Salvia columbariae contains tanshinones

Salvia columbariae (chia) was examined and found to contain miltionone II, cryptotanshinone and tanshinone IIA. These compounds may be of interest in the treatment of stroke and heart attack.

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Analytical Methods
HPLC and UV spectra of extracts. The extracts were injected (50-100 l) onto an HPLC system with a Supelcosil LC-18T column. The mobile phase was 80% methanol, 20% water flowing at 1 ml/min. UV spectra were collected with a photodiode array detector.
HPLC-Mass Spectrometry of extracts. The extracts were submitted to the California Institute of Technology, Regional Mass Spectrometry Facility (Dr Mona Shagholi). The extracts were injected onto an HPLC-MS system with an Eclipse   XDB-C18 column and were developed at 1 ml/min in 80/20 methanol/water containing 1% formic acid.

Identification of Tanshinones
The root extracts were found to contain three major peaks on the HPLC system as visualized at 254 nm. The retention times were 4.2, 6.9 and 10.2 min. The UV spectra of each peak were similar with maxima at about 250 and 300 nm. The HPLC conditions were chosen based on the chromatography of tanshinones (5). The retention times were similar to published retention times for tanshinones (5). The UV spectra were similar to published spectra for miltionones, cryptotanshinone and related compounds (4). The extinction coefficients of tanshinone IIA are lamba max MeoH nm (log epsilon): 220 (4.46), 250 (4.40) and 269 (4.44), (9). Based on the similar UV spectra and similar chromophores of the three compounds, the extinction coefficients are probably similar for each. The HPLC peaks for the three compounds integrated as follows: miltionone II (see below), 4.2 min 25.2%, cryptotanshinone, 6.9 min 69% and tanshinone IIA, 10.2 min 5.8%. The amounts of each compound in the 50 ml extract were: miltionone II, 0.7 mol, cryptotanshinone, 2 mol, and tanshinone IIA, 0.2 mol. An HPLC-MS chromatogram is shown in Figure 3.

Results
Chia was grown at the Rancho Santa Ana Botanic Garden, Claremont, CA. When the plants were about 1 m tall, the entire plants were harvested. At this time, they were in flower with large seed clusters. The plants were put into plastic bags and stored in a freezer. Chia can be difficult to grow to maturity. The seeds sprout sometimes abundantly, but die quickly if not in the correct environment. The seeds were found to grow best in full shade, with plenty of water, good drainage and the application of lime (a few grams) when the plants are about 2 cm tall. Transplanting the seedlings into lime-containing soil resulted in the loss of most plants.
Chia was found to contain 17.5 mol of tanshinone IIA per kg of root material (Table 1). This is three-fold less than is found in dan shen (5). However, chia contains nearly fivefold more cryptotanshinone than is found in dan shen (5). Cryptotanshinone is a precursor for tanshinone IIA and is converted into tanshinone IIA in the liver (10). This implies that chia contains 192.5 mol/kg of active tanshinones. Dan shen contains 91 mol/kg of active tanshinones.

Discussion
The presence of tanshinone IIA and similar compounds in chia could explain the historical use of this plant, to 'wake the dead, or the nearly dead' such as with stroke and heart attack patients. Tanshinones have a range of pharmacological activities including inhibition of clotting (6), vasodilatation (7) and inhibition of NO synthase (11). All of these activities are potentially beneficial in stroke. Stroke is frequently caused by blood clots that dislodge from one location and travel in the blood system until they lodge in small cerebral arteries. This causes brain ischemia and usually stimulates more clotting in the area. Vasodilatation and inhibition of clotting may help dislodge and dissolve the clot. NO synthase is known to become activated in ischemia and can generate NO that damages DNA leading to cell death. Inhibition of NO synthase may protect neurons from DNA damage and cell death.
Chia contains some of the same compounds found in dan shen, including tanshinone IIA. In China, tanshinone IIA is available as a purified sulfonate salt for use in stroke, heart attack and angina patients (3,12). Although, tanshinone IIA is regarded as the active agent in chia, it is also recognized that cryptotanshinone is a precursor to tanshinone IIA in the body (10). While tanshinone IIA is very rapidly cleared from the body by hepatic metabolism (10), cryptotanshinone is oxidized in the liver to make tanshinone IIA. Therefore, tanshinone IIA levels may be higher and stay higher for a longer time period after cryptotanshinone than after tanshinone IIA administration. Chia contains more cryptotanshinone and less tanshinone IIA than dan shen (Table 1). Chia contains two times more active tanshinones than does dan shen. This implies that chia may be superior to dan shen for use as a delivery agent or precursor for tanshinone IIA. It may be of interest to test dan shen and chia extracts to see which plant extract produces higher plasma levels of tanshinone IIA and better protection from infarction.