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Antioxidant capacity of food has come to be shown in terms of oxygen radical absorbance capacity (ORAC) mainly on vegetables or fruit. However, the evaluation of Kampo in terms of ORAC has not yet been accomplished. It is important that such an investigation is also conducted for Kampo medicine. We measured the ORAC value of almost all the available Kampo formulas used in the Japanese National health insurance system and examined the ORAC value both for the daily prescription, and also the crude herb ingredients. The ORAC value of Kampo medicine ranged 4.65–5913 units/day. The ORAC value was high in Kampo formulas including Rhei Rhizoma, and was relatively high in Kampo formulas including anti-inflammatory herbs other than Rhei Rhizoma. The ORAC value was also high in Kampo formulas including crude herbs that have relaxation effects. The ORAC value of a crude herb would seem to not be fixed but be dependent on combination with other crude herbs from the comparison of different herbs added to the basic Kampo medicine. These results suggest variability and complexity of the antioxidant capacity of Kampo medicine within the similar range of food. On the other hand, investigation of the compound changes of various crude herbs with ORAC may lead to the elucidation of the action mechanism of Kampo medicine.

Active oxygen is important for defending the human body against bacteria or other foreign bodies. However, when active oxygen becomes superabundant, it harms the body. Thus it is erased by antioxidants in the elimination system of the living body in order to maintain the health condition. However, in modern society, aging, carcinogenesis and arteriosclerosis are thought to be caused by production of excess active oxygen through damage by various factors such as ultraviolet rays, smoking, exhaust gas, air pollution and stress. It has become important to take food and supplements including antioxidants to prevent oxidative toxicity. Oxygen radical absorbance capacity (ORAC) is one method to measure antioxidant capacity.

In the US, 1992, ORAC was developed in National Institute on Aging by Guohua Cao [

Although it is said that Kampo medicine has antioxidant capacity examined by electron spin resonance spectrometry, several investigations have shown the capacity of only a few kinds of Kampo formulas [

Among various Kampo medicines available in Japan, 141 kinds of extract formulas, one kind of herbal powder formula and one kind of ointment formula used in the Japanese health insurance system were investigated. All medicines examined in this study were obtained from Japanese pharmaceutical companies whose manufacturing is governed by the regulations of the Pharmaceutical Affairs Law, and strictly controlled by other government regulations including Good Manufacturing Practice. As a result, products are assured of quality and safety at the highest level.

ORAC is the direct capacity of chain-breaking antioxidant based on the hydrogen atom transfer mechanism. In this assay system,

We measured the ORAC of Kampo formulas with the method of Huang et al. [

ORAC is usually expressed as the capacity of 1 g in each object. However, we examined the ORAC unit for a daily dose, because doses are different for each Kampo formula.

We investigated the characteristics of Kampo formulas with high ORAC value.

Kampo formulas can be modified to form new formulas when other herbs are added to them. Comparing the ORAC values between the basic and the modified formulas, we can examine the characteristics of the additional herbs.

When we calculated the ORAC value of Kampo formula per daily dose, it ranged from 4.65 to 5913.

The ORAC value of Kampo formulas containing Rhei Rhizoma (junchoto, tokakujokito, bofutsushosan, choijokito, daiokanzoto, jizusoippo, tsudosan, san'oshashinto, mashiningan, daijokito, keishikashakuyakudaioto, inchinkoto) was high (Table

The 50 highest ORAC value Kampo formulas.

