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Table 3. Contd.<br />

Xu et al. 2441<br />

s27 Beisanjia, Fushun, Liaoning 1.25 0.03 6.65 - 0.30 8.22<br />

s28 Hongtoushan, Fushun, Liaoning 1.24 - 4.44 - 0.13 5.80<br />

s29 Dongling, Shenyang, Liaoning 1.90 0.02 1.90 - - 3.82<br />

s30 Hunhe, Shenyang, Liaoning 1.71 0.03 2.12 - 0.08 3.94<br />

s31 Hunhepu, Shenyang, Liaoning 2.67 0.08 4.10 - 0.30 7.16<br />

s32 Baiqingzai, Shenyang, Liaoning 3.02 0.03 2.92 - 0.15 6.12<br />

s33 Cuigangzi, Heishan, Liaoning 0.55 - 2.45 - 0.21 3.22<br />

s34 Qianxing, Heishan, Liaoning 0.79 0.01 2.90 - 0.24 3.95<br />

s35 Zhangjiahuang, Heishan, Liaoning 2.34 - 2.12 - 1.10 5.56<br />

s36 Lijiazhen, Heishan, Liaoning 1.25 0.03 3.00 0.32 0.24 4.84<br />

s37 Yonglingzhen, Xinbin, Liaoning 1.61 0.02 3.50 - 0.10 5.22<br />

s38 Qianjincun, Xinbin, Liaoning 0.77 - 4.14 - 0.16 5.06<br />

s39 Guchengzhen, Huairen, Liaoning 1.03 - 3.17 - 0.29 4.49<br />

s40 Shuanglingzicun, Huairen, liaoning 2.08 - 3.37 - 0.11 5.56<br />

s41 Heigouxiang, Huairen, Liaoning 1.14 0.04 4.93 - 0.74 6.86<br />

s42 Xiaoshizhen, Benxi, Liaoning 1.73 0.03 6.34 - 0.20 8.30<br />

s43 Kazuo, Chaoyang, Liaoning 1.15 0.05 3.61 - 0.13 4.95<br />

s44 Zhongtunxiang, Jinzhou, Liaoning 1.42 0.03 7.31 - 0.40 9.16<br />

s45 Dawaxian, Panjin, Liaoning 2.37 0.09 6.64 - 0.78 9.87<br />

s46 Sanshijiazizhen, Lingyuan, Liaoning 0.74 0.16 2.78 - 1.12 4.78<br />

s47 Qianshan, Anshan, Liaoning 0.57 - 6.44 - 0.37 7.38<br />

s48 Heishankexiang, Huludao, Liaoning 2.66 0.03 4.86 - 0.16 7.72<br />

s49 Yangmadianzixiang, Huludao, Liaoning 0.85 0.04 9.86 - 0.20 10.95<br />

s50 Yingkou, Liaoning 0.94 0.03 2.37 - 0.16 3.50<br />

s51 Gaizhou, Yingkou, Liaoning 0.35 0.02 2.70 - 0.14 3.21<br />

s52 Xiongyue, Yingkou, Liaoning 0.34 - 8.12 - 0.12 8.57<br />

s53 Donggang, Liaoning 1.44 0.09 8.88 - 0.29 10.70<br />

s54 Pulandian, Dalian, Liaoning 1.04 0.05 3.90 - 0.11 5.09<br />

s55 Yuerjiancun, Datong, Shanxi 0.61 0.14 9.42 - 0.75 10.91<br />

s56 Shikuxiang, Lucheng, Shanxi 1.07 0.07 4.71 - 0.69 6.55<br />

s57 Qumenzhen, Yiyuan, Shandong 0.08 - 0.62 - 0.70 1.40<br />

s58 Tiefou, Huaibei, Anhui 0.13 - 0.84 - 0.23 1.20<br />

s59 Fulizhen, Shuzhou, Anhui 0.96 - 3.58 0.31 0.25 5.10<br />

s60 Fulizhen, Shuzhou, Anhui 0.61 - 3.55 - 0.60 4.76<br />

PS: “-” represented the contents had not been detected.<br />

directly analyzed to decrease assay variability, and five<br />

active ingredients showed good resolution and high<br />

recovery.<br />

Method validation<br />

Linearity<br />

The five calibration curves in the concentration ranges of<br />

Luteoloside, Luteolin, Physalin A, Physalin P and<br />

Physalin O were 0.532~2.66, 0.04~0.2, 0.48~2.4,<br />

0.04~0.2 and 0.36~1.8 μg/mL, respectively.<br />

The regression equations and coefficients were: y =<br />

236.44x + 198.43, r = 0.9997; y = 371.34x + 95.48,<br />

r = 0.9995; y = 76.337x + 25.217, r = 0.9996; y = 79.788x<br />

– 8.81, r = 0.9997; y = 104.53x + 38.59 and r = 0.9997,<br />

respectively, where y is the peak area of the five active<br />

ingredients and x is the concentration of the five active<br />

ingredients.<br />

Recovery<br />

For a validation of the extraction recoveries of the five<br />

active ingredients, the analysis for each active ingredient<br />

was carried out in five replicates. The results showed that<br />

the mean extraction recoveries were acceptable,<br />

suggesting that there was negligible loss during the<br />

calyces extraction (Table 1).

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