.2	> 5,000
Ojeog-san	B	04	56.250	21.00	KIT	1220.6	> 2,000
Ijung-tang²⁵⁾	B	19	26.250	24.80	KIT	672.7	> 2,000
Ijin-tang²⁶⁾	B	08	20.625	18.50	KIT	394.3	> 2,000
Insampaedok-san	D	14	45.000	24.30	KIT	1130.0	> 5,000
Jaeumganghwa-tang	B	01	43.125	20.80	KIT	926.9	> 5,000
Palmul-tang²⁷⁾	B	05	36.000	25.90	KIT	963.5	> 5,000
Pyeongwi-san²⁸⁾	D	18	26.250	23.40	KIT	634.7	> 2,000
Hyangsapyeongwi-san	B	51	33.750	18.75	KIT	653.9	> 5,000
Hyeonggaeyeongyo-tang	B	60	33.440	25.93	KIT	896.0	> 5,000
Hwangryeonhaedok-tang	B	20	18.000	17.10	KIT	318.1	> 2,000

ALD (approximate lethal dose), B (Bang-Yak-Hap-Pyun), BW (body weight), D (Dong-Ui-Bo-Gam), GLP (good laboratory practice), KE (KIOM (Korea Institute of Oriental Medicine) evidence-based medicine), KIT (Korea Institute of Toxicology), NIKOM (National Institute for Korean Medicine Development), U (Ui-Hak-Ip-Mun)

Table 2

Acute Oral Toxicity of 26 Kinds of Herbal Formula in Dispensing Instructions

Herbal formula	Reference	Extract code (KE)	Single-serving size (g)	Yield of extract (%)	GLP organizations	Human equivalent dose for rats (mg/kg)	ALD (mg/kg)
Goakhyangjeonggi-san	D	32	33.750	12.89	KIT	449.5	> 5,000
Gwibi-tang²⁹⁾	D	22	43.000	24.30	KIT	1079.7	> 2,000
Danggwisu-san	D	33	30.375	16.91	KIT	530.8	> 5,000
Danggwijakyak-san	K	75	22.000	24.13	KIT	548.6	> 5,000
Daegunjoong-tang	K	65	8.000	12.13	KIT	100.3	> 5,000
Dokhwalgisaeng-tang	D	74	31.800	22.27	NIKOM	731.8	> 5,000
Bangpungtongseong-san³⁰⁾	D	23	44.125	17.70	KIT	807.0	> 2,000
Baenongsangeup-tang	K	68	18.000	26.46	KIT	492.2	> 5,000
Buhnsimgi-eum	B	54	47.250	14.57	KIT	711.4	> 5,000
Sagunja-tang³¹⁾	D	24	18.750	22.30	KIT	432.1	> 5,000
Samul-tang³²⁾	D	25	18.750	33.30	KIT	645.2	> 5,000
Samryeongbaekchul-san	D	39	91.875	14.10	KIT	1338.6	> 5,000
Sogeonjung-tang	D	63	39.800	17.74	KIT	729.6	> 5,000
Sopung-san	T	71	36.000	17.44	NIKOM	648.8	> 5,000
Sibjeondaebo-tang¹⁹⁾	D	03	45.000	22.90	KIT/KTR	1064.9	> 5,000
Ssanghwa-tang³³⁾	D	21	37.500	25.50	KIT	988.1	> 2,000
Yeonggyechulgam-tang	D	48	22.500	12.73	KIT	296.0	> 5,000
Oyaksungi-san	D	27	46.125	24.40	KIT	1163.0	> 2,000
Onkyeong-tang	D	41	45.000	30.60	KIT	1422.9	> 5,000
Yongdamsagan-tang	D	49	26.500	18.53	KIT	507.4	> 5,000
Wiryeong-tang	D	50	41.250	12.50	KIT	532.8	> 5,000
Yukmijihwang-tang²¹⁾	U	07	46.875	27.00	KIT	1307.8	> 2,000
Jagamcho-tang	D	73	43.020	26.12	NIKOM	1161.1	> 5,000
Chokyungjongok-tang	B	64	45.730	24.02	KIT	1135.0	> 5,000
Cheongsangbangpung-tang	M	69	27.460	28.20	KIT	800.2	> 5,000
Chengsimyeunja-eum	D	43	31.875	13.03	KIT	429.2	> 5,000