Kampo formula | ORAC | ||
---|---|---|---|

Unit (extact) | Unit (formula) | Unit (formula/ day) | |

tsudosan | 1314.06 | 788.44 | 5913.27 |

daijokito | 1825.25 | 730.10 | 5475.76 |

mashiningan | 2387.72 | 716.32 | 5372.37 |

daiokanzoto | 2919.12 | 583.82 | 4378.69 |

tokiinshi | 867.44 | 578.29 | 4337.19 |

daisaikotokyodaio | 759.70 | 481.14 | 4330.30 |

junchoto | 855.02 | 570.01 | 4275.11 |

keigairengyoto | 930.50 | 558.30 | 4187.25 |

saikoseikanto | 879.31 | 556.90 | 4176.74 |

kakkonkajutsubuto | 802.34 | 513.50 | 3851.23 |

chikujountanto | 672.94 | 493.49 | 3701.18 |

kakkontokasenkyushin'i | 920.78 | 491.08 | 3683.11 |

san'oshashinto | 2100.18 | 490.04 | 3675.31 |

bofutsushosan | 798.67 | 479.20 | 3594.02 |

seijobofuto | 753.58 | 477.27 | 3579.52 |

tokakujokito | 1179.57 | 471.83 | 3538.71 |

choijokito | 2830.01 | 471.67 | 3537.52 |

kakkonto | 925.17 | 462.58 | 3469.38 |

keishikashakuyakudaioto | 856.27 | 456.68 | 3425.09 |

jidabokuippo | 1512.87 | 453.86 | 3403.95 |

daisaikoto | 755.43 | 453.26 | 3399.43 |

kososan | 1695.17 | 452.04 | 3390.33 |

shomakakkonto | 1505.41 | 451.62 | 3387.18 |

saikanto | 676.92 | 451.28 | 3384.61 |

nyoshinsan | 749.25 | 449.55 | 3371.63 |

senkyuchachosan | 1024.73 | 444.05 | 3330.37 |

inchinkoto | 2218.58 | 443.72 | 3327.86 |

saibokuto | 653.80 | 435.87 | 3269.02 |

yokukansankachimpihange | 700.31 | 420.19 | 3151.40 |

saikokeishikankyoto | 868.91 | 405.49 | 3041.18 |

jinsoin | 758.19 | 404.37 | 3032.76 |

shin’iseihaito | 666.29 | 399.77 | 2998.28 |

shinpito | 1089.68 | 399.55 | 2996.61 |

boiogito | 788.37 | 394.19 | 2956.40 |

nijutsuto | 582.32 | 388.22 | 2911.62 |

seishinrenshiin | 579.39 | 386.26 | 2896.93 |

otsujito | 723.38 | 385.81 | 2893.54 |

hangeshashinto | 642.44 | 385.46 | 2890.96 |

ogonto | 715.29 | 381.49 | 2861.18 |

heiisan | 863.41 | 374.15 | 2806.09 |

orengedokuto | 1847.38 | 369.48 | 2771.07 |

tokishigyakukagoshuyushokyoto | 679.20 | 362.24 | 2716.78 |

gorinsan | 542.33 | 361.55 | 2711.66 |

kyukikyogaito | 440.86 | 293.91 | 2645.16 |

jizusoippo | 866.79 | 346.72 | 2600.38 |

daiobotanpito | 742.25 | 346.38 | 2597.87 |

saikokaryukotsuboreito | 567.35 | 340.41 | 2553.07 |

shofusan | 634.90 | 338.62 | 2539.62 |

goshakusan | 631.23 | 336.66 | 2524.91 |

bukuryoingohangekobokuto | 559.71 | 335.83 | 2518.72 |

The ORAC value of Kampo formulas containing anti-inflammatory herbs other than Rhei Rhizoma (saikokeishikankyoto, keigairengyoto, seijobofuto, jinsoin,nyoshinsan, kososan, saikanto, saikoseikanto, chikujountan-to, saibokuto, san'oshashinto, senkyuchachosan, inchinkoto) was also high (Table

The ORAC value of Kampo formulas that relax the muscles (kakkonto, kakkontokasenkyushin’i, jinsoin, saibokuto, shomakakkonto, tsudosan, daijokito, mashiningan) was relatively high (Table

The ORAC value was extremely high in tsudosan, mashiningan, daijokito. For these Kampo medicines, Magnoliae Cortex for relaxing muscle tonus is included rather than Rhei Rhizoma. In other words, the more a Kampo medicine contains herbs of a high ORAC value, the higher the overall ORAC value (Table

In the Kampo formula series based on keishikashakuyakuto, it was recognized that Rhei Rhizoma has an influence on high antioxidant capacity, which Paeoniae Radix, Astragali Radix, Angelicae Radix, Saccharum Granorum did not have (Table

The ORAC value of Kampo formulas according to the additional herbs added to keishikashakuyakuto (base formula).