ALD (approximate lethal dose), B (Bang-Yak-Hap-Pyun), BW (body weight), D (Dong-Ui-Bo-Gam), GLP (good laboratory practice), K (Keum-Kyue-Yo-Rak), KE (KIOM (Korea Institute of Oriental Medicine) evidence-based medicine), KIT (Korea Institute of Toxicology), KTR (Korea Testing and Research Institute), M (Man-Byeong-Ye-Chun), NIKOM (National Institute for Korean Medicine Development), T (text of Herbal Formula), U (Ui-Hak-Sim-O)

Table 3

Acute Oral Toxicity of 23 Kinds of the Rest Herbal Formula

Herbal formula	Reference	Extract code (KE)	Single-serving size (g)	Yield of extract (%)	GLP organizations	Human equivalent dose for rats (mg/kg)	ALD (mg/kg)
Gamisoyo-san	D	45	35.250	19.41	KIT	707.0	> 5,000
Gamiondam-tang	B	66	29.750	22.55	KIT	693.2	> 5,000
Gyejibokryeong-hwan	D	31	18.750	11.82	KIT	229.0	> 5,000
Gyeji-tang	D	46	30.000	9.75	KIT	302.3	> 5,000
Daeyoung-jeon	B	53	80.625	27.57	KIT	2296.9	> 5,000
Mahwang-tang	D	47	38.500	4.52	KIT	179.8	> 5,000
Maekmundong-tang	B	37	43.125	9.26	KIT	412.6	> 5,000
Sanjoin-tang	K	90	56.25	16.80	KIT	976.5	> 5,000
Sokyunghwalhyeol-tang	M	79	44.500	26.54	NIKOM	1220.4	> 5,000
Siryung-tang	B	89	27.54	21.43	KIT	609.9	> 5,000
Youngseonjetong-eum	B	76	33.300	26.72	NIKOM	919.4	> 5,000
Oryung-san³⁴⁾	D	17	28.125	22.70	KIT	659.7	> 2,000
Jagyakgamcho-tang	D	42	22.500	19.35	KIT	449.9	> 5,000
Jodeung-san	M	78	33.000	20.13	NIKOM	686.4	> 5,000
Cheonwangbosim-dan	D	34	48.750	21.30	KIT	1073.0	> 5,000
Takrisodok-eum	B	70	63.750	24.67	NIKOM	1625.1	> 5,000
Palmijihwang-hwan	D	35	101.250	24.79	KIT	2593.7	> 5,000
Paljung-san	D	56	36.000	9.34	KIT	347.4	> 5,000
Hyangsayukgunja-tang	D	36	43.125	12.95	KIT	577.1	> 5,000
Hyangso-san	D	44	33.750	13.41	KIT	467.7	> 5,000
Hyulbuchukeo-tang	T	77	76.000	26.72	NIKOM	2098.4	> 5,000
Gongjin-dan	B	92	6.887	-	NIKOM	711.7	> 5,000
Kyuongok-go	B	93	23.000	-	KCL	2376.7	> 5,000

ALD (approximate lethal dose), B (Bang-Yak-Hap-Pyun), BW (body weight), D (Dong-Ui-Bo-Gam), GLP (good laboratory practice), K (Keum-Kyue-Yo-Rak), KCL (Korea Conformity Laboratories), KE (KIOM (Korea Institute of Oriental Medicine) evidence-based medicine), KIT (Korea Institute of Toxicology), M (Man-Byeong-Ye-Chun), NIKOM (National Institute for Korean Medicine Development), T (text of Herbal Formula)