Kampo formula | Additional herbs added to base formula | ORAC value of Kampo formula per daily dose ( |
---|---|---|

keishikashakuyakuto | 1796.7 | |

keishikashakuyakutd aioto | Rhei Rhizoma | 3425.1 |

shokenchuto | Saccharum Granorum | 1840.2 |

ogikenchuto | Saccharum Granorum + Astragali Radix | 2174.04 |

tokikenchuto | Angelicae Radix | 1923 |

Difference of the ORAC value of Kampo formulas according to the additional herbs added to the basic Kampo formula. Major change patterns were classified into six groups in this study. Group A, which had little change of ORAC value, consisted of shokenchuto and tokikenchuto shown in Table

In the Kampo formula series based on rokumigan, it was found that Cinnamomi Cortex, Aconiti Tuber, Achyranthis Radix, Plantaginis Semen did not have an influence on antioxidant capacity (Table

The ORAC value of Kampo formulas according to the additional herbs added to rokumigan (base formula).

Kampo formula | Additional herbs added to base formula | ORAC value of Kampo formula per daily dose ( |
---|---|---|

rokumigan | 1842.98 | |

hachimijiogan | Cinnamomi Cortex+ Aconiti Tuber | 1050 |

goshajinkigan | Cinnamomi Cortex + Aconiti Tuber+ Achyranthis Radix + Plantaginis Semen | 1097.03 |

In the Kampo formula series based on keishito, strong antioxidant capacity was not recognized by the addition of Fossilia Ossis Mastodi, Ostreae Testa, Atractylodis Lanceae Rhizoma, Aconiti Tuber, Astragali Radix. Additional Magnoliae Cortex or Puerariae Radix deteriorated the antioxidant capacity. However, strong antioxidant capacity was identified by addition of the combination of Ephedrae Herba and Puerariae Radix. The addition of Evodiae Fructus, Angelicae Radix, Akebiae Caulis and Asiasari Radix showed little antioxidant capacity (Table

The ORAC value of Kampo formulas according to the additional herbs added to keishito (base formula).

Kampo formula | Additional herbs added to base formula | ORAC value of Kampo formula per daily dose ( |
---|---|---|

keishito | 1804.28 | |

keishikaryukotsuboreito | Fossilia Ossis Mastodi+Ostreae Testa | 1246.65 |

keishikajutsubuto | Atractylodis Lanceae Rhizoma + Aconiti Tuber | 1751.1 |

keishikaryojutsubuto | Atractylodis Lanceae Rhizoma + Aconiti Tuber + Poria | 1710.15 |

keishikaogito | Astragali Radix | 538.2 |

keishikakakkonto | Puerariae Radix | 609.9 |

keishikakobokukyoninto | Magnoliae Cortex + Armeniacae Semen | 1374.45 |

kakkonto | Puerariae Radix + Ephedrae Herba | 3469.35 |

kakkontokasenkyushin’i | Puerariae Radix + Ephedrae Herba + Cnidii Rhizoma + Magnoliae Flos | 3683.1 |

tokishigyakukagosh uyushokyoto | Angelicae Radix + Evodiae Fructus + Akebiae Caulis + Asiasari Radix | 2716.8 |

keishikashakuyakuto | Paeoniae Radix^{a} | 1796.7 |

^{
a}Keishikashakuyakuto contains more Paeoniae Radix than keishito.

The present study showed following results. The ORAC value in a daily dose of Kampo formula in Japan was 4.65–5913. The ORAC value in the dose was high in Kampo formulas including Rhei Rhizoma, and was relatively high in Kampo formula including anti-inflammatory herbs other than Rhei Rhizoma. The ORAC value was also high in Kampo formula including crude herbs that have relaxation effects. The change in antioxidant capacity from the addition of a crude herb varied with different base formulas.

Although it is said that Kampo medicine has high antioxidant capacity, the ORAC value taken in on one day by Kampo medicine did not seem to be high compared to a meal judging from this ORAC measurement results. Actually, the quantity of ORAC which we can take in with food (vegetables, fruit) on one day is 3264 ORAC units in total. This referred to the National Health and Nutrition Survey in Japan [

Because Rhei Rhizoma, a component herb in many Kampo formulas, has anti-inflammatory action, and anti-tumor effect, it may be possible that the ORAC value of Kampo formulas including Rhei Rhizoma is high. For example, in examination using KHC rabbit, which is a model of the familial hypercholesterolemia, the development of arteriosclerosis was inhibited by the dosage of choijokito of 1 g kg^{−1} of 24 weeks [

Kampo formulas which contain herbs that have anti-inflammation action except Rhei Rhizoma had high ORAC value, which were not as high as that of Rhei Rhizoma. Judging from the crude herb constitution of a high ORAC value Kampo formula, Coptidis Rhizoma, Scutellariae Radix, Bupleuri Radix, Gardeniae Fructus, Phellodendri Cortex, Forsythiae Fructus, Arctii Fructus, and Menthae Herba are thought to be candidates as anti-inflammatory herb. Kampo formulas that have anti-inflammatory effect are generally classified to two groups, one is the group that contains Bupleuri Radix and Scutellariae Radix, and the other is the group that contains Coptidis Rhizoma, Scutellariae Radix, Phellodendri Cortex and Asiasari Radix, in addition to Kampo formulas containing Rhei Rhizoma. And the investigations about the antioxidant capacity have been accomplished according to such a classification. For example, regarding the former group, Sakaguchi et al. [_{3}-induced epilepsy, and also inhibited hippocampal neuronal death in gerbils with transient ischemia.

In Kampo formulas including crude herbs such as Magnoliae Cortex, Puerariae Radix, which have muscle relaxation effects, ORAC measurement showed a high value in this investigation. And there are some reports to suggest antioxidant effect of these medicines. About Magnoliae Cortex, Son et al. [

Our data suggest that the effects of Kampo medicine come from the composition and interaction of the ingredients during the boiling process, and not simply the aggregate effects of each individual component. In other words, the effect of the added crude herb is not a simple extension of the base formula characteristic; rather the addition of a crude herb creates a new Kampo formula with new characteristics. When we compared rokumigan as hachimijiogan and goshajinkigan, which contain additional Cinnamomi Cortex and Aconiti Tuber to rokumigan, the ORAC value of hachimijiogan and goshajinkigan, the new formulas were lower than that of rokumigan. On the other hand, when we compared keishito as keishikajutsubuto, the formula to which Atractylodis Lanceae Rhizoma and Aconiti Tuber are added, the ORAC value of keishito was the same as that of keishikajutsubuto. These findings may show discrepancy in the function of Aconiti Tuber. The ORAC value of keishikakakkonto, which contains additional Puerariae Radix to keisihito, was lower than that of keisihito. The ORAC value of kakkonto, which contains additional Puerariae Radix and Ephedrae Herba to keisihito, was higher than that of keisihito. These finding may suggest that Kampo medicine has complicated characteristics that cannot be assumed from each simple herb. And it is expected that the functions of herbs might be clarified by investigating changes of complicated herbal formulas with the ORAC method.

In this discussion, the antioxidant capacity of Kampo formulas was confirmed both

Special Co-ordination Fund for Promoting Science and Technology.

The authors are grateful to Mr Anthony Todd for reviewing the manuscript